U.S. patent application number 15/605856 was filed with the patent office on 2018-04-26 for compounds and methods for kinase modulation, and indications therefor.
The applicant listed for this patent is Plexxikon Inc.. Invention is credited to Dean R. Artis, Gideon Bollag, Ryan Bremer, Hanna Cho, Samuel J. Gillette, Gaston Habets, Klaus-Peter Hirth, Prabha N. Ibrahim, Shumeye Mamo, Marika Nespi, Shenghua Shi, Wayne Spevak, Yoshisa Suzuki, James Tsai, Brian West, Guoxian Wu, Chao Zhang, Jiazhong Zhang, Hongyao Zhu, Yong-Liang Zhu, Rebecca Zuckerman.
Application Number | 20180111929 15/605856 |
Document ID | / |
Family ID | 37033831 |
Filed Date | 2018-04-26 |
United States Patent
Application |
20180111929 |
Kind Code |
A1 |
Ibrahim; Prabha N. ; et
al. |
April 26, 2018 |
COMPOUNDS AND METHODS FOR KINASE MODULATION, AND INDICATIONS
THEREFOR
Abstract
Compounds active on protein kinases are described, as well as
methods of using such compounds to treat diseases and conditions
associated with aberrant activity of protein kinases.
Inventors: |
Ibrahim; Prabha N.;
(Mountain View, CA) ; Artis; Dean R.; (Kensington,
CA) ; Bremer; Ryan; (Oakland, CA) ; Habets;
Gaston; (Pleasant Hill, CA) ; Mamo; Shumeye;
(Oakland, CA) ; Nespi; Marika; (Berkeley, CA)
; Zhang; Chao; (Moraga, CA) ; Zhang; Jiazhong;
(Foster City, CA) ; Zhu; Yong-Liang; (Fremont,
CA) ; Zuckerman; Rebecca; (Alameda, CA) ;
West; Brian; (San Francisco, CA) ; Suzuki;
Yoshisa; (San Pablo, CA) ; Tsai; James;
(Vallejo, CA) ; Hirth; Klaus-Peter; (San
Francisco, CA) ; Bollag; Gideon; (Orinda, CA)
; Spevak; Wayne; (Berkeley, CA) ; Cho; Hanna;
(Oakland, CA) ; Gillette; Samuel J.; (Oakland,
CA) ; Wu; Guoxian; (Foster City, CA) ; Zhu;
Hongyao; (Berkeley, CA) ; Shi; Shenghua; (San
Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Plexxikon Inc. |
Berkeley |
CA |
US |
|
|
Family ID: |
37033831 |
Appl. No.: |
15/605856 |
Filed: |
May 25, 2017 |
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14798167 |
Jul 13, 2015 |
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15605856 |
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13243748 |
Sep 23, 2011 |
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14798167 |
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12906980 |
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13243748 |
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11473347 |
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60692960 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/437 20130101;
A61P 13/08 20180101; A61P 25/28 20180101; A61P 19/02 20180101; A61P
9/00 20180101; A61P 25/06 20180101; A61P 17/06 20180101; A61P 35/00
20180101; C07C 37/62 20130101; A61P 21/04 20180101; A61P 37/06
20180101; C07D 209/08 20130101; A61P 7/02 20180101; A61P 43/00
20180101; A61P 9/10 20180101; A61P 17/02 20180101; C07C 45/00
20130101; A61P 15/08 20180101; C07C 39/27 20130101; A61P 19/10
20180101; A61P 27/02 20180101; A61P 37/00 20180101; C07C 47/565
20130101; A61P 29/00 20180101; A61K 31/5377 20130101; A61P 1/00
20180101; A61P 25/14 20180101; A61P 35/02 20180101; C07D 471/04
20130101; A61P 3/06 20180101; A61P 3/14 20180101; A61P 37/02
20180101; A61P 25/00 20180101; A61P 3/04 20180101; C07C 45/71
20130101; C07C 45/673 20130101; A61P 1/18 20180101; A61P 1/04
20180101; A61P 31/16 20180101; A61P 9/04 20180101; A61P 13/12
20180101; A61P 31/04 20180101; A61K 31/496 20130101; A61P 27/16
20180101; A61P 3/10 20180101; A61P 7/00 20180101; A61P 1/16
20180101; A61P 11/00 20180101; A61P 11/06 20180101; C07C 47/575
20130101; A61P 37/08 20180101; A61P 17/00 20180101; A61P 25/16
20180101; C07C 45/00 20130101; C07C 47/565 20130101; C07C 45/673
20130101; C07C 47/575 20130101; C07C 45/71 20130101; C07C 47/575
20130101; C07C 37/62 20130101; C07C 39/27 20130101 |
International
Class: |
C07D 471/04 20060101
C07D471/04; A61K 31/5377 20060101 A61K031/5377; C07C 37/62 20060101
C07C037/62; A61K 31/437 20060101 A61K031/437; A61K 31/496 20060101
A61K031/496; C07C 45/00 20060101 C07C045/00; C07C 45/67 20060101
C07C045/67; C07C 45/71 20060101 C07C045/71; C07C 47/565 20060101
C07C047/565; C07C 47/575 20060101 C07C047/575; C07D 209/08 20060101
C07D209/08; C07C 39/27 20060101 C07C039/27 |
Claims
1. A compound of Formula III having the following structure:
##STR03475## all salts, prodrugs, tautomers, and isomers thereof,
wherein: Q has a structure selected from the group consisting of
##STR03476## in which ##STR03477## indicates the attachment point
of Q to A of Formula III; Z.sub.2 is N or CR.sup.12; Z.sub.4 is N
or CR.sup.14; Z.sub.5 is N or CR.sup.15; Z.sub.6 is N or CR.sup.16;
L.sub.2 is selected from the group consisting of
--(CR.sup.10R.sup.11).sub.p--NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--O--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(O)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(S)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O).sub.2--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(O)NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(S)NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O).sub.2NR.sup.25--(CR.sup.10R.sup.11).sub-
.q--,
--(CR.sup.10R.sup.11).sub.p--NR.sup.25C(O)--(CR.sup.10R.sup.11).sub.-
q--
--(CR.sup.10R.sup.11).sub.p--NR.sup.25C(S)--(CR.sup.10R.sup.11).sub.q--
-, and
--(CR.sup.10R.sup.11).sub.p--NR.sup.25S(O).sub.2--(CR.sup.10R.sup.1-
1).sub.q--; p and q are independently 0, 1, or 2 provided, however,
that at least one of p and q is 0; s is 1 or 2; X is O or S; A is
selected from the group consisting of --O--, --S--,
--CR.sup.aR.sup.b--, --NR.sup.1--, --C(O)--, --C(S)--, --S(O)--,
and --S(O).sub.2--; R.sup.a and R.sup.b at each occurrence are
independently selected from the group consisting of hydrogen,
fluoro, --OH, --NH.sub.2, lower alkyl, lower alkoxy, lower
alkylthio, mono-alkylamino, di-alkylamino, and --NR.sup.8R.sup.9,
wherein the alkyl chain(s) of lower alkyl, lower alkoxy, lower
alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro; or R.sup.a and R.sup.b combine to form a 3-7 membered
monocyclic cycloalkyl or 5-7 membered monocyclic heterocycloalkyl,
wherein the monocyclic cycloalkyl or monocyclic heterocycloalkyl
are optionally substituted with one or more substituents selected
from the group consisting of halogen, --OH, --NH.sub.2, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino;
R.sup.1 is selected from the group consisting of hydrogen, lower
alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
--C(O)R.sup.7, --C(S)R.sup.7, --S(O).sub.2R.sup.7, --C(O)NHR.sup.7,
--C(S)NHR.sup.7, and --S(O).sub.2NHR.sup.7, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein the alkyl chain(s) of lower alkoxy,
lower alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro, further provided that when R.sup.1 is lower alkyl, any
substitution on the lower alkyl carbon bound to the N of
--NR.sup.1-- is fluoro, and wherein cycloalkyl, heterocycloalkyl,
aryl or heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; R.sup.7 is selected from the group consisting of
lower alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, lower alkylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, provided, however, that any
substitution of the alkyl carbon bound to the N of --C(O)NHR.sup.7,
--C(S)NHR.sup.7 or --S(O).sub.2NHR.sup.7 is fluoro, wherein the
alkyl chain(s) of lower alkoxy, lower alkylthio, mono-alkylamino,
or di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro, and wherein
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more substituents selected from the group
consisting of halogen, --OH, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; R.sup.4,
R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15, R.sup.16, R.sup.42,
R.sup.43, R.sup.45, R.sup.46 and R.sup.47 are independently
selected from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, --CN,
--NO.sub.2, --CR.sup.aR.sup.bR.sup.26, and -LR.sup.26; L at each
occurrence is independently selected from the group consisting of
-(alk).sub.a-S-(alk).sub.b-, -(alk).sub.a-O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(O)-(alk).sub.b-, -(alk).sub.a-C(S)-(alk).sub.b-,
-(alk).sub.a-S(O)-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-OC(O)-(alk).sub.b-, -(alk).sub.a-C(O)O-(alk).sub.b-,
-(alk).sub.a-OC(S)-(alk).sub.b-, -(alk).sub.a-C(S)O-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-, and
-(alk).sub.a-NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-; a and b are
independently 0 or 1; alk is C.sub.1-3 alkylene or C.sub.1-3
alkylene substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkyl,
lower alkoxy, lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein lower alkyl or the alkyl chain(s) of
lower alkoxy, lower alkylthio, mono-alkylamino or di-alkylamino are
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution of the
alkyl chain carbon bound to O of alkoxy, S of thioalkyl or N of
mono- or di-alkylamino is fluoro; R.sup.25 at each occurrence is
independently selected from the group consisting of hydrogen,
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; R.sup.26
at each occurrence is independently selected from the group
consisting of hydrogen, provided, however, that hydrogen is not
bound to any of S(O), S(O)2, C(O) or C(S) of L, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.26 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of L, optionally substituted lower
alkynyl, provided, however, that when R.sup.26 is optionally
substituted lower alkynyl, no alkyne carbon thereof is bound to N,
S, O, S(O), S(O).sub.2, C(O) or C(S) of L, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; R.sup.10
and R.sup.11 at each occurrence are independently selected from the
group consisting of hydrogen, fluoro, lower alkyl, and lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; or any two of R.sup.10 and R.sup.11 on the same or
adjacent carbon atoms combine to form a 3-7 membered monocyclic
cycloalkyl or 5-7 membered monocyclic heterocycloalkyl, and any
others of R.sup.10 and R.sup.11 are independently selected from the
group consisting of hydrogen, fluoro, lower alkyl, and lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, and wherein the monocyclic cycloalkyl or
monocyclic heterocycloalkyl are optionally substituted with one or
more substituents selected from the group consisting of halogen,
--OH, --NH.sub.2, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; R.sup.8 and R.sup.9 combine with the nitrogen
to which they are attached to form a 5-7 membered heterocycloalkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkyl,
fluoro substituted lower alkyl, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, and fluoro substituted lower
alkylthio; R.sup.17 is selected from the group consisting of
hydrogen, halogen, optionally substituted lower alkyl and
--OR.sup.18; R.sup.31 and R.sup.33 are independently selected from
the group consisting of optionally substituted aryl, optionally
substituted heteroaryl, optionally substituted cycloalkyl, and
optionally substituted heterocycloalkyl; R.sup.36 is selected from
the group consisting of substituted methyl, optionally substituted
C.sub.2-6 alkyl, optionally substituted lower alkenyl, provided,
however, that when R.sup.36 is optionally substituted lower
alkenyl, no alkene carbon thereof is bound to the S(O).sub.2of
S(O).sub.2R.sup.36, optionally substituted lower alkynyl, provided,
however, that when R.sup.36 is optionally substituted lower
alkynyl, no alkyne carbon thereof is bound to the S(O).sub.2of
S(O).sub.2R.sup.36, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, and --NR.sup.19R.sup.20;
R.sup.19, R.sup.20, R.sup.34, R.sup.35, R.sup.37, and R.sup.38 are
independently selected from the group consisting of hydrogen,
optionally substituted lower alkyl, optionally substituted lower
alkenyl, provided, however, that when R.sup.19, R.sup.20, R.sup.34,
R.sup.35, R.sup.37, or R.sup.38 is optionally substituted lower
alkenyl, no alkene carbon thereof is bound to the N of
NR.sup.19R.sup.20, NR.sup.34R.sup.35 or NR.sup.37R.sup.38,
optionally substituted lower alkynyl, provided, however, that when
R.sup.19, R.sup.20, R.sup.34, R.sup.35, R.sup.37, or R.sup.38 is
optionally substituted lower alkynyl, no alkyne carbon thereof is
bound to the N of NR.sup.19R.sup.20, NR.sup.34R.sup.35 or
NR.sup.37R.sup.38, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl and
optionally substituted heteroaryl; or R.sup.34 and R.sup.35
together with the nitrogen to which they are attached form
optionally substituted 5-7 membered heterocycloalkyl or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl; or
R.sup.37 and R.sup.38 together with the nitrogen to which they are
attached form optionally substituted 5-7 membered heterocycloalkyl
or optionally substituted 5 or 7 membered nitrogen containing
heteroaryl; R.sup.32 is selected from the group consisting of
hydrogen, optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl, and
--OR.sup.18; R.sup.82 is selected from hydrogen or lower alkyl; and
R.sup.18 is hydrogen or optionally substituted lower alkyl;
provided, however, that the compound is not
3-{3-[2-(tetrahydropyran-2-yloxy)-ethoxy]-benzyl}-5-thiophen-3-yl-1H-pyrr-
olo[2,3-b]pyridine, which has the structure ##STR03478## or
4-(4-methyl-piperazin-1-ylmethyl)-N-[4-(1H-pyrrolo[2,3-b]pyridin-3-ylmeth-
yl)-phenyl]-benzamide, which has the structure ##STR03479##
2. The compound according to claim 1 having structure of Formula
IIIa: ##STR03480##
3. The compound according to claim 2 having structure of Formula
IIIb: ##STR03481## wherein: A.sub.1 is --O--,
--CR.sup.40R.sup.41--, --C(O)--or --NR.sup.48--; Z.sub.12 is N or
CR.sup.52; Z.sub.16 is N or CR.sup.56; R.sup.40 and R.sup.41 are
independently selected from the group consisting of hydrogen,
fluoro, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; or R.sup.40 and R.sup.41 combine to form a 3-7
membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl, wherein the monocyclic cycloalkyl or monocyclic
heterocycloalkyl is optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; L.sub.3 is selected from the group consisting of
--NR.sup.48--, --S--, --O--, --NR.sup.48CH(R.sup.49)--,
--SCH(R.sup.49)--, --OCH(R.sup.49)--, --C(O)NR.sup.48--,
--S(O).sub.2NR.sup.48--, --CH(R.sup.49)NR.sup.48--,
--CH(R.sup.49)O--, --CH(R.sup.49)S--, --NR.sup.48C(O)--, and
--NR.sup.48S(O).sub.2--; R.sup.53 and R.sup.55 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl and lower alkoxy, wherein the alkyl chain of lower alkyl or
lower alkoxy is optionally substituted with fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino or cycloalkylamino, provided, however, that any
substitution on the alkyl carbon bound to the --O-- of lower alkoxy
is fluoro; R.sup.52 and R.sup.56 are independently selected from
the group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy; R.sup.49 is selected from the group consisting of hydrogen,
lower alkyl, and fluoro substituted lower alkyl; Cy is selected
from the group consisting of aryl, heteroaryl, cycloalkyl, and
heterocycloalkyl; R.sup.39 is selected from the group consisting of
hydrogen, halogen, lower alkyl, lower alkoxy, aryl, heteroaryl, and
NR.sup.50R.sup.51, wherein the alkyl chain of lower alkyl or lower
alkoxy is optionally, substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino
and cycloalkylamino, and wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
R.sup.50 is hydrogen or lower alkyl optionally substituted with one
or more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino; R.sup.51 is
aryl or heteroaryl, wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
R.sup.23 at each occurrence is independently selected from the
group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2, --C(O)OH,
--S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --NR.sup.48C(O)R.sup.57,
--NR.sup.48S(O).sub.2R.sup.57, --S(O).sub.2R.sup.57,
--C(O)R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57,
--S(O).sub.2NR.sup.48R.sup.57, halogen, lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl as R.sup.23, or as substituents of lower alkyl, are
optionally substituted with one or more substituents selected from
the group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2,
--C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.58,
--SR.sup.58, --NR.sup.48R.sup.58, --NR.sup.48C(O)R.sup.58,
--NR.sup.48S(O).sub.2R.sup.58, --S(O).sub.2R.sup.58,
--C(O)R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58,
--S(O).sub.2NR.sup.48R.sup.58, halogen, lower alkyl, fluoro
substituted lower alkyl, and cycloalkylamino; R.sup.57 is selected
from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.58 at each occurrence is independently selected from the
group consisting of lower alkyl, heterocycloalkyl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and t is 0, 1,
2, or 3.
4. The compound according to laim 2 having structure of Formula
IIIp: ##STR03482## all salts, prodrugs, tautomers, and isomers
thereof, wherein: A.sub.1 is --O--, --CR.sup.40R.sup.41--,
--C(O)--or --NR.sup.48--; Z.sub.22 is N or CR.sup.62; Z.sub.26 is N
or CR.sup.66; r is 0, 1, or 2; R.sup.40 and R.sup.41 are
independently selected from the group consisting of hydrogen,
fluoro, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; or R.sup.40 and R.sup.41 combine to form a 3-7
membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl, wherein the monocyclic cycloalkyl or monocyclic
heterocycloalkyl is optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; R.sup.62, R.sup.63, R.sup.65 and R.sup.66 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl and lower alkoxy, wherein the alkyl chain of
lower alkyl or lower alkoxy is optionally substituted with one or
more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; Cy is selected from the group
consisting of aryl, heteroaryl, cycloalkyl, and heterocycloalkyl;
R.sup.39 is selected from the group consisting of hydrogen,
halogen, lower alkyl, lower alkoxy, aryl, heteroaryl, and
NR.sup.50R.sup.51, wherein the alkyl chain of lower alkyl or lower
alkoxy is optionally substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino
and cycloalkylamino, and wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
R.sup.50 is hydrogen or lower alkyl optionally substituted with one
or more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino; R.sup.51 is
aryl or heteroaryl, wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
R.sup.23 at each occurrence is independently selected from the
group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2, --C(O)OH,
--S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --NR.sup.48C(O)R.sup.57,
--NR.sup.48S(O).sub.2R.sup.57, --S(O).sub.2R.sup.57,
--C(O)R.sup.57, --C(O)()R.sup.57, --C(O)NR.sup.48R.sup.57,
--S(O).sub.2NR.sup.48R.sup.57, halogen, lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl as R.sup.23, or as substituents of lower alkyl, are
optionally substituted with one or more substituents selected from
the group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2,
--C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.58,
--SR.sup.58, --NR.sup.48R.sup.58, --NR.sup.48C(O)R.sup.58,
--NR.sup.48S(O).sub.2R.sup.58, --S(O).sub.2R.sup.58,
--C(O)R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58,
--S(O).sub.2NR.sup.48R.sup.58, halogen, lower alkyl, fluoro
substituted lower alkyl, and cycloalkylamino; R.sup.57 is selected
from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.58 at each occurrence is independently selected from the
group consisting of lower alkyl, heterocycloalkyl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and t is 0, 1,
2, or 3.
5. The compound according to claim 1 having structure of Formula
IIIc: ##STR03483##
6. The compound according to claim 1 having structure of Formula
IIIo: ##STR03484##
7. The compound according to claim 6 having structure of Formula
IIId: ##STR03485## wherein: r is 0, 1, or 2.
8. The compound according to claim 7 of structure of Formula IIIe:
##STR03486## wherein: A.sub.1 is --O--, --CR.sup.40R.sup.41--,
--C(O)--or --NR.sup.48--; Z.sub.22 is N or CR.sup.62; Z.sub.24 is N
or CR.sup.64; Z.sub.25 is N or CR.sup.65; Z.sub.26 is N or
CR.sup.66; r is 0, 1, or 2; R.sup.40 and R.sup.41 are independently
selected from the group consisting of hydrogen, fluoro, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino; or
R.sup.40 and R.sup.41 combine to form a 3-7 membered monocyclic
cycloalkyl or 5-7 membered monocyclic heterocycloalkyl, wherein the
monocyclic cycloalkyl or monocyclic heterocycloalkyl is optionally
substituted with one or more substituents selected from the group
consisting of halogen, --OH, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; R.sup.62,
R.sup.64, R.sup.65 and R.sup.66 are independently selected from the
group consisting of hydrogen, halogen, lower alkyl and lower
alkoxy, wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro; Cy is
selected from the group consisting of aryl, heteroaryl, cycloalkyl,
and heterocycloalkyl; R.sup.39 is selected from the group
consisting of hydrogen, halogen, lower alkyl, lower alkoxy, aryl,
heteroaryl, and NR.sup.50R.sup.51, wherein the alkyl chain of lower
alkyl or lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, and wherein aryl and heteroaryl
are optionally substituted with one or more independent
substituents R.sup.23; R.sup.50 is hydrogen or lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino; R.sup.51 is aryl or heteroaryl, wherein aryl and
heteroaryl are optionally substituted with one or more independent
substituents R.sup.23; R.sup.23 at each occurrence is independently
selected from the group consisting of --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.23, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.57 is selected from the group consisting of lower alkyl,
cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein lower
alkyl is optionally substituted with one or more substituents
selected from the group consisting of fluoro, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, provided, however, that any
substitution of the alkyl carbon bound to O, S, or N of
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, or --S(O).sub.2NR.sup.48R.sup.57 is
fluoro, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or
as substituents of lower alkyl are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.58 at each occurrence is independently selected from the
group consisting of lower alkyl, heterocycloalkyl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and t is 0, 1,
2, or 3.
9. The compound according to claim 6 having structure of Formula
IIIq: ##STR03487## all salts, prodrugs, tautomers, and isomers
thereof, wherein: A.sub.1 is --O--, --CR.sup.40R.sup.41--,
--C(O)--or --NR.sup.48--; Z.sub.12 is N or CR.sup.52; Z.sub.14 is N
or CR.sup.54; Z.sub.15 is N or CR.sup.55; Z.sub.16 is N or
CR.sup.56; R.sup.40 and R.sup.41 are independently selected from
the group consisting of hydrogen, fluoro, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; or R.sup.40
and R.sup.41 combine to form a 3-7 membered monocyclic cycloalkyl
or 5-7 membered monocyclic heterocycloalkyl, wherein the monocyclic
cycloalkyl or monocyclic heterocycloalkyl is optionally substituted
with one or more substituents selected from the group consisting of
halogen, --OH, --NH.sub.2, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; L.sub.3 is selected from the
group consisting of --NR.sup.48--, --S--, --O--,
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, --OCH(R.sup.49)--,
--C(O)NR.sup.48--, --S(O).sub.2NR.sup.48--,
--CH(R.sup.49)NR.sup.48--, --CH(R.sup.49)O--, --CH(R.sup.49)S--,
--NR.sup.48C(O)--, and --NR.sup.48S(O).sub.2--; R.sup.54 and
R.sup.55 are independently selected from the group consisting of
hydrogen, halogen, lower alkyl and lower alkoxy, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino or cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; R.sup.52 and R.sup.56 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy; R.sup.49 is selected from the
group consisting of hydrogen, lower alkyl, and fluoro substituted
lower alkyl; Cy is selected from the group consisting of aryl,
heteroaryl, cycloalkyl, and heterocycloalkyl; R.sup.39 is selected
from the group consisting of hydrogen, halogen, lower alkyl, lower
alkoxy, aryl, heteroaryl, and NR.sup.50R.sup.51, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
one or more substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, and wherein
aryl and heteroaryl are optionally substituted with one or more
independent substituents R.sup.23; R.sup.50 is hydrogen or lower
alkyl optionally substituted with one or more substituents selected
from the group consisting of fluoro, --OH, --NH.sub.2, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino; R.sup.51 is aryl or heteroaryl, wherein aryl and
heteroaryl are optionally substituted with one or more independent
substituents R.sup.23; R.sup.23 at each occurrence is independently
selected from the group consisting of --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.4SR.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.23, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.57 is selected from the group consisting of lower alkyl,
cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein lower
alkyl is optionally substituted with one or more substituents
selected from the group consisting of fluoro, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, provided, however, that any
substitution of the alkyl carbon bound to O, S, or N of
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, or --S(O).sub.2NR.sup.48R.sup.57 is
fluoro, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or
as substituents of lower alkyl are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
R.sup.58 at each occurrence is independently selected from the
group consisting of lower alkyl, heterocycloalkyl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and t is 0, 1,
2, or 3.
10. The compound according to claim 1 having structure of Formula
IIIf: ##STR03488##
11. The compound according to claim 10 having structure of Formula
IIIg: ##STR03489## wherein: A.sub.1 is --O--,
--CR.sup.40R.sup.41--, --C(O)--or --NR.sup.48--; Z.sub.32 is N or
CR.sup.72; Z.sub.34 is N or CR.sup.74; Z.sub.35 is N or CR.sup.75;
Z.sub.36 is N or CR.sup.76; X is O or S; R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; R.sup.40 and
R.sup.41 are independently selected from the group consisting of
hydrogen, fluoro, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; or R.sup.40 and R.sup.41 combine to form a 3-7
membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl, wherein the monocyclic cycloalkyl or monocyclic
heterocycloalkyl is optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; R.sup.72, R.sup.74, R.sup.75 and R.sup.76 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl and lower alkoxy, wherein the alkyl chain of
lower alkyl and lower alkoxy is optionally substituted with one or
more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; R.sup.67 is selected from the
group consisting of hydrogen, halogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; one of R.sup.60 and R.sup.61 is lower alkyl,
fluoro substituted lower alkyl, or --(CH.sub.2).sub.0-2R.sup.70,
and the other of R.sup.60 and R.sup.61 is hydrogen or lower alkyl;
or R.sup.60 and R.sup.61 together with the nitrogen to which they
are attached form optionally substituted 5-7 membered
heterocycloalkyl or optionally substituted 5 or 7 membered nitrogen
containing heteroaryl; R.sup.68 is selected from the group
consisting of optionally substituted lower alkyl, optionally
substituted lower alkenyl, provided, however, that when R.sup.68 is
optionally substituted lower alkenyl, no alkene carbon thereof is
bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of --OR.sup.68,
--SR.sup.68, --NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; R.sup.69 is hydrogen or
optionally substituted lower alkyl; R.sup.70 is selected from the
group consisting of optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; and R.sup.82 is hydrogen or
lower alkyl.
12. The compound according to claim 1 having structure of Formula
IIIh: ##STR03490##
13. The compound according to claim 12 having structure of Formula
IIIi: ##STR03491## wherein: A.sub.1 is --O--,
--CR.sup.40R.sup.41--, --C(O)--or --NR.sup.48--; X is O or S;
R.sup.48 at each occurrence is independently hydrogen or lower
alkyl; R.sup.40 and R.sup.41 are independently selected from the
group consisting of hydrogen, fluoro, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; or R.sup.40
and R.sup.41 combine to form a 3-7 membered monocyclic cycloalkyl
or 5-7 membered monocyclic heterocycloalkyl, wherein the monocyclic
cycloalkyl or monocyclic heterocycloalkyl is optionally substituted
with one or more substituents selected from the group consisting of
halogen, --OH, --NH.sub.2, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; R.sup.67 is selected from the
group consisting of hydrogen, halogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; one of R.sup.60 and R.sup.61 is lower alkyl,
fluoro substituted lower alkyl, or --(CH.sub.2).sub.0-2R.sup.70 and
the other of R.sup.60 and R.sup.61 is hydrogen or lower alkyl; or
R.sup.60 and R.sup.61 together with the nitrogen to which they are
attached form optionally substituted 5-7 membered heterocycloalkyl
or optionally substituted 5 or 7 membered nitrogen containing
heteroaryl; R.sup.68 is selected from the group consisting of
optionally substituted lower alkyl, optionally substituted lower
alkenyl, provided, however, that when R.sup.68 is optionally
substituted lower alkenyl, no alkene carbon thereof is bound to N,
S, O, S(O), S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2) --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; R.sup.69 is hydrogen or
optionally substituted lower alkyl; and R.sup.70 is selected from
the group consisting of optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl.
14. The compound according to claim 1 having structure of Formula
IIIj: ##STR03492##
15. The compound according to claim 14 having structure of Formula
IIIk: ##STR03493## wherein: A.sub.1 is --O--,
--CR.sup.40R.sup.41--, --C(O)--or --NR.sup.48--; R.sup.81 is
selected from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; R.sup.71 and R.sup.78 are independently
selected from the group consisting of hydrogen, halogen, C.sub.1-3
alkyl, and fluoro substituted C.sub.1-3 alkyl; R.sup.77 is selected
from the group consisting of substituted methyl, optionally
substituted C.sub.2-6 alkyl, optionally substituted aryl,
optionally substituted heteroaryl, and --NR.sup.79R.sup.80, wherein
methyl is substituted with one or more substituents selected from
the group consisting of optionally substituted aryl and optionally
substituted heteroaryl; R.sup.68 is selected from the group
consisting of optionally substituted lower alkyl, optionally
substituted lower alkenyl, provided, however, that when R.sup.68 is
optionally substituted lower alkenyl, no alkene carbon thereof is
bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of --OR.sup.68,
--SR.sup.68, --NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; R.sup.69 is hydrogen or
optionally substituted lower alkyl; and R.sup.79 and R.sup.80 are
independently hydrogen or optionally substituted lower alkyl, or
R.sup.79 and R.sup.80 combine with the nitrogen to which they are
attached to form optionally substituted 5-7 membered
heterocycloalkyl.
16. The compound according to claim 15 having structure of Formula
IIIm: ##STR03494## wherein: R.sup.81 is selected from the group
consisting of hydrogen, halogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; R.sup.83 is selected from the group
consisting of hydrogen, fluoro and chloro; R.sup.112 is selected
from the group consisting of optionally substituted C.sub.2-6alkyl,
optionally substituted aryl, optionally substituted heteroaryl, and
--NR.sup.79R.sup.80; R.sup.68 is selected from the group consisting
of optionally substituted lower alkyl, optionally substituted lower
alkenyl, provided, however, that when R.sup.68 is optionally
substituted lower alkenyl, no alkene carbon thereof is bound to N,
S, O, S(O), S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2) --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; R.sup.69 is selected from the
group consisting of hydrogen and optionally substituted lower
alkyl; and R.sup.79 and R.sup.80 are independently hydrogen or
optionally substituted lower alkyl, or R.sup.79 and R.sup.80
combine with the nitrogen to which they are attached to form
optionally substituted 5-7 membered heterocycloalkyl.
17. The compound according to claim 1 having structure of Formula
IIIn: ##STR03495##
18. A compound of Formula II having the following structure:
##STR03496## all salts, prodmgs, tautomers, and isomers thereof,
wherein: R.sup.4, R.sup.5, and R.sup.6 are independently selected
from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, --CN,
--NO.sub.2, --CR.sup.aR.sup.bR.sup.26, and -LR.sup.26; R.sup.12,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are independently
selected from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, --CN,
--NO.sub.2, --CR.sup.aR.sup.bR.sup.24, and -LR.sup.24; A is
selected from the group consisting of --O--, --S--,
--CR.sup.aR.sup.b--, --NR.sup.1--, --C(O)--, --C(S)--, --S(O)--,
and --S(O).sub.2--; R.sup.a and R.sup.b at each occurrence are
independently selected from the group consisting of hydrogen,
fluoro, --OH, --NH.sub.2, lower alkyl, lower alkoxy, lower
alkylthio, mono-alkylamino, di-alkylamino, and --NR.sup.8R.sup.9,
wherein the alkyl chain(s) of lower alkyl, lower alkoxy, lower
alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro; or R.sup.a and R.sup.b combine to form a 3-7 membered
monocyclic cycloalkyl or 5-7 membered monocyclic heterocycloalkyl,
wherein the monocyclic cycloalkyl or monocyclic heterocycloalkyl
are optionally substituted with one or more substituents selected
from the group consisting of halogen, --OH, --NH.sub.2, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino;
R.sup.1 is selected from the group consisting of hydrogen, lower
alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
--C(O)R.sup.7, --C(S)R.sup.7, --S(O).sub.2R.sup.7, --C(O)NHR.sup.7,
--C(S)NHR.sup.7, and --S(O).sub.2NHR.sup.7, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein the alkyl chain(s) of lower alkoxy,
lower alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro, further provided that when R.sup.1 is lower alkyl, any
substitution on the lower alkyl carbon bound to the N of
--NR.sup.1-- is fluoro, and wherein cycloalkyl, heterocycloalkyl,
aryl or heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; R.sup.7 is selected from the group consisting of
lower alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, lower alkylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, provided, however, that any
substitution of the alkyl carbon bound to the N of --C(O)NHR.sup.7,
--C(S)NHR.sup.7 or --S(O).sub.2NHR.sup.7 is fluoro, wherein the
alkyl chain(s) of lower alkoxy, lower alkylthio, mono-alkylamino,
or di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro, and wherein
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more substituents selected from the group
consisting of halogen, --OH, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; L at each
occurrence is independently selected from the group consisting
of-(alk).sub.a-S-(alk).sub.b-, -(alk).sub.a-O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(O)-(alk).sub.b-, -(alk).sub.a-C(S)-(alk).sub.b-,
-(alk).sub.a-S(O)-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-OC(O)-(alk).sub.b-, -(alk).sub.a-C(O)O-(alk).sub.b-,
-(alk).sub.a-OC(S)-(alk).sub.b-, -(alk).sub.a-C(S)O-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-, -(alk).sub.a-NR.sup.25
S(O).sub.2-(alk).sub.b-, -(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-, and
-(alk).sub.a-NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-; a and b are
independently 0 or 1; alk is C.sub.1-3 alkylene or C.sub.1-3
alkylene substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkyl,
lower alkoxy, lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein lower alkyl or the alkyl chain(s) of
lower alkoxy, lower alkylthio, mono-alkylamino or di-alkylamino are
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution of the
alkyl chain carbon bound to O of alkoxy, S of thioalkyl or N of
mono- or di-alkylamino is fluoro; R.sup.8 and R.sup.9 combine with
the nitrogen to which they are attached to form a 5-7 membered
heterocycloalkyl optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, and
fluoro substituted lower alkylthio; R.sup.25 at each occurrence is
independently selected from the group consisting of hydrogen,
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; and
R.sup.24 and R.sup.26 at each occurrence are independently selected
from the group consisting of hydrogen, provided, however, that
hydrogen is not bound to any of S(O), S(O).sub.2, C(O) or C(S) of
L, optionally substituted lower alkyl, optionally substituted lower
alkenyl, provided, however, that when R.sup.24 or R.sup.26 is
optionally substituted lower alkenyl, no alkene carbon thereof is
bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of L, optionally
substituted lower alkynyl, provided, however, that when R.sup.24 or
R.sup.26 is optionally substituted lower alkynyl, no alkyne carbon
thereof is bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of L,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, and optionally
substituted heteroaryl, provided, however that the at least one of
R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is -LR.sup.24,
wherein R.sup.24 is substituted methyl, optionally substituted
C.sub.2-6 alkyl, optionally substituted lower alkenyl, optionally
substituted lower alkynyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl or optionally substituted heteroaryl, further provided,
however, that R.sup.13 is not ##STR03497## and that R.sup.14 is not
##STR03498## wherein ##STR03499## indicates the bond to the phenyl
ring.
19. The compound according to claim 18 wherein R.sup.24 is
optionally substituted C.sub.2-6 alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, or substituted
methyl, wherein methyl is substituted with optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl.
20. The compound of claim 19, wherein R.sup.5 is other than
hydrogen.
21.-39. (canceled)
Description
RELATED PATENT APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 12/906,980 filed Oct. 18, 2010, which is a divisional of U.S.
application Ser. No. 11/473,347 filed Jun. 21, 2006, now issued as
U.S. Pat. No. 7,836,288, which claims the benefit under 35 U.S.C.
.sctn. 119(e) of U.S. Provisional Application No. 60/692,960, filed
Jun. 22, 2005, and U.S. Provisional Application No. 60/731,528,
filed Oct. 28, 2005, which are incorporated herein by reference in
their entireties and for all purposes.
FIELD OF THE INVENTION
[0002] The present invention relates to kinases and compounds which
modulate kinases, and uses therefore. Particular embodiments
contemplate disease indications which are amenable to treatment by
modulation of kinase activity by the compounds of the present
invention.
BACKGROUND OF THE INVENTION
[0003] The information provided herein is intended solely to assist
the understanding of the reader. None of the information provided
nor references cited is admitted to be prior art to the present
invention. Each of the references cited herein is incorporated in
its entirety.
[0004] Receptor protein kinases regulate key signal transduction
cascades that control or are involved in the control of a plethora
of physiological functions including cellular growth and
proliferation, cell differentiation, cellular development, cell
division, cell adhesion, stress response, short-range
contact-mediated axonal guidance, transcription regulation,
aberrant mutagenesis, angiogenesis, abnormal endothelial cell-cell
or cell-matrix interactions during vascular development,
inflammation, lymphohematopoietic stem cell activity, protective
immunity against specific bacteria, allergic asthma, aberrant
tissue-specific responses to the activation of the JNK signal
transduction pathway, cell transformation, memory, apoptosis,
competitive activity-dependent synapse modification at the
neuromuscular synapse, immunological mediation of disease, and
calcium regulation.
[0005] Specific disease states associated with aberrant regulation
of protein kinases include, for example without limitation,
acrocephalo-syndactyly type I, acute myeloid leukemia, AIDS-induced
non-Hodgkin's lymphoma, Alzheimer's disease, amyotrophic lateral
sclerosis, arthritis, asthma, atherosclerosis, atopic dermatitis,
autoimmune diseases, bacterial infection, bladder cancer, cancer of
the breast, cancer of the central nervous system, cancer of the
colon, cancer of the endometrium, cancer of the fallopian tube,
cancer of the gastrointestinal tract, cancer of the ovary, heart
failure, chronic myeloid leukemia, colon carcinoma, colorectal
cancer, chronic obstructive pulmonary disease (COPD), Crouzon
Syndrome, diabetes, diabetic nephropathy, emphysema, endometriosis,
epidermoid cancer, fibrotic disorders, gastrointestinal stromal
tumor (GIST), glomerulonephritis, Graves' disease, head injury,
hepatocellular carcinoma, Hirschsprung's disease, human gliomas,
immunodeficiency diseases, inflammatory disorders, ischemic stroke,
Jackson-Weiss syndrome, leiomyosarcoma, leukemias, lupus nephritis,
malignant melanoma, malignant nephrosclerosis, mastocytosis, mast
cell tumors, melanoma of the colon, MEN2 syndromes, metabolic
disorders, migraine, multiple sclerosis, myeloproliferative
disorders, nephritis, neurodegenerative diseases, neurotraumatic
diseases, non small cell lung cancer, organ transplant rejection,
osteoporosis, pain, Parkinson's disease, Pfeiffer Syndrome,
polycystic kidney disease, primary lymphoedema, prostate cancer,
psoriasis, vascular restenosis, rheumatoid arthritis, dermal and
tissue scarring, selective T-cell defect (STD), severe combined
immunodeficiency (SCID), small cell lung cancer, spinal cord
injury, squamous cell carcinoma, systemic lupus erythematosis,
testicular cancer, thrombotic microangiopathy syndromes, Wegener's
granulomatosis, X-linked agammaglobulinemia, viral infection,
diabetic retinopathy, alopecia, erectile dysfunction, macular
degeneration, chronic lymphocytic leukemia (CLL), myelodysplastic
syndrome (MDS), neurofibromatosis, and tuberous sclerosis.
Accordingly, there is a need in the art for additional compounds
and methods of use thereof for the modulation of receptor protein
kinases.
[0006] This application is related to the following published
patent applications: WO 2004024895, US 20040142864, WO 2004078923,
US 20050170431, WO 2005028624, US 20050164300, and WO 2005062795,
each of which are hereby incorporated by reference herein in their
entireties including all specifications, figures, and tables, and
for all purposes.
SUMMARY OF THE INVENTION
[0007] The present invention concerns compounds active on protein
kinases in general, including, but not limited to, Abl, Akt1, Akt2,
Akt3, ALK, Alk5, B-Raf, Brk, Btk, Cdk2, CDK4, CDK5, CDK6, CHK1,
c-Raf-1, Csk, EGFR, EphA1, EphA2, EphB2, EphB4, Erk2, Fak, FGFR1,
FGFR2, FGFR3, FGFR4, Flt1, Flt3, Flt4, Fms, Frk, Fyn, Gsk3.alpha.,
Gsk3.beta., HCK, Her2/Erbb2, Her4/Erbb4, IGF1R, IKK beta, Irak4,
Itk, Jak1, Jak2, Jak3, Jnk1, Jnk2, Jnk3, Kdr, Kit, LCK, MAP2K1,
MAP2K2, MAP4K4, MAPKAPK2, Met, Mnk1, MLK1, p38, PDGFRA, PDGFRB,
PDPK1, Pim1, Pim2, Pim3, PKC alpha, PKC beta, PKC theta, Plk1,
Pyk2, Ret, ROCK1, ROCK2, Ron, Src, Stk6, Syk, TEC, Tie2, TrkA, Yes,
and/or Zap70, including any mutations of these kinases, and the use
thereof in treating disease and conditions associated with
regulation of the activity of the kinase. In particular, the
invention concerns compounds of Formula I as described below. Thus,
the invention provides novel use of compounds for therapeutic
methods involving modulation of protein kinases, as well as novel
compounds that can be used for therapeutic methods involving
modulation of protein kinases.
[0008] The compounds of Formula I have the following structure:
##STR00001##
all salts, prodrugs, tautomers, and isomers thereof, [0009]
wherein: [0010] R.sup.2, R.sup.4, R.sup.5, and R.sup.6 are
independently selected from the group consisting of hydrogen,
halogen, optionally substituted lower alkyl, optionally substituted
lower alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --NO.sub.2, --CR.sup.aR.sup.bR.sup.26, and --LR.sup.26;
[0011] R.sup.3 is selected from the group consisting of hydrogen,
halogen, optionally substituted lower alkyl, optionally substituted
lower alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --NO.sup.2, --CR.sup.aR.sup.bR.sup.26, -LR.sup.26 and
--A--Ar-L.sub.1-R.sup.24; [0012] A is selected from the group
consisting of --O--, --S--, --CR.sup.aR.sup.b--, --NR.sup.1--,
--C(O)--, --C(S)--, --S(O)--, and --S(O).sub.2--; [0013] R.sup.1 is
selected from the group consisting of hydrogen, lower alkyl,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, --C(O)R.sup.7,
--C(S)R.sup.7, --S(O).sub.2R.sup.7, --C(O)NHR.sup.7,
--C(S)NHR.sup.7, and --S(O).sub.2NHR.sup.7, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein the alkyl chain(s) of lower alkoxy,
lower alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro, further provided, however, that when R.sup.1 is lower
alkyl, any substitution on the lower alkyl carbon bound to the N of
--NR.sup.1-- is fluoro, and wherein cycloalkyl, heterocycloalkyl,
aryl or heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0014] R.sup.7 is selected from the group
consisting of lower alkyl, cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, lower alklylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, provided, however, that any
substitution of the alkyl carbon bound to the N of --C(O)NHR.sup.7,
--C(S)NHR.sup.7 or --S(O).sub.2NHR.sup.7 is fluoro, wherein the
alkyl chain(s) of lower alkoxy, lower alkylthio,mono-alkylamino, or
di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro, and wherein
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more substituents selected from the group
consisting of halogen, --OH, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino; [0015] Ar is
selected from the group consisting of optionally substituted
arylene and optionally substituted heteroarylene; [0016] L at each
occurrence is independently selected from the group consisting of
-(alk).sub.a-S-(alk).sub.b-, -(alk).sub.a-O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(O)-(alk).sub.b-, -(alk).sub.a-C(S)-(alk).sub.b-,
-(alk).sub.a-S(O)-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-OC(O)-(alk).sub.b-, -(alk).sub.a-C(O)O-(alk).sub.b-,
-(alk).sub.a-OC(S)-(alk).sub.b-, -(alk).sub.a-C(S)O-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-, and
-(alk).sub.a-NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-; [0017] a
and b are independently 0 or 1; [0018] alk is C.sub.1-3 alkylene or
C.sub.1-3 alkylene substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkyl, lower alkoxy, lower alkylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, wherein lower alkyl or the
alkyl chain(s) of lower alkoxy, lower alkylthio, mono-alkylamino or
di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro; [0019] L.sub.1
is --(CR.sup.aR.sup.b).sub.v-- or L, wherein v is 1, 2, or 3;
[0020] wherein R.sup.a and R.sup.b at each occurrence are
independently selected from the group consisting of hydrogen,
fluoro, --OH, --NH.sub.2, lower alkyl, lower alkoxy, lower
alklylthio, mono-alkylamino, di-alkylamino, and --NR.sup.8R.sup.9,
wherein the alkyl chain(s) of lower alkyl, lower alkoxy, lower
alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro; or [0021] any two of R.sup.a and R.sup.b on the same or
different carbons combine to form a 3-7 membered monocyclic
cycloalkyl or 5-7 membered monocyclic heterocycloalkyl and any
others of R.sup.a and R.sup.b are independently selected from the
group consisting of hydrogen, fluoro, --OH, --NH.sub.2, lower
alkyl, lower alkoxy, lower alklylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, wherein the alkyl chain(s) of
lower alkyl, lower alkoxy, lower alkylthio, mono-alkylamino, or
di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro, and wherein the
3-7 membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl are optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0022] R.sup.8 and R.sup.9 combine with the
nitrogen to which they are attached to form a 5-7 membered
heterocycloalkyl optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, and
fluoro substituted lower alkylthio; [0023] R.sup.25 at each
occurrence is independently selected from the group consisting of
hydrogen, optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, and optionally substituted heteroaryl;
and [0024] R.sup.24 and R.sup.26 at each occurrence are
independently selected from the group consisting of hydrogen,
provided, however, that hydrogen is not bound to any of S(O),
S(O).sub.2, C(O) or C(S) of L or L.sub.1, optionally substituted
lower alkyl, optionally substituted lower alkenyl, provided,
however, that when R.sup.24 or R.sup.26 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of L or L.sub.1, optionally substituted
lower alkynyl, provided, however, that when R.sup.24 or R.sup.26 is
optionally substituted lower alkynyl, no alkyne carbon thereof is
bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of L or L.sub.1,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, and optionally
substituted heteroaryl.
[0025] The description above of substituents in Formula I includes
descriptions of each combination of the specified substituents,
R.sup.2, R.sup.3, R.sup.4, R.sup.5 and R.sup.6. In some
embodiments, at least one of R.sup.2, R.sup.3, R.sup.4, R.sup.5 and
R.sup.6 is other than hydrogen.
[0026] In some embodiments involving compounds of Formula I,
R.sup.3 and R.sup.6 are hydrogen, or R.sup.3 and R.sup.5 are
hydrogen, or R.sup.2 and R.sup.4 are hydrogen, or R.sup.2 and
R.sup.3 are hydrogen, or R.sup.3 and R.sup.6 are hydrogen, or
R.sup.3 and R.sup.5 are hydrogen, or R.sup.3 and R.sup.4 are
hydrogen, or R.sup.4 and R.sup.6 are hydrogen, or R.sup.4 and
R.sup.5 are hydrogen, or R.sup.5 and R.sup.6 are hydrogen, wherein
the substitutions at the other positions are non-hydrogen. In some
embodiments, R.sup.2, R.sup.3 and R.sup.4 are hydrogen, or R.sup.2,
R.sup.3 and R.sup.5 are hydrogen, or R.sup.2, R.sup.3 and R.sup.6
are hydrogen, or R.sup.2, R.sup.4 and R.sup.6 are hydrogen, or
R.sup.2, R.sup.4 and R.sup.6 are hydrogen, or R.sup.2, R.sup.5 and
R.sup.6 are hydrogen, or R.sup.3, R.sup.4 and R.sup.5 are hydrogen,
or R.sup.3, R.sup.4 and R.sup.6 are hydrogen, or R.sup.3, R.sup.5
and R.sup.6 are hydrogen, or R.sup.4, R.sup.5 and R.sup.6 are
hydrogen, wherein the substitutions at the other positions are
non-hydrogen. In some embodiments, the compounds are
mono-substituted with non-hydrogen at one of R.sup.2, R.sup.3,
R.sup.4, R.sup.5 or R.sup.6 (i.e. hydrogen at the other four
positions). In some embodiments, compounds of Formula I have
non-hydrogen substitution at R.sup.3; non-hydrogen substitution at
R.sup.4; non-hydrogen substitution at R.sup.3; non-hydrogen
substitution at R.sup.3 and R.sup.4; non-hydrogen substitution at
R.sup.3 and R.sup.5. In some embodiments, the substitutions as
listed are the only substitutions; the substitutions as listed are
combined with R.sup.2 and R.sup.6 as H; the substitutions as listed
are combined with substitution at one other of the substitution
positions shown in Formula I. The compounds of Formula I, and all
sub-embodiments detailed herein, may be used to treat a subject
suffering from or at risk for any of the protein kinase mediated
diseases or conditions contemplated herein.
[0027] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ia:
##STR00002##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 is a bond, --CR.sup.aR.sup.b--, -A-Ar-L.sub.1-, or L, and
Y.sup.5 is a bond, --CR.sup.aR.sup.b--, or L, and each R.sup.27 is
independently halogen, provided that Y.sup.3 or Y.sup.5 is a bond,
or R.sup.26, provided, however, that neither of Y.sup.3R.sup.27 and
Y.sup.5R.sup.27 are hydrogen, wherein R.sup.a, R.sup.b, L, L.sub.1,
A, Ar and R.sup.26 are as defined with reference to Formula I.
[0028] In some embodiments of compounds of Formula Ia, Y.sup.3 and
Y.sup.5 are bonds. In some embodiments, Y.sup.3 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.3 and Y.sup.5 are independently L. In some embodiments,
Y.sup.3 and Y.sup.5 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.3 is a bond, and Y.sup.5 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.3 is a bond, and Y.sup.5 is L. In
some embodiments, Y.sup.3 is a bond, and Y.sup.5 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.5 is a bond, and
Y.sup.3 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.5
is a bond, and Y.sup.3 is L. In some embodiments, Y.sup.5 is a
bond, and Y.sup.3 is --CR.sup.aR.sup.b--.
[0029] In some embodiments of any of the above embodiments of
compounds of Formula Ia, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3 or Y.sup.5 is a bond.
In some embodiments, Y.sup.5 is a bond, --CR.sup.aR.sup.b--, or L,
Y.sup.3 is --CR.sup.aR.sup.b--, --O--, --S--, --NR.sup.25--,
--C(O)--, --C(S)--, --S(O)--, or --S(O).sub.2--, wherein R.sup.25
is as defined for Formula I, and each R.sup.27 is independently
R.sup.26 or Y.sup.5R.sup.27 is halogen; in further embodiments,
Y.sup.3 is --CR.sup.aR.sup.b-- or --C(O)--; in further embodiments,
--CR.sup.3R.sup.b-- is --CH.sub.2--. In some embodiments, Y.sup.3
is a bond, --CR.sup.aR.sup.b--, or L, R.sup.27 bound to Y.sup.5 is
R.sup.26 or Y.sup.5R.sup.27 is halogen, Y.sup.3 is
--CR.sup.aR.sup.b-- or --C(O)--, and R.sup.27 bound to Y.sup.3 is
optionally substituted aryl or optionally substituted
heteroaryl.
[0030] In some embodiments of any of the above embodiments of
compounds of Formula Ia, Y.sup.3R.sup.27 is -A-Ar-L.sub.1-R.sup.24,
wherein A, Ar, and L.sub.1 are as defined for Formula I, and
R.sup.24 is substituted methyl, optionally substituted C.sub.2-6
alkyl, optionally substituted lower alkenyl, provided, however,
that when R.sup.24 is optionally substituted lower alkenyl, no
alkene carbon thereof is bound to N, S, O, S(O), S(O).sub.2, C(O)
or C(S) of L.sub.1, optionally substituted lower alkynyl, provided,
however, that when R.sup.24 is optionally substituted lower
alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of L.sub.1, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl, provided,
however, that -A-Ar-L.sub.1-R.sup.24 is not
##STR00003##
wherein
##STR00004##
indicates the point of attachment to the 3 position of the
azaindole ring; in further embodiments, R.sup.24 is optionally
substituted C.sub.2-6 alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, or substituted methyl,
wherein methyl is substituted with optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl; in further
embodiments, when Ar is optionally substituted heteroarylene, the
heteroarylene ring is not a five or six membered ring having the
structure
##STR00005##
wherein
##STR00006##
indicates the point of attachment to A and
##STR00007##
indicates the point of attachment to L.sub.1, and wherein the
indicated N is either .dbd.N-- or --N.dbd., and wherein each Q is
independently a heteroaryl ring atom that may be optionally
substituted. The term "heteroaryl ring atom" refers to any atom
that can be part of a heteroaryl ring structure (i.e., C, N, O, or
S).
[0031] In some embodiments of any of the above embodiments of
compounds of Formula Ia, Y.sup.3 and Y.sup.5 are independently
--O--, --S--, --CR.sup.aR.sup.b--, --NR.sup.25--, --C(O)--,
--C(S)--, --S(O)--, or --S(O).sub.2--, where R.sup.a, R.sup.b and
R.sup.25 are as defined in Formula I, and each R.sup.27 is
independently optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, Y.sup.3 is --O--, --NR.sup.25--, or
--S(O).sub.2--, preferably wherein R.sup.25 is hydrogen or lower
alkyl, and R.sup.27 bound to Y.sup.3 is optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl, preferably
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, Y.sup.3 is --CR.sup.aR.sup.b--, or
--C(O)--, preferably --CH.sub.2-- or --C(O)--, Y.sup.5 is --O--,
--NR.sup.25--, or --S(O).sub.2--, preferably --NR.sup.25--, wherein
R.sup.25 is hydrogen or lower alkyl, and each R.sup.27 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, preferably optionally substituted aryl, or
optionally substituted heteroaryl, provided, however, that
Y.sup.3R.sup.27 is not
##STR00008##
wherein
##STR00009##
indicates the bond to the 3 position of the 7-azaindole ring, and
Y.sup.5R.sup.7 is not
##STR00010##
wherein
##STR00011##
indicates the bond to the 5 position of the 7-azaindole ring, i.e.
the compound is not
(3-hydroxy-phenyl)-(5-phenylamino-1H-pyrrol[2,3-b]pyridin-3-yl)-methanone-
; in further embodiments, when R.sup.27 bound to Y.sup.3 is
optionally substituted heteroaryl, the heteroaryl ring is not a
five or six membered ring having the structure
##STR00012##
wherein
##STR00013##
indicates the point of attachment to Y.sup.3, and wherein the
indicated N is either .dbd.N-- or --N.dbd., and wherein each Q is
independently a heteroaryl ring atom that may be appropriately
optionally substituted and wherein R.sup.200 is other than
hydrogen. The term "other than hydrogen" and like terms refer to
substituents contemplated herein which are not hydrogen. For
example without limitation, if substituent R.sup.ex were defined as
selected from the group consisting of hydrogen and optionally
substituted lower alkyl, then the phrase "R.sup.ex is other than
hydrogen" would contemplate only optionally substituted lower
alkyl, i.e., all options of the substituent, excluding
hydrogen.
[0032] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ib:
##STR00014##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 and Y.sup.4 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 , independently halogen, provided that
Y.sup.3 or Y.sup.4 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.3R.sup.27 and Y.sup.4R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0033] In some embodiments of compounds of Formula Ib, Y.sup.3 and
Y.sup.4 are bonds. In some embodiments, Y.sup.3 and Y.sup.4 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.3 and Y.sup.4 are independently L. In some embodiments,
Y.sup.3 and Y.sup.4 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.3 is a bond and Y.sup.4 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.3 is a bond and Y.sup.4 is L. In
some embodiments, Y.sup.3 is a bond and Y.sup.4 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.4 is a bond, and
Y.sup.3 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.4
is a bond and Y.sup.3 is L. In some embodiments, Y.sup.4 is a bond
and Y.sup.3 is --CR.sup.aR.sup.b--.
[0034] In some embodiments of any of the above embodiments of
compounds of Formula Ib, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3 or Y.sup.4 is a bond.
In some embodiments, Y.sup.4 is a bond, --CR.sup.aR.sup.b--, or L,
Y.sup.3 is --CR.sup.aR.sup.b--, --O--, --S--, --NR.sup.25--,
--C(O)--, --C(S)--, S(O)--, or --S(O).sub.2--, wherein R.sup.25 is
as defined for Formula I, and R.sup.27 is independently R.sup.26 or
Y.sup.4R.sup.27 is halogen; in further embodiments, Y.sup.3 is
--CR.sup.aR.sup.b-- or --C(O)--; in further embodiments,
--CR.sup.aR.sup.b-- is --CH.sub.2--. In some embodiments, Y.sup.4
is a bond, --CR.sup.aR.sup.b--, or L, R.sup.27 bound to Y.sup.4 is
R.sup.26 or Y.sup.4R.sup.27 is halogen, Y.sup.3 is
--CR.sup.aR.sup.b-- or --C(O)--, and R.sup.27 bound to Y.sup.3 is
optionally substituted aryl or optionally substituted
heteroaryl.
[0035] In some embodiments of any of the above embodiments of
compounds of Formula Ib, Y.sup.3 and Y.sup.4 are independently
--O--, --S--, --CR.sup.aR.sup.b--, --NR.sup.25--, --C(O)--,
--C(S)--, --S(O)--, or --S(O).sub.2--, where R.sup.a, R.sup.b and
R.sup.25 are as defined in Formula I, and each R.sup.27 is
independently optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in a further embodiment Y.sup.4 is --O--, --NR.sup.2513 , or
--S(O).sub.2--, preferably wherein R.sup.25 is hydrogen or lower
alkyl, and R.sup.27 bound to Y.sup.3 is optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl, preferably
optionally substituted aryl, or optionally substituted heteroaryl;
in a further embodiment Y.sup.3 is --CR.sup.aR.sup.b--, or
--C(O)--, preferably --CH.sub.2-- or --C(O)--, Y.sup.4 is --O--,
--NR.sup.25--, or --S(O).sub.2--, preferably --NR.sup.25--, wherein
R.sup.25 is hydrogen or lower alkyl, and each R.sup.27 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, preferably optionally substituted aryl, or
optionally substituted heteroaryl.
[0036] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ic:
##STR00015##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 and Y.sup.6 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.3 or Y.sup.6 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.3R.sup.27 and Y.sup.6R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0037] In some embodiments of compounds of Formula Ic, Y.sup.3 and
Y.sup.6 are bonds. In some embodiments, Y.sup.3 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.3 and Y.sup.6 are independently L. In some embodiments,
Y.sup.3 and Y.sup.6 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.3 is a bond, and Y.sup.6 is --CR.sup.3R.sup.b--
or L. In some embodiments, Y.sup.3 is a bond, and Y.sup.6 is L. In
some embodiments, Y.sup.3 is a bond, and Y.sup.6 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.6 is a bond, and
Y.sup.3 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.6
is a bond, and Y.sup.3 is L. In some embodiments, Y.sup.6 is a
bond, and Y.sup.3 is --CR.sup.aR.sup.b--.
[0038] In some embodiments of any of the above embodiments of
compounds of Formula Ic, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3 or Y.sup.6 is a
bond.
[0039] In some embodiments of any of the above embodiments of
compounds of Formula Ia, Y.sup.3 and Y.sup.6 are independently
--O--, --S--, --CR.sup.3R.sup.b--, --NR.sup.25--, --C(O)--,
--C(S)--, --S(O)--, or --S(O).sub.2--, where R.sup.a, R.sup.b and
R.sup.25 are as defined in Formula I, and each R.sup.27 is
independently optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, Y.sup.6 is --O--, --NR.sup.25--, or
--S(O).sub.2--, preferably wherein R.sup.25 is hydrogen or lower
alkyl, and R.sup.27 bound to Y.sup.3 is optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl, preferably
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, Y.sup.3 is --CR.sup.3R.sup.b--, or
--C(O)--, preferably --CH.sub.2-- or --C(O)--, Y.sup.6 is --O--,
--NR.sup.25--, or --S(O).sub.2--, preferably --NR.sup.25--, wherein
R.sup.25 is hydrogen or lower alkyl, and each R.sup.27 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, preferably optionally substituted aryl, or
optionally substituted heteroaryl.
[0040] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Id:
##STR00016##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 and Y.sup.2 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.3 or Y.sup.2 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.2R.sup.27 and Y.sup.3R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0041] In some embodiments of compounds of Formula Id, Y.sup.3 and
Y.sup.2 are bonds. In some embodiments, Y.sup.3 and Y.sup.2 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.3 and Y.sup.2 are independently L. In some embodiments,
Y.sup.3 and Y.sup.2 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.3 is a bond, and Y.sup.2 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.3 is a bond, and Y.sup.2 is L. In
some embodiments, Y.sup.3 is a bond, and Y.sup.2 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.2 is a bond, and
Y.sup.3 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.2
is a bond, and Y.sup.3 is L. In some embodiments, Y.sup.2 is a
bond, and Y.sup.3 is --CR.sup.aR.sup.b--.
[0042] In some embodiments of any of the above embodiments of
compounds of Formula Id, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3 or Y.sup.2 is a
bond.
[0043] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ie:
##STR00017##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.4 and Y.sup.2 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.4 or Y.sup.2 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.2R.sup.27 and Y.sup.4R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0044] In some embodiments of compounds of Formula Ie, Y.sup.4 and
Y.sup.2 are bonds. In some embodiments, Y.sup.4 and Y.sup.2 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.4 and Y.sup.2 are independently L. In some embodiments,
Y.sup.4 and Y.sup.2 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.4 is a bond, and Y.sup.2 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.4 is a bond, and Y.sup.2 is L. In
some embodiments, Y.sup.4 is a bond, and Y.sup.2 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.2 is a bond and
Y.sup.4 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.2
is a bond, and Y.sup.4 is L. In some embodiments, Y.sup.2 is a
bond, and Y.sup.4 is --CR.sup.aR.sup.b--.
[0045] In some embodiments of any of the above embodiments of
compounds of Formula Id, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.4 or Y.sup.2 is a
bond.
[0046] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
If:
##STR00018##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.5 and Y.sup.2 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.5 or Y.sup.2 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.2R.sup.27 and Y.sup.5R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0047] In some embodiments of compounds of Formula If, Y.sup.5 and
Y.sup.2 are bonds. In some embodiments, Y.sup.5 and Y.sup.2 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.5 and Y.sup.2 are independently L. In some embodiments,
Y.sup.5 and Y.sup.2 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.5 is a bond, and Y.sup.2 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.5 is a bond, and Y.sup.2 is L. In
some embodiments, Y.sup.5 is a bond, and Y.sup.2 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.2 is a bond, and
Y.sup.5 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.2
is a bond, and Y.sup.5 is L. In some embodiments, Y.sup.2 is a
bond, and Y.sup.5 is --CR.sup.aR.sup.b--.
[0048] In some embodiments of any of the above embodiments of
compounds of Formula If, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.5 or Y.sup.2 is a
bond.
[0049] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ig:
##STR00019##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.6 and Y.sup.2 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.6 or Y.sup.2 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.2R.sup.27 and Y.sup.6R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0050] In some embodiments of compounds of Formula Ig, Y.sup.6 and
Y.sup.2 are bonds. In some embodiments, Y.sup.6 and Y.sup.2 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.6 and Y.sup.2 are independently L. In some embodiments,
Y.sup.6 and Y.sup.2 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.6 is a bond, and Y.sup.2 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.6 is a bond, and Y.sup.2 is L. In
some embodiments, Y.sup.6 is a bond, and Y.sup.2 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.2 is a bond, and
Y.sup.6 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.2
is a bond, and Y.sup.6 is L. In some embodiments, Y.sup.2 is a
bond, and Y.sup.6 is --CR.sup.aR.sup.b--.
[0051] In some embodiments of any of the above embodiments of
compounds of Formula Ig, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.6 or Y.sup.2 is a
bond.
[0052] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ih:
##STR00020##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.6 and Y.sup.4 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.6 or Y.sup.4 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.4R.sup.27 and Y.sup.6R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0053] In some embodiments of compounds of Formula Ih, Y.sup.6 and
Y.sup.4 are bonds. In some embodiments, Y.sup.6 and Y.sup.4 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.6 and Y.sup.4 are independently L. In some embodiments,
Y.sup.6 and Y.sup.4 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.6 is a bond, and Y.sup.4 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.6 is a bond, and Y.sup.4 is L. In
some embodiments, Y.sup.6 is a bond, and Y.sup.4 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.4 is a bond and
Y.sup.6 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.4
is a bond, and Y.sup.6 is L. In some embodiments, Y.sup.4 is a
bond, and Y.sup.6 is --CR.sup.aR.sup.b--.
[0054] In some embodiments of any of the above embodiments of
compounds of Formula Ih, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.6 or Y.sup.4 is a
bond.
[0055] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ii:
##STR00021##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.6 and Y.sup.5 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.6 or Y.sup.5 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.5R.sup.27 and Y.sup.6R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0056] In some embodiments of compounds of Formula Ii, Y.sup.6 and
Y.sup.5 are bonds. In some embodiments, Y.sup.6 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.6 and Y.sup.5 are independently L. In some embodiments,
Y.sup.6 and Y.sup.5 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.6 is a bond, and Y.sup.5 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.6 is a bond, and Y.sup.5 is L. In
some embodiments, Y.sup.6 is a bond, and Y.sup.5 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.5 is a bond, and
Y.sup.6 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.5
is a bond, and Y.sup.6 is L. In some embodiments, Y.sup.5 is a
bond, and Y.sup.6 is --CR.sup.aR.sup.b--.
[0057] In some embodiments of any of the above embodiments of
compounds of Formula Ii, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.6 or Y.sup.5 is a
bond.
[0058] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ij:
##STR00022##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.4 and Y.sup.5 are independently a bond, --CR.sup.aR.sup.b--,
or L, and each R.sup.27 is independently halogen, provided that
Y.sup.4 or Y.sup.5 is a bond, or R.sup.26, provided, however, that
neither of Y.sup.4R.sup.27 and Y.sup.5R.sup.27 are hydrogen,
wherein R.sup.a, R.sup.b, L and R.sup.26 are as defined with
reference to Formula I.
[0059] In some embodiments of compounds of Formula Ij, Y.sup.4 and
Y.sup.5 are bonds. In some embodiments, Y.sup.4 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments,
Y.sup.4 and Y.sup.5 are independently L. In some embodiments,
Y.sup.4 and Y.sup.5 are independently --CR.sup.aR.sup.b--. In some
embodiments, Y.sup.4 is a bond, and Y.sup.5 is --CR.sup.aR.sup.b--
or L. In some embodiments, Y.sup.4 is a bond, and Y.sup.5 is L. In
some embodiments, Y.sup.4 is a bond, and Y.sup.5 is
--CR.sup.aR.sup.b--. In some embodiments, Y.sup.5 is a bond, and
Y.sup.4 is --CR.sup.aR.sup.b-- or L. In some embodiments, Y.sup.5
is a bond, and Y.sup.4 is L. In some embodiments, Y.sup.5 is a
bond, and Y.sup.4 is --CR.sup.aR.sup.b--.
[0060] In some embodiments of any of the above embodiments of
compounds of Formula Ij, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.4 or Y.sup.5 is a
bond.
[0061] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
Ik:
##STR00023##
all salts, prodrugs, tautomers, and isomers thereof, wherein
R.sup.26 is as defined in Formula I, and Y is selected from the
group consisting of a bond; --CR.sup.aR.sup.b-- and L, where
R.sup.a, R.sup.b and L are as defined with reference to Formula I,
provided that YR.sup.26 is not hydrogen.
[0062] In some embodiments of compounds of Formula Ik, Y is
-(alk).sub.a-S-(alk).sub.b-, -(alk).sub.a-O-(alk).sub.b-,
-(alk).sub.a-OC(O)-(alk).sub.b-, -(alk).sub.a-C(O)O-(alk).sub.b-,
-(alk).sub.a-OC(S)-(alk).sub.b-, -(alk).sub.a-C(S)O-(alk).sub.b-,
-(alk).sub.a-C(O)-(alk).sub.b-, -(alk).sub.a-C(S)-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O)-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25S(O).sub.2-(alk).sub.b-, or
-(alk).sub.a-NR.sup.25 S(O).sub.2NR.sup.25-(alk).sub.b-, wherein
alk, a, b, and R.sup.25 are as defined for Formula I.
[0063] In some embodiments of compounds of Formula Ik, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--C(O)O-(alk).sub.b-, --OC(S)-(alk).sub.b-, --C(S)O-(alk).sub.b-,
--C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)O(alk)h-,
--NR.sup.25C(S)O-(alk).sub.b-, --NR.sup.25S(O).sub.2-(alk).sub.b-,
or --NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0064] In some embodiments of compounds of Formula Ik, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--OC(S)-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --S(O)-(alk).sub.b-,
--S(O).sub.2-(alk).sub.b-, S(O).sub.2NR.sup.25-(alk).sub.b-,
--NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0065] In some embodiments of compounds of Formula Ik, R.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl. In some embodiments, Y is --O--, --S--,
--NR.sup.25--, --C(O)--, --C(S)--, --S(O)--, or --S(O).sub.2-- and
R.sup.26 is optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl. In some embodiments, Y is
--NR.sup.25--, preferably wherein R.sup.25 is hydrogen or lower
alkyl, preferably wherein Y is --NH--; in further embodiments,
R.sup.26 is optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, lower alkyl is substituted with optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, heteroaryl is monocyclic. In some
embodiments, Y is --NH--; in further embodiments, R.sup.26 is
substituted phenyl or optionally substituted heteroaryl, provided
that heteroaryl is monocyclic.
[0066] In some embodiments of any of the above embodiments of
compounds of Formula Ik, YR.sup.26 is not optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl,
--C.ident.CCH.sub.2N(CH.sub.3).sub.2, --C(O)H,
--CH.sub.2N(CH.sub.3).sub.2, --C(O)OCH.sub.3, --CH.sub.2OH, --OH,
--OCH.sub.3, --NHCH.sub.2CH.dbd.CH.sub.2, --NHCH.sub.3,
--NHCH.sub.2CH.sub.3, --NHphenyl,
##STR00024##
wherein
##STR00025##
indicates the bond attached to the 5-position of the 7-azaindole
ring; and when Y is --O-- or --NR.sup.25--, then R.sup.26 is
not
##STR00026##
which is optionally substituted, wherein
##STR00027##
indicates the bond attached to Y; and when Y is --O--, then
R.sup.26 is not
##STR00028##
which is optionally substituted, wherein
##STR00029##
indicates the bond attached Y.
[0067] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
Im:
##STR00030##
all salts, prodrugs, tautomers, and isomers thereof, wherein
R.sup.26 and Y are as defined for Formula Ik.
[0068] In some embodiments of compounds of Formula Im, Y is
--OC(O)-(alk).sub.b-, --C(O)O(alk).sub.b-, --OC(S)-(alk).sub.b-,
--C(S)O-(alk).sub.b-, --C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0069] In other embodiments of compounds of Formula Im, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--C(O)O-(alk).sub.b-, --OC(S)-(alk).sub.b-, --C(S)O-(alk).sub.b-,
--C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.a--,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0070] In other embodiments of compounds of Formula Im, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(Q)-(alk).sub.b-,
--OC(S)-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0071] In some embodiments of compounds of Formula Im, R.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl. In some embodiments, Y is --O--, --S--,
--NR.sup.25--, --C(O)--, --C(S)--, --S(O)--, or --S(O).sub.2-- and
R.sup.26 is optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl. In some embodiments, Y is
--NR.sup.25--, preferably wherein R.sup.25 is hydrogen or lower
alkyl, preferably wherein Y is --NH--; in further embodiments,
R.sup.26 is optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, lower alkyl is substituted with optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, heteroaryl is monocyclic.
[0072] In some embodiments of any of the above embodiments of
compounds of Formula Im, when Y is --CH.sub.2NH--, then R.sup.26 is
not optionally substituted thiophene or optionally substituted
pyridine; when Y is --O-- or --NH--, then R.sup.26 is not
optionally substituted bicyclic heteroaryl; when Y is --O--, then
R.sup.26 is not optionally substituted phenyl; when Y is --NH-- or
--N(CH.sub.3)-- and R.sup.26 is substituted phenyl, the phenyl is
not substituted by halogen ortho to Y and optionally substituted
amine para to Y, Y is not --NH--C(O)-- or --C(O)--NH--, and
YR.sup.26 is not optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
--CH.dbd.CH.sub.2, --C(O)OH, --C(O)OCH.sub.3, --C(O)OtBu, --OH,
--OCH.sub.3, --NHCH.sub.2CH.dbd.CH.sub.2, --N(CH.sub.3).sub.2,
--NH.sub.2, --NHCH.sub.2C(O)OCH.sub.2CH.sub.3,
--N(CH.sub.3)phenyl,
##STR00031##
wherein
##STR00032##
indicates the bond attached to the 4-position of the 7-azaindole
ring.
[0073] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
In:
##STR00033##
all salts, prodrugs, tautomers, and isomers thereof, wherein
R.sup.26 and Y are as defined for Formula Ik.
[0074] In other embodiments of compounds of Formula In, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--C(O)O-(alk).sub.b-, --OC(S)-(alk).sub.b-, --C(S)O-(alk).sub.b-,
--C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.a--,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25 C(O)NR.sup.25-(alk).sub.b,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0075] In other embodiments of compounds of Formula In, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--OC(S)-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --S(O)-(alk).sub.b-,
--S(O).sub.2-(alk).sub.b-, --S(O).sub.2NR.sup.25-(alk).sub.b-,
--NR.sup.25-(alk).sub.b-, --NR--C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0076] In some embodiments of compounds of Formula In, R.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl. In some embodiments, Y is --NR.sup.25--,
preferably wherein R.sup.25 is hydrogen or lower alkyl, preferably
wherein Y is --NH--; in further embodiments, R.sup.26 is optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl; in further embodiments,
lower alkyl is substituted with optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl.
[0077] In some embodiments of any of the above embodiments of
compounds of Formula In, compounds are excluded where Y is --O--
and R.sup.26 is optionally substituted
##STR00034##
wherein
##STR00035##
indicates the bond attached to Y; and where YR.sup.26 is
--NH.sub.2, --CH.sub.3, --OC(O)phenyl,
##STR00036##
wherein
##STR00037##
indicates the bond attached to the 6-position of the 7-azaindole
ring.
[0078] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
Io:
##STR00038##
all salts, prodrugs, tautomers, and isomers thereof, wherein
R.sup.26 and Y are as defined for Formula Ik.
[0079] In other embodiments of compounds of Formula Io, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--C(O)O-(alk).sub.b-, --OC(S)-(alk).sub.b-, --C(S)O-(alk).sub.b-,
--C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.2-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0080] In other embodiments of compounds of Formula Io, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--OC(S)-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --S(O)-(alk).sub.b-,
--S(O).sub.2-(alk).sub.b-, --S(O).sub.2NR.sup.25-(alk).sub.b-,
--NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0081] In some embodiments of compounds of Formula Io, R.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl. In some embodiments, Y is --NR.sup.25--,
preferably wherein R.sup.25 is hydrogen or lower alkyl, preferably
wherein Y is --NH--; in further embodiments, R.sup.26 is optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl; in further embodiments,
lower alkyl is substituted with optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl.
[0082] In some embodiments of any of the above embodiments of
compounds of Formula Io, compounds are excluded when --YR.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, optionally
substituted heteroaryl, --CH.sub.2NR'R'', NR'R'' is optionally
substituted heterocycloalkyl or optionally substituted heteroaryl,
--C(O)NR'R'', wherein NR'R'' is optionally substituted
heterocycloalkyl or optionally substituted heteroaryl, or R' is H
and R'' is optionally substituted cycloalkyl, optionally
substituted aryl or optionally substituted heteroaryl, optionally
substituted --CH.dbd.CH.sub.2, --CH.sub.3,
--CH.sub.2CH.sub.2NHCH.sub.3, --CH.sub.2CH(NH.sub.2)C(O)OH,
--CH.sub.2CH(NH.sub.2)C(O)OCH.sub.3,
--CH.sub.2CH(C(O)OH)NHCH.sub.3, --C(O)C(O)OCH.sub.3,
--CH.sub.2C(O)OCH.sub.2CH.sub.3, --CH.sub.2C(O)NH.sub.2,
--CH.sub.2CN, --NH.sub.2, --N(CH.sub.3).sub.2, --SCH.sub.3,
--N.dbd.C(CH.sub.3)NHOAc, --C(O)CCl.sub.3, --C(O)OCH.sub.3,
--C(O)CH.sub.2Br, --C(O)NH.sub.2, --C(S)NH.sub.2,
--CH.sub.2NH-thiophene, wherein thiophene is optionally
substituted,
##STR00039##
wherein the phenyl ring is optionally substituted,
##STR00040## ##STR00041## ##STR00042##
wherein the ring is optionally substituted at D.sub.1, D.sub.2 and
D.sub.3,
##STR00043##
wherein the ring is optionally substituted at D.sub.1, D.sub.2 and
D.sub.3,
##STR00044## ##STR00045## ##STR00046##
##STR00047##
when R is H, F, Cl, CH.sub.3, CF.sub.3 or OCH.sub.3,
##STR00048##
when R is H, 4-F, 4-CF.sub.3 or 3-F, or
##STR00049##
when R' is OCH.sub.3 or CH.sub.3 and R is CF.sub.3, or when R' is
CH.sub.3 and R is Cl, wherein
##STR00050##
indicates the bond attached to the 3-position of the 7-azaindole
ring.
[0083] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
Ip:
##STR00051##
all salts, prodrugs, tautomers, and isomers thereof, wherein
R.sup.26 and Y are as defined for Formula Ik.
[0084] In other embodiments of compounds of Formula Ip, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--C(O)O-(alk).sub.b-, --OC(S)-(alk).sub.b-, --C(S)O-(alk).sub.b,
--C(O)-(alk).sub.b-, --C(S)-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O)-(alk).sub.b-, --S(O).sub.2-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0085] In other embodiments of compounds of Formula Ip, Y is
--S-(alk).sub.b-, --O-(alk).sub.b-, --OC(O)-(alk).sub.b-,
--OC(S)-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.2-(alk).sub.b-, --S(O)-(alk).sub.b-,
--S(O).sub.2-(alk).sub.b-, --S(O).sub.2NR.sup.25-(alk).sub.b-,
--NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-, --NR.sup.25
C(S)NR.sup.25-((alk).sub.b-, --NR.sup.25C(O)O-(alk).sub.b-,
--NR.sup.25C(S)O-(alk).sub.b-, --NR.sup.25S(O).sub.2-(alk).sub.b,
or --NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein alk, b and
R.sup.25 are as defined for Formula I.
[0086] In some embodiments of compounds of Formula Ip, R.sup.26 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl. In some embodiments, Y is --NR.sup.25--,
preferably wherein R.sup.25 is hydrogen or lower alkyl, preferably
wherein Y is --NH--; in further embodiments, R.sup.26 is optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl; in further
embodiments, lower alkyl is substituted with optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl.
[0087] In some embodiments of any of the above embodiments of
compounds of Formula Ip above, compounds are excluded when
YR.sup.26 is --CH.sub.3, optionally substituted aryl, optionally
substituted cycloalkyl,
##STR00052##
wherein
##STR00053##
indicates the bond attached to the 2-position of the 7-azaindole
ring.
[0088] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iq:
##STR00054##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.3 and Y.sup.4 are independently a bond,
--CR.sup.aR.sup.b-- or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.4 is a bond, or
R.sup.26 provided, however, that none of Y.sup.2R.sup.27,
Y.sup.3R.sup.27, and Y.sup.4R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0089] In some embodiments of compounds of Formula Iq, Y.sup.2,
Y.sup.3 and Y.sup.4 are bonds. In some embodiments, Y.sup.2,
Y.sup.3 and Y.sup.4 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.2, Y.sup.3 and Y.sup.4 are independently L.
In some embodiments, one of Y.sup.2, Y.sup.3 and Y.sup.4 is a bond,
and the others are independently --CR.sup.3R.sup.b-- or L. In some
embodiments, one of Y.sup.2, Y.sup.3 and Y.sup.4 is a bond, and the
others are independently L. In some embodiments, two of Y.sup.2,
Y.sup.3 and Y.sup.4 are bonds, and the other is --CR.sup.aR.sup.b--
or L. In some embodiments, two of Y.sup.2, Y.sup.3 and Y.sup.4 are
bonds and the other is L.
[0090] In some embodiments of any of the above embodiments of
compounds, of Formula Iq, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.4
is a bond.
[0091] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ir:
##STR00055##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.3 and Y.sup.5 are independently a bond,
--CR.sup.aR.sup.b-- or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.5 is a bond, or
R.sup.26, provided, however, that none of Y.sup.2R.sup.27,
Y.sup.3R.sup.27, and Y.sup.5R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0092] In some embodiments of compounds of Formula Ir, Y.sup.2,
Y.sup.3 and Y.sup.5 are bonds. In some embodiments, Y.sup.2,
Y.sup.3 and Y.sup.5 are independently --CR.sup.aR.sup.b-- or L. In.
some embodiments, Y.sup.2, Y.sup.3 and Y.sup.5 are independently L.
In some embodiments, any one of Y.sup.2, Y.sup.3 and Y.sup.5 is a
bond, and the remaining of Y.sup.2, Y.sup.3 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.2, Y.sup.3 and Y.sup.5 is a bond, and the remaining of
Y.sup.2, Y.sup.3 and Y.sup.5 are independently L. In some
embodiments, any two of Y.sup.2, Y.sup.3 and Y.sup.5 are bonds, and
the remaining of Y.sup.2, Y.sup.3 and Y.sup.5 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two, of Y.sup.2,
Y.sup.3 and Y.sup.5 are bonds, and the remaining of Y.sup.2,
Y.sup.3 and Y.sup.5 is L.
[0093] In some embodiments of any of the above embodiments of
compounds of Formula Ir, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.5
is a bond.
[0094] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Is:
##STR00056##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.3 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.2R.sup.27,
Y.sup.3R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0095] In some embodiments of compounds of Formula Is, Y.sup.2,
Y.sup.3 and Y.sup.6 are bonds. In some embodiments, Y.sup.2,
Y.sup.3 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.2, Y.sup.3 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.2, Y.sup.3 and Y.sup.6 is a
bond, and the remaining of Y.sup.2, Y.sup.3 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.2, Y.sup.3 and Y.sup.6 is a bond, and the remaining of
Y.sup.2, Y.sup.3 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.2, Y.sup.3 and Y.sup.6 are bonds, and
the remaining of Y.sup.2, Y.sup.3 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.2
Y.sup.3 and Y.sup.6 are bonds, and the remaining of Y.sup.2,
Y.sup.3 and Y.sup.6 is L.
[0096] In some embodiments of any of the above embodiments of
compounds of Formula Is, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.3 or Y.sup.6
is a bond.
[0097] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
It:
##STR00057##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.4 and Y.sup.5 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.4 or Y.sup.5 is a bond, or
R.sup.26, provided, however, that none of Y.sup.2R.sup.27,
Y.sup.4R.sup.27, and Y.sup.5R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0098] In some embodiments of compounds of Formula It, Y.sup.2,
Y.sup.4 and Y.sup.5 are bonds. In some embodiments, Y.sup.2,
Y.sup.4 and Y.sup.5 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.2, Y.sup.4 and Y.sup.5 are independently L.
In some embodimcnts,any one of Y.sup.2, Y.sup.4 and Y.sup.5 is a
bond, and the remaining of Y.sup.2, Y.sup.4 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.2, Y.sup.4 and Y.sup.5 is a bond, and the remaining of
Y.sup.2, Y.sup.4 and Y.sup.5 are independently L. In some
embodiments, any two of Y.sup.2, Y.sup.4 and Y.sup.5 are bonds, and
the remaining of Y.sup.2, Y.sup.4 and Y.sup.5 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.2,
Y.sup.4 and Y.sup.5 are bonds, and the remaining of Y.sup.2,
Y.sup.4 and Y.sup.5 is L.
[0099] In some embodiments of any of the above embodiments of
compounds of Formula It, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.4 or Y.sup.5
is a bond.
[0100] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iu:
##STR00058##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.4 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.4 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.2R.sup.27,
Y.sup.4R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0101] In some embodiments of compounds of Formula Iu, Y.sup.2,
Y.sup.4 and Y.sup.6 are bonds. In some embodiments, Y.sup.2,
Y.sup.4 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.2, Y.sup.4 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.2, Y.sup.4 and Y.sup.6 is a
bond, and the remaining of Y.sup.2, Y.sup.4 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.2, Y.sup.4 and Y.sup.6 is a bond, and the remaining of
Y.sup.2, Y.sup.4 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.2, Y.sup.4 and Y.sup.6 are bonds, and
the remaining of Y.sup.2, Y.sup.4 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.2,
Y.sup.4 and Y.sup.6 are bonds, and the remaining of Y.sup.2,
Y.sup.4 and Y.sup.6 is L.
[0102] In some embodiments of any of the above embodiments of
compounds of Formula Iu, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.4 or Y.sup.6
is a bond.
[0103] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iv:
##STR00059##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.2 Y.sup.5 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.2, Y.sup.5 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.2R.sup.27,
Y.sup.5R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0104] In some embodiments of compounds of Formula Iv, Y.sup.2,
Y.sup.5 and Y.sup.6 are bonds. In some embodiments, Y.sup.2,
Y.sup.5 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.2, Y.sup.3 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.2, Y.sup.5 and Y.sup.6 is a
bond, and the remaining of Y.sup.2, Y.sup.5 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.2, Y.sup.5 and Y.sup.6 is a bond, and the remaining of
Y.sup.2, Y.sup.5 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.2, Y.sup.5 and Y.sup.6 are bonds, and
the remaining of Y.sup.2, Y.sup.5 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.2,
Y.sup.5 and Y.sup.6 are bonds, and the remaining of Y.sup.2,
Y.sup.5 and Y.sup.6 is L.
[0105] In some embodiments of any of the above embodiments of
compounds of Formula Iv, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.2, Y.sup.5 or Y.sup.6
is a bond.
[0106] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iw:
##STR00060##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 Y.sup.4 and Y.sup.5 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.3, Y.sup.4 or Y.sup.5 is a bond, or
R.sup.26, provided, however, that none of Y.sup.3R.sup.27,
Y.sup.4R.sup.27, and Y.sup.5R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0107] In some embodiments of compounds of Formula Iw, Y.sup.3,
Y.sup.4 and Y.sup.5 are bonds. In some embodiments, Y.sup.3,
Y.sup.4 and Y.sup.5 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.3, Y.sup.4 and Y.sup.5 are independently L.
In some embodiments, any one of Y.sup.3, Y.sup.4 and Y.sup.5 is a
bond, and the remaining of Y.sup.3, Y.sup.4 and Y.sup.5 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.3, Y.sup.4 and Y.sup.5 is a bond, and the remaining of
Y.sup.3, Y.sup.4 and Y.sup.5 are independently L. In some
embodiments, any two of Y.sup.3, Y.sup.4 and Y.sup.5 are bonds, and
the remaining of Y.sup.3, Y.sup.4 and Y.sup.5 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.3,
Y.sup.4 and Y.sup.5 are bonds, and the remaining of Y.sup.3,
Y.sup.4 and Y.sup.5 is L.
[0108] In some embodiments of any of the, above embodiments of
compounds of Formula Iw, each R.sup.27 is independently Optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3, Y.sup.4 or Y.sup.5
is a bond.
[0109] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Ix:
##STR00061##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 Y.sup.4 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.3, Y.sup.4 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.3R.sup.27,
Y.sup.4R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0110] In some embodiments of compounds of Formula Ix, Y.sup.3,
Y.sup.4 and Y.sup.6 are bonds. In some embodiments, Y.sup.3,
Y.sup.4 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.3, Y.sup.4 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.3, Y.sup.4 and Y.sup.6 is a
bond, and the remaining of Y.sup.3, Y.sup.4 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.3, Y.sup.4 and Y.sup.6 is a bond, and the remaining of
Y.sup.3, Y.sup.4 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.3, Y.sup.4 and Y.sup.6 are bonds, and
the remaining of Y.sup.3, Y.sup.4 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.3,
Y.sup.4 and Y.sup.6 are bonds, and the remaining of Y.sup.3,
Y.sup.4 and Y.sup.6 is L.
[0111] In some embodiments of any of the above embodiments of
compounds of Formula Ix, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3, Y.sup.4 or Y.sup.6
is a bond.
[0112] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iy:
##STR00062##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.3 Y.sup.5 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.3, Y.sup.5 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.3R.sup.27,
Y.sup.5R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0113] In some embodiments of compounds of Formula Iy, Y.sup.3,
Y.sup.5 and Y.sup.6 are bonds. In some embodiments, Y.sup.3,
Y.sup.5 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.3, Y.sup.5 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.3, Y.sup.5 and Y.sup.6 is a
bond, and the remaining of Y.sup.3, Y.sup.5 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.3, Y.sup.5 and Y.sup.6 is a bond, and the remaining of
Y.sup.3, Y.sup.5 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.3, Y.sup.5 and Y.sup.6 are bonds, and
the remaining of Y.sup.3, Y.sup.5 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.3,
Y.sup.5 and Y.sup.6 are bonds, and the remaining of Y.sup.3,
Y.sup.5 and Y.sup.6 is L.
[0114] In some embodiments of any of the above embodiments of
compounds of Formula Iy, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.3, Y.sup.5 or Y.sup.6
is a bond.
[0115] In some embodiments, the compound of Formula I has a
structure according to the following sub-generic structure Formula
Iz:
##STR00063##
all salts, prodrugs, tautomers, and isomers thereof, wherein
Y.sup.4 Y.sup.5 and Y.sup.6 are independently a bond,
--CR.sup.aR.sup.b--, or L, and each R.sup.27 is independently
halogen, provided that Y.sup.4, Y.sup.5 or Y.sup.6 is a bond, or
R.sup.26, provided, however, that none of Y.sup.4R.sup.27,
Y.sup.5R.sup.27, and Y.sup.6R.sup.27 are hydrogen, wherein R.sup.a,
R.sup.b, L and R.sup.26 are as defined with reference to Formula
I.
[0116] In some embodiments of compounds of Formula Iz, Y.sup.4,
Y.sup.5 and Y.sup.6 are bonds. In some embodiments, Y.sup.4,
Y.sup.5 and Y.sup.6 are independently --CR.sup.aR.sup.b-- or L. In
some embodiments, Y.sup.4, Y.sup.5 and Y.sup.6 are independently L.
In some embodiments, any one of Y.sup.4, Y.sup.3 and Y.sup.6 is a
bond, and the remaining of Y.sup.4, Y.sup.5 and Y.sup.6 are
independently --CR.sup.aR.sup.b-- or L. In some embodiments, any
one of Y.sup.4, Y.sup.5 and Y.sup.6 is a bond, and the remaining of
Y.sup.4, Y.sup.5 and Y.sup.6 are independently L. In some
embodiments, any two of Y.sup.4, Y.sup.5 and Y.sup.6 are bonds, and
the remaining of Y.sup.4, Y.sup.5 and Y.sup.6 is
--CR.sup.aR.sup.b-- or L. In some embodiments, any two of Y.sup.4,
Y.sup.5 and Y.sup.6 are bonds, and the remaining of Y.sup.4,
Y.sup.5 and Y.sup.6 is L.
[0117] In some embodiments, of any of the above embodiments of
compounds of Formula Iz, each R.sup.27 is independently optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.27 is halogen, provided that Y.sup.4, Y.sup.5 of Y.sup.6
is a bond.
[0118] The compounds of Formulae Ia-Iz, and all sub-embodiments
detailed herein, may be used to treat a subject suffering from or
at risk for any of the protein kinase mediated diseases or
conditions contemplated herein.
[0119] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
II:
##STR00064##
all salts, prodrugs, tautomers and isomers thereof, [0120] wherein:
[0121] A, R.sup.4, R.sup.5 and R.sup.6 are as defined with
reference to Formula I; and [0122] R.sup.12, R.sup.13, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, --CN, --NO.sub.2,
--CR.sup.aR.sup.bR.sup.24, and -LR.sup.24, where L and R.sup.24 are
as defined for Formula I.
[0123] In some embodiments of compounds of Formula II, at least one
of R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is other
than hydrogen. In some embodiments, at least two of R.sup.12,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are other than hydrogen.
In some embodiments, at least three of R.sup.12, R.sup.13,
R.sup.14, R.sup.15 and R.sup.16 are other than hydrogen. In some
embodiments, at least four of R.sup.12, R.sup.13, R.sup.14,
R.sup.15 and R.sup.16 are other than hydrogen. In some embodiments,
R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are other than
hydrogen.
[0124] In some embodiments of compounds of Formula II, one of
R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is other than
hydrogen, and the others of R.sup.12, R.sup.13, R.sup.14, R.sup.15
and R.sup.16 are each independently hydrogen, halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In some
embodiments, R.sup.12 is other than hydrogen and R.sup.13,
R.sup.14, R.sup.15 and R.sup.16 are each independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, R.sup.13 is other than
hydrogen, and R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are each
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.14 is other than hydrogen, and R.sup.12, R.sup.13, R.sup.15
and R.sup.16 are each independently hydrogen, halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio.
[0125] In some embodiments of compounds of Formula II, any two of
R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are
independently other than hydrogen, and the others of R.sup.12,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are each independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.12 and
R.sup.13 are each independently other than hydrogen, and R.sup.14,
R.sup.15 and R.sup.16 are each independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.12 and R.sup.14 are each
independently other than hydrogen, and R.sup.13, R.sup.15 and
R.sup.16 are each independently hydrogen, halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In some
embodiments, R.sup.12 and R.sup.15 are each independently other
than hydrogen, and R.sup.13, R.sup.14 and R.sup.16 are each
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.12 and R.sup.16 are each independently other than hydrogen,
and R.sup.13, R.sup.14 and R.sup.15 are each independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.13 and
R.sup.14 are each independently other than hydrogen, and R.sup.12,
R.sup.15 and R.sup.16 are each independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.13 and R.sup.15 are each
independently other than hydrogen, and R.sup.12, R.sup.14 and
R.sup.16 are each independently hydrogen, halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio.
[0126] In some embodiments of any of the above embodiments of
compounds of Formula II, wherein any of R.sup.12, R.sup.13,
R.sup.14, R.sup.15, and R.sup.16 is designated as other than
hydrogen, each such R.sup.12, R.sup.13, R.sup.14, R.sup.15, or
R.sup.16 is independently -LR.sup.24.
[0127] In some embodiments of any of the above embodiments of
compounds of Formula II, R.sup.5 is other than hydrogen, and
R.sup.4 and R.sup.6 are hydrogen, or R.sup.4 is other than
hydrogen, and R.sup.5 and R.sup.6 are hydrogen, or R.sup.6 is other
than hydrogen, and R.sup.4 and R.sup.5 are hydrogen, or R.sup.4 and
R.sup.5 are other than hydrogen, and R.sup.6 is hydrogen, or
R.sup.4 and R.sup.6 are other than hydrogen, and R.sup.5 is
hydrogen, or R.sup.3 and R.sup.6 are other than hydrogen, and
R.sup.4 is hydrogen, or R.sup.4, R.sup.5 and R.sup.6 are other than
hydrogen.
[0128] In some embodiments of compounds of Formula II, the
following compounds are excluded: [0129] R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are hydrogen
and A is --CH.sub.2--, --S-- or --S(O).sub.2--; [0130] R.sup.4,
R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are
hydrogen, R.sup.5 is --Br or thiophen-3-yl, and A is --C(O)--;
[0131] R.sup.4 is 3,5 di-fluorophenyl, --NH.sub.2, or --NO.sub.2,
R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, and A is --C(O)--; [0132] R.sup.4 is
NO.sub.2, R.sup.5 is Br, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, and A is --C(O)--; [0133] R.sup.5 is --Br,
R.sup.4, R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, and A is --S(O)--; [0134] R.sup.12 is
--CH.sub.3 or --F, R.sup.6, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, and A is --C(O)--; [0135] R.sup.12 is --OH,
R.sup.4, R.sup.6, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are
hydrogen, R.sup.5 is thiophen-2-yl, and A is --C(O)--; [0136]
R.sup.12 is --CF.sub.3, R.sup.4, R.sup.5, R.sup.6, R.sup.13,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--S(O).sub.2--; [0137] R.sup.13 is --OH or --OCH.sub.3, R.sup.4,
R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen,
and A is --CH.sub.2--; [0138] R.sup.13 is --OCH.sub.3, R.sup.5,
R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen,
R.sup.4 is --Br, and A is --CH.sub.2--; [0139] R.sup.13 is --OH or
--OCH.sub.3, R.sup.4, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, R.sup.5 is thiophen-2-yl, and A is --CHOH--;
[0140] R.sup.13 is
##STR00065##
[0140] wherein
##STR00066##
indicates the bond to the phenyl ring, R.sup.4, R.sup.6, R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, R.sup.5 is
thiophen-3-yl, and A is --CH.sub.2--; [0141] R.sup.13 is --F, --OH
or --OCH.sub.3, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16
are hydrogen, and A is --C(O)--; [0142] R.sup.13 is --NO.sub.2,
--NH.sub.2, or
##STR00067##
[0142] wherein
##STR00068##
indicates the, bond to the phenyl ring, R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--C(O)--; [0143] R.sup.13 is --F, --Cl, or --CF.sub.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are
hydrogen, and A is --S(O).sub.2--; [0144] R.sup.13 and R.sup.14 are
--OH, R.sup.4, R.sup.6, R.sup.12, R.sup.15 and R.sup.16 are
hydrogen, R.sup.5 is thiophen-3-yl, and A is --CH.sub.2--; [0145]
R.sup.14 is --OH or --OCH.sub.3, R.sup.4, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen, R.sup.5 is
thioplien-2-yl, and A is --C(O)--; [0146] R.sup.14 is --OCH.sub.3,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 are hydrogen, and A is --C(O)--; [0147] R.sup.14 is --Cl,
R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen,
and A is --C(O)--; [0148] R.sup.14 is
##STR00069##
[0148] wherein
##STR00070##
indicates the bond to the phenyl ring, R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, and A is
--CH.sub.2--; [0149] R.sup.14 is --F, R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, and A is
--S-- or --S(O).sub.2--; [0150] R.sup.14 is --CH.sub.3, R.sup.5,
R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen,
R.sup.4 is 3-(hydroxymethyl)phenyl, and A is --S--; [0151] R.sup.12
and R.sup.16 are --F, R.sup.5, R.sup.6, R.sup.13, R.sup.14 and
R.sup.15 are hydrogen, R.sup.4 is 3,5 difluorophenyl, and A is
--C(O)--; [0152] R.sup.12 is --Cl, R.sup.13 is --Cl, R.sup.6,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is --C(O)--;
[0153] R.sup.12 is --F, R.sup.13 is --F, R.sup.5, R.sup.6,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, R.sup.4 is 3,5
difluorophenyl, and A is --C(O)--; [0154] R.sup.12 is --F, R.sup.13
is --OH or --OCH.sub.3, R.sup.4, R.sup.6, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, R.sup.5 is thiophen-2-yl, and A is --C(O)--;
[0155] R.sup.12 is --F, R.sup.13 is --OCH.sub.3, R.sup.4, R.sup.6,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, R.sup.5 is --Br, and
A is --C(O)--; [0156] R.sup.12 and R.sup.14 are --F, R.sup.5,
R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, R.sup.4 is
3,5 difluorophenyl, and A is --C(O)--; [0157] R.sup.12 is
--CH.sub.3, R.sup.15 is --F, R.sup.6, R.sup.13, R.sup.14 and
R.sup.16 are hydrogen and A is --C(O)--; [0158] R.sup.12 is --F,
R.sup.15 is --Cl, R.sup.4, R.sup.6, R.sup.13, R.sup.14 and R.sup.16
are hydrogen, R is thiophen-2-yl and A is --C(O)--; [0159] R.sup.12
and R.sup.15 are --F, R.sup.6, R.sup.13, R.sup.14 and R.sup.16 are
hydrogen, and A is --C(O)--; [0160] R.sup.12 is halogen, R.sup.15
is --OH or --OCH.sub.3, R.sup.6, R.sup.13, R.sup.14 and R.sup.16
are hydrogen and A is --C(O)--; [0161] R.sup.12 is --F, R.sup.15 is
--NHS(O).sub.2CH.sub.3, R.sup.4, R.sup.5, R.sup.6, R.sup.13,
R.sup.14 and R.sup.16 are hydrogen and A is --C(O)--; [0162]
R.sup.13 and R.sup.15 are --OCH.sub.3, R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.14 and R.sup.16 are hydrogen, and A is
--CH.sub.2--; [0163] R.sup.13 and R.sup.15 are --Cl, R R.sup.5,
R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are hydrogen, and A is
--S(O).sub.2--; [0164] R.sup.13 is --OH and R.sup.15 is --OH or
--OCH.sub.3, R.sup.4, R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are
hydrogen, R.sup.5 is thiophen-3-yl, and A is --CH.sub.2--; [0165]
R.sup.4, R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, A is --S(O).sub.2--, and R.sup.5 is
##STR00071##
[0165] wherein
##STR00072##
indicates the bond to the 5-position of the 7-azaindole ring;
[0166] R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, A is --S(O).sub.2--, and R.sup.4 is,
##STR00073##
[0166] wherein
##STR00074##
indicates the bond to the 4-position of the 7-azaindole ring;
[0167] R.sup.4, R.sup.5, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, A is --S(O).sub.2--, and R.sup.6 is
##STR00075##
[0167] wherein
##STR00076##
indicates the bond to the 6-position of the 7-azaindole ring;
[0168] R.sup.13 is --CN, R.sup.4, R.sup.6, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are hydrogen, A is --S(O).sub.2--, and
R.sup.5 is
##STR00077##
[0168] wherein
##STR00078##
indicates the bond to the 5-position of the 7-azaindole ring;
[0169] R.sup.12 is --Cl, R.sup.14 is --F or hydrogen, R.sup.4,
R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, A is
--S(O).sub.2--, and R.sup.5 is
##STR00079##
[0169] wherein
##STR00080##
indicates the bond to the 5-position of the 7-azaindole ring;
[0170] R.sup.14 is --NH.sub.2, R.sup.4, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen, A is --S(O).sub.2--,
and R.sup.5 is
##STR00081##
[0170] wherein
##STR00082##
indicates the bond to the 5-position of the 7-azaindole ring;
[0171] R.sup.6, R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16
are hydrogen, A is --S(O).sub.2--, R.sup.4 is --Cl, and R.sup.5
is
##STR00083##
[0171] wherein
##STR00084##
indicates the bond to the 5-position of the 7-azaindole ring;
[0172] R.sup.13 is --F, R.sup.4, R.sup.6, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are hydrogen, A is --CH.sub.2--, and R.sup.5
is 3-hydroxy-phenyl; [0173] R.sup.14 is --N(CH.sub.3).sub.2,
R.sup.4, R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are
hydrogen, A is --CH.sub.2--, and R.sup.5 is 3-hydroxy-phenyl;
[0174] R.sup.14 is hydrogen or --Br, R.sup.4, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen, A is --S--, and
R.sup.5 is 3-hydroxy-phenyl; and [0175] R.sup.4, R.sup.6, R.sup.12,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, A is
--C(O)--, and R.sup.5 is 3-hydroxy-phenyl.
[0176] In some embodiments of compounds of Formula II, at least one
of R.sup.12, R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is
-LR.sup.24, wherein R.sup.24 is substituted methyl, optionally
substituted C.sub.2-6 alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.24 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of L, optionally substituted lower
alkynyl, provided, however, that when R.sup.24 is optionally
substituted lower alkynyl, no alkyne carbon thereof is bound to N,
S, O, S(O), S(O).sub.2, C(O) or C(S) of L, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl, provided,
however, that R.sup.13 is not
##STR00085##
and that R.sup.14 is not
##STR00086##
wherein
##STR00087##
indicates the bond to the phenyl ring; in further embodiments,
R.sup.24 is optionally substituted C.sub.2-6 alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl, or
substituted methyl, wherein methyl is substituted with optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl;
in further embodiments, R.sup.5 is other than hydrogen; in further
embodiments, R.sup.4 and R.sup.6 are hydrogen.
[0177] In some embodiments of compounds of Formula II, R.sup.12 is
other than hydrogen. In some embodiments, R.sup.12 is -LR.sup.24.
In some embodiments, R.sup.12 is other than hydrogen and R.sup.13,
R.sup.14, R.sup.15 and R.sup.16 are each independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, R.sup.12 is -LR.sup.24 and
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.12 is
-LR.sup.24, any three of R.sup.13, R.sup.14, R.sup.15 and R.sup.16
are independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and the remaining of
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is hydrogen. In some
embodiments, R.sup.12 is -LR.sup.24, any two of R.sup.13, R.sup.14,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio and the remaining of R.sup.13, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen. In some embodiments, R.sup.12 is -LR.sup.24,
any one of R.sup.13, R.sup.14, R.sup.15 and R.sup.16 is hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining of R.sup.13, R.sup.14, R.sup.15
and R.sup.16 are hydrogen. In some embodiments, R.sup.12 is
-LR.sup.24, and R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are
hydrogen. In some embodiments, R.sup.12 is other than hydrogen, and
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are hydrogen.
[0178] In other embodiments of compounds of Formula II, wherein
R.sup.12 is other than hydrogen, when R.sup.13, R.sup.14, R.sup.15
and R.sup.16 are hydrogen and A is --C(O)--, then R.sup.12 is not
--CH.sub.3, --F, or --OH; and when R.sup.4, R.sup.5, R.sup.6,
R.sup.13, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--S(O).sub.2--, then R.sup.12 is not --CF.sub.3; and when R.sup.12
and R.sup.16 are --F, R.sup.5 , R.sup.6, R.sup.13, R.sup.14 and
R.sup.15 are hydrogen, and A is --C(O)--, R.sup.4 is not 3,5
difluorophenyl; and when R.sup.12 is halogen, R.sup.6, R.sup.14,
R.sup.15 and R.sup.16 are hydrogen, and A is --C(O)--, R.sup.13 is
not halogen, --OH, or --OCH.sub.3; and when R.sup.12 and R.sup.14
are --F, R.sup.5, R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are
hydrogen, and A is --C(O)--, R.sup.4 is not 3,5 difluorophenyl; and
when R.sup.15 is halogen, --OH or --OCH.sub.3, R.sup.6, R.sup.13,
R.sup.14 and R.sup.16 are hydrogen, and A is --C(O)--, R.sup.12 is
not halogen; and when R.sup.15 is --F, R.sup.6, R.sup.13, R.sup.14
and R.sup.16 are hydrogen and A is --C(O)--, R.sup.12 is not
--CH.sub.3; and when R.sup.12 is --F, R.sup.4, R.sup.5, R.sup.6,
R.sup.13, R.sup.14 and R.sup.16 are hydrogen and A is --C(O)--,
R.sup.15 is not --NHS(O).sub.2CH.sub.3; and when R.sup.12 is --Cl,
R.sup.14 is --F or hydrogen, R.sup.4, R.sup.6, R.sup.13, R.sup.15
and R.sup.16 are hydrogen, and A is --S(O).sub.2--, R.sup.5 is
not
##STR00088##
wherein
##STR00089##
indicates the bond to the 5-position of the 7-azaindole ring.
[0179] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.12 is other than hydrogen,
R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6 are
hydrogen; or R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen; or R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen; or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen; or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen; or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen; or R.sup.4, R.sup.5
and R.sup.6 are all other than hydrogen.
[0180] In some embodiments of compounds of Formula II, R.sup.13 is
other than hydrogen. In some embodiments, R.sup.13 is -LR.sup.24.
In some embodiments, R.sup.13 is other than hydrogen and R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are each independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, R.sup.13 is -LR.sup.24 and
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are each independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.13 is
-LR.sup.24, any three of R.sup.12, R.sup.14, R.sup.15 and R.sup.16
are independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and the remaining of
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 is hydrogen. In some
embodiments, R.sup.13 is -LR.sup.24, any two of R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio and the the remaining of R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen. In some embodiments, R.sup.13 is -LR.sup.24,
any one of R.sup.12, R.sup.14, R.sup.15 and R.sup.16 is
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio, and the remaining of
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some
embodiments, R.sup.13 is -LR.sup.24, and R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, R.sup.13
is other than hydrogen, and R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen.
[0181] In other embodiments of compounds of Formula II, wherein
R.sup.13 is other than hydrogen, when R.sup.4, R.sup.6, R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is --CH.sub.2--
or --CH(OH)--, then R.sup.13 is not --OH or --OCH.sub.3; and when
R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are
hydrogen, and A is --CH.sub.2--, then R.sup.13 is not --OCH.sub.3;
and when R.sup.13 is
##STR00090##
wherein
##STR00091##
indicates the bond to the phenyl ring, R.sup.4, R.sup.6, R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--CH.sub.2--, then R.sup.5 is not thiophen-3-yl; and when R.sup.6,
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--C(O)--, then R.sup.13 is not --F, --OH, or --OCH.sub.3; and when
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen, and A is --C(O)--, then R.sup.13 is not
--NO.sub.2, --NH.sub.2, or
##STR00092##
wherein
##STR00093##
indicates the bond to the phenyl ring; and when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen,
and A is --S(O).sub.2--, then R.sup.13 is not --F, --Cl, or
--CF.sub.3; and when R.sup.13 and R.sup.14 are --OH, R.sup.4,
R.sup.6, R.sup.12, R.sup.15 and R.sup.16 are hydrogen and A is
--CH.sub.2--, R.sup.5 is not thiophen-3-yl; and when R.sup.6,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen and A is --C(O)--,
then R.sup.13 and R.sup.12 are not both --Cl; and when R.sup.12 and
R.sup.13 are both --F, R.sup.5, R.sup.6, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen and A is --C(O)--, then R.sup.4 is not
3,5-difluorophenyl; and when R.sup.13 is --OH or --OCH.sub.3,
R.sup.12 is --F, R.sup.4, R.sup.6, R.sup.14, R.sup.15 and R.sup.16
are hydrogen, and A is --C(O)--, then R.sup.5 is not thiophen-2-yl;
and when R.sup.13 is --OCH.sub.3, R.sup.12 is --F, R.sup.4,
R.sup.6, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, and A is
--C(O)--, then R.sup.5 is not --Br; and when R.sup.6, R.sup.12,
R.sup.14 and R.sup.15 are hydrogen, R.sup.16 is --CH.sub.3 and A is
--C(O)--, then R.sup.13 is not --F; and when R.sup.13 is --Cl,
R.sup.16 is --F, R.sup.4, R.sup.6, R.sup.12, R.sup.14 and R.sup.15
are hydrogen, and A is --C(O)--, then R.sup.5 is not thiophen-2-yl;
and when R.sup.6, R.sup.12, R.sup.14 and R.sup.15 are hydrogen and
A is --C(O)--, then R.sup.13 and R.sup.16 are not both --F; and
when R.sup.6, R.sup.12, R.sup.14 and R.sup.15 are hydrogen,
R.sup.16 is halogen, and A is --C(O)--, then R.sup.13 is not --OH
or --OCH.sub.3; and when R.sup.4, R.sup.5, R.sup.6, R.sup.12,
R.sup.14 and R.sup.15 and hydrogen, R.sup.13 is
--NHS(O).sub.2CH.sub.3, and A is --C(O)--, then R.sup.16 is not
--F; and when R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14 and
R.sup.16 are hydrogen, and A is --CH.sub.2--, then R.sup.13 and
R.sup.15 are not both --OCH.sub.3; and when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are hydrogen, and A is
--S(O).sub.2--, then R.sup.13 and R.sup.15 are not both --Cl; and
when R.sup.13 is --OH, R.sup.15 is --OH or --OCH.sub.3, R.sup.4,
R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are hydrogen, and A is
--CH.sub.2--, then R.sup.5 is not thiophen-3-yl; and when R.sup.13
is --F, R.sup.4, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and R.sup.16
are hydrogen, and A is --CH.sub.2--, then R.sup.5 is not
3-hydroxy-phenyl; and when R.sup.13 is --CN, R.sup.4, R.sup.6,
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, A is
--S(O).sub.2--, then R.sup.5 is not
##STR00094##
wherein
##STR00095##
indicates the bond to the 5-position of the 7-azaindole ring.
[0182] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.13 is other than hydrogen,
R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6 are
hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0183] In some embodiments of compounds of Formula II, R.sup.14 is
other than hydrogen. In some embodiments, R.sup.14 is -LR.sup.24.
In some embodiments, R.sup.14 is other than hydrogen and R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are each independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, R.sup.14 is -LR.sup.24, and
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.14 is
-LR.sup.24, any three of R.sup.12, R.sup.13, R.sup.15 and R.sup.16
are independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio, and the remaining of
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 is hydrogen. In some
embodiments, R.sup.14 is -LR.sup.24, any two of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio, and the remaining of R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 are hydrogen. In some embodiments, R.sup.14 is -LR.sup.24,
any one of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio, and the remaining of
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen. In some
embodiments, R.sup.14 is -LR.sup.24, and R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, R.sup.14
is other than hydrogen, and R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 are hydrogen.
[0184] In other embodiments of compounds of Formula II, wherein
R.sup.14 is other than hydrogen, when R.sup.4, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen, R.sup.5 is
thiophen-2-yl and A is --C(O)--, then R.sup.14 is not --OH, or
--OCH.sub.3; and when R.sup.4, R.sup.5, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen and A is --C(O)--,
then R.sup.14 is not --OCH.sub.3; and when R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen and A is --C(O)--,
then R.sup.14 is not --Cl; and when R.sup.4, R.sup.5, R.sup.6,
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen and A is
--CH.sub.2--, then R.sup.14 is not
##STR00096##
wherein
##STR00097##
indicates the bond to the phenyl ring; and when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen and
A is --S--or --S(O).sub.2--, then R.sup.14 is not --F; and when
R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are
hydrogen, R.sup.4 is 3-(hydroxymethyl)phenyl and A is --S--, then
R.sup.14 is not --CH.sub.3; and when R.sup.5, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen, R.sup.4 is
3,5-difluorophenyl and A is --C(O)--, R.sup.12 and R.sup.14 are not
both --F; and when R.sup.14 is --N(CH.sub.3).sub.2, R.sup.4,
R.sup.6, R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen,
and A is --CH.sub.2--, then R.sup.5 is not 3-hydroxy-phenyl; and
when R.sup.14 is --Br, R.sup.4, R.sup.6, R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen, and A is --S--, then R.sup.5 is
not 3-hydroxy-phenyl; and when R.sup.12 is --Cl, R.sup.14 is --F,
R.sup.4, R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, and
A is --S(O).sub.2--, then R.sup.5 is not
##STR00098##
wherein
##STR00099##
indicates the bond to the 5-position of the 7-azaindole ring; and
when R.sup.14 is --NH.sub.2, R.sup.4, R.sup.6, R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen, and A is --S(O).sub.2--, then
R.sup.5 is not
##STR00100##
wherein
##STR00101##
indicates the bond to the 5-position of the 7-azaindole ring; and
when R.sup.4, R.sup.6, R.sup.12, R.sup.15 and R.sup.16 are
hydrogen; R.sup.13 and R.sup.14 are --OH, and A is --CH.sub.2--,
then R.sup.5 is not thiophen-3-yl.
[0185] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.14 is other than hydrogen,
R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6 are
hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0186] In some embodiments of compounds of Formula II, R.sup.12 and
R.sup.16 are other than hydrogen. In some embodiments, R.sup.12 and
R.sup.16 are independently -LR.sup.24. In some embodiments,
R.sup.12 and R.sup.16 are other than hydrogen and R.sup.13,
R.sup.14 and R.sup.15 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.12 and R.sup.16 are
independently -LR.sup.24 and R.sup.13, R.sup.14 and R.sup.15 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.12 and R.sup.16 are independently -LR.sup.24, any two of
R.sup.13, R.sup.14 and R.sup.15 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio, and the remaining of R.sup.13, R.sup.14 and
R.sup.15 is hydrogen. In some embodiments, R.sup.12 and R.sup.16
are independently -LR.sup.24, any one of R.sup.13, R.sup.14 and
R.sup.15 is independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio, and the
remaining of R.sup.13, R.sup.14 and R.sup.15 are hydrogen. In some
embodiments, R.sup.12 and R.sup.16 are independently -LR.sup.24,
and R.sup.13, R.sup.14 and R.sup.15 are hydrogen. In some
embodiments, R.sup.12 and R.sup.16 are other than hydrogen, and
R.sup.13, R.sup.14 and R.sup.15 are hydrogen. In other embodiments,
wherein R.sup.12 and R.sup.16 are other than hydrogen, when
R.sup.5, R.sup.6, R.sup.13, R.sup.14 and R.sup.15 are hydrogen and
A is --C(O)--, then R.sup.12 and R.sup.16 are not both --F.
[0187] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.12 and R.sup.16 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0188] In some embodiments of compounds of Formula II, R.sup.12 and
R.sup.13 are other than hydrogen. In some embodiments, R.sup.12 and
R.sup.13 are independently -LR.sup.24. In some embodiments,
R.sup.12 and R.sup.13 are other than hydrogen and R.sup.14,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.12 and R.sup.13 are
independently -LR.sup.24, and R.sup.14, R.sup.13 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.12 and R.sup.13 are independently -LR.sup.24, any two of
R.sup.14, R.sup.15 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio, and the remaining of R.sup.14, R.sup.15 and
R.sup.16 is hydrogen. In some embodiments, R.sup.12 and R.sup.13
are independently -LR.sup.24, any one of R.sup.14, R.sup.15 and
R.sup.16 is independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio, and the
remaining of R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some
embodiments, R.sup.12 and R.sup.13 are independently -LR.sup.24,
and R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some
embodiments, R.sup.12 and R.sup.13 are other than hydrogen and
R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In other embodiments,
wherein R.sup.12 and R.sup.13 are other than hydrogen, when
R.sup.6, R.sup.14, R.sup.15 and R.sup.16 are hydrogen, A is
--C(O)--, and R.sup.13 is halogen, --OH, or --OCH.sub.3, then
R.sup.12 is not halogen. In alternate embodiments, when R.sup.12
and R.sup.13 are other than hydrogen, R.sup.6, R.sup.14, R.sup.15
and R.sup.16 are hydrogen and A is --C(O)--, then both R.sup.12 and
R.sup.13 are not halogen; and when R.sup.12 and R.sup.13 are other
than hydrogen, R.sup.4, R.sup.6, R.sup.14, R.sup.15 and R.sup.16
are hydrogen, and A is --C(O)--, R.sup.12 is --F and R.sup.13 is
--OH or --OCH.sub.3, then R.sup.5 is not --Br or thiophen-2-yl.
[0189] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.12 and R.sup.13 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0190] In some embodiments of compounds of Formula II, R.sup.12 and
R.sup.14 are other than hydrogen. In some embodiments, R.sup.12 and
R.sup.14 are independently -LR.sup.24. In some embodiments,
R.sup.12 and R.sup.14 are other than hydrogen and R.sup.13,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.12 and R.sup.14 are
independently -LR.sup.24, and R.sup.13, R.sup.15 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.12 and R.sup.14 are independently -LR.sup.24, and any one of
R.sup.13, R.sup.15 and R.sup.16 is independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio, and the remaining of R.sup.13, R.sup.15 and R.sup.16 is
hydrogen. In some embodiments, R.sup.12 and R.sup.14 are
independently -LR.sup.24, and any two of R.sup.13, R.sup.15 and
R.sup.16 are independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio and the
remaining of R.sup.13, R.sup.15 and R.sup.16 is hydrogen. In some
embodiments, R.sup.12 and R.sup.14 are independently -LR.sup.24 and
R.sup.13, R.sup.15 and R.sup.16 are hydrogen. In some embodiments,
R.sup.12 and R.sup.14 are other than hydrogen and R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, wherein
R.sup.12 and R.sup.14 are other than hydrogen, when R.sup.5,
R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, A is
--C(O)--, and R.sup.4 is 3,5 difluorophenyl, then R.sup.12 and
R.sup.14 are not both --F; and when R.sup.12 is --Cl, R.sup.14 is
--F, R.sup.4, R.sup.6, R.sup.13, R.sup.15 and R.sup.16 are
hydrogen, and A is --S(O).sub.2--, then R.sup.5 is not
##STR00102##
wherein
##STR00103##
indicates the bond to the 5-position of the 7-azaindole ring.
[0191] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.12 and R.sup.14 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0192] In some embodiments of compounds of Formula II, R.sup.12 and
R.sup.13 are other than hydrogen. In some embodiments, R.sup.12 and
R.sup.13 are independently -LR.sup.24. In some embodiments,
R.sup.12 and R.sup.15 are other than hydrogen and R.sup.13,
R.sup.14 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.12 and R.sup.15 are
independently -LR.sup.24 and R.sup.13, R.sup.14 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.12 and R.sup.15 are independently -LR.sup.24, any two of
R.sup.13, R.sup.14 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining of R.sup.13, R.sup.14 and
R.sup.16 is hydrogen. In some embodiments, R.sup.12 and R.sup.15
are independently -LR.sup.24, any one of R.sup.13, R.sup.14 and
R.sup.16 is hydrogen, halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio and the remaining of R.sup.13,
R.sup.14 and R.sup.16 are hydrogen. In some embodiments, R.sup.12
and R.sup.15 are independently -LR.sup.24 and R.sup.13, R.sup.14
and R.sup.16 are hydrogen. In some embodiments, R.sup.12 and
R.sup.15 are other than hydrogen and R.sup.13, R.sup.14 and
R.sup.16 are hydrogen. In other embodiments, wherein R.sup.12 and
R.sup.15 are other than hydrogen, when R.sup.6, R.sup.13, R.sup.14
and R.sup.16 are hydrogen and A is --C(O)--, then R.sup.12 and
R.sup.15 are not both halogen; and when R.sup.6, R.sup.13, R.sup.14
and R.sup.16 are hydrogen, A is --C(O)--, and R.sup.12 is
--CH.sub.3, then R.sup.15 is not --F; and when R.sup.6, R.sup.13,
R.sup.14 and R.sup.16 are hydrogen, A is --C(O)--, and R.sup.12 is
halogen, then R.sup.15 is not --OH or --OCH.sub.3; and when
R.sup.4, R.sup.5, R.sup.6, R.sup.13, R.sup.14 and R.sup.16 are
hydrogen, A is --C(O)--, and R.sup.12 is --F then R.sup.15 is not
NHS(O).sub.2CH.sub.3.
[0193] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.12 and R.sup.15 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0194] In some embodiments of compounds of Formula II, R.sup.13 and
R.sup.14 are other than hydrogen. In some embodiments, R.sup.13 and
R.sup.14 are independently -LR.sup.24. In some embodiments,
R.sup.13 and R.sup.14 are other than hydrogen and R.sup.12,
R.sup.15 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R.sup.13 and R.sup.14 are
independently -LR.sup.24 and R.sup.12, R.sup.15 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
R.sup.13 and R.sup.14 are independently -LR.sup.24, any two of
R.sup.12, R.sup.15 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining of R.sup.12, R.sup.15 and
R.sup.16 is hydrogen. In some embodiments, R.sup.13 and R.sup.14
are independently -LR.sup.24, any one of R.sup.12, R.sup.15 and
R.sup.16 is hydrogen, halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio and the remaining of R.sup.12,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, R.sup.13
and R.sup.14 are independently -LR.sup.24 and R.sup.12, R.sup.15
and R.sup.16 are hydrogen. In some embodiments, R.sup.13 and
R.sup.14 are other than hydrogen and R.sup.12, R.sup.15 and
R.sup.16 are hydrogen. In other embodiments, wherein R.sup.13 and
R.sup.14 are other than hydrogen, when R.sup.4, R.sup.6, R.sup.12,
R.sup.15 and R.sup.16 are hydrogen, A is --CH.sub.2--, and R.sup.5
is thiophen-3-yl, then R.sup.13 and R.sup.14 are not both --OH.
[0195] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.13 and R.sup.14 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0196] In some embodiments of compounds of Formula II, R.sup.13 and
R.sup.15 are other than hydrogen. In some embodiments, R.sup.13 and
R.sup.15 are independently -LR.sup.24. In some embodiments,
R.sup.13 and R.sup.15 are other than hydrogen and R.sup.12 ,
R.sup.14 and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, R and R are independently
-LR.sup.24 and R.sup.12, R.sup.14 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, R.sup.13and
R.sup.15 are independently -LR.sup.24, two of R.sup.12, R.sup.14
and R.sup.16 are independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio and the
remaining of R.sup.12, R.sup.14 and R.sup.16 is hydrogen. In some
embodiments, R.sup.13 and R.sup.15 are independently -LR.sup.24,
one of R.sup.12, R.sup.14 and R.sup.16 is hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio and the remaining of R.sup.12, R.sup.14 and R.sup.16 are
hydrogen. In some embodiments, R.sup.13 and R.sup.15 are
independently -LR.sup.24 and R.sup.12, R.sup.14 and R.sup.15 are
hydrogen, some embodiments, R.sup.13 and R.sup.15 are other than
hydrogen and R.sup.12, R.sup.14 and R.sup.16 are hydrogen. In other
embodiments, wherein R.sup.13 and R.sup.15 are other than hydrogen,
when R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are
hydrogen and A is --CH.sub.2--, then R.sup.13 and R.sup.15 are not
both --OCH.sub.3; and when R.sup.4, R.sup.5, R.sup.6, R.sup.12,
R.sup.14 and R.sup.16 are hydrogen and A is --S(O).sub.2--, then
R.sup.13 and R.sup.15 are not both --Cl; and when R.sup.4, R.sup.6,
R.sup.12, R.sup.14 and R.sup.16 are hydrogen, A is --CH.sub.2--,
R.sup.13 is --OH and R.sup.15 is --OH or --OCH.sub.3, then R.sup.5
is not thiophen-3-yl.
[0197] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.13 and R.sup.15 is other than
hydrogen, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0198] In some embodiments of compounds of Formula II, R.sup.13 is
-LR.sup.24, provided, however, that when A is --CH.sub.2--,
--CH(OH)-- or --C(O)--, and R.sup.12, R.sup.14, R.sup.15, and
R.sup.16 are hydrogen, then R.sup.13 is not --OH, --OCH.sub.3,
or
##STR00104##
wherein
##STR00105##
indicates the bond to the phenyl ring; and when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are hydrogen
and A is --C(O)--, R.sup.13 is not NH.sub.2, or
##STR00106##
wherein
##STR00107##
indicates the bond to the phenyl ring; and when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are hydrogen and A is
--CH.sub.2--, R.sup.13 and R.sup.15 are not both --OCH.sub.3; and
when R.sup.4, R.sup.6, R.sup.12, R.sup.14 and R.sup.16 are
hydrogen, A is --CH.sub.2--, R.sup.5 is thiophen-3-yl, R.sup.13 is
--OH, R.sup.15 is not --OH or --OCH.sub.3; and when R.sup.4,
R.sup.6, R.sup.12, R.sup.15 and R.sup.16 are hydrogen, A is
--CH.sub.2--, and R.sup.3 is thiophen-3-yl, R.sup.13 and R.sup.14
are not both --OH; and when R.sup.4, R.sup.6, R.sup.14, R.sup.15
and R.sup.16 are hydrogen and A is --C(O)--, R.sup.12 is --F,
R.sup.5 is --Br or thiophen-2-yl, R.sup.13 is not --OH or
--OCH.sub.3; and when R.sup.12, R.sup.14, and R.sup.15 are
hydrogen, A is --C(O)--, and R.sup.16 is halogen, R.sup.13 is not
--OH or --OCH.sub.3; and when R.sup.4, R.sup.3, R.sup.6, R.sup.12,
R.sup.14, and R.sup.15 are hydrogen, A is --C(O)--, and R.sup.16 is
--F, R.sup.13 is not --NHS(O).sub.2CH.sub.3. In further
embodiments, R.sup.13 is -LR.sup.24 and L is
--NHS(O).sub.2CH.sub.2--, --O--, or --O--CHO.sub.2--, R.sup.24 is
not H, or when L is --NHS(O).sub.2-- or --O--, R.sup.24 is not
CH.sub.3. In further embodiments, where R.sup.13 is -LR.sup.24, one
of R.sup.12, R.sup.14, R.sup.15, and R.sup.16 is other than
hydrogen and the remaining of R.sup.12, R.sup.14, R.sup.15, and
R.sup.16 are hydrogen. In further embodiments, where R.sup.13 is
-LR.sup.24, two of R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are
independently other than hydrogen and the remaining two of
R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are hydrogen. In some
embodiments where R.sup.13 is -LR.sup.24, R.sup.14 and R.sup.15 are
hydrogen, and at least one of R.sup.12 and R.sup.16 is other than
hydrogen. In further embodiments, where R.sup.13 is -LR.sup.24,
R.sup.14 and R.sup.15 are hydrogen, and at least one of R.sup.12
and R.sup.16 is other than hydrogen, L is
-(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-or
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25S(O).sub.2-(alk).sub.b-, or
-(alk).sub.a-NR.sup.25S(O).sub.2NR.sup.23-(alk).sub.b-, wherein a,
b, R.sup.25 and alk are as defined for Formula I.
[0199] In further embodiments of compounds of Formula II, where
R.sup.13 is -LR.sup.24, R.sup.14 and R.sup.15 are hydrogen, and at
least one of R.sup.12 and R.sup.16 is other than hydrogen, L is
--NR.sup.25--, --NR.sup.25-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-,
or--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-.
[0200] In further embodiments of compounds of Formula II, where
R.sup.13 is -LR.sup.24, R.sup.14 and R.sup.15 are hydrogen, and at
least one of R.sup.12 and R.sup.16 is other than hydrogen, L is
--NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-. In further embodiments
where R.sup.13 is -LR.sup.24, R.sup.14 and R.sup.15 are hydrogen,
and at least one of R.sup.12 and R.sup.16 is other than hydrogen, L
is --NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-. In further embodiments
where R.sup.13 is -LR.sup.24, R.sup.14 and R.sup.15 are hydrogen,
and at least one of R.sup.12 and R.sup.16 is other than hydrogen, L
is --NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--OC(O)NR.sup.25-(alk).sub.b-, or --OC(S)NR.sup.25-(alk).sub.b-. In
some embodiments of any of the above embodiments where R.sup.13 is
-LR.sup.24, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen, or R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen, or R.sup.6 is other than hydrogen and R.sup.4
and R.sup.5 are hydrogen, or R.sup.4 and R.sup.5 are other than
hydrogen and R.sup.6 is hydrogen, or R.sup.4 and R.sup.6 are other
than hydrogen and R.sup.5 is hydrogen, or R.sup.5 and R.sup.6 are
other than hydrogen and R.sup.4 is hydrogen, or R.sup.4, R.sup.5
and R.sup.6 are other than hydrogen.
[0201] In some embodiments of compounds of Formula II, A is
--CR.sup.aR.sup.b-- or --C(O)--, R.sup.13 is -LR.sup.24, R.sup.14
and R.sup.15 are hydrogen, R.sup.12 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio, wherein one of R.sup.12 and R.sup.16
is other than hydrogen, and wherein L is --NR.sup.25-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein b and R.sup.25
are as defined for Formula I, alk is C.sub.1-3 alkylene optionally
substituted with fluoro or optionally fluoro substituted lower
alkyl, and R.sup.24 is hydrogen, provided, however, that said
hydrogen would not be attached to S(O).sub.2--, optionally fluoro
substituted lower alkyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio, and
R.sup.a and R.sup.b are independently hydrogen-halogen, optionally
fluoro substituted lower alkyl or optionally fluoro substituted
lower alkoxy, provided, however, that when R.sup.4, R.sup.5,
R.sup.6, R.sup.12, R.sup.14, and R.sup.16 are hydrogen, A is
--C(O)--, and R.sup.16 is --F, then R.sup.13 is not
--NHS(O).sub.2CH.sub.3. In other embodiments, R.sup.12 and R.sup.16
are independently halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio. In other embodiments, A is
--CH.sub.2--. In other embodiments, A is --C(O)--. In other
embodiments, R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen. In other embodiments, A is --CH.sub.2--, R.sup.4 is
other than hydrogen and R.sup.5 and R.sup.6 are hydrogen. In other
embodiments, A is --C(O)--, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In other embodiments, R.sup.5 is
other than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In other
embodiments, A is --CH.sub.2--, R.sup.5 is other than hydrogen and
R.sup.4 and R.sup.6 are hydrogen. In other embodiments, A is
--C(O)--, R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6
are hydrogen. In other embodiments, R.sup.6 is other than hydrogen
and R.sup.4 and R.sup.5 are hydrogen. In other embodiments, A is
--CH.sub.2--, R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen. In other embodiments, A is --C(O)--, R.sup.6
is other than hydrogen and R.sup.4 and R.sup.5 are hydrogen. In
other embodiments, R.sup.4 and R.sup.5 are other than hydrogen and
R.sup.6 is hydrogen. In other embodiments, R.sup.4 and R.sup.6 are
other than hydrogen, and R.sup.5 is hydrogen. In other embodiments,
R.sup.4 and R.sup.5 are hydrogen, and R.sup.6 is other than
hydrogen. In other embodiments, R.sup.4, R.sup.5, and R.sup.6 are
other than hydrogen.
[0202] In some embodiments of compounds of Formula II, A is
--CR.sup.aR.sup.b-- or --C(O)--, R.sup.13 is -LR.sup.24, R.sup.14
and R.sup.15 are hydrogen, R.sup.12 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio, wherein one of R.sup.12 and R.sup.16
is other than hydrogen, and wherein L is --NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)-(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein b and R.sup.25
are as defined for Formula I, alk is C.sub.1-3 alkylene optionally
substituted with fluoro or optionally fluoro substituted lower
alkyl, R.sup.24 is hydrogen provided, however, that said hydrogen
would not be attached to S(O).sub.2--, optionally fluoro
substituted lower alkyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, and optionally fluoro substituted lower alkylthio, R.sup.a
and R.sup.b are independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, or optionally fluoro substituted lower
alkoxy, one of R.sup.4 and R.sup.5 is other than hydrogen and
R.sup.6 is hydrogen. In other embodiments, R.sup.16 is hydrogen. In
other embodiments, R.sup.12 is hydrogen. In other embodiments,
R.sup.12 and R.sup.16 are independently halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In other
embodiments, A is --CH.sub.2--. In other embodiments, A is
--C(O)--. In other embodiments, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In other embodiments, A is
--CH.sub.2--, R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen. In other embodiments, A is --C(O)--, R.sup.4
is other than hydrogen and R.sup.5 and R.sup.6 are hydrogen. In
other embodiments, R.sup.5 is other than hydrogen and R.sup.4 and
R.sup.6 are hydrogen. In other embodiments, A is --CH.sub.2--,
R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6 are
hydrogen. In other embodiments, --C(O)--, R.sup.5 is other than
hydrogen and R.sup.4 and R.sup.6 are hydrogen. In other
embodiments, R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5
are hydrogen. In other embodiments, A is --CH.sub.2--, R.sup.6 is
other than hydrogen and R.sup.4 and R.sup.5 are hydrogen. In other
embodiments, A is --C(O)--, R.sup.6 is other than hydrogen and
R.sup.4 and R.sup.5 are hydrogen. In other embodiments, R.sup.4 and
R.sup.5 are other than hydrogen and R.sup.6 is hydrogen. In other
embodiments, R.sup.4 and R.sup.6 are other than hydrogen and
R.sup.5 is hydrogen. In other embodiments, R.sup.4 and R.sup.5 are
hydrogen and R.sup.6 is other than hydrogen. In other embodiments,
R.sup.4, R.sup.5, and R.sup.6 are other than hydrogen.
[0203] In some embodiments of compounds of Formula II, A is
--CR.sup.aR.sup.b-- or --C(O)--, R.sup.13 is -LR.sup.24, R.sup.12
is halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio, one of R.sup.14, R.sup.15 and R.sup.16 are
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the other two are hydrogen, L is
--NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein b and R.sup.25
are as defined for Formula I, alk is C.sub.1-3 alkylene optionally
substituted with fluoro or optionally fluoro substituted lower
alkyl, R.sup.24 is hydrogen provided, however, that said hydrogen
would not be attached to S(O) or S(O).sub.2, optionally substituted
lower alkyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl, R.sup.a and R.sup.b are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, or optionally fluoro substituted lower alkoxy, one of
R.sup.4 and R.sup.5 is other than hydrogen and R.sup.6 is hydrogen.
In other embodiments, R.sup.4 is hydrogen and R.sup.5 is other than
hydrogen. In other embodiments, R.sup.5 is hydrogen and R.sup.4 is
other than hydrogen. In other embodiments, A is --CH.sub.2--,
R.sup.5 is hydrogen and R.sup.4 is other than hydrogen. In other
embodiments, A is --C(O)--, R.sup.3 is hydrogen and R.sup.4 is
other than hydrogen. In other embodiments, A is --CH.sub.2--,
R.sup.4 is hydrogen and R.sup.5 is other than hydrogen. In other
embodiments, A is --C(O)--, R.sup.4 is hydrogen and R.sup.5 is
other than hydrogen.
[0204] In some embodiments of compounds of Formula II, A is
--CR.sup.aR.sup.b-- or --C(O)--, R.sup.13 is -LR.sup.24, R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio, where at least one of R.sup.12, R.sup.14, R.sup.15
and R.sup.16 is other than hydrogen, L is
--NR.sup.25C(O)--(alk).sub.b-, --NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein b and R.sup.25
are as defined for Formula I, alk is C.sub.1-3 alkylene optionally
substituted with fluoro or optionally fluoro substituted lower
alkyl, R.sup.24 is hydrogen provided, however, that said hydrogen
would not be attached to S(O).sub.2, optionally substituted lower
alkyl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, R.sup.a and R.sup.b are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl, or
optionally fluoro substituted lower alkoxy, provided, however, that
when R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, and R.sup.15
are hydrogen, A is --C(O)--, and R.sup.16 is --F, then R.sup.13 is
not --NHS(O).sub.2CH.sub.3. In further embodiments, R.sup.12 is
other than hydrogen and R.sup.14, R.sup.15 and R.sup.16 are
hydrogen. In further embodiments, R.sup.16 is other than hydrogen
and R.sup.12, R.sup.14 and R.sup.15 are hydrogen. In further
embodiments, at least two of R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are other than hydrogen. In further embodiments, R.sup.12
and R.sup.16 are other than hydrogen and R.sup.14 and R.sup.15 are
hydrogen. In further embodiments, R.sup.5 is other than hydrogen
and R.sup.4 and R.sup.6 are hydrogen. In further embodiments,
R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6 are
hydrogen.
[0205] In some embodiments of compounds of Formula II, A is
--CR.sup.aR.sup.b-- or --C(O)--, R.sup.13 is -LR.sup.24, R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio, where at least one of R.sup.12, R.sup.14, R.sup.15
and R.sup.16 is other than hydrogen, L is
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--OC(O)NR.sup.25-(alk).sub.b-, or --OC(S)NR.sup.25-(alk).sub.b-,
wherein b and R.sup.25 are as defined for Formula I, alk is
C.sub.1-3 alkylene optionally substituted with fluoro or optionally
fluoro substituted lower alkyl, R.sup.24 is hydrogen, optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl, R.sup.a and R.sup.b are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, and optionally fluoro substituted lower alkoxy. In
further embodiments, R.sup.12 is other than hydrogen and R.sup.14,
R.sup.15 and R.sup.16 are hydrogen. In further embodiments,
R.sup.16 is other than hydrogen and R.sup.12, R.sup.15 and R.sup.16
are hydrogen. In further embodiments, at least two of R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are other than hydrogen. In further
embodiments, R.sup.12 and R.sup.16 are other than hydrogen and
R.sup.14 and R.sup.15 are hydrogen. In further embodiments, R.sup.5
is other than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In
further embodiments, R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen.
[0206] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.13 is -LR.sup.24, -LR.sup.24
is not --NH.sub.2, --OH, --OCH.sub.3, --NHS(O).sub.2CH.sub.3,
##STR00108##
wherein
##STR00109##
indicates the bond to the phenyl ring.
[0207] In some embodiments of compounds of Formula II, R.sup.14 is
-LR.sup.24, provided, however, that when R.sup.4, R.sup.6,
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, R.sup.5 is
hydrogen or thiophen-2-yl and A is --C(O)--, then R.sup.14 is not
--OH, or --OCH.sub.3; and when R.sup.4, R.sup.5, R.sup.6, R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen and A is --CH.sub.2--,
R.sup.14 is not
##STR00110##
wherein
##STR00111##
indicates the bond to the phenyl ring, when R.sup.4, R.sup.6,
R.sup.12, R.sup.15 and R.sup.16 are hydrogen, A is --CH.sub.2--,
and R.sup.5 is thiophen-3-yl, R.sup.13 and R.sup.14 are not both
--OH; and when R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 are hydrogen and A is --S--, R.sup.14 is not --CH.sub.3;
and when R.sup.14 is --N(CH.sub.3).sub.2, R.sup.4, R.sup.6,
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen, and A is
--CH.sub.2--, then R.sup.5 is not 3-hydroxy-phenyl; and when
R.sup.14 is --NH.sub.2, R.sup.4, R.sup.6, R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen, and A is --S(O).sub.2--, then
R.sup.5 is not
##STR00112##
wherein
##STR00113##
indicates the bond to the 5-position of the 7-azaindole ring. In
further embodiments, when L is --O--, R.sup.24 is not H or
CH.sub.3; and when L is --OCH.sub.2--, R.sup.24 is not H. In some
embodiments, one of R.sup.12, R.sup.13, R.sup.15, and R.sup.16 is
other than hydrogen and the others are hydrogen. In some
embodiments, at least one of R.sup.12, R.sup.13, R.sup.15, and
R.sup.16 are other than hydrogen. In some embodiments, two of
R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are other than hydrogen
and the others are hydrogen. In some embodiments, at least two of
R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are other than hydrogen.
In a some embodiments, at least three of R.sup.12, R.sup.13,
R.sup.15, and R.sup.16 are other than hydrogen. In some
embodiments, R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are other
than hydrogen. In some embodiments, one of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 is hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio, and the
others of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen.
In some embodiments, any two of R.sup.12, R.sup.13, R.sup.15 and
R.sup.16 is hydrogen, halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio and the remaining of R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen. In some embodiments,
R.sup.14 is --OR.sup.24, where R.sup.24 is optionally substituted
C.sub.2-6 alkyl, optionally substituted lower alkenyl, optionally
substituted lower alkynyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl, or R.sup.14 is
--O-alk-R.sup.24, where R.sup.24 is optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl. In some
embodiments, R.sup.14 is --SR.sup.24, where R.sup.24 is hydrogen,
optionally substituted lower alkyl, optionally substituted lower
alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.14 is --S--alk-R.sup.24, where R.sup.24 is optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl.
In some embodiments, R.sup.14 is --NHR.sup.24, where R.sup.24 is
hydrogen, optionally substituted lower alkyl, optionally
substituted lower alkenyl, optionally substituted lower alkynyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, or R.sup.14 is --NH-alk-R.sup.24, where
R.sup.24 is optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl. In some embodiments, A is
--CH.sub.2-- or --C(O)--. In some embodiments, R.sup.5 is other
than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In some
embodiments, R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen. In some embodiments, R.sup.6 is other than hydrogen
and R.sup.4 and R.sup.5 are hydrogen. In some embodiments, A is
--CH.sub.2--, R.sup.5 is other than hydrogen and R.sup.4 and
R.sup.6 are hydrogen. In some embodiments, A is --CH.sub.2--,
R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6 are
hydrogen. In some embodiments, A is --CH.sub.2--, R.sup.6 is other
than hydrogen and R.sup.4 and R.sup.5 are hydrogen. In some
embodiments, A is --C(O)--, R.sup.5 is other than hydrogen and
R.sup.4 and R.sup.6 are hydrogen. In some embodiments, A is
--C(O)--, R.sup.4 is other than hydrogen and R.sup.5 and R.sup.6
are hydrogen. In some embodiments, A is --C(O)--, R.sup.6 is other
than hydrogen and R.sup.4 and R.sup.5 are hydrogen.
[0208] In some embodiments of compounds of Formula II, R.sup.14 is
--OR.sup.24 where R.sup.24 is optionally substituted C.sub.2-6
alkyl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, or R.sup.14 is --SR.sup.24 where R.sup.24
is hydrogen, optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or R.sup.14 is --NHR.sup.24, where R.sup.24 is hydrogen, optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl. In some embodiments,
any one of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 is hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining of R.sup.12, R.sup.13, R.sup.15
and R.sup.16 are hydrogen. In some embodiments, two of R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining two of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, R.sup.12
is hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and R.sup.13, R.sup.15 and R.sup.16 are
hydrogen. In some embodiments, A is --CH.sub.2-- or --C(O)--. In
some embodiments, R.sup.5 is other than hydrogen and R.sup.4 and
R.sup.6 are hydrogen. In some embodiments, R.sup.4 is other than
hydrogen and R.sup.5 and R.sup.6 are hydrogen. In some embodiments,
R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5 are
hydrogen. In some embodiments, A is --CH.sub.2--, R.sup.5 is other
than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In some
embodiments, A is --CH.sub.2--, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In some embodiments, A is
--CH.sub.2--, R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen. In some embodiments, A is --C(O)--, R.sup.5
is other than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In
some embodiments, A is --C(O)--, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In some embodiments, A is
--C(O)--, R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5
are hydrogen.
[0209] In some embodiments of compounds of Formula II, R.sup.14 is
-LR.sup.24, where L is --NR.sup.25-(alk).sub.b-,
--C(O)NR.sup.25-(alk).sub.b-, --OC(O)NR.sup.25-(alk).sub.b-,
--OC(S)NR.sup.25-(alk).sub.b-, --C(S)NR.sup.25-(alk).sub.b-,
--S(O).sub.2NR.sup.25-(alk).sub.b-, --NR.sup.25C(O)-(alk).sub.b-,
--NR.sup.25C(S)-(alk).sub.b-,
--NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(S)NR.sup.25-(alk).sub.b-,
--NR.sup.25C(O)O-(alk).sub.b-, --NR.sup.25C(S)O-(alk).sub.b-,
--NR.sup.25S(O).sub.2-(alk).sub.b-, or
--NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-, wherein b, alk and
R.sup.25 are as defined for Formula I, and R.sup.24 is optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl.
In some embodiments, any one of R.sup.12, R.sup.13, R.sup.15, and
R.sup.16 is other than hydrogen and the remaining of R.sup.12,
R.sup.13, R.sup.15, and R.sup.16 are hydrogen. In some embodiments,
at least one of R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are
other than hydrogen. In some embodiments, any two of R.sup.12,
R.sup.13, R.sup.15, and R.sup.16 are other than hydrogen and the
remaining of R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are
hydrogen. In some embodiments, at least two of R.sup.12, R.sup.13,
R.sup.15, and R.sup.16 are other than hydrogen. In a some
embodiments, at least three of R.sup.12, R.sup.13, R.sup.15, and
R.sup.16 are other than hydrogen. In a some embodiments, R.sup.12,
R.sup.13, R.sup.15, and R.sup.16 are other than hydrogen. In some
embodiments, any one of R.sup.12, R.sup.13, R.sup.15 and R.sup.16
is hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio, and the remaining of R.sup.12,
R.sup.13, R.sup.15, and R.sup.16 are hydrogen. In some embodiments,
two of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the others of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, A is
selected from --CH.sub.2-- and --C(O)--. In some embodiments,
R.sup.5 is other than hydrogen and R.sup.4 and R.sup.6 are
hydrogen. In some embodiments, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In some embodiments, R.sup.6 is
other than hydrogen and R.sup.4 and R.sup.5 are hydrogen. In some
embodiments, A is --CH.sub.2--, R.sup.5 is other than hydrogen and
R.sup.4 and R.sup.6 are hydrogen. In some embodiments, A is
--CH.sub.2--, R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen. In some embodiments, A is --CH.sub.2--,
R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5 are
hydrogen. In some embodiments, A is --C(O)--, R.sup.5 is other than
hydrogen and R.sup.4 and R.sup.6 are hydrogen. In some embodiments,
A is --C(O)--, R.sup.4 is other than hydrogen and R.sup.5 and
R.sup.6 are hydrogen. In some embodiments, A is --C(O)--, R.sup.6
is other than hydrogen and R.sup.4 and R.sup.5 are hydrogen.
[0210] In some embodiments of any of the above embodiments of
compounds of Formula II wherein R.sup.14 is -LR.sup.24, -LR.sup.24
is not --NH.sub.2, --OH, --OCH.sub.3, --N(CH.sub.3).sub.2, or
##STR00114##
wherein
##STR00115##
indicates the bond to the phenyl ring.
[0211] In some embodiments of compounds of Formula II, R.sup.13 and
R.sup.15 are halogen, optionally fluoro substituted lower alkyl,
--OR.sup.24, where R.sup.24 is optionally substituted C.sub.2-6
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl, --O-alk-R.sup.24, where R.sup.24
is optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, --SR.sup.24, where R.sup.24 is hydrogen,
optionally substituted lower alkyl, optionally substituted lower
alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl,
--S--alk-R.sup.24, where R.sup.24 is optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl,
--NHR.sup.24, where R.sup.24 is hydrogen, optionally substituted
lower alkyl, optionally substituted lower alkenyl, optionally
substituted lower alkynyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl, or --NH--alk-R.sup.24,
where R.sup.24 is optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, or
optionally substituted heteroaryl. In some embodiments, at least
one of R.sup.12, R.sup.14 and R.sup.16 is other than hydrogen. In
some embodiments, at least two of R.sup.12, R.sup.14 and R.sup.16
are other than hydrogen. In some embodiments, R.sup.12, R.sup.14
and R.sup.16 are other than hydrogen. In some embodiments, any one
of R.sup.12, R.sup.14 and R.sup.16 is hydrogen, halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio and the
remaining of R.sup.12, R.sup.14 and R.sup.16 are hydrogen. In some
embodiments, any two of R.sup.12, R.sup.14 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and the remaining of
R.sup.12, R.sup.14 and R.sup.16 is hydrogen. In some embodiments,
R.sup.12, R.sup.14 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, A is --CH.sub.2-- or
--C(O)--. In some embodiments, R.sup.5 is other than hydrogen and
R.sup.4 and R.sup.6 are hydrogen. In some embodiments, R.sup.4 is
other than hydrogen and R.sup.5 and R.sup.6 are hydrogen. In some
embodiments, R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5
are hydrogen. In some embodiments, A is --CH.sub.2--, R.sup.5 is
other than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In some
embodiments, A is --CH.sub.2--, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In some embodiments, A is
--CH.sub.2--, R.sup.6 is other than hydrogen and R.sup.4 and
R.sup.5 are hydrogen. In some embodiments, A is --C(O)--, R.sup.5
is other than hydrogen and R.sup.4 and R.sup.6 are hydrogen. In
some embodiments, A is --C(O)--, R.sup.4 is other than hydrogen and
R.sup.5 and R.sup.6 are hydrogen. In some embodiments, A is
--C(O)--, R.sup.6 is other than hydrogen and R.sup.4 and R.sup.5
are hydrogen.
[0212] The compounds of Formula II, and all sub-embodiments
detailed herein, may be used to treat a subject suffering from or
at risk for any of the protein kinase mediated diseases or
conditions contemplated herein.
[0213] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structure Formula
IIa:
##STR00116##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.16 and R.sup.24 are as
defined for Formula II, provided, however, that when A is C(O),
R.sup.4, R.sup.5, R.sup.6, and R.sup.12 are hydrogen and R.sup.16
is fluoro, then R.sup.24 is not CH.sub.3.
[0214] In some embodiments of compounds of Formula IIa, A is
--CH.sub.2-- or --C(O)--.
[0215] In other embodiments of compounds of Formula IIa, A is
--CH.sub.2-- or --C(O)--and R.sup.12 and R.sup.16 are independently
hydrogen, halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio; in further embodiments, R.sup.12 and
R.sup.16 are independently hydrogen, halogen, optionally fluoro
substituted C.sub.1-3alkyl, optionally fluoro substituted
C.sub.1-3alkoxy, or optionally fluoro substituted
C.sub.1-3alkylthio. In some embodiments, A is --CH.sub.2-- or
--C(O)--, and R.sup.12 and R.sup.16 are independently halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio; in further embodiments, R.sup.12 and R.sup.16 are
independently halogen, optionally fluoro substituted
C.sub.1-3alkyl, optionally fluoro substituted C.sub.1-3alkoxy, or
optionally fluoro substituted C.sub.1-3alkylthio.
[0216] In other embodiments of compounds of Formula IIa, A is
--CH.sub.2--, R.sup.12 is hydrogen and R.sup.16 is halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio; in further embodiments, R.sup.16 is halogen, optionally
fluoro substituted C.sub.1-3alkyl, optionally fluoro substituted
C.sub.1-3alkoxy, or optionally fluoro substituted
C.sub.1-3alkylthio. In some embodiments, A is --CH.sub.2--,
R.sup.16 is hydrogen and R.sup.12 is halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio; in
further embodiments, R.sup.12 is halogen, optionally fluoro
substituted C.sub.1-3alkyl, optionally fluoro substituted
C.sub.1-3alkoxy, or optionally fluoro substituted
C.sub.1-3alkylthio.
[0217] In some embodiments of compounds of Formula IIa, A is
--CH.sub.2-- and R.sup.12 and R.sup.16 are independently halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio; in further embodiments, R.sup.12 and R.sup.16 are
independently halogen, optionally fluoro substituted
C.sub.1-3alkyl, optionally fluoro substituted C.sub.1-3alkoxy, and
optionally fluoro substituted C.sub.1-3alkylthio.
[0218] In other embodiments of compounds of Formula IIa, A is
--C(O)--, R.sup.12 is hydrogen and R.sup.16 is halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio; in
further embodiments, R.sup.16 is halogen, optionally fluoro
substituted C.sub.1-3alkyl, optionally fluoro substituted
C.sub.1-3alkoxy, or optionally fluoro substituted
C.sub.1-3alkylthio. In some embodiments, A is --C(O)--, R.sup.16 is
hydrogen and R.sup.12 is halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio; in further
embodiments, R.sup.12 is halogen, optionally fluoro substituted
C.sub.1-3alkyl, optionally fluoro substituted C.sub.1-3alkoxy, or
optionally fluoro substituted C.sub.1-3alkylthio.
[0219] In some embodiments of compounds of Formula IIa, A is
--C(O)--and R.sup.12 and R.sup.16 are independently halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio; in further embodiments R.sup.12 and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
C.sub.1-3alkyl, optionally fluoro substituted C.sub.1-3alkoxy, or
optionally fluoro substituted C.sub.1-3alkylthio.
[0220] In some embodiments of any of the above embodiments of
compounds of Formula IIa, R.sup.4 and R.sup.5 are hydrogen, or
R.sup.5 and R.sup.6 are hydrogen, or R.sup.4 and R.sup.5 are
hydrogen.
[0221] In some embodiments of any of the above embodiments of
compounds of Formula IIa, R.sup.24 is substituted methyl,
optionally substituted C.sub.2-6alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl; in further
embodiments, R.sup.24 is C.sub.2-6lower alkyl, aryl or heteroaryl,
wherein aryl and heteroaryl are optionally substituted with
halogen, lower alkyl or lower alkoxy; in further embodiments,
R.sup.24 is n-propyl, i-propyl, or phenyl, wherein phenyl is
optionally substituted with halogen, lower alkyl or lower
alkoxy.
[0222] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structure Formula
IIb:
##STR00117##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15 and
R.sup.16, are as defined for Formula II, R.sup.28 is hydrogen,
lower alkyl, or lower alkyl substituted with fluoro, hydroxyl,
lower alkoxy, thiol, lower alkylthio, or --NR.sup.8R.sup.9, wherein
R.sup.8 and R.sup.9 are as defined for Formula I, provided,
however, that when R.sup.28 is substituted lower alkyl, any
substitution of the alkyl carbon bound to the nitrogen of NR.sup.28
is fluoro, and R.sup.29 is hydrogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, provided, however,
that nitrogen of NR.sup.29 is not bound to any alkene carbon
thereof, optionally substituted lower alkynyl, provided, however,
that nitrogen of NR.sup.29 is not bound to any alkyne carbon
thereof, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, or R.sup.28 and R.sup.29 combine with the
nitrogen to which they are attached form optionally substituted 5-7
membered heterocycloalkyl or optionally substituted 5 or 7 membered
nitrogen containing heteroaryl. In some embodiments of compounds of
Formula IIb, A is --CH.sub.2-- or --C(O)--, R.sup.12, R.sup.14,
R.sup.15, and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio, R.sup.28 is hydrogen or lower alkyl and R.sup.29 is
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, or R.sup.28 and R.sup.29 combine with the
nitrogen to which they are attached form optionally substituted 5-7
membered heterocycloalkyl or optionally substituted 5 or 7 membered
nitrogen containing heteroaryl.
[0223] In some embodiments of compounds of Formula IIb, A is
--CH.sub.2--, one of R.sup.12, R.sup.14, R.sup.15 and R.sup.16 is
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining of R.sup.12, R.sup.14, R.sup.15
and R.sup.16 are hydrogen.
[0224] In other embodiments of compounds of Formula IIb, A is
--CH.sub.2--, R.sup.12, R.sup.14 and R.sup.15 are hydrogen and
R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio.
[0225] In other embodiments of compounds of Formula IIb, A is
--CH.sub.2--, R.sup.14, R.sup.15 and R.sup.16 are hydrogen and
R.sup.12 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio.
[0226] In some embodiments of compounds of Formula IIb, A is
--CH.sub.2--, two of R.sup.12, R.sup.14, R.sup.13 and R.sup.16 are
independently halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the other two of R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen.
[0227] In some embodiments of compounds of Formula IIb, A is
--CH.sub.2--, R.sup.14 and R.sup.15 are hydrogen and R.sup.12 and
R.sup.16 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio.
[0228] In some embodiments of compounds of Formula IIb, A is
--C(O)--, one of R.sup.12, R.sup.14, R.sup.15 and R.sup.16 is
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the others of R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 are hydrogen.
[0229] In some embodiments of compounds of Formula IIb, A is
--C(O)--, R.sup.12, R.sup.14 and R.sup.15 are hydrogen and R.sup.16
is halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio.
[0230] In some embodiments of compounds of Formula IIb, A is
--C(O)--, R.sup.14, R.sup.15 and R.sup.16 are hydrogen and R.sup.12
is halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio.
[0231] In some embodiments of compounds of Formula IIb, A is
--C(O)--, two of R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are
independently halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the other two of R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen.
[0232] In some embodiments of compounds of Formula IIb, A is
--C(O)--, R.sup.14 and R.sup.15 are hydrogen and R.sup.12 and
R.sup.16 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio.
[0233] In some embodiments of any of the above embodiments of
compounds of Formula IIb, R.sup.4 and R.sup.6 are hydrogen, or
R.sup.5 and R.sup.6 are hydrogen, or R.sup.4 and R.sup.5 are
hydrogen. In some embodiments of any of the above embodiments of
compounds of Formula IIb, R.sup.28 and R.sup.29 are both lower
alkyl, or R.sup.28 and R.sup.29 combine with the nitrogen to which
they are attached to form optionally substituted 5-7 membered
heterocycloalkyl, further wherein the heterocycloalkyl is
pyrrolidine, piperidine, piperazine or morpholine.
[0234] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structures
Formulae IIc, IId, IIe, IIf, or IIg:
##STR00118##
all salts, prodrugs, tautomers and isomers thereof, wherein A, X,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15, R.sup.16
and R.sup.24 are as defined for Formula II. In some embodiments of
compounds of Formulae IIc, IId, IIe, IIf, or IIg, A is --CH.sub.2--
or --C(O)--. In some embodiments, any one of R.sup.12, R.sup.14,
R.sup.15, and R.sup.16 is other than hydrogen. In some embodiments,
any two of R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are other
than hydrogen. In some embodiments, any three of R.sup.12,
R.sup.14, R.sup.15, and R.sup.16 are other than hydrogen. In some
embodiments, R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are other
than hydrogen. In some embodiments, A is --CH.sub.2-- or
--C(O)--and R.sup.12, R.sup.14, R.sup.15, and R.sup.16 are
independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
A is --CH.sub.2--, any one of R.sup.12, R.sup.14, R.sup.15 and
R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the remaining of R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some embodiments,
A is --CH.sub.2--, R.sup.12, R.sup.14 and R.sup.15 are hydrogen and
R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, A is
--CH.sub.2--, R.sup.14, R.sup.15 and R.sup.16 are hydrogen and
R.sup.12 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, A is
--CH.sub.2--, any two of R.sup.12, R.sup.14, R.sup.15 and R.sup.16
are independently halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio and the remaining two of
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some
embodiments, A is --CH.sub.2--, R.sup.14 and R.sup.15 are hydrogen
and R.sup.12 and R.sup.16 are independently halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In some
embodiments, A is --C(O)--, any one of R.sup.12, R.sup.14, R.sup.15
and R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the remaining of R.sup.12,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen. In some embodiments,
A is --C(O)--, R.sup.12, R.sup.14 and R.sup.15 are hydrogen and
R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, A is --C(O)--,
R.sup.14, R.sup.15 and R.sup.16 are hydrogen and R.sup.12 is
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, A is --C(O)--, any two of
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are independently
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining two of R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, A is
--C(O)--, R.sup.14 and R.sup.15 are hydrogen and R.sup.12 and
R.sup.16 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments
of any of the above embodiments of compounds of Formula IIc, IId,
IIe, IIf, and IIg, R.sup.4 and R.sup.6 are hydrogen or R.sup.5 and
R.sup.6 are hydrogen, or R.sup.4 and R.sup.5 are hydrogen.
[0235] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structure Formula
IIh:
##STR00119##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
R.sup.3, R.sup.12, R.sup.13, R.sup.15, R.sup.16 and R.sup.24 are as
defined for Formula II, and Z is --O--, --S--or --NH--. In some
embodiments of compounds of Formula IIh, R.sup.24 is hydrogen,
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl. In some
embodiments, R.sup.24 is hydrogen, optionally substituted lower
alkyl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl and at least one of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 is halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and the others of
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen; in further
embodiments, R.sup.13 is halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and R.sup.12,
R.sup.15 and R.sup.16 are hydrogen; in further embodiments, A is
--CH.sub.2-- or --C(O)--.
[0236] In some embodiments of compounds of Formula IIh, Z is --O--
or --NH--, R.sup.24 is hydrogen, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, or lower alkyl optionally
substituted with optionally substituted aryl or optionally
substituted heteroaryl, and at least one of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 is halogen, optionally fluoro substituted
lower alkyl, or optionally fluoro substituted lower alkoxy and the
others of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are hydrogen;
in further embodiments, R.sup.13 is halogen, optionally fluoro
substituted lower alkyl, or optionally fluoro substituted lower
alkoxy and R.sup.12, R.sup.15 and R.sup.16 are hydrogen; in further
embodiments, A is --CH.sub.2-- or --C(O)--.
[0237] In some embodiments of compounds of Formula IIh, Z is --O--
or --NH--, R.sup.24 is hydrogen, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, or lower alkyl optionally
substituted with optionally substituted aryl or optionally
substituted heteroaryl, R.sup.13 is halogen, optionally fluoro
substituted lower alkyl, or optionally fluoro substituted lower
alkoxy and one of R.sup.12, R.sup.15 and R.sup.16 is halogen,
optionally fluoro substituted lower alkyl, or optionally fluoro
substituted lower alkoxy and the others of R.sup.12, R.sup.15 and
R.sup.16 are hydrogen; in further embodiments, A is --CH.sub.2-- or
--C(O)--.
[0238] In some embodiments of compounds of Formula IIh, Z is --O--
or --NH--, R.sup.24 is hydrogen, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, or lower alkyl optionally
substituted with optionally substituted aryl or optionally
substituted heteroaryl, R.sup.13 is halogen, optionally fluoro
substituted lower alkyl, or optionally fluoro substituted lower
alkoxy and two of R.sup.12, R.sup.15 and R.sup.16 are independently
halogen, optionally fluoro substituted lower alkyl, or optionally
fluoro substituted lower alkoxy and the other of R.sup.12, R.sup.15
and R.sup.16 is hydrogen; in further embodiments, A is --CH.sub.2--
or --C(O)--.
[0239] In some embodiments of compounds of Formula IIh, Z is --O--
or --NH--, R.sup.24 is hydrogen, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, or lower alkyl optionally
substituted with optionally substituted aryl or optionally
substituted heteroaryl, R.sup.12, R.sup.13, R.sup.15 and R.sup.16
are independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, or optionally fluoro substituted lower alkoxy; in
further embodiments, A is --CH.sub.2-- or --C(O)--.
[0240] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structures
Formulae IIi, IIj, IIk, IIm, or IIn:
##STR00120##
all salts, prodrugs, tautomers and isomers thereof, wherein A, X,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.13, R.sup.15, R.sup.16
and R.sup.24 are as defined for Formula II. In some embodiments of
compounds of Formulae IIi, IIj, IIk, IIm, or IIn, A is --CH.sub.2--
or --C(O)--. In some embodiments, one of R.sup.12, R.sup.13,
R.sup.15, and R.sup.16 is other than hydrogen. In some embodiments,
two of R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are other than
hydrogen. In one embodiment, three of R.sup.12, R.sup.13, R.sup.15,
and R.sup.16 are other than hydrogen. In some embodiments,
R.sup.12, R.sup.13, R.sup.15, and R.sup.16 are other than
hydrogen.
[0241] In some embodiments of compounds of Formulae IIi, IIj, IIk,
IIm, or IIn, A is --CH.sub.2-- or --C(O)--, R.sup.12, R.sup.13,
R.sup.15, and R.sup.16 are independently hydrogen, halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, A is --CH.sub.2--, any one of
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 is halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio and the
remaining of R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are
hydrogen. In some embodiments, A is --CH.sub.2--, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen and R.sup.12 is halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, A is --CH.sub.2--, R.sup.12,
R.sup.15 and R.sup.16 are hydrogen and R.sup.13 is halogen,
optionally fluoro substituted lower alkyl, optionally fluoro
substituted lower alkoxy, or optionally fluoro substituted lower
alkylthio. In some embodiments, A is --CH.sub.2--, any two of
R.sup.12, R.sup.13, R.sup.13 and R.sup.16 are independently
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining two of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, A is
--CH.sub.2--, R.sup.15 and R.sup.16 are hydrogen and R.sup.12 and
R.sup.13 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
A is --CH.sub.2--, R.sup.13 and R.sup.16 are hydrogen and R.sup.12
and R.sup.15 are independently halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In some
embodiments, A is --CH.sub.2--, R.sup.13 and R.sup.15 are hydrogen
and R.sup.12 and R.sup.16 are independently halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio. In some
embodiments, A is --CH.sub.2--, R.sup.12 and R.sup.16 are hydrogen
and R.sup.13 and R.sup.15 are independently halogen, optionally
fluoro substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio.
[0242] In some embodiments of compounds of Formulae IIi, IIj, IIk,
IIm, or IIn, A is --C(O)--, any one of R.sup.12, R.sup.13, R.sup.15
and R.sup.16 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio and the remaining of R.sup.12,
R.sup.13, R.sup.15 and R.sup.16 are hydrogen. In some embodiments,
A is --C(O)--, R.sup.13, R.sup.15 and R.sup.16 are hydrogen and
R.sup.12 is halogen, optionally fluoro substituted lower alkyl,
optionally fluoro substituted lower alkoxy, or optionally fluoro
substituted lower alkylthio. In some embodiments, A is --C(O)--,
R.sup.12, R.sup.15 and R.sup.16 are hydrogen and R.sup.13 is
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio. In some embodiments, A is --C(O)--, two of
R.sup.12, R.sup.13, R.sup.15 and R.sup.16 are independently
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and the remaining two of R.sup.12, R.sup.13,
R.sup.15 and R.sup.16 are hydrogen. In some embodiments, A is
--C(O)--, R.sup.15 and R.sup.16 are hydrogen and R.sup.12 and
R.sup.13 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
A is --C(O)--, R.sup.13 and R.sup.16 are hydrogen and R.sup.12and
R.sup.15 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
A is --C(O)--, R.sup.13 and R.sup.15 are hydrogen and R.sup.12 and
R.sup.16 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments,
A is --C(O)--, R.sup.12 and R.sup.16 are hydrogen and R.sup.13 and
R.sup.15 are independently halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio. In some embodiments
of any of the above embodiments of compounds of Formula IIa,
R.sup.4 and R.sup.6 are hydrogen or R.sup.5 and R.sup.6 are
hydrogen, or R.sup.4 and R.sup.5 are hydrogen.
[0243] In some embodiments, compounds of Formula II have the
structure according to the following sub-generic structure Formula
IIo:
##STR00121##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
R.sup.5, R.sup.12, R.sup.14, R.sup.16 and R.sup.24 are as defined
for Formula II, where each R.sup.24 is selected independently, and
each Z is independently --O--, --S--or --NH--. In some embodiments
of compounds of Formula IIo, each R.sup.24 is independently
hydrogen, optionally substituted lower alkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, or optionally substituted heteroaryl.
In some embodiments, each R.sup.24 is independently hydrogen,
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, or optionally substituted heteroaryl and
R.sup.12, R.sup.14 and R.sup.16 are hydrogen.
[0244] In some embodiments of compounds of Formula IIo, each
R.sup.24 is independently hydrogen, optionally substituted lower
alkyl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl and at least one of R.sup.12, R.sup.14 and
R.sup.16 is hydrogen, halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio, and the others of R.sup.12,
R.sup.14 and R.sup.16 are hydrogen; in further embodiments,
R.sup.12 is hydrogen, halogen, optionally fluoro substituted lower
alkyl, optionally fluoro substituted lower alkoxy, or optionally
fluoro substituted lower alkylthio and R.sup.14 and R.sup.16 are
hydrogen; in further embodiments, A is --CH.sub.2-- or
--C(O)--.
[0245] In some embodiments of compounds of Formula IIo, each
R.sup.24 is independently hydrogen, optionally substituted lower
alkyl, optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl and at least two of R.sup.12, R.sup.14 and
R.sup.16 are independently hydrogen, halogen, optionally fluoro
substituted lower alkyl, optionally fluoro substituted lower
alkoxy, or optionally fluoro substituted lower alkylthio, and the
other of R.sup.12, R.sup.14 and R.sup.16 is hydrogen; in further
embodiments, R.sup.12 and R.sup.16 are independently hydrogen,
halogen, optionally fluoro substituted lower alkyl, optionally
fluoro substituted lower alkoxy, or optionally fluoro substituted
lower alkylthio and R.sup.14 is hydrogen; in further embodiments, A
is --CH.sub.2-- or --C(O)--.
[0246] In some embodiments of compounds of Formula IIo, L is --O--
or --NH--, each R.sup.24 is independently hydrogen, optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, or optionally substituted heteroaryl, R.sup.12 and R.sup.16
are independently hydrogen, halogen, optionally fluoro substituted
lower alkyl, optionally fluoro substituted lower alkoxy, or
optionally fluoro substituted lower alkylthio and R.sup.14 is
hydrogen; in further embodiments, A is --CH.sub.2-- or
--C(O)--.
[0247] The compounds of Formulae IIa-IIo, and all sub-embodinients
detailed herein, may be used to treat a subject suffering from or
at risk for any of the protein kinase mediated diseases or
conditions contemplated herein.
[0248] In some embodiments, compounds of Formula I have the
structure according to the following sub-generic structure Formula
III:
##STR00122##
all salts, prodrugs, tautomers, and isomers thereof, [0249]
wherein: [0250] Q has a structure selected from the group
consisting of
##STR00123##
[0250] in which
##STR00124##
indicates the attachment point of Q to A of Formula III; [0251]
Z.sub.2 is N or CR.sup.12; Z.sub.4 is N or CR.sup.14; Z.sub.5 is N
or CR.sup.15; Z.sub.6 is N or CR.sup.16; [0252] L.sub.2 is selected
from the group consisting of
--(CR.sup.10R.sup.11).sub.p--NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--O--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--, --C(O)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(S)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O)--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O).sub.2--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(O)NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--C(S)NR.sup.25--(CR.sup.10R.sup.11).sub.q--,
--(CR.sup.10R.sup.11).sub.p--S(O).sub.2NR.sup.25--(CR.sup.10R.sup.11).sub-
.q--,
--(CR.sup.10R.sup.11).sub.p--NR.sup.25C(O)--(CR.sup.10R.sup.11).sub.-
q--,
--(CR.sup.10R.sup.11).sub.p--NR.sup.25C(S)--(CR.sup.10R.sup.11).sub.q-
--, and
--(CR.sup.10R.sup.11).sub.p--NR.sup.25S(O).sub.2--(CR.sup.10R.sup.-
11).sub.q--; [0253] p and q are independently 0, 1, or 2 provided,
however, that at least one of p and q is 0; [0254] s is 1 or 2;
[0255] X is O or S; [0256] A is selected from the group consisting
of --O--, --S--, --CR.sup.aR.sup.b--, --NR.sup.1--, --C(O)--,
--C(S)--, --S(O)--, and --S(O).sub.2--; [0257] R.sup.a and R.sup.b
at each occurrence are independently selected from the group
consisting of hydrogen, fluoro, --OH, --NH.sub.2, lower alkyl,
lower alkoxy, lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein the alkyl chain(s) of lower alkyl, lower
alkoxy, lower alkylthio, mono-alkylamino, or di-alkylamino are
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, provided, however, that any substitution of the
alkyl chain carbon bound to O of alkoxy, S of thioalkyl or N of
mono- or di-alkylamino is fluoro; or [0258] R.sup.a and R.sup.b
combine to form a 3-7 membered monocyclic cycloalkyl or 5-7
membered monocyclic heterocycloalkyl, wherein the monocyclic
cycloalkyl or monocyclic heterocycloalkyl are optionally
substituted with one or more substituents selected from the group
consisting of halogen, --OH, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkyl amino; [0259]
R.sup.1 is selected from the group consisting of hydrogen, lower
alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
--C(O)R.sup.7, --C(S)R.sup.7, --S(O).sub.2R.sup.7, --C(O)NHR.sup.7,
--C(S)NHR.sup.7, and --S(O).sub.2NHR.sup.7, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
lower alkylthio, mono-alkylamino, di-alkylamino, and
--NR.sup.8R.sup.9, wherein the alkyl chain(s) of lower alkoxy,
lower alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro, further provided that when R.sup.1 is lower alkyl, any
substitution on the lower alkyl carbon bound to the N of
--NR.sup.1-is fluoro, and wherein cycloalkyl, heterocycloalkyl,
aryl or heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0260] R.sup.7 is selected from the group
consisting of lower alkyl, cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, lower alkylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, provided, however, that any
substitution of the alkyl carbon bound to the N of
--C(O)NHR.sup.7--, --C(S)NHR.sup.7 or --S(O).sub.2NHR.sup.7 is
fluoro, wherein the alkyl chain(s) of lower alkoxy, lower
alkylthio, mono-alkylamino, or di-alkylamino are optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the alkyl chain carbon
bound to O of alkoxy, S of thioalkyl or N of mono- or di-alkylamino
is fluoro, and wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with one or more substituents
selected from the group consisting of halogen, --OH, --NH.sub.2,
lower alkyl, fluoro substituted lower alkyl, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino;
[0261] R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15,
R.sup.16, R.sup.42, R.sup.43, R.sup.45, R.sup.46 and R.sup.47 are
independently selected from the group consisting of hydrogen,
halogen, optionally substituted lower alkyl, optionally substituted
lower alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --NO.sub.2, --CR.sup.aR.sup.bR.sup.26, and -LR.sup.26; [0262]
L at each occurrcnce is independently selected from the group
consisting of -(alk).sub.a-S-(alk).sub.b-,
-(alk).sub.a-O-(alk).sub.b-, -(alk).sub.a-NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(O)-(alk).sub.b-, -(alk).sub.a-C(S)-(alk).sub.b-,
-(alk).sub.a-S(O)-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-OC(O)-(alk).sub.b-, -(alk).sub.a-C(O)O-(alk).sub.b-,
-(alk).sub.a-OC(S)-(alk).sub.b-, -(alk).sub.a-C(S)O-(alk).sub.b-,
-(alk).sub.a-C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-C(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-S(O).sub.2NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)-(alk).sub.b-,
-(alk).sub.a-NR.sup.25S(O).sub.2-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)O-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)O-(alk).sub.b-,
-(alk).sub.a-OC(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-OC(S)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(O)NR.sup.25-(alk).sub.b-,
-(alk).sub.a-NR.sup.25C(S)NR.sup.25-(alk).sub.b-, and
-(alk).sub.a-NR.sup.25S(O).sub.2NR.sup.25-(alk).sub.b-; [0263] a
and b are independently 0 or 1; [0264] alk is C.sub.1-3 alkylene or
C.sub.1-3 alkylene substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkyl, lower alkoxy, lower alkylthio, mono-alkylamino,
di-alkylamino, and --NR.sup.8R.sup.9, wherein lower alkyl or the
alkyl chain(s) of lower alkoxy, lower alkylthio, mono-alkylamino or
di-alkylamino are optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, provided, however, that any
substitution of the alkyl chain carbon bound to O of alkoxy, S of
thioalkyl or N of mono- or di-alkylamino is fluoro; [0265] R.sup.25
at each occurrcnce is independently selected from the group
consisting of hydrogen, optionally substituted lower alkyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, and optionally
substituted heteroaryl; [0266] R.sup.26 at each occurrence is
independently selected from the group consisting of hydrogen,
provided, however, that hydrogen is not bound to any of S(O),
S(O).sub.2, C(O) or C(S) of L, optionally substituted lower alkyl,
optionally substituted lower alkenyl, provided, however, that when
R.sup.26 is optionally substituted lower alkenyl, no alkene carbon
thereof is bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of L,
optionally substituted lower alkynyl, provided, however, that when
R.sup.26 is optionally substituted lower alkynyl, no alkyne carbon
thereof is bound to N, S, O, S(O), S(O).sub.2, C(O) or C(S) of L,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, and optionally
substituted heteroaryl; [0267] R.sup.10 and R.sup.11 at each
occurrence are independently selected from the group consisting of
hydrogen, fluoro, lower alkyl, and lower alkyl optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino; or
[0268] any two of R.sup.10 and R.sup.11 on the same or adjacent
carbon atoms combine to form a 3-7 membered monocyclic cycloalkyl
or 5-7 membered monocyclic heterocycloalkyl, and any others of
R.sup.10 and R.sup.11 are independently selected from the group
consisting of hydrogen, fluoro, lower alkyl, and lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino, and wherein the monocyclic cycloalkyl or
monocyclic heterocycloalkyl are optionally substituted with one or
more substituents selected from the group consisting of halogen,
--OH, --NH.sub.2, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; [0269] R.sup.8 and R.sup.9 combine with the
nitrogen to which they are attached to form a 5-7 membered
heterocycloalkyl optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, and
fluoro substituted lower alkylthio; [0270] R.sup.17 is selected
from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl and --OR.sup.18; [0271] R.sup.31 and
R.sup.33 are independently selected from the group consisting of
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted cycloalkyl, and optionally substituted
heterocycloalkyl; [0272] R.sup.36 is selected from the group
consisting of substituted methyl, optionally substituted
C.sub.2-6alkyl, optionally substituted lower alkenyl, provided,
however, that when R.sup.36 is optionally substituted lower
alkenyl, no alkene carbon thereof is bound to the S(O).sub.2of
S(O).sub.2R.sup.36, optionally substituted lower alkynyl, provided,
however, that when R.sup.36 is optionally substituted lower
alkynyl, no alkyne carbon thereof is bound to the S(O).sub.2of
S(O).sub.2R.sup.36, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, and [0273] R.sup.19, R.sup.20,
R.sup.34, R.sup.35, R.sup.37, and R.sup.38 are independently
selected from the group consisting of hydrogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.19, R.sup.20, R.sup.34,
R.sup.35, R.sup.37, or R.sup.38 is optionally substituted lower
alkenyl, no alkene carbon thereof is bound to the N of
NR.sup.19R.sup.20, NR.sup.34R.sup.35 or NR.sup.37R.sup.38,
optionally substituted lower alkynyl, provided, however, that when
R.sup.19, R.sup.20, R.sup.34, R.sup.35, R.sup.37, or R.sup.38 is
optionally substituted lower alkynyl, no alkyne carbon thereofis
bound to the N of NR.sup.19R.sup.20, NR.sup.34R.sup.35 or
NR.sup.37R.sup.38, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl and
optionally substituted heteroaryl; or [0274] R.sup.34 and R.sup.35
together with the nitrogen to which they are attached form
optionally substituted 5-7 membered heterocycloalkyl or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl; or
[0275] R.sup.37 and R.sup.38 together with the nitrogen to which
they are attached form optionally substituted 5-7 membered
heterocycloalkyl or optionally substituted 5 or 7 membered nitrogen
containing heteroaryl; [0276] R.sup.32 is selected from the group
consisting of hydrogen, optionally substituted lower alkyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, optionally
substituted heteroaryl, and --OR.sup.18; [0277] R.sup.82 is
selected from hydrogen or lower alkyl; and [0278] R.sup.18 is
hydrogen or optionally substituted lower alkyl; [0279] provided,
however, that the compound is not
3-{3-[2-(tetrahydropyran-2-yloxy)-ethoxy]-benzyl}-5-thiophen-3-yl-1H-pyrr-
olo[2,3-b]pyridine, which has the structure
[0279] ##STR00125## [0280]
4-(4-Methyl-piperazin-1-ylmethyl)-N-[4-(1H-pyrrolo[2,3-b]pyridin-3-ylmeth-
yl)-phenyl]-benzamide, which has the structure
##STR00126##
[0281] The compounds of Formula III, and all sub-embodiments
detailed herein, may be used to treat a subject suffering from or
at risk for any of the protein kinase mediated diseases or
conditions contemplated herein.
[0282] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIa:
##STR00127##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
L.sub.2, Z.sub.2, Z.sub.6, R.sup.4, R.sup.5, R.sup.6, R.sup.15,
R.sup.17 and R.sup.31 are as defined for Formula III.
[0283] In some embodiments of compounds of Formula IIIa, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
more preferably --CH.sub.2--, R.sup.17 is selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro, and
R.sup.15 is selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy.
[0284] In some embodiments of compounds of Formula IIIa, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
R.sup.17 is selected from the group consisting of hydrogen,
halogen, lower alkyl and lower alkoxy, wherein the alkyl chain of
lower alkyl or lower alkoxy is optionally substituted with one or
more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro, R.sup.15 is selected from the
group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy, Z.sub.2 is N or CR.sup.12, Z.sub.6 is N or CR.sup.16,
R.sup.12 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, and fluoro substituted lower alkoxy, and
R.sup.5 is selected from the group consisting of hydrogen, halogen,
lower alkyl, lower alkoxy, optionally substituted aryl, optionally
substituted heteroaryl, and NR.sup.21R.sup.22, wherein R.sup.21 is
hydrogen or lower alkyl, and R.sup.22 is hydrogen, lower alkyl,
optionally substituted aryl or optionally substituted heteroaryl,
and wherein the alkyl chain of R.sup.5, R.sup.21 or R.sup.22, when
lower alkyl, or the alkyl chain of lower alkoxy is optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino and cycloalkylamino.
[0285] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIb:
##STR00128##
all salts, prodrugs, tautomers, and isomers thereof, [0286]
wherein: [0287] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--
or --NR.sup.48--; [0288] Z.sub.12 is N or CR.sup.52; [0289]
Z.sub.16 is N or CR.sup.56; [0290] R.sup.40 and R.sup.41 are
independently selected from the group consisting of hydrogen,
fluoro, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio/mono-alkylamino, di-alkylamino, and
cycloalkylamino; or [0291] R.sup.40 and R.sup.41 combine to form a
3-7 membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl, wherein the monocyclic cycloalkyl or monocyclic
heterocycloalkyl is optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0292] L.sub.3 is selected from the group
consisting of --NR.sup.48--, --S--, --O--,
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, --OCH(R.sup.49)--,
--C(O)NR.sup.48--, --S(O).sub.2NR.sup.48--,
--CH(R.sup.49)NR.sup.48--, --CH(R.sup.49)O--, --CH(R.sup.49)S--,
--NR.sup.48C(O)--, and --NR.sup.48S(O).sub.2--; [0293] R.sup.53 and
R.sup.55 are independently selected from the group consisting of
hydrogen, halogen, lower alkyl and lower alkoxy, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino or cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; [0294] R.sup.52 and R.sup.56 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy; [0295] R.sup.49 is selected
from the group consisting of hydrogen, lower alkyl, and fluoro
substituted lower alkyl; [0296] Cy is selected from the group
consisting of aryl, heteroaryl; cycloalkyl, and heterocycloalkyl;
[0297] R.sup.39 is selected from the group consisting of hydrogen,
halogen, lower alkyl, lower alkoxy, aryl, heteroaryl, and
NR.sup.50R.sup.51, wherein the alkyl chain of lower alkyl or lower
alkoxy is optionally substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino
and cycloalkylamino, and wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
[0298] R.sup.50 is hydrogen or lower alkyl optionally substituted
with one or more substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, tower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino; [0299] R.sup.51
is aryl or heteroaryl, wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
[0300] R.sup.23 at each occurrence is independently selected from
the group consisting of --OH, NH.sub.2, --CN, --NO.sub.2, --C(O)OH,
--S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --NR.sup.48C(O)R.sup.57,
--NR.sup.48S(O).sub.2R.sup.57, --S(O).sub.2R.sup.57,
--C(O)R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57,
--S(O).sub.2NR.sup.48R.sup.57, halogen, lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl as R.sup.23, or as substituents of lower alkyl, are
optionally substituted with one or more substituents selected from
the group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2,
--C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.58,
--SR.sup.58, --NR.sup.48R.sup.58, --NR.sup.48C(O)R.sup.58,
--NR.sup.48S(O).sub.2R.sup.58, --S(O).sub.2R.sup.58,
--C(O)R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58,
--S(O).sub.2NR.sup.48R.sup.58, halogen, lower alkyl, fluoro
substituted lower alkyl, and cycloalkylamino; [0301] R.sup.57 is
selected from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0302] R.sup.58 at each occurrence is independently selected from
the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48SR.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0303] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0304] t
is 0, 1, 2, or 3.
[0305] In some embodiments of compounds of Formula IIIb, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --CH.sub.2--. In some embodiments,
A.sub.1 is --CR.sup.40R.sup.41-- or --C(O)--, preferably
--CH.sub.2-- or --C(O)--, more preferably --CH.sub.2--, and
R.sup.53 and R.sup.55 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, and fluoro substituted lower alkoxy. In
some embodiments, L.sub.3 is --NR.sup.48CH(R.sup.49)--,
--SCH(R.sup.49)--, or --OCH(R.sup.49)--, preferably
--OCH(R.sup.49)--. In some embodiments, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --CH.sub.2--, and L.sub.3 is
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, or --OCH(R.sup.49)--,
preferably --OCH(R.sup.49)--.
[0306] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIp:
##STR00129##
all salts, prodrugs, tautomers, and isomers thereof, [0307]
wherein: [0308] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--
or --NR.sup.48--; [0309] Z.sub.22 is N or CR.sup.62; [0310]
Z.sub.26 is N or CR.sup.66; [0311] r is 0, 1, or 2; [0312] R.sup.40
and R.sup.41 are independently selected from the group consisting
of hydrogen, fluoro, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; or [0313] R.sup.40 and R.sup.41 combine to
form a 3-7 membered monocyclic cycloalkyl or 5-7 membered
monocyclic heterocycloalkyl, wherein the monocyclic cycloalkyl or
monocyclic heterocycloalkyl is optionally substituted with one or
more substituents selected from the group consisting of halogen,
--OH, --NH.sub.2, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; [0314] R.sup.62, R.sup.63, R.sup.65 and
R.sup.66 are independently selected from the group consisting of
hydrogen, halogen, lower alkyl and lower alkoxy, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
one or more substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; [0315] Cy is selected from the
group consisting of aryl, heteroaryl, cycloalkyl, and
heterocycloalkyl; [0316] R.sup.39 is selected from thegroup
consisting of hydrogen, halogen, lower alkyl, lower alkoxy, aryl,
heteroaryl, and NR.sup.50R.sup.51, wherein the alkyl chain of lower
alkyl or lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, and wherein aryl and heteroaryl
are optionally substituted with one or more independent
substituents R.sup.23; [0317] R.sup.50 is hydrogen or lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino; [0318] R.sup.51 is aryl or heteroaryl, wherein
aryl and heteroaryl are optionally substituted with one or more
independent substituents R.sup.23; [0319] R.sup.23 at each
occurrence is independently selected from the group consisting of
--OH, --NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.4,R.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl,-atyl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.23, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58;
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0320] R.sup.57 is selected from the group consisting of lower
alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein
lower alkyl is optionally substituted with one or more substituents
selected from the group consisting of fluoro, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, provided, however, that any
substitution of the alkyl carbon bound to O, S, or N of
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, or --S(O).sub.2NR.sup.48R.sup.57 is
fluoro, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or
as substituents of lower alkyl are optionally substituted with one
or more:substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)OR.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0321] R.sup.58 at each occurrence is independently selected from
the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0322] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0323] t
is 0, 1, 2, or 3.
[0324] In some embodiments of compounds of Formula IIIp, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--. In some embodiments, A.sub.1 is --CR.sup.40R.sup.41-- or
--C(O)--, preferably --CH.sub.2-- or --C(O)--, and R.sup.62,
R.sup.64, R.sup.65 and R.sup.66 are independently selected from the
group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy.
[0325] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIc:
##STR00130##
all salts, prodrugs, tautomers and isomers thereof, wherein A, s,
Z.sub.2, Z.sub.6, R.sup.4, R.sup.5, R.sup.6, R.sup.15, R.sup.17,
and R.sup.32 are as defined for Formula III.
[0326] In some embodiments of compounds of Formula IIIc, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
more preferably --CH.sub.2--, R.sup.17 is selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro, and
R.sup.15 is selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy.
[0327] In some embodiments of compounds of Formula IIIc, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
R.sup.17 is selected from the group consisting of hydrogen,
halogen, lower alkyl and lower alkoxy, wherein the alkyl chain of
lower alkyl or lower alkoxy is optionally substituted with one or
more substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy,
lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro, R.sup.15 is selected from the
group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy, Z.sub.2 is N or CR.sup.12, Z.sub.6 is N or CR.sup.16,
R.sup.12 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, and fluoro substituted lower alkoxy, and
R.sup.5 is selected from the group consisting of hydrogen, halogen,
lower alkyl, lower alkoxy, optionally substituted aryl, optionally
substituted heteroaryl, and --NR.sup.21R.sup.22, wherein R.sup.21
is hydrogen or lower alkyl, and R.sup.22 is hydrogen, lower alkyl,
optionally substituted aryl or optionally substituted heteroaryl,
and wherein the alkyl chain of R.sup.5, R.sup.21 or R.sup.22, when
lower alkyl, or the alkyl chain of lower alkoxy is optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino and cycloalkylamino,
further wherein R.sup.32 is optionally substituted lower alkyl or
--OR.sup.18, where R.sup.18 is as defined for Formula III.
[0328] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIn:
##STR00131##
all salts, prodrugs, tautomers and isomers thereof, wherein A, s,
Z.sub.2, Z.sub.6, R.sup.4, R.sup.5, R.sup.6, R.sup.15, and R.sup.32
are as defined for Formula III.
[0329] In some embodiments of compounds of Formula IIIn, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
more preferably --CH.sub.2--, and R.sup.15 is selected from the
group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy.
[0330] In some, embodiments of compounds of Formula IIIc, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
R.sup.15 is selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy, Z.sub.2 is N or CR.sup.12,
Z.sub.6 is N or CR.sup.16, R.sup.12 and R.sup.16 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy, and R.sup.5 is selected from the group
consisting of hydrogen, halogen, lower alkyl, lower alkoxy,
optionally substituted aryl, optionally substituted heteroaryl, and
--NR.sup.21R.sup.22, wherein R.sup.21 is hydrogen or lower alkyl,
and R.sup.22 is hydrogen, lower alkyl, optionally substituted aryl
or optionally substituted heteroaryl, and wherein the alkyl chain
of R.sup.5, R.sup.21 or R.sup.22, when lower alkyl, or the alkyl
chain of lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, further wherein R.sup.32 is
optionally substituted lower alkyl or --OR.sup.18, where R.sup.18
is, as defined for Formula III.
[0331] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIo:
##STR00132##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
L.sub.2, Z.sub.2, Z.sub.4, Z.sub.5, Z.sub.6, R.sup.4, R.sup.5,
R.sup.6, and R.sup.33 are as defined for Formula III.
[0332] In some embodiments of compounds of Formula IIIo, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, and R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkylcarbon bound to the --O-- of lower alkoxy is fluoro.
[0333] In some embodiments of compounds of Formula IIIo, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro, and
R.sup.5 is selected from the group consisting of hydrogen, halogen,
lower alkyl, lower alkoxy, optionally substituted aryl, optionally
substituted heteroaryl, and NR.sup.21R.sup.22, wherein R.sup.21 is
hydrogen or lower alkyl, and R.sup.22 is hydrogen, lower alkyl,
optionally substituted aryl or optionally substituted heteroaryl,
and wherein the alkyl chain of R.sup.5, R.sup.21 or R.sup.22, when
lower alkyl, or the alkyl chain of lower alkoxy is optionally
substituted with one or more substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino and cycloalkylamino.
[0334] In some embodiments of compounds of Formula IIIo, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, and fluoro substituted lower alkoxy, and
R.sup.5 is; selected from the group consisting of hydrogen,
halogen, lower alkyl, lower alkoxy, optionally substituted aryl,
optionally substituted heteroaryl, and NR.sup.21R.sup.22, wherein
R.sup.21 is hydrogen or lower alkyl, and R.sup.22 is hydrogen,
lower alkyl, optionally substituted aryl or optionally substituted
heteroaryl, and wherein the alkyl chain of R.sup.5, R.sup.21 or
R.sup.22, when lower alkyl, or the alkyl chain of lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino.
[0335] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIq:
##STR00133##
all salts, prodrugs, tautomers, and isomers thereof, [0336]
wherein: [0337] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--
or --NR.sup.48--; [0338] Z.sub.12 is N or CR.sup.52; [0339]
Z.sub.14 is N or CR.sup.54; [0340] Z.sub.15 is N or CR.sup.55;
[0341] Z.sub.16 is N or CR.sup.56; [0342] R.sup.40 and R.sup.41 are
independently selected from the group consisting of hydrogen,
fluoro, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; or [0343] R.sup.40 and R.sup.41 combine to form a
3-7 membered monocyclic cycloalkyl or 5-7 membered monocyclic
heterocycloalkyl, wherein the monocyclic cycloalkyl or monocyclic
heterocycloalkyl is optionally substituted with one or more
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio; fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0344] L.sub.3 is selected from the group
consisting of --NR.sup.48--, --S--, --O--,
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, --OCH(R.sup.49)--,
--C(O)NR.sup.48--, --S(O).sub.2NR.sup.48--,
--CH(R.sup.49)NR.sup.48--, --CH(R.sup.49)O--, --CH(R.sup.49)S--,
--NR.sup.48C(O)--, and --NR.sup.48S(O).sub.2--; [0345] R.sup.54 and
R.sup.55 are independently selected from the group consisting of
hydrogen, halogen, lower alkyl and lower alkoxy, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
fluoro, --OH, --NH.sub.2 lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino or cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; [0346] R.sup.52 and R.sup.56 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy; [0347] R.sup.49 is selected
from the group consisting of hydrogen, lower alkyl, and fluoro
substituted lower alkyl; [0348] Cy is selected from the group
consisting of aryl, heteroaryl, cycloalkyl, and heterocycloalkyl;
[0349] R.sup.39 is selected from the group consisting of hydrogen,
halogen, lower alkyl, lower alkoxy, aryl, heteroaryl, and
NR.sup.50R.sup.51, wherein the alkyl chain of lower alkyl or lower
alkoxy is optionally substituted with one or more substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino
and cycloalkylamino, and wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
[0350] R.sup.50 is hydrogen or lower alkyl optionally substituted
with one or more substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino; [0351] R.sup.51
is aryl or heteroaryl, wherein aryl and heteroaryl are optionally
substituted with one or more independent substituents R.sup.23;
[0352] R.sup.23 at each occurrence is independently selected from
the group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2,
--C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.57,
--SR.sup.57, --NR.sup.48R.sup.57, --NR.sup.48C(O)R.sup.57,
--NR.sup.48S(O).sub.2R.sup.57, --S(O).sub.2R.sup.57,
--C(O)R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57,
--S(O).sub.2NR.sup.48R.sup.57, halogen, lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl as R.sup.23, or as substituents of lower alkyl, are
optionally substituted with one or more substituents selected from
the group consisting of --OH, --NH.sub.2, --CN, --NO.sub.2,
--C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.58,
--SR.sup.58, --NR.sup.48R.sup.58, --NR.sup.48C(O)R.sup.58,
--NR.sup.48S(O).sub.2R.sup.58, --S(O).sub.2R.sup.58,
--C(O)R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58,
--S(O).sub.2NR.sup.48R.sup.58, halogen, lower alkyl, fluoro
substituted lower alkyl, and cycloalkylamino; [0353] R.sup.57 is
selected from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0354] R.sup.58 at each occurrence is independently selected from
the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0355] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0356] t
is 0, 1, 2, or 3.
[0357] In some embodiments of compounds of Formula IIIq, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --CH.sub.2--. In some embodiments, A is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --CH.sub.2--, and R.sup.54 and R.sup.55
are independently selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy. In some embodiments, L.sub.3
is --NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, or
--OCH(R.sup.49)--, preferably --OCH(R.sup.49)--. In some
embodiments, A.sub.1 is --CR.sup.40R.sup.41-- or --C(O)--,
preferably --CH.sub.2-- or --C(O)--, more preferably --CH.sub.2--,
and L.sub.3 is --NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, or
--OCH(R.sup.49)--, preferably --OCH(R.sup.49)--.
[0358] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIId:
##STR00134##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
Z.sub.2, Z.sub.4, Z.sub.5, Z.sub.6, R.sup.4, R.sup.5, R.sup.6,
R.sup.10 , R.sup.11 and R.sup.33 are as defined for Formula III,
and r is 0, 1, or 2.
[0359] In some embodiments of compounds of Formula IIId, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, and R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro.
[0360] In some embodiments of compounds of Formula IIId, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O--of lower alkoxy is fluoro, R.sup.10
and R.sup.11 are independently selected from the group consisting
of hydrogen, fluoro, lower alkyl, and fluoro substituted lower
alkyl, and R.sup.5 is selected from the group consisting of
hydrogen, halogen, lower alkyl, lower alkoxy, optionally
substituted aryl, optionally substituted heteroaryl, and
NR.sup.2lR.sup.22, wherein R.sup.21 is hydrogen or lower alkyl, and
R.sup.22 is hydrogen, lower alkyl, optionally substituted aryl or
optionally substituted heteroaryl, and wherein the alkyl chain of
R.sup.5, R.sup.21 or R.sup.22, when lower alkyl, or the alkyl chain
of lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino.
[0361] In some embodiments of compounds of Formula IIId, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or OR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, and fluoro substituted lower alkoxy,
R.sup.10 and R.sup.11 are independently selected from the group
consisting of hydrogen, fluoro, lower alkyl, and fluoro substituted
lower alkyl, and R.sup.5 is selected from the group consisting of
hydrogen, halogen, lower alkyl, lower alkoxy, optionally
substituted aryl, optionally substituted heteroaryl, and
NR.sup.21R.sup.22, wherein R.sup.21 is hydrogen or lower alkyl, and
R.sup.22 is hydrogen, lower alkyl, optionally substituted aryl or
optionally substituted heteroaryl, and wherein the alkyl chain of
R.sup.5, R.sup.21 or R.sup.22, when lower alkyl, or the alkyl chain
of lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino.
[0362] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIe:
##STR00135##
all salts, prodrugs, tautomers, and isomers thereof, [0363]
wherein: [0364] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--or
--NR.sup.48--; [0365] Z.sub.22 is N or CR.sup.62; [0366] Z.sub.24
is N or CR.sup.64; [0367] Z.sub.25 is N or CR.sup.65; [0368]
Z.sub.26 is N or CR.sup.66; [0369] r is 0, 1, or 2; [0370] R.sup.40
and R.sup.41 are independently selected from the group consisting
of hydrogen, fluoro, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; or [0371] R.sup.40 and R.sup.41 combine to
form a 3-7 membered monocyclic cycloalkyl or 5-7 membered
monocyclic heterocycloalkyl, wherein the monocyclic cycloalkyl or
monocyclic heterocycloalkyl is optionally substituted with one or
more substituents selected from the group consisting of halogen,
--OH, --NH.sub.2, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino; [0372] R.sup.62, R.sup.64, R.sup.65 and
R.sup.66 are independently selected from the group consisting of
hydrogen, halogen, lower alkyl and lower alkoxy, wherein the alkyl
chain of lower alkyl or lower alkoxy is optionally substituted with
one or more substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino and cycloalkylamino, provided,
however, that any substitution on the alkyl carbon bound to the
--O-- of lower alkoxy is fluoro; [0373] Cy is selected from the
group consisting of aryl, heteroaryl, cycloalkyl, and
heterocycloalkyl; [0374] R.sup.39 is selected from the group
consisting of hydrogen, halogen, lower alkyl, lower alkoxy, aryl,
heteroaryl, and NR.sup.50R.sup.51, wherein the alkyl chain of lower
alkyl or lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, and wherein aryl and heteroaryl
are optionally substituted with one or more independent
substituents R.sup.23; [0375] R.sup.50 is hydrogen or lower alkyl
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino; [0376] R.sup.51 is aryl or heteroaryl, wherein
aryl and heteroaryl are optionally substituted with one or more
independent substituents R.sup.23; [0377] R.sup.23 at each
occurrence is independently selected from the group consisting of
--OH, --NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.23, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0378] R.sup.57 is selected from the group consisting of lower
alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein
lower alkyl is optionally substituted with one or more substituents
selected from the group consisting of fluoro, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, provided, however, that any
substitution of the alkyl carbon bound to O, S, or N of
--OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, or --S(O).sub.2NR.sup.48R.sup.57 is
fluoro, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or
as substituents of lower alkyl are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0379] R.sup.58 at each occurrence is independently selected from
the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0380] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0381] t
is 0, 1, 2, or 3.
[0382] In some embodiments of compounds of Formula IIIe, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--. In some embodiments, A.sub.1 is --CR.sup.40R.sup.41-- or
--C(O)--, preferably --CH.sub.2-- or --C(O)--, and R.sup.62,
R.sup.64, R.sup.65 and R.sup.66 are independently selected from the
group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy.
[0383] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIf:
##STR00136##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
Z.sub.2, Z.sub.4, Z.sub.5, Z.sub.6, X, R.sup.4, R.sup.5, R.sup.6,
R.sup.34, R.sup.35 and R.sup.82 are as defined for Formula III.
[0384] In some embodiments of compounds of Formula IIIf, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, and R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro.
[0385] In some embodiments of compounds of Formula IIIf, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or CR.sup.14, Z.sub.5 is N
or CR.sup.15, Z.sub.6 is N or CR.sup.16, R.sup.12, R.sup.14,
R.sup.15 and R.sup.16 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl or lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro, and one
of R.sup.34 and R.sup.35 is selected from the group consisting of
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl and optionally substituted heteroaryl, and the
other of R.sup.34 and R.sup.35 is hydrogen or lower alkyl, or
R.sup.34 and R.sup.35 together with the nitrogen to which they are
attached form optionally substituted 5-7 membered heterocycloalkyl
or optionally substituted 5 or 7 membered nitrogen containing
heteroaryl.
[0386] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIg:
##STR00137##
all salts, prodrugs, tautomers, and isomers thereof, [0387]
wherein: [0388] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--or
--NR.sup.48--; [0389] Z.sub.32 is N or CR.sup.72; [0390] Z.sub.34
is N or CR.sup.74; [0391] Z.sub.35 is N or CR.sup.75; [0392]
Z.sub.36 is N or CR.sup.76; [0393] X is O or S; [0394] R.sup.48 at
each occurrence is independently hydrogen or lower alkyl; [0395]
R.sup.40 and R.sup.41 are independently selected from the group
consisting of hydrogen, fluoro, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; or [0396] R.sup.40 and R.sup.41
combine to form a 3-7 membered monocyclic cycloalkyl or 5-7
membered monocyclic heterocycloalkyl, wherein the monocyclic
cycloalkyl or monocyclic heterocycloalkyl is optionally substituted
with one or more substituents selected from the group consisting of
halogen, --OH, --NH.sub.2, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; [0397] R.sup.72, R.sup.74,
R.sup.75 and R.sup.76 are independently selected from the group
consisting of hydrogen, halogen, lower alkyl and lower alkoxy,
wherein the alkyl chain of lower alkyl and lower alkoxy is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, --OH, --NH.sub.2, lower alkoxy,
fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino and
cycloalkylamino, provided, however, that any substitution on the
alkyl carbon bound to the --O-- of lower alkoxy is fluoro; [0398]
R.sup.67 is selected from the group consisting of hydrogen,
halogen, optionally substituted lower alkyl, optionally substituted
lower alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--OH, --NH.sub.2, --CN, --NO.sub.2,--C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.69, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; [0399] one of R.sup.60 and R.sup.61 is lower
alkyl, fluoro substituted lower alkyl, or
--(CH.sub.2).sub.0-2R.sup.70, and the other of R.sup.60 and
R.sup.61 is hydrogen or lower alkyl; [0400] or R.sup.60 and
R.sup.61 together with the nitrogen to which they are attached form
optionally substituted 5-7 membered heterocycloalkyl or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl; [0401]
R.sup.68 is selected from the group consisting of optionally
substituted lower alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O; S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NR.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; [0402] R.sup.69 is hydrogen or
optionally substituted lower alkyl; [0403] R.sup.70 is selected
from the group consisting of optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl; and [0404] R.sup.82 is
hydrogen or lower alkyl.
[0405] In some embodiments of compounds of Formula IIIg, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--.
[0406] In some embodiments of compounds of Formula IIIg, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, and R.sup.67 is selected from the group consisting of
hydrogen, halogen, optionally substituted lower alkyl, optionally
substituted lower alkenyl, optionally substituted lower alkynyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, optionally
substituted heteroaryl, --CN, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--OR.sup.68, --SR.sup.68, --NR.sup.69R.sup.68, --C(O)R.sup.68,
--C(S)R.sup.68, --C(O)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69R.sup.68, --NR.sup.69S(O)
, --S(O)R.sup.68, and --S(O).sub.2R.sup.68.
[0407] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIh:
##STR00138##
all salts, prodrugs, tautomers and isomers thereof, wherein A, X,
R.sup.4, R.sup.5, R.sup.6, R.sup.37, R.sup.38, R.sup.42, R.sup.43,
R.sup.45, R.sup.46, and R.sup.47 are as defined for Formula
III.
[0408] In some embodiments of compounds of Formula IIIh, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--, and
R.sup.42, R.sup.43, R.sup.45, R.sup.46 and R.sup.47 are
independently selected from the group consisting of hydrogen,
halogen, --OH, --CN, --NO.sub.2, --NH.sub.2, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkyl,
mono-alkylamino, di-alkylamino, and cycloalkylamino, further
wherein at least one of, at least two of, at least three of, at
least four of, or preferably all of R.sup.42, R.sup.43, R.sup.45,
R.sup.46 and R.sup.47 are hydrogen.
[0409] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIi:
##STR00139##
all salts, prodrugs, tautomers, and isomers thereof, [0410]
wherein: [0411] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--
or --NR.sup.48--; [0412] X is O or S; [0413] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; [0414]
R.sup.40 and R.sup.41 are independently selected from the group
consisting of hydrogen, fluoro, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; or [0415] R.sup.40 and R.sup.41
combine to form a 3-7 membered monocyclic cycloalkyl or 5-7
membered monocyclic heterocycloalkyl, wherein the monocyclic
cycloalkyl or monocyclic heterocycloalkyl is optionally substituted
with one or more substituents selected from the group consisting of
halogen, --OH, --NH.sub.2, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; [0416] R.sup.67 is selected
from the group consisting of hydrogen, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted heteroaryl, --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2, --NR.sup.69S(O).sub.2NR.sup.68,
--S(O)R.sup.68, and --S(O).sub.2R.sup.68; [0417] one of R.sup.60
and R.sup.61 is lower alkyl, fluoro substituted lower alkyl, or
--(CH.sub.2).sub.0-2R.sup.70 and the other of R.sup.60 and R.sup.61
is hydrogen or lower alkyl; [0418] or R.sup.60 and R.sup.61
together with the nitrogen to which they are attached form
optionally substituted 5-7 membered heterocycloalkyl or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl; [0419]
R.sup.68 is selected from the group consisting of optionally
substituted lower alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; [0420] R.sup.69 is hydrogen or
optionally substituted lower alkyl; and [0421] R.sup.70 is selected
from the group consisting of optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl.
[0422] In some embodiments of compounds of Formula IIIi, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--.
[0423] In some embodiments of compounds of Formula IIIi, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, and R.sup.69 is selected from the group consisting of
hydrogen, halogen, optionally substituted lower alkyl, optionally
substituted lower alkenyl, optionally substituted lower alkynyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, optionally
substituted heteroaryl, --CN, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--OR.sup.68, --SR.sup.68, --NR.sup.69R.sup.68, --C(O)R.sup.68,
--C(S)R.sup.68, --C(O)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69S(O).sub.2R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68.
[0424] In some embodiments of compounds of Formula IIIi, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --C(O)--, R.sup.62 is selected from the
group consisting of hydrogen, halogen, lower alkyl, lower alkoxy,
optionally substituted aryl, optionally substituted heteroaryl, and
NR.sup.21R.sup.22, wherein R.sup.21 is hydrogen or lower alkyl, and
R.sup.22 is hydrogen, lower alkyl, optionally substituted aryl or
optionally substituted heteroaryl, and wherein the alkyl chain of
R.sup.62, R.sup.21 or R.sup.22, when lower alkyl, or the alkyl
chain of lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, and one of R.sup.60 and R.sup.61
is lower alkyl or fluoro substituted lower alkyl, and the other of
R.sup.60 and R.sup.61 is hydrogen or lower alkyl. In some
embodiments, A.sub.1 is --C(O)--, R.sup.67 is optionally
substituted aryl or optionally substituted heteroaryl, and one of
R.sup.60 and R.sup.61 is lower alkyl or fluoro substituted lower
alkyl, and the other of R.sup.60 and R.sup.61 is hydrogen or lower
alkyl.
[0425] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIj:
##STR00140##
all salts, prodrugs, tautomers and isomers thereof, wherein A,
R.sup.4, R.sup.5, R.sup.6, R.sup.12, R.sup.14, R.sup.15, R.sup.16,
and R.sup.36 are as defined for Formula III.
[0426] In some embodiments of compounds of Formula IIIj, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--, and
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl and lower alkoxy, wherein the alkyl chain of lower alkyl or
lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, provided, however, that any
substitution on the alkyl carbon bound to the --O-- of lower alkoxy
is fluoro, preferably wherein R.sup.14 and R.sup.15 are hydrogen,
more preferably wherein R.sup.12 is fluoro, R.sup.16 is hydrogen,
fluoro or chloro, and R.sup.14 and R.sup.15 are hydrogen.
[0427] In some embodiments of compounds of Formula IIIj, R.sup.4
and R.sup.6 are hydrogen, A is --O--, --CR.sup.aR.sup.b--,
--NR.sup.1--, or --C(O)--, preferably --CH.sub.2-- or --C(O)--,
R.sup.12, R.sup.14, R.sup.15 and R.sup.16 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl and lower alkoxy, wherein the alkyl chain of lower alkyl or
lower alkoxy is optionally substituted with one or more
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro, substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino and cycloalkylamino, provided, however, that any
substitution on the alkyl carbon bound to the --O-- of lower alkoxy
is fluoro, preferably wherein R.sup.14 and R.sup.15 are hydrogen,
more preferably wherein R.sup.12 is fluoro, R.sup.16 is hydrogen,
fluoro or chloro, and R.sup.14 and R.sup.15 are hydrogen, and
R.sup.36 is selected from the group consisting of optionally
substituted C.sub.2-6 alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, and --NR.sup.19R.sup.20,
where R.sup.19 and R.sup.20 are as defined for Formula III, further
wherein one of R.sup.19 and R.sup.20 is hydrogen or optionally
substituted lower alkyl; and the other of R.sup.19 and R.sup.20 is
selected from the group consisting of hydrogen, optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl and optionally substituted heteroaryl.
[0428] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIk:
##STR00141##
all salts, prodrugs, tautomers and isomers thereof, [0429] wherein:
[0430] A.sub.1 is --O--, --CR.sup.40R.sup.41--, --C(O)--or
--NR.sup.48--; [0431] R.sup.81 is selected from the group
consisting of hydrogen, halogen, optionally substituted lower
alkyl, optionally substituted lower alkenyl, optionally substituted
lower alkynyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, --OH, --NH.sub.2, --CN,
--NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2,
--C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2,
--NR.sup.69C(S)NR.sup.69R.sup.68,--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; [0432] R.sup.71 and R.sup.78 are
independently selected from the group consisting of hydrogen,
halogen, C.sub.1-3 alkyl, and fluoro substituted C.sub.1-3 alkyl;
[0433] R.sup.77 is selected from the group consisting of
substituted methyl, optionally substituted C.sub.2-6 alkyl,
optionally substituted aryl, optionally substituted heteroaryl, and
--NR.sup.79R.sup.80, wherein methyl is substituted with one or more
substituents selected from the group consisting of optionally
substituted aryl and optionally substituted heteroaryl; [0434]
R.sup.68 is selected from the group consisting of optionally
substituted lower alkyl, optionally substituted lower alkenyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkenyl, no alkene carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68)
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup. ,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; [0435] R.sup.69 is hydrogen or
optionally substituted lower alkyl; and [0436] R.sup.79 and
R.sup.80 are independently hydrogen or optionally substituted lower
alkyl, or R.sup.79 and R.sup.80 combine with the nitrogen to which
they are attached to form optionally substituted 5-7 membered
heterocycloalkyl.
[0437] In some embodiments of compounds of Formula IIIk, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --C(O)--.
[0438] In some embodiments of compounds of Formula IIIk, A.sub.1 is
--CR.sup.40R.sup.41-- or --C(O)--, preferably --CH.sub.2-- or
--C(O)--, more preferably --C(O)--, and R.sup.81 is selected from
the group consisting of hydrogen, halogen, optionally substituted
lower alkyl, optionally substituted lower alkenyl, optionally
substituted lower alkynyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, --CN,
--S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68; --C(S)R.sup.68,
--C(O)NR.sup.69R.sup.68, --S(O).sub.2NR.sup.69R.sup.68,
--NR.sup.69C(O)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68;
--S(O)R.sup.68, and --S(O).sub.2R.sup.68.
[0439] In some embodiments, compounds of Formula III have the
structure according to the following sub-generic structure Formula
IIIm:
##STR00142##
all salts, prodrugs, tautomers and isomers thereof, [0440] wherein:
[0441] R.sup.81 is selected from the group consisting of hydrogen,
halogen, optionally substituted lower alkyl, optionally substituted
lower alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--OH, --NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --C(S)NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68,
--C(S)NR.sup.69R.sup.68,--S(O).sub.2NR.sup.69R.sup.68,
--NR.sup.69C(O)R.sup.68, --NR.sup.69C(S)R.sup.68,
--NR.sup.69S(O).sub.2R.sup.68, --NR.sup.69C(O)NH.sub.2,
--NR.sup.69C(O)NR.sup.69R.sup.68, --NR.sup.69C(S)NH.sub.2,
--NR.sup.69C(S)NR.sup.69R.sup.68, --NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, and
--S(O).sub.2R.sup.68; [0442] R.sup.83 is selected from the group
consisting of hydrogen, fluoro and chloro; [0443] R.sup.112 is
selected from the group consisting of optionally substituted
C.sub.2-6 alkyl, optionally substituted aryl, optionally
substituted heteroaryl, and --NR.sup.79R.sup.80; [0444] R.sup.68 is
selected from the group consisting of optionally substituted lower
alkyl, optionally substituted lower alkenyl, provided, however,
that when R.sup.68 is optionally substituted lower alkenyl, no
alkene carbon thereof is bound to N, S, O, S(O), S(O).sub.2, C(O)
or C(S) of --OR.sup.68, --SR.sup.68, --NR.sup.69R.sup.68,
--C(O)R.sup.68, --C(S)R.sup.68, --C(O)OR.sup.68,
--C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.2,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O)R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted lower alkynyl,
provided, however, that when R.sup.68 is optionally substituted
lower alkynyl, no alkyne carbon thereof is bound to N, S, O, S(O),
S(O).sub.2, C(O) or C(S) of --OR.sup.68, --SR.sup.68,
--NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)OR.sup.68, --C(O)NR.sup.69R.sup.68, --C(S)NR.sup.69R.sup.68,
--S(O).sub.2NR.sup.69R.sup.68, --NR.sup.69C(O)R.sup.68,
--NR.sup.69C(S)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--NR.sup.69C(O)NH.sub.2, --NR.sup.69C(O)NR.sup.69R.sup.68,
--NR.sup.69C(S)NH.sub.2, --NR.sup.69C(S)NR.sup.69R.sup.68,
--NR.sup.69S(O).sub.2NH.sub.3,
--NR.sup.69S(O).sub.2NR.sup.69R.sup.68, --S(O).sub.2R.sup.68, or
--S(O).sub.2R.sup.68, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl; [0445] R.sup.69 is selected from
the group consisting of hydrogen and optionally substituted lower
alkyl; and [0446] R.sup.79 and R.sup.80 are independently hydrogen
or optionally substituted lower alkyl, or R.sup.79 and R.sup.80
combine with the nitrogen to which they are attached to form
optionally substituted 5-7 membered heterocycloalkyl.
[0447] In some embodiments of compounds of Formula IIIm, R.sup.81
is selected from the group consisting of hydrogen, halogen,
optionally substituted lower alkyl, optionally substituted lower
alkenyl, optionally substituted lower alkynyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --S(O).sub.2NH.sub.2, --C(O)NH.sub.2, --OR.sup.68,
--SR.sup.68, --NR.sup.69R.sup.68, --C(O)R.sup.68, --C(S)R.sup.68,
--C(O)NR.sup.69R.sup.68, --S(O).sub.2NR.sup.69R.sup.68,
--NR.sup.69C(O)R.sup.68, --NR.sup.69S(O).sub.2R.sup.68,
--S(O)R.sup.68, and --S(O).sub.2R.sup.68.
[0448] The compounds of Formulae IIIa-IIIq, and all sub-embodiments
detailed herein, may be used to treat a subject suffering from or
at risk for any of the protein kinase mediated diseases or
conditions contemplated herein.
[0449] In one aspect, the present invention includes compounds that
are useful as intermediates in the preparation of compounds of
Formula III, the compounds having a structure selected from the
group consisting of Formula IV, Formula V, Formula VI, Formula VII,
Formula VIII, and Formula IX as follows:
##STR00143## [0450] wherein: [0451] Z.sub.2, Z.sub.4, Z.sub.5,
Z.sub.6, L.sub.2, X, s, R.sup.15, R.sup.17, R.sup.31, R.sup.32,
R.sup.33, R.sup.37, R.sup.38, R.sup.42, R.sup.45, R.sup.46, and
R.sup.47 are as defined for Formula III; [0452] R.sup.108 is
selected from the group consisting of --C(O)R.sup.84, --CH.sub.2I,
--CH.sub.2Cl, --CH.sub.2Br, --CH.sub.2OH, and
--CH.sub.2OS(O).sub.2R.sup.109; [0453] R.sup.109 is selected from
the group consisting of lower alkyl and aryl; [0454] R.sup.84 is
selected from the group consisting of hydrogen, lower alkoxy, --OH,
and --Cl; [0455] R.sup.85 is selected from the group consisting of
hydrogen, a nitrogen protecting group, --S(O).sub.2R.sup.87,
--C(O)NR.sup.88R.sup.89, and --C(S)NR.sup.88R.sup.89; [0456]
R.sup.86 is selected from the group consisting of hydrogen, lower
alkyl, and a nitrogen protecting group; [0457] R.sup.87 is selected
from the group consisting of optionally substituted lower alkyl,
optionally substituted lower alkenyl, provided, however, that when
R.sup.87 is optionally substituted lower alkenyl, no alkene carbon
thereof is bound to S(O).sub.2, optionally substituted lower
alkynyl, provided, however, that when R.sup.87 is optionally
substituted lower alkynyl, no alkyne carbon thereof is bound to
S(O).sub.2, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, and --NR.sup.90R.sup.91; and
[0458] R.sup.88, R.sup.89, R.sup.90 and R.sup.91 are independently
selected from the group consisting of hydrogen, optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl and optionally substituted heteroaryl; or [0459] R.sup.88 and
R.sup.89 together with the nitrogen to which they are attached form
optionally substituted 5-7 membered heterocycloalkyl or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl.
[0460] In some embodiments of compounds of Formulae IV, V, VI, VII,
or VIII, R.sup.108 is --C(O)R.sup.84, preferably wherein R.sup.84
is hydrogen. In some embodiments of compounds of Formulae IV, V,
VI, VII, or VIII, Z.sub.2 is N or CR.sup.12, Z.sub.4 is N or
CR.sup.14, Z.sub.5 is N or CR.sup.15, and Z.sub.6 is N or CR.sup.16
and R.sup.12, R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.42,
R.sup.43, R.sup.45, R.sup.46 and R.sup.47 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy.
[0461] In some embodiments, compounds of Formula IV have the
structure according to the following sub-generic structure Formula
IVa:
##STR00144## [0462] wherein: [0463] R.sup.84 is selected from the
group consisting of hydrogen, lower alkoxy, --OH, and --Cl; [0464]
R.sup.92, R.sup.93, R.sup.95, and R.sup.96 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy; [0465] L.sub.4 is selected from the group
consisting of --NR.sup.48--, --S--, --O--,
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, --OCH(R.sup.49)--,
--C(O)NR.sup.48--, --S(O).sub.2NR.sup.48--,
--CH(R.sup.49)NR.sup.48--, --CH(R.sup.49)O--, --CH(R.sup.49)S--,
--NR.sup.48C(O)--, and --NR.sup.48S(O).sub.2--; [0466] Cy is
selected from the group consisting of cycloalkyl, heterocycloalkyl,
aryl and heteroaryl; [0467] R.sup.97 at each occurrence is
independently selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.97, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0468] R.sup.49 is selected from the group consisting of hydrogen,
lower alkyl, and fluoro substituted lower alkyl; [0469] R.sup.57 is
selected from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
and [0470] R.sup.58 at each occurrence is independently selected
from the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0471] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0472] u
is 0, 1, 2, or 3.
[0473] In one embodiment of compounds of Formula IVa, at least two
of R.sup.92, R.sup.93, R.sup.95 and R.sup.96 are hydrogen. In one
embodiment, at least two of R.sup.92, R.sup.93, R.sup.95 and
R.sup.96 are hydrogen, L.sub.4 is --NR.sup.48CH(R.sup.49)--,
--SCH(R.sup.49)--, or --OCH(R.sup.49)--, preferably L.sub.4 is
--OCH.sub.2--, Cy is aryl or heteroaryl, and each R.sup.97 is
independently selected from the group consisting of halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy.
[0474] In some embodiments, compounds of Formula IV have the
structure according to the following sub-generic structure Formula
IVb:
##STR00145## [0475] wherein R.sup.84, R.sup.92, R.sup.93, R.sup.95,
R.sup.96, R.sup.97, Cy and u are as defined for Formula IVa and r
is 0, 1 or 2.
[0476] In some embodiments of compounds of Formula IVb, at least
two of R.sup.92, R.sup.93, R.sup.95 and R.sup.96 are hydrogen. In
some embodiments, at least two of R.sup.92, R.sup.93, R.sup.95 and
R.sup.96 are hydrogen, Cy is aryl or heteroaryl, and each R.sup.97
is independently selected from the group consisting of halogen,
lower alkyl, fluoro substituted lower alkyl, lower alkoxy, and
fluoro substituted lower alkoxy.
[0477] In some embodiments, compounds of Formula V have the
structure according to the following sub-generic structure Formula
Va:
##STR00146## [0478] wherein: [0479] R.sup.84 is selected from the
group consisting of hydrogen, lower alkoxy, --OH, and --Cl; [0480]
R.sup.92, R.sup.93, R.sup.95, and R.sup.96 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy; [0481] R.sup.98 is selected from the
group consisting of hydrogen, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, and fluoro substituted lower alkoxy; and
[0482] s is 0, 1, or 2;
[0483] In some embodiments, compounds of Formula VI have the
structure according to the following sub-generic structure Formula
VIa:
##STR00147## [0484] wherein: [0485] R.sup.84 is selected from the
group consisting of hydrogen, lower alkoxy, --OH, and --Cl; [0486]
R.sup.92, R.sup.94, R.sup.95, and R.sup.96 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy; [0487] L.sub.4 is selected from the group
consisting of --NR.sup.48--, --S--, --O--,
--NR.sup.48CH(R.sup.49)--, --SCH(R.sup.49)--, --OCH(R.sup.49)--,
--C(O)NR.sup.48--, --S(O).sub.2NR.sup.48--,
--CH(R.sup.49)NR.sup.48--, --CH(R.sup.49)O--, --CH(R.sup.49)S--,
--NR.sup.48C(O)--, and --NR.sup.48S(O).sub.2--; [0488] Cy is
selected from the group consisting of cycloalkyl, heterocycloalkyl,
aryl and heteroaryl; [0489] R.sup.97 at each occurrence is
independently selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.57, --SR.sup.57, --NR.sup.48R.sup.57,
--NR.sup.48C(O)R.sup.57, --NR.sup.48S(O).sub.2R.sup.57,
--S(O).sub.2R.sup.57, --C(O)R.sup.57, --C(O)OR.sup.57,
--C(O)NR.sup.48R.sup.57, --S(O).sub.2NR.sup.48R.sup.57, halogen,
lower alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl,
wherein lower alkyl is optionally substituted with one or more
substituents selected from the group consisting of fluoro, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino,
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl as R.sup.97, or
as substituents of lower alkyl, are optionally substituted with one
or more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0490] R.sup.49 is selected from the group consisting of hydrogen,
lower alkyl, and fluoro substituted lower alkyl; [0491] R.sup.57 is
selected from the group consisting of lower alkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more substituents selected from
the group consisting of fluoro, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.57, --SR.sup.57,
--NR.sup.48R.sup.57, --C(O)OR.sup.57, --C(O)NR.sup.48R.sup.57, or
--S(O).sub.2NR.sup.48R.sup.57 is fluoro, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl as R.sup.57 or as
substituents of lower alkyl are optionally substituted with one or
more substituents selected from the group consisting of --OH,
--NH.sub.2, --CN, --NO.sub.2, --C(O)OH, --S(O).sub.2NH.sub.2,
--C(O)NH.sub.2, --OR.sup.58, --SR.sup.58, --NR.sup.48R.sup.58,
--NR.sup.48C(O)R.sup.58, --NR.sup.48S(O).sub.2R.sup.58,
--S(O).sub.2R.sup.58, --C(O)R.sup.58, --C(O)OR.sup.58,
--C(O)NR.sup.48R.sup.58, --S(O).sub.2NR.sup.48R.sup.58, halogen,
lower alkyl, fluoro substituted lower alkyl, and cycloalkylamino;
[0492] R.sup.58 at each occurrence is independently selected from
the group consisting of lower alkyl, heterocycloalkyl and
heteroaryl, wherein lower alkyl is optionally substituted with one
or more substituents selected from the group consisting of fluoro,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and cycloalkylamino, provided, however, that any substitution of
the alkyl carbon bound to O, S, or N of --OR.sup.58, --SR.sup.58,
--NR.sup.48R.sup.58, --C(O)OR.sup.58, --C(O)NR.sup.48R.sup.58, or
--S(O).sub.2NR.sup.48R.sup.58 is fluoro; [0493] R.sup.48 at each
occurrence is independently hydrogen or lower alkyl; and [0494] u
is 0, 1, 2 or 3.
[0495] In some embodiments of compounds of Formula VIa, at least
two of R.sup.92, R.sup.94, R.sup.95 and R.sup.96 are hydrogen. In
some embodiments, at least two of R.sup.92, R.sup.94, R.sup.95 and
R.sup.96 are hydrogen, L.sub.4 is --NR.sup.48CH(R.sup.49)--,
--SCH(R.sup.49)--, or --OCH(R.sup.49)--, preferably L.sub.4 is
--OCH.sub.2--, Cy is aryl or heteroaryl, and each R.sup.97 is
independently selected from the group consisting of halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy.
[0496] In some embodiments, compounds of Formula VI have the
structure according to the following sub-generic structure Formula
VIb:
##STR00148## [0497] wherein R.sup.84, R.sup.92, R.sup.94, R.sup.95,
R.sup.96, R.sup.97, Cy and u are as defined for Formula VIa and r
is 0, 1 or 2.
[0498] In some embodiments of compounds of Formula VIb, at least
two of R.sup.92, R.sup.94, R.sup.95 and R.sup.96 are hydrogen. In
some embodiments, at least two of R.sup.92, R.sup.94, R.sup.95 and
R.sup.96 are hydrogen, Cy is aryl or heteroaryl, and each R.sup.97
is independently selected from the group consisting of halogen,
lower alkyl, fluoro substituted lower alkyl, lower alkoxy, and
fluoro substituted lower alkoxy.
[0499] In some embodiments, compounds of Formula VII have the
structure according to the following sub-generic structure Formula
VIIa:
##STR00149## [0500] wherein: [0501] R.sup.84 is selected from the
group consisting of hydrogen, lower alkoxy, --OH, and --Cl; [0502]
R.sup.92, R.sup.94, R.sup.95, and R.sup.96 are independently
selected from the group consisting of hydrogen, halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy; [0503] R.sup.99 is selected from the
group consisting of optionally substituted lower alkyl, optionally
substituted aryl, optionally substituted heteroaryl, and
--NR.sup.79R.sup.80; and [0504] R.sup.79 and R.sup.80 are
independently hydrogen or optionally substituted lower alkyl, or
R.sup.79 and R.sup.80 combine with the nitrogen to which they are
attached to form optionally substituted 5-7 membered
heterocycloalkyl.
[0505] In some embodiments of compounds of Formula VIIa, one of
R.sup.92 and R.sup.96 is selected from the group consisting of
hydrogen, halogen, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, and fluoro substituted lower alkoxy, and the other of
R.sup.92 and R.sup.96 is selected from the group consisting of
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy; in further embodiments, one of
R.sup.92 and R.sup.96 is hydrogen, fluoro or chloro, and the other
of R.sup.92 and R.sup.96 is fluoro or chloro; in further
embodiments, R.sup.92 is fluoro and R.sup.96 is hydrogen, fluoro,
or chloro; in further embodiments, R.sup.92 and R.sup.96 are both
fluoro.
[0506] In some embodiments of compounds of Formula VIIa, R.sup.94
and R.sup.95 are hydrogen; in further embodiments, one of R.sup.92
and R.sup.96 is selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy and the other of R.sup.92 and
R.sup.96 is selected from the group consisting of halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy; in further embodiments, one of R.sup.92
and R.sup.96 is selected from hydrogen, fluoro or chloro and the
other of R.sup.92 and R.sup.96 is selected from fluoro or chloro;
in further embodiments, R.sup.92 is fluoro and R.sup.96 is selected
from hydrogen, fluoro, or chloro; in further embodiments, R.sup.92
and R.sup.96 are both fluoro.
[0507] In some embodiments, compounds of Formula VII have the
structure according to the following sub-generic structure Formula
VIIb:
##STR00150## [0508] wherein: [0509] X is O or S; [0510] R.sup.84 is
selected from the group consisting of hydrogen, lower alkoxy, --OH,
and --Cl; [0511] R.sup.92, R.sup.94, R.sup.95, and R.sup.96 are
independently selected from the group consisting of hydrogen,
halogen, lower alkyl, fluoro substituted lower alkyl, lower alkoxy,
and fluoro substituted lower alkoxy; [0512] R.sup.82 is hydrogen or
lower alkyl; [0513] one of R.sup.100 and R.sup.101 is lower alkyl,
fluoro substituted lower alkyl, or --(CH.sub.2).sub.0-2R.sup.70,
and the other of R.sup.100 and R.sup.101 is hydrogen or lower
alkyl; or [0514] R.sup.100 and R.sup.101 together with the nitrogen
to which they are attached form optionally substituted 5-7 membered
heterocycloalkyl or optionally substituted 5 or 7 membered nitrogen
containing heteroaryl; and [0515] R.sup.70 is selected from the
group consisting of optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl.
[0516] In some embodiments of compounds of Formula VIIb, at least
two of R.sup.92, R.sup.94, R.sup.95 and R.sup.96 are hydrogen. In
some embodiments, at least two of R.sup.92, R.sup.94, R.sup.95 and
R.sup.96 are hydrogen, and R.sup.70 is aryl or heteroaryl, wherein
aryl and heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy.
[0517] In some embodiments, compounds of Formula VIII have the
structure according to the following sub-generic structure Formula
VIIIa:
##STR00151## [0518] wherein: [0519] X is O or S; [0520] R.sup.84,
is selected from the group consisting of hydrogen, lower alkoxy,
--OH, and --Cl; [0521] R.sup.102, R.sup.103, R.sup.105, R.sup.106,
and R.sup.107 are independently selected from the group consisting
of hydrogen, halogen, lower alkyl, fluoro substituted lower alkyl,
lower alkoxy, and fluoro substituted lower alkoxy; [0522] one of
R.sup.100 and R.sup.101 is lower alkyl, fluoro substituted lower
alkyl, or --(CH.sub.2).sub.0-2R.sup.70 and the other of R.sup.100
and R.sup.101 is hydrogen or lower alkyl; [0523] or R.sup.100 and
R.sup.101 together with the nitrogen to which they are attached
form optionally substituted 5-7 membered heterocycloalkyl or
optionally substituted 5 or 7 membered nitrogen containing
heteroaryl; and [0524] R.sup.70 is selected from the group
consisting of optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl, and
optionally substituted heteroaryl.
[0525] In some embodiments of compounds of Formula VIIIa, at least
two, also at least three, also at least four, or all of R.sup.102,
R.sub.103, R.sup.105, R.sup.106, and R.sup.107 are hydrogen. In
some embodiments, at least two, also at least three, also at least
four, or all of R.sup.102, R.sup.103, R.sup.105, R.sup.106, and
R.sup.107 are hydrogen, and R.sup.70 is aryl or heteroaryl, wherein
aryl and heteroaryl are optionally substituted with one or more
substituents selected from the group consisting of halogen, lower
alkyl, fluoro substituted lower alkyl, lower alkoxy, and fluoro
substituted lower alkoxy.
[0526] In some embodiments, compounds of Formula IX have the
structure according to the following sub-generic structure Formula
IXa:
##STR00152## [0527] wherein: [0528] R.sup.84 is selected from the
group consisting of hydrogen, lower alkoxy, --OH, and --Cl; [0529]
R.sup.92, R.sup.95, and R.sup.96 are independently selected from
the group consisting of hydrogen, halogen, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, and fluoro substituted lower
alkoxy; [0530] R.sup.98 is selected from the group consisting of
hydrogen, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, and fluoro substituted lower alkoxy; and [0531] s is 0, 1,
or 2;
[0532] In some embodiments of any of the above embodiments of
compounds of Formula IVa, IVb, Va, VIa, VIb, VIIa, VIIb, VIIIa, or
IXa, R.sup.84 is hydrogen.
[0533] In some embodiments of the above compounds, compounds are
excluded where N (except where N is a heteroaryl ring atom), O, or
S is bound to a carbon that is also bound to N (except where N is a
heteroaryl ring atom), O, or S; or where N (except where N is a
heteroaryl ring atom), O, C(S), C(O), or S(O).sub.n (n is 0-2) is
bound to an alkene carbon of an alkenyl group or bound to an alkyne
carbon of an alkynyl group; accordingly, in some embodiments
compounds which include linkages such as the following are excluded
from the present invention: --NR--CH.sub.2--NR--,
--O--CH.sub.2--NR--, --S--CH.sub.2--NR--, --NR--CH.sub.2--O--,
--O--CH.sub.2--O--, --S--CH.sub.2--O--, --NR--CH.sub.2--S--,
--O--CH.sub.2--S--, --S--CH.sub.2--S--, --NR--CH.dbd.CH--,
--CH.dbd.CH--NR--, --NR--C.ident.C--, --C.ident.C--NR--,
--O--CH.dbd.CH--, --CH.dbd.CH--O--, --O--C.ident.C--,
--C.ident.C--O--, --S(O).sub.0-2--CH.dbd.CH--,
--CH.dbd.CH--S(O).sub.0-2--, --S(O).sub.0-2--C.ident.C--,
--C.ident.C--S(O).sub.0-2--, --C(O)--CH.dbd.CH--,
--CH.dbd.CH--C(O)--, --C.ident.C--C(O)--, or --C(O)C.ident.C--,
--C(S)--CH.dbd.CH--, --CH.dbd.CH--C(S)--, --C.ident.C--C(S)--, or
--C(S)--C.ident.C--.
[0534] In reference to compounds herein, specification of a
compound or group of compounds includes pharmaceutically acceptable
salts of such compound(s) unless clearly indicated to the contrary,
prodrug(s), and all stereoisomers. In reference to compositions,
kits, methods of use, etc. of compounds of Formula I described
herein, it is understood that a compound of Formula I includes
compounds of Formulae Ia-Iz, and all sub-embodiments thereof,
compounds of Formula II, including Formulae IIa-IIo, and all
sub-embodiments thereof, and compounds of Formula III, including
Formulae IIIa-IIIq, and all sub-embodiments thereof, unless
indicated otherwise. In reference to compositions, kits, methods of
use, etc. of compounds of Formula II described herein, it is
understood that this includes compounds of Formulae IIa-IIo, and
all sub-embodiments thereof, unless indicated otherwise. In
reference to compositions, kits, methods of use, etc. of compounds
of Formula III described herein, it is understood that this
includes compounds of Formulae IIIa-IIIq, and all sub-embodiments
thereof, unless indicated otherwise.
[0535] In one aspect, the invention provides methods for treating a
protein kinase mediated disease or condition in an animal subject,
wherein the method involves administering to the subject an
effective amount of a compound of Formula I. The terms "treat,"
"therapy," and like terms refer to the administration of material,
e.g., compound of Formula I, in an amount effective to prevent,
alleviate, or ameliorate one or more symptoms of a disease or
condition, i.e., indication, and/or to prolong the survival of the
subject being treated. The term "protein kinase mediated disease or
condition" refers to a disease or condition in which the biological
function of a protein kinase affects the development and/or course
of the disease or condition, and/or in which modulation of the
protein kinase alters the development, course, and/or symptoms of
the disease or condition. A protein kinase mediated disease or
condition includes a disease or condition for which modulation
provides a therapeutic benefit, e.g. wherein treatment with protein
kinase inhibitors, including compounds described herein, provides a
therapeutic benefit to the subject suffering from or at risk of the
disease or condition. In one aspect, the method involves
administering to the subject an effective amount of a compound of
Formula I in combination with one or more other therapies for the
disease or condition.
[0536] In one aspect, the invention provides methods for treating a
protein kinase mediated disease or condition in an animal subject,
wherein the method involves administering to the subject an
effective, amount of a compound of any one or more of Formula Ia
through Formula Iz, and all sub-embodiments thereof.
[0537] In another aspect, the invention provides methods for
treating a protein kinase mediated disease or condition in an
animal subject, wherein the method involves administering to the
subject an effective amount of a compound of Formula II.
[0538] In another aspect, the invention provides methods for
treating a protein kinase mediated disease or condition in an
animal subject, wherein the method involves administering to the
subject an effective amount of a compound of any one or more of
Formula IIa through Formula IIo, and all sub-embodiments
thereof.
[0539] In another aspect, the invention provides methods for
treating a protein kinase mediated disease or condition in an
animal subject, wherein the method involves administering to the
subject an effective amount of a compound of Formula III.
[0540] In another aspect, the invention provides methods for
treating a protein kinase mediated disease or condition in an
animal subject, wherein the method involves administering to the
subject an effective amount of a compound of any one or more of
Formula IIIa through Formula IIIq, and all sub-embodiments
thereof.
[0541] In one aspect, the invention provides methods for treating a
Raf protein kinase mediated disease or condition in an animal
subject, wherein the method involves administering to the subject
an effective amount of a compound of Formula I. The terms "Raf
protein kinase mediated disease or condition," "Raf mediated
disease or condition," and the like refer to a disease or condition
in which the biological function of a Raf kinase, including any
mutations thereof, affects the development and/or course of the
disease or condition, and/or in which modulation of the Raf protein
kinase alters the development, course, and/or symptoms of the
disease or condition. The Raf protein kinase includes, but is not
limited to, B-Raf, mutations of B-Raf, c-Raf-1 and mutations of
c-Raf-1. In some embodiments, the Raf protein kinase is B-Raf
mutation V600E. In further embodiments, the disease or condition is
a cancer that is amenable to treatment by an inhibitor of the V600E
mutant B-Raf. A Raf protein kinase mediated disease or condition
includes a disease or condition for which Raf inhibition provides a
therapeutic benefit, e.g. wherein treatment with Raf inhibitors,
including compounds described herein, provides a therapeutic
benefit to the subject suffering from or at risk of the disease or
condition. In one aspect, the method involves administering to the
subject an effective amount of a compound of Formula I in
combination with one or more other therapies for the disease or
condition.
[0542] In one aspect, the invention provides methods for treating a
Raf protein kinase mediated disease or condition in an animal
subject, wherein the method involves administering to the subject
an effective amount of a compound of Formula IIa or III. In one
aspect, the method involves administering to the subject an
effective amount of a compound of Formula IIa or III in combination
with one or more other therapies for the disease, further wherein
the compound is of Formula IIIj, IIIk, or IIIm. The Raf protein
kinase includes, but is not limited to, B-Raf, mutations of B-Raf,
c-Raf-1 and mutations of c-Raf-1. In some embodiments, the Raf
protein kinase is B-Raf mutation V600E. In further embodiments, the
disease or condition is a cancer that is amenable to treatment by
an inhibitor of the V600E mutant B-Raf.
[0543] In one aspect, the invention provides methods for treating a
Fms protein kinase mediated disease or condition in an animal
subject, wherein the method involves administering to the subject
an effective amount of a compound of Formula I. The terms "Fms
protein kinase mediated disease or condition," "Fms mediated
disease or condition," and the like refer to a disease or condition
in which the biological function of a Fms protein kinase, including
any mutations thereof, affects the development and/or course of the
disease or condition, and/or in which modulation of Fms alters the
development, course, and/or symptoms of the disease or condition. A
Fms mediated disease or condition includes a disease or condition
for which Fms inhibition provides a therapeutic benefit, e.g.
wherein treatment with Fms inhibitors, including compounds
described herein, provides a therapeutic benefit to the subject
suffering from or at risk of the disease or condition. In one
aspect, the method involves administering to the subject an
effective amount of a compound of Formula I in combination with one
or more other therapies for the disease or condition.
[0544] In one aspect, the invention provides methods for treating a
Kit protein kinase mediated disease or condition in an animal
subject, wherein the method involves administering to the subject
an effective amount of a compound of Formula I. The terms "Kit
mediated disease or condition," "Kit protein kinase mediated
disease or condition," and the like refer to a disease or condition
in which the biological function of a Kit protein kinase, including
any mutation thereof, affects the development and/or course of the
disease or condition, and/or in which modulation of Kit alters the
development, course, and/or symptoms of the disease or condition. A
Kit mediated disease or condition includes a disease or condition
for which Kit inhibition provides a therapeutic benefit, e.g.
wherein treatment with Kit inhibitors, including compounds
described herein, provides a therapeutic benefit to the subject
suffering from or at risk of the disease or condition. In one
aspect, the method involves administering to the subject an
effective amount of a compound of Formula I in combination with one
or more other therapies for the disease or condition.
[0545] In one aspect, the invention provides methods for treating a
Jnk protein kinase mediated disease or condition in an animal
subject, wherein the method involves administering to the subject
an effective amount of a compound of Formula I. The terms "Jnk
mediated disease or condition," "Jnk protein kinase mediated
disease or condition," and the like refer to a disease or condition
in which the biological function of a Jnk kinase, e.g. Jnk1, Jnk2,
Jnk3, or any mutation thereof, affects the development and/or
course of the disease or condition, and/or in which modulation of
the Jnk kinase alters the development, course, and/or symptoms of
the disease or condition. A Jnk mediated disease or condition
includes a disease or condition for which Jnk inhibition provides a
therapeutic benefit, e.g. wherein treatment with Jnk inhibitors,
including compounds described herein, provides a therapeutic
benefit to the subject suffering from or at risk of the disease or
condition. In one aspect, the method involves administering to the
subject an effective amount of a compound of Formula I in
combination with one or more other therapies for the disease or
condition. The Jnk protein kinase includes, but is not limited to,
Jnk1, Jnk2, or Jnk3.
[0546] In some embodiments, a compound of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
as determined in a generally accepted kinase activity assay. In
some embodiments, a compound of any of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of Abl, Akt1, Akt2, Akt3, ALK, Alk5, B-Raf, Brk, Btk,
Cdk2, CDK4, CDK5, CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2,
EphB2, EphB4, Erk2, Fak, FGFR1, FGFR2, FGFR3, FGFR4, Flt1, Flt3,
Flt4, Fms, Frk, Fyn, Gsk3.alpha., Gsk3.beta., HCK, Her2/Erbb2,
Her4/Erbb4, IGF1R, IKK beta; Irak4, Itk, Jak1, Jak2, Jak3, Jnk1,
Jnk2, Jnk3, Kdr, Kit, LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2, Met,
Mnk1, MLK1, p38, PDGFRA, PDGFRB, PDPK1, Pim1, Pim2, Pim3, PKC
alpha, PKC beta, PKC theta, Plk1, Pyk2, ROCK1, ROCK2, Ron, Src,
Stk6, Syk, TEC, Tie2, TrkA, Yes, and Zap70, including any mutations
thereof.
[0547] In some embodiments, a compound of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of Abl, Akt 1, Akt2, Akt3, ALK, Alk5, B-Raf, Btk, Cdk2,
CDK4, CDK5, CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2, EphB2,
EphB4, Erk2, Fak, Fms, Fyn, Gsk3.alpha., Gsk3.beta., HCK,
Her2/Erbb2, Her4/Erbb4, IGF1R, IKK beta, Irak4, Itk, Jak1, Jak2,
Jak3, Jnk1, Jnk2, Jnk3, Kit, LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2,
Met, Mnk1, MLK1, p38, PDPK1, Pim1, Pim2, Pim3, PKC alpha, PKC beta,
PKC theta, Plk1, Pyk2, Ron, Src, Stk6, Syk, TEC, Tie2, TrkA, Yes,
and Zap70, including any mutations thereof.
[0548] In some embodiments, a compound of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of Abl, B-Raf, Btk, c-Raf-1, EGFR, EphB2, Erk2, Fak,
FGFR1, Flt1, Flt3, Flt4, Fms, Irak4, Jnk1, Jnk2, Jnk3, Kdr, Kit,
MAP2K1, MAPKAP kinase 2, Met, p38, PDGFRB, Pim1, PKC theta, Pyk2,
Ret, Src, Stk6, Yes, and Zap70, including any mutations
thereof.
[0549] In some embodiments, a compound of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of Abl, B-Raf, Btk, c-Raf-1, EGFR, EphB2, Erk2, Fak,
Fms, Irak4, Jnk1, Jnk2, Jnk3, Kit, MAP2K1, MAPKAP kinase 2, Met,
p38, Pim1, PKC theta, Pyk2, Src, Stk6, Yes, and Zap70, including
any mutations thereof.
[0550] In some embodiments, a compound of Formula I will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of B-Raf, B-Raf V600E mutant, c-Raf-1, FGFR1, FGFR2,
FGFR3, FGFR4, Jnk1, Jnk2, Jnk3, Met, Pim1, Pim2, Pim3, Pyk2, Kdr
and Ret, including any mutations thereof.
[0551] In some embodiments, a compound of Formula III will have an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
with respect to at least one kinase selected from the group
consisting of B-Raf, c-Raf-1, Fms, Jnk1, Jnk2, Jnk3, and Kit, and
any mutations thereof. In some embodiments, a compound of any of
Formula III will have an IC.sub.50 of less than 500 nm, less than
100 nM, less than 50 nM, less than 20 nM, less than 10 nM, less
than 5 nM, or less than 1 nM with respect to at least one kinase
selected from the group consisting of B-Raf, B-Raf V600E mutant,
c-Raf-1, Fms, Jnk1, Jnk2, Jnk3, and Kit, preferably B-Raf, B-Raf
V600E mutant or c-Raf-1.
[0552] In some embodiments, a compound of Formula III is an
inhibitor of a Raf kinase and has an IC.sub.50 of less than 500 nm,
less than 100 nM, less than 50 nM, less than 20 nM, less than 10
nM, less than 5 nM, or less than 1 nM as determined in a generally
accepted Raf kinase activity assay. In some embodiments, a compound
of Formula III will have an IC.sub.50 of less than 500 nm, less
than 100 nM, less than 50 nM, less than 20 nM, less than 10 nM,
less than 5 nM, or less than 1 nM with respect to B-Raf, c-Raf-1,
or B-Raf V600E mutant. In some embodiments, a compound of Formula
III will selectively inhibit one Raf kinase relative to one or more
other Raf kinases. In some embodiments, the compound of Formula III
will selectively inhibit a mutation of the Raf kinase relative to
the wild type kinase, for example B-Raf V600E relative to wild type
B-Raf.
[0553] In some embodiments, a compound of Formula III is an
inhibitor of a Fms kinase and has an IC.sub.50 of less than 500 nm,
less than 100 nM, less than 50 nM, less than 20 nM, less than 10
nM, less than 5 nM, or less than 1 nM as determined in a generally
accepted Fms kinase activity assay. In some embodiments, a compound
of Formula III will selectively inhibit Fms kinase relative to Kit
kinase.
[0554] In some embodiments, a compound of Formula III is an
inhibitor of a Kit kinase and has an IC.sub.50 of less than 500 nm,
less than 100 nM, less than 50 nM, less than 20 nM, less than 10
nM, less than 5 nM, or less than 1 nM as determined in a generally
accepted Kit kinase activity assay.
[0555] In some embodiments, a compound of Formula III is an
inhibitor of a Jnk kinase and has an IC.sub.50 of less than 500 nm,
less than 100 nM, less than 50 nM, less than 20 nM, less than 10
nM, less than 5 nM, or less than 1 nM as determined in a generally
accepted Jnk kinase activity assay. In some embodiments, a compound
of Formula III is an inhibitor of a Jnk1 kinase and has an
IC.sub.50 of less than 500 nm, less than 100 nM, less than 50 nM,
less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM
as determined in a generally accepted Jnk1 kinase activity assay.
In some embodiments, a compound of Formula III is an inhibitor of a
Jnk2 kinase and has an IC.sub.50 of less than 500 nm, less than 100
nM, less than 50 nM, less than 20 nM, less than 10 nM, less than 5
nM, or less than 1 nM as determined in a generally accepted Jnk2
kinase activity assay. In some embodiments, a compound of Formula
III is an inhibitor of a Jnk3 kinase and has an IC.sub.50 of less
than 500 nm, less than 100 nM, less than 50 nM, less than 20 nM,
less than 10 nM, less than 5 nM, or less than 1 nM as determined in
a generally accepted Jnk3 kinase activity assay. In some
embodiments, a compound of Formula III will selectively inhibit one
Jnk kinase relative to one or more other Jnk kinases, such as
selectively inhibiting Jnk 1 relative to Jnk 2 and/or Jnk3,
selectively inhibiting Jnk2 relative to Jnk3 and/or Jnk1, or
selectively inhibiting Jnk3 relative to Jnk1 and/or Jnk 2.
[0556] Further to any of the above mentioned embodiments, a
compound of the invention will also inhibit the effects of a
mutation of the kinase, including, but net limited to, a mutation
that is related to a disease state, such as a cancer. For example,
B-Raf V600E mutant is present in a high percentage of some cancers,
such as melanoma, and compounds of the invention will inhibit the
kinase activity of this mutant.
[0557] Further to any of the above embodiments, a compound of the
invention may selectively inhibit one kinase relative to one or
more other kinases, where preferably inhibition is selective with
respect to any of the other kinases, whether a kinase discussed
herein, or other kinases. In some embodiments, the compound may
selectively inhibit the effects of a mutation of the kinase
relative to the wild type kinase, for example B-Raf V600E relative
to wild type B-Raf. In some embodiments, the compound may
selectively inhibit Fms relative to Kit. Selective inhibition of
one kinase relative to another is such that the IC.sub.50 for the
one kinase may be at least about 2-fold, also 5-fold, also 10-fold,
also 20-fold, also 50-fold, or at least about 100-fold less than
the IC.sub.50 for any of the other kinases as determined in a
generally accepted kinase activity assay.
[0558] In another aspect, the invention provides compositions that
include a therapeutically effective amount of a compound of Formula
I and at least one pharmaceutically acceptable carrier, excipient,
and/or diluent. The composition can include a plurality of
different pharmacologically active compounds, which can include a
plurality of compounds of Formula I. In another aspect, the
composition can include one or more compounds of Formula I, Formula
II, or Formula III along with one or more compounds that are
therapeutically effective for the same disease indication. In one
aspect, the composition includes one or more compounds of Formula
I, Formula II, or Formula III along with one or more compounds that
are therapeutically effective for the same disease indication,
wherein the compounds have a synergistic effect on the disease
indication.
[0559] In another aspect, the invention provides compositions that
include a therapeutically effective amount of at least one compound
of Formula III and at least one pharmaceutically acceptable
carrier, excipient, and/or diluent. The composition can include a
plurality of different pharmacologically active compounds, which
can include a plurality of compounds of Formula III, or can include
at least one compound of Formula III along with at least one
compound that is therapeutically effective for the same disease
indication. In one aspect, the at least one compound of Formula III
and the at least one compound that is therapeutically effective for
the same disease indication have a synergistic effect on the
disease indication. In one aspect, the composition includes one or
more compounds of Formula III effective in treating a cancer and
one or more other compounds that are effective in treating the
cancer, further wherein the compounds are synergistically effective
in treating the cancer.
[0560] In another aspect, the invention provides a method for
modulating the activity of a protein kinase selected from the group
consisting of Abl, Akt1, Akt2, Akt3, ALK, Alk5, B-Raf, Brk, Btk,
Cdk2, CDK4, CDK5, CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2,
EphB2, EphB4, Erk2, Fak, FGFR1, FGFR2, FGFR3, FGFR4, Flt1, Flt3,
Flt4, Fms, Frk, Fyn, Gsk3.alpha., Gsk3.beta., HCK, Her2/Erbb2,
Her4/Erbb4, IGF1R, IKK beta, Irak4, Itk, Jak1, Jak2, Jak3, Jnk1,
Jnk2, Jnk3, Kdr, Kit, LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2, Met,
Mnk1, MLK1, p38, PDGFRA, PDGFRB, PDPK1, Pim1, Pim2, Pim3, PKC
alpha, PKC beta, PKC theta, Plk1, Pyk2, ROCK1, ROCK2, Ron, Src,
Stk6, Syk, TEC, Tie2, TrkA, Yes, or Zap70 by contacting the protein
kinase with an effective amount of a compound of Formula I.
[0561] In another aspect, the invention provides methods for
treating a protein kinase mediated disease or condition in an
animal subject, wherein the method involves administering to the
subject an effective amount of a composition including a compound
of Formula I.
[0562] In one aspect, the invention provides methods for treating a
disease or condition mediated by a protein kinase selected from the
group consisting of Abl, Akt 1, Akt2, Akt3, ALK, Alk5, B-Raf, Btk,
Cdk2, CDK4, CDK5, CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2,
EphB2, EphB4, Erk2, Fak, FGFR1, FGFR2, FGFR3, FGFR4, Flt1, Flt3,
Flt4, Fms, Fyn, Gsk3.alpha., Gsk3.beta., HCK, Her2/Erbb2,
Her4/Erbb4, IGF1R, IKK beta, Irak4, Itk, Jak1, Jak2, Jak3, Jnk1,
Jnk2, Jnk3, Kdr, Kit, LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2, Met,
Mnk1, MLK1, p38, PDGFRA, PDGFRB, PDPK1, Pim1, Pim2, Pim3, PKC
alpha, PKC beta, PKC theta, Plk1, Pyk2, ROCK1, ROCK2, Ron, Src,
Stk6, Syk, TEC, Tie2, TrkA, Yes, and Zap70 by administering to the
subject an effective amount of a composition including a compound
of Formula I.
[0563] In one aspect, the invention provides methods for treating a
disease or condition mediated by a protein kinase selected from the
group consisting of Abl, Akt 1, Akt2, Akt3, ALK, Alk5, B-Raf, Btk,
Gdk2, CDK4, CDK5, CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2,
EphB2, EphB4, Erk2, Fak, Fms, Fyn, Gsk3.alpha., Gsk3.beta., HCK,
Her2/Erbb2, Her4/Erbb4, IGFIR, IKK beta, Irak4, Itk, Jak1, Jak2,
Jak3, Jnk1, Jnk2, Jnk3, Kit, LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2,
Met, Mnk1, MLK1, p38, PDPK1, Pim1, Pim2, Pim3, PKC alpha, PKC beta,
PKC theta, Plk1, Pyk2, Ron, Src, Stk6, Syk, TEC, Tie2, TrkA, Yes,
and Zap70 by administering to the subject an effective amount of a
composition including a compound of Formula I.
[0564] In one aspect, the invention provides methods for treating a
disease or condition mediated by a protein kinase selected from the
group consisting of Abl, B-Raf, Btk, c-Raf-1, EGFR; EphB2, Erk2,
Fak, FGFR1, Flt1, Flt3, Flt4, Fms, Irak4, Jnk1, Jnk2, Jnk3, Kdr,
Kit, MAP2K1, MAPKAPK2, Met, p38, PDGFRB, Pim1, PKC theta, Pyk2,
Ret, Src, Stk6, Yes, and Zap70 by administering to the subject an
effective amount of a composition including a compound of Formula
I.
[0565] In one aspect, the invention provides methods for treating a
disease or condition mediated by a protein kinase selected from the
group consisting of Abl, B-Raf, Btk, c-Raf-1, EGFR, EphB2, Erk2,
Fak, Fms, Irak4, Jnk1, Jnk2, Jnk3, Kit, MAP2K1, MAPKAPK2, Met, p38,
Pim1, PKC theta, Pyk2, Src, Stk6, Yes, and Zap70 by administering
to the subject an effective amount of a composition including a
compound of Formula I.
[0566] In one aspect, the invention provides methods for treating a
disease or Condition mediated by a protein kinase selected from the
group consisting of B-Raf, B-Raf V600E mutant, c-Raf-1, FGFR1,
FGFR2, FGFR3, FGFR4, Jnk1, Jnk2, Jnk3, Met, Pim1, Pim2, Pim3, Pyk2,
Kdr and Ret by administering to the subject an effective amount of
a composition including a compound of Formula I.
[0567] In one aspect, the invention provides methods for treating a
disease or condition mediated by a protein kinase selected from the
group consisting of B-Raf, c-Raf-1, Fms, Jnk1, Jnk2, Jnk3, and Kit,
and any mutations thereof, by administering to the subject an
effective amount of a composition including a compound of Formula
III.
[0568] In one aspect, the invention provides methods for treating a
disease or condition mediated by B-Raf, c-Raf-1, or B-Raf V600E by
administering to the subject an effective amount of a composition
including a compound of Formula II or Formula III, where in further
embodiments, the compound is of Formula IIa or Formula III. In one
aspect, the invention provides methods for treating a disease or
condition mediated by B-Raf, c-Raf-1, or B-Raf V600E by
administering to the subject an effective amount of a composition
including a compound of Formula IIIj, IIIk, or IIIm in combination
with one or more other suitable therapies for treating the disease.
In one aspect, the invention provides methods for treating a cancer
mediated by B-Raf V600E mutant by administering to the subject an
effective amount of a composition of Formula IIIj, IIIk, or IIIm in
combination with one or more suitable anticancer therapies, such as
one or more chemotherapeutic drugs.
[0569] In one aspect, the invention provides a method of treating a
cancer by administering to the subject an effective amount of a
composition including a compound of Formula I, or where the
compound is of Formula III, or where the compound is of Formula
IIIj, IIIk, or IIIm, in combination with one or more other
therapies or medical procedures effective in treating the cancer.
Other therapies or medical procedures include suitable anticancer
therapy (e.g. drug therapy, vaccine therapy, gene therapy,
photodynamic therapy) or medical procedure (e.g. surgery, radiation
treatment, hyperthermia heating, bone marrow or stem cell
transplant). In one aspect, the one or more suitable anticancer
therapies or medical procedures is selected from treatment with a
chemotherapeutic agent (e.g. chemotherapeutic drug), radiation
treatment (e.g. x-ray, .gamma.-ray, or electron, proton, neutron,
or a particle beam), hyperthermia heating (e.g. microwave,
ultrasound, radiofrequency ablation), Vaccine therapy (e.g. AFP
gene hepatocellular carcinoma vaccine, AFP adenoviral vector
vaccine, AG-858, allogeneic GM-CSF-secretion breast cancer vaccine,
dendritic cell peptide vaccines), gene therapy (e.g. Ad5CMV-p53
vector, adenovector encoding MDA7, adenovirus 5-tumor necrosis
factor alpha), photodynamic therapy (e.g. aminolevulinic acid,
motexafin lutetium), surgery, and bone marrow and stem cell
transplantation.
[0570] In a preferred embodiment, the invention provides a method
of treating a cancer by administering to the subject an effective
amount of a composition including a compound of Formula I, or
wherein the compound is of Formula III, or wherein the compound is
of Formula IIIj, IIIk, or IIIm, in combination with one or more
suitable chemotherapeutic agents. In one aspect, the one or more
suitable chemotherapeutic agents is selected from an alkylating
agent, including, but not limited to, adozelesin, altretamine,
bizelesin, busulfan, carboplatin, carboquone, carmustine,
chlorambucil, cisplatin, cyclophosphamide, dacarbazine,
estramustine, fotemustine, hepsulfam, ifosfamide, improsulfan,
irofulven, lomustine, mechlorethamine, melphalan, oxaliplatin,
piposulfan, semustine, streptozocin, temozolomide, thiotepa, and
treosulfan; an antibiotic, including, but not limited to,
bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin,
idarubicin, menogaril, mitomycin, mitoxantrone, neocarzinostatin,
pentostatin, and plicamycin; an antimetabolite, including, but not
limited to, azacitidine, capecitabine, cladribine, clofarabine,
cytarabine, decitabine, floxuridine, fludarabine, 5-fluorouracil,
florafur, gemcitabine, hydroxyurea, mercaptopurine, methotrexate,
nelarabine, pemetrexed, raltitrexed, thioguanine, and trimetrexate;
an immunotherapy, including, but not limited to, alemtuzumab,
bevacizumab, cetuximab, galiximab, gemtuzumab, panitumumab,
pertuzumab, rituximab, tositumomab, trastuzumab, and 90 Y
ibritumomab tiuxetan; a hormone or hormone antagonist, including,
but not limited to, anastrozole, androgens, buserelin,
diethylstilbestrol, exemestane, flutamide, fulvestrant, goserelin,
idoxifene, letrozole, leuprolide, magestrol, raloxifene, tamoxifen,
and toremifene; a taxane, including, but not limited to, DJ-927,
docetaxel, TPI 287, paclitaxel and DHA-paclitaxel; a retinoid,
including, but not limited to, alitretinoin, bexarotene,
fenretinide, isotretinoin, and tretinoin; an alkaloid, including,
but not limited to, etoposide, homoharringtonine, teniposide,
vinblastine, vincristine, vindesine, and vinorelbine; an
antiangiogenic agent, including, but not limited to, AE-941
(GW786034, Neovastat), ABT-510, 2-methoxyestradiol, lenalidomide,
and thalidomide; a topoisomerase inhibitor, including, but not
limited to, amsacrine, edotecarin, exatecan, irinotecan (also
active metabolite SN-38 (7-ethyl-10-hydroxy-camptothecin)),
rubitecan, topotecan, and 9-aminocamptothecin; a kinase inhibitor,
including, but not limited to, erlotinib, gefitinib, flavopiridol,
imatinib mesylate, lapatinib, sorafenib, sunitinib malate, AEE-788,
AG-013736, AMG 706, AMN107, BMS-354825, BMS-599626, UCN-01
(7-hydroxystaurosporine), and vatalanib; a targeted signal
transduction inhibitor including, but not limited to bortezomib,
geldanamycin, and rapamycin; a biological response modifier,
including, but not limited to, imiquimod, interferon-.alpha., and
interleukin-2; and other chemotherapeutics, including, but not
limited to 3-AP (3-amino-2-carboxyaldehyde thiosemicarbazone),
aminoglutethimide, asparaginase, bryostatin-1, cilengitide, E7389,
ixabepilone, procarbazine, sulindac, temsirolimus, tipifarnib.
Preferably, the method of treating a cancer involves administering
to the subject an effective amount of a composition of Formula I
(more preferably Formula III, and even more preferably Formulae
IIIj, IIIk, or IIIm) in combination with a chemotherapeutic agent
selected from 5-fluorouracil, carboplatin, dacarbazine, gefitinib,
oxaliplatin, paclitaxel, SN-38, temozolomide, vinblastine,
bevacizumab, cetuximab, or erlotinib.
[0571] In another aspect, the invention provides a method of
treating or prophylaxis of a disease or condition in a mammal, by
administering to the mammal a therapeutically effective amount of a
compound of Formula I, a prodrug of such compound, or a
pharmaceutically acceptable salt of such compound or prodrug. The
compound can be alone or can be part of a composition. In another
aspect, the invention provides a method of treating or prophylaxis
of a disease or condition in a mammal, by administering to the
mammal a therapeutically effective amount of a compound of Formula
III, a prodrug of such compound, or a pharmaceutically acceptable
salt of such compound or prodrug in combination with one or more
other suitable therapies for the disease or condition.
[0572] In another aspect, the invention provides compositions that
include a therapeutically effective amount of a compound of Formula
III and at least one pharmaceutically acceptable carrier,
excipient, and/or diluent. The composition can include a plurality
of different pharmacologically active compounds, which can include
a plurality of compounds of Formula III.
[0573] In a related aspect, the invention provides kits that
include a composition as described herein. In some embodiments, the
composition is packaged, e.g., in a vial, bottle, flask, which may
be further packaged, e.g., within a box, envelope, or bag; the
composition is approved by the U.S. Food and Drug Administration or
similar regulatory agency for administration to a mammal, e.g., a
human; the composition is approved for administration to a mammal,
e.g., a human, for a protein kinase mediated disease or condition;
the invention kit includes written instructions for use and/or
other indication that the composition is suitable or approved for
administration to a mammal, e.g., a human, for a protein
kinase-mediated disease or condition; and the composition is
packaged in unit dose or single dose form, e.g., single dose pills,
capsules, or the like.
[0574] In aspects involving treatment or prophylaxis of a disease
or condition with the compounds of Formula I, the disease or
condition is, for example without limitation, neurologic diseases
such as ischemic stroke, cerebrovascular ischemia, multi-infarct
dementia, head injury, spinal cord injury, Alzheimer's disease
(AD), Parkinson's disease, amyotrophic lateral sclerosis, dementia,
senile chorea, and Huntington's disease; neoplastic diseases and
associated complications, including chemotherapy-induced hypoxia,
gastrointestinal stromal tumors (GISTs), prostate tumors, mast cell
tumors (including canine mast cell tumors), acute myeloid leukemia,
acute lymphocytic leukemia, chronic myeloid leukemia, chronic
lymphocytic leukemia, multiple myeloma, melanoma, mastocytosis,
gliomas, glioblastoma, astrocytoma, neuroblastoma, sarcomas (e.g.
sarcomas of neuroectodermal origin, leiomyosarcoma), carcinomas
(e.g. lung, breast, pancreatic, colon, hepatocellular, renal,
female genital tract, squamous cell, carcinoma in situ), lymphoma
(e.g. histiocytic lymphoma, non-Hodgkin's lymphoma), MEN2
syndromes, neurofibromatosis (including Schwann cell neoplasia),
myelodysplastic syndrome, leukemia, tumor angiogenesis, and cancers
of the thyroid, liver, bone, skin, brain, central nervous system,
pancreas, lung (e.g. small cell lung cancer, non small cell lung
cancer), breast, colon, bladder, prostate, gastrointestinal tract,
endometrium, fallopian tube, testes and ovary; pain of neuropathic
or inflammatory origin, including acute pain, chronic pain, and
migraine; cardiovascular diseases including heart failure, cardiac
hypertrophy, thrombosis (e.g. thrombotic microangiopathy
syndromes), atherosclerosis, reperfusion injury and ischemia (e.g.
cerebrovascular ischemia, liver ischemia); inflammation including,
but not limited to, polycystic kidney disease (PKD), age-related
macular degeneration, rheumatoid arthritis, allergic rhinitis,
inflammatory bowel disease (IBD), ulcerative colitis, Crohn's
disease, systemic lupus erythematosis, Sjogren's Syndrome,
Wegener's granulomatosis, psoriasis, scleroderma, chronic
thyroiditis, Grave's disease, myasthenia gravis, multiple
sclerosis, osteoarthritis, endometriosis, scarring (e.g. dermal,
tissue), vascular restenosis, fibrotic disorders,
hypereosinophilia, CNS inflammation, pancreatitis, nephritis,
atopic dermatitis, and hepatitis; immunodeficiency diseases (e.g.
severe combined immunodeficiency (SCID)), organ transplant
rejection, graft versus host disease; renal or prostatic diseases
including diabetic nephropathy, nephrosclerosis,
glomerulonephritis, interstitial nephritis, Lupus nephritis,
prostate hyperplasia, chronic renal failure, tubular necrosis,
diabetes-associated renal complications, and hypertrophy; metabolic
diseases including type 1 diabetes, type 2 diabetes, metabolic
syndrome, obesity, hepatic steatosis, insulin resistance,
hyperglycemia, lipolysis and obesity; infection, including, but not
limited to Helicobacter pylori and Influenza virus, fever, sepsis;
pulmonary diseases including chronic obstructive pulmonary disease
(COPD), acute respiratory distress syndrome (ARDS), asthma,
allergy, bronchitis, emphysema, and pulmonary fibrosis; genetic
developmental diseases such as Noonan's syndrome, Crouzon syndrome,
acrocephalo-syndactyly type 1, Pfeiffer's syndrome, Jackson-Weiss
syndrome, Costello syndrome, (faciocutaneoskeletal syndrome),
leopard syndrome, cardio-faciocutaneous syndrome (CFG) and neural
crest syndrome abnormalities causing cardiovascular, skeletal,
intestinal, skin, hair and endocrine diseases; disorders of bone
structure or mineralization, including osteoporosis, increased risk
of fracture, hypercalcemia, and bone metastases; Grave's disease;
Hirschsprung's disease; lymphoedema; selective T-cell defect (STD);
X-linked agammaglobulinemia; diabetic retinopathy; alopecia;
erectile dysfunction; and tuberous sclerosis.
[0575] In a related aspect, compounds of Formula I, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a B-Raf-mediated disease or
condition, selected from the group consisting of neurologic
diseases such as ischemic stroke, multi-infarct dementia, head
injury, spinal cord injury, Alzheimer's disease (AD), Parkinson's
disease; neoplastic diseases including, but not limited to,
melanoma, glioma, sarcoma, carcinoma (e.g. lung, breast,
pancreatic, renal), lymphoma (e.g. histiocytic lymphoma) and cancer
of the thyroid, lung (e.g. small cell lung cancer), liver, breast,
ovary and colon, neurofibromatosis, myelodysplastic syndrome,
leukemia, tumor angiogenesis; pain of neuropathic or inflammatory
origin, including acute pain, chronic pain, and migraine;
cardiovascular diseases including heart failure, cardiac
hypertrophy, thrombosis (e.g. thrombotic microangiopathy
syndromes), atherosclerosis, reperfusion injury; inflammation
including, but not limited to, psoriasis, polycystic kidney disease
(PKD), arthritis and autoimmune diseases and conditions,
osteoarthritis, endometriosis, scarring, vascular restenosis,
fibrotic disorders, rheumatoid arthritis, inflammatory bowel
disease (IBD); immunodeficiency diseases, organ transplant
rejection, graft versus host disease; renal or prostatic diseases
including diabetic nephropathy, nephrosclerosis,
glomerulonephritis, prostatehyperplasia; metabolic disorders,
obesity; infection, including, but not limited to Helicobacter
pylori and Influenza virus, fever, sepsis; pulmonary diseases
including chronic obstructive pulmonary disease (COPD) and acute
respiratory distress syndrome (ARDS); genetic developmental
diseases such as Noonan's syndrome, Costello syndrome,
(faciocutaneoskeletal syndrome), leopard syndrome,
cardio-faciocutaneous syndrome (CFC), and neural crest syndrome
abnormalities causing cardiovascular, skeletal, intestinal, skin,
hair and endocrine diseases.
[0576] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a c-Raf-1-mediated disease or
condition selected from the group consisting of colorectal,
ovarian, lung and renal cell carcinoma, acute myeloid leukemia,
myelodysplastic syndromes, tumor angiogenesis, and neuroendocrine
tumors such as medullary thyroid cancer, carcinoid, small cell lung
cancer and pheochromocytoma.
[0577] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Fms-mediated disease or condition
selected from the group consisting of immune disorders, including
rheumatoid arthritis, systemic lupus erythematosis (SLE), Wegener's
granulomatosis, and transplant rejection, inflammatory diseases
including Chronic Obstructive Pulmonary Disease (COPD), emphysema,
and atherosclerosis, metabolic disorders, including insulin
resistance, hyperglycemia, and lipolysis, disorders of bone
structure or mineralization, including osteoporosis, increased risk
of fracture, hypercalcemia, and bone metastases, kidney diseases,
including nephritis (e.g. glomerulonephritis, interstitial
nephritis, Lupus nephritis), tubular necrosis, diabetes-associated
renal complications, and hypertrophy and cancers, including
multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia
(CML), breast cancer, and ovarian cancer.
[0578] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Jnk-mediated disease or condition
selected from the group consisting of metabolic diseases including
type 1 diabetes, type 2 diabetes, metabolic syndrome, obesity, and
hepatic steatosis; cardiovascular diseases such as atherosclerosis,
ischemia (e.g. cerebrovascular ischemia, liver ischemia),
reperfusion injury, cardiac hypertrophy; renal diseases such as
chronic renal failure; neoplastic diseases and associated
complications, including chemotherapy-induced hypoxia, prostate
tumors, myeloid leukemia and cancers of the liver, bone, skin,
brain, pancreas, lung breast, colon, prostate and ovary; transplant
rejection, pain of neuropathic or inflammatory origin including
acute and chronic pain; inflammatory and autoimmune diseases
including age-related macular degeneration, rheumatoid arthritis,
inflammatory bowel disease, ulcerative colitis, Crohn's disease,
systemic lupus erythematosis, Sjogren's Syndrome, psoriasis,
scleroderma, chronic thyroiditis, Grave's disease, myasthenia
gravis, and multiple sclerosis, and inflammation in other organs
including CNS inflammation, pancreatitis; nephritis, atopic
dermatitis, and hepatitis; airway inflammatory diseases such as
asthma, allergy, bronchitis, pulmonary fibrosis, chronic
obstructive pulmonary disease; neurologic diseases such as stroke,
cerebrovascular ischemia, neurodegenerative diseases such as
Parkinson's disease, Alzheimer's disease, amyotrophic lateral
sclerosis, dementia, senile chorea, head and spinal cord trauma,
and Huntington's disease.
[0579] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Jnk1-mediated disease or
condition selected from the group consisting of type 1 diabetes,
type 2 diabetes, metabolic syndrome, obesity and hepatic
steatosis.
[0580] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Jnk2-mediated disease or
condition, such as atherosclerosis.
[0581] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Jnk3-mediated disease or
condition selected from the group consisting of inflammatory
diseases including autoimmune diseases such as rheumatoid
arthritis, inflammatory bowel syndrome, Crohn's disease, systemic
lupus erythematosis, Sjogren's Syndrome, psoriasis and multiple
sclerosis, airway inflammatory diseases such as asthma, allergy,
pulmonary fibrosis, and chronic obstructive pulmonary disease, and
inflammation in other organs, such as CNS inflammation,
pancreatitis, nephritis, and hepatitis; neurologic diseases such as
stroke, cerebrovascular ischemia, and neurodegenerative diseases
such as Parkinson's disease, Alzheimer's disease, and Huntington's
disease; and neoplastic diseases such as prostate tumors and
myeloid leukemia.
[0582] In a related aspect, compounds of Formula III, further where
the compound is of Formula III, can be used in the preparation of a
medicament for the treatment of a Kit-mediated disease or condition
selected from the group consisting of malignancies, including mast
cell tumors, small cell lung cancer, testicular cancer,
gastrointestinal stromal tumors (GISTs), glioblastoma, astrocytoma,
neuroblastoma, carcinomas of the female genital tract, sarcomas of
neuroectodermal origin, colorectal carcinoma, carcinoma in situ,
Schwann cell neoplasia associated with neurofibromatosis, acute
myelocytic leukemia, acute lymphocytic leukemia, chronic
myelogenous leukemia, mastocytosis, melanoma, and canine mast cell
tumors, and inflammatory diseases, including asthma, rheumatoid
arthritis, allergic rhinitis, multiple sclerosis, inflammatory
bowel syndrome, transplant rejection, and hypereosinophilia.
[0583] The compounds of Formula I with kinase activity IC.sub.50
less than 10 nM as determined in a standard assay described herein
can be used to treat protein kinase mediated diseases and
conditions related to the following protein kinases, for example
without limitation: [0584] Abl, related to chronic myeloid leukemia
(CML), acute lymphoblastic leukemia (ALL) and acute myelogenous
leukemia (AML); [0585] Akt1, related to gastric, prostate,
colorectal, ovarian, pancreatic and breast cancer, glioblastoma and
leukemia, as well as schizophrenia and bipolar disorders, and also
use in combination with other chemotherapeutic drugs; [0586] Akt2,
related to hyperglycemia due to peripheral insulin resistance and
nonsuppressible hepatic glucose production accompanied by
inadequate compensatory hyperinsulinemia, also related to
pancreatic, ovarian and breast cancer; [0587] Akt3, related to
melanoma, prostate and breast cancer; [0588] ALK, related to
non-Hodgkin lymphomas such as diffuse large B-cell lymphoma and
anaplastic large cell lymphoma; [0589] Alk5, related to pancreatic
and biliary cancers, and cutaneous T-cell lymphoma; [0590] B-Raf,
related to neurologic diseases such as ischemic stroke,
multi-infarct dementia, head injury, spinal cord injury,
Alzheimer's disease (AD), Parkinson's disease; neoplastic diseases
including, but not limited to, melanoma, glioma, sarcoma, carcinoma
(e.g. lung, breast, pancreatic, renal), lymphoma (e.g. histiocytic
lymphoma) and cancer of the thyroid, lung (e.g. small cell lung
cancer), liver, breast, ovary and colon, neurofibromatosis,
myelodysplastic syndrome, leukemia, tumor angiogenesis; pain of
neuropathic or inflammatory origin, including acute pain, chronic
pain, and migraine; cardiovascular diseases including heart
failure, cardiac hypertrophy, thrombosis (e.g. thrombotic
microangiopathy syndromes), atherosclerosis, reperfusion injury;
inflammation including, but not limited to, psoriasis, polycystic
kidney disease (PKD), arthritis and autoimmune diseases and
conditions, osteoarthritis, endometriosis, scarring, vascular
restenosis, fibrotic disorders, rheumatoid arthritis, inflammatory
bowel disease (IBD); immunodeficiency diseases, organ transplant
rejection, graft versus host disease; renal or prostatic diseases
including diabetic nephropathy, nephrosclerosis,
glomerulonephritis, prostate hyperplasia; metabolic disorders,
obesity; infection, including, but not limited to Helicobacter
pylori and Influenza virus, fever, sepsis; pulmonary diseases
including chronic obstructive pulmonary disease (COPD) and acute
respiratory distress syndrome (ARDS); genetic developmental
diseases such as Noonan's syndrome, Costello syndrome,
(faciocutaneoskeletal syndrome), leopard syndrome,
cardio-faciocutaneous syndrome (CFC), and neural crest syndrome
abnormalities causing cardiovascular, skeletal, intestinal, skin,
hair and endocrine diseases; [0591] c-Raf-1, related to colorectal,
ovarian, lung and renal cell carcinoma, acute myeloid leukemia,
myelodysplastic syndromes, tumor angiogenesis, and neuroendocrine
tumor's such as medullary thyroid cancer, carcinoid, small cell
lung cancer and pheochromocytoma; [0592] Brk, related to breast and
colon cancer, and head and neck squamous cell carcinoma; [0593]
Btk, related to X-linked agammaglobulinemia, acute lymphocytic
leukemia, autoimmune diseases such as multiple sclerosis, systemic
lupus erythematous, rheumatoid arthritis, and Graves' disease,
immune suppression in organ transplant, and drug sensitivity of
B-lineage cells; [0594] Cdk2, related to prostate, breast,
colorectal and ovarian cancer; [0595] Cdk4, related to glioblastoma
(e.g. glioblastoma multiforme), anaplastic astrocytoma, and breast
cancer; [0596] Cdk5, related to Alzheimer's disease, amyotrophic
lateral sclerosis and Lewy body disease; [0597] Cdk6, related to
glioblastoma multiforme, non-Hodgkin's lymphoma, splenic marginal
zone lymphoma, T-cell lymphoblastic lymphoma (T-LBL) and T-cell
acute lymphoblastic leukemia (T-ALL); [0598] CHK1, related to DNA
damage repair, sensitizes cells to chemotherapeutic agents; [0599]
Csk, related to colon and pancreatic carcinomas and autoimmune
pathology such as type 1 diabetes, rheumatoid arthritis and
systemic lupus erythematosus; [0600] EGFR, related to breast,
colorectal, bladder, prostate and non small cell lung cancer,
squamous cell carcinomas of the head and neck cancer, oral cavity,
and esophagus, and glioblastoma multiforme; [0601] EphA1, related
to head and neck squamous cell carcinoma, hepatoma and lung cancer,
[0602] EphA2, related to aberrant short-range contact-mediated
axonal guidance, bladder, breast, prostate, colon, skin, cervical,
ovarian, pancreatic and lung cancers, and metastatic melanoma;
[0603] EphB2, related to angiogenesis disorder (e.g. ocular
angiogenesis disease such as retinopathy), and cancer (e.g.
glioblastoma, breast and liver cancer); [0604] EphB4, related to
colorectal cancer (CRC), head and neck squamous cell carcinoma, and
tumours of the prostate, breast, endometrium, and bladder; [0605]
Erk2, related to aberrant proliferation, differentiation,
transcription regulation and development, and may be useful in
treating inflammation, for example inflammation associated with
Lyme neuroborreliosis, and in treating cancers, such as gastric
cancer; [0606] Fak, related to colon and breast tumors, and is also
related to esophageal squamous cell carcinoma, melanoma, anaplastic
astrocytoma, glioblastoma, ductal carcinoma in situ, prostate and
hepatocellular carcinoma, and tumor metastases, and may also
provide synergistic effects when used with other chemotherapeutic
drugs; [0607] FGFR1, related to 8p11 myeloproliferative syndrome;
[0608] FGFR2, related to Crouzon Syndrome, Jackson-Weiss Syndrome,
Apert Syndrome, craniosynostosis, Pfeiffer Syndrome, acrocephalo
syndactyly type V, and Beare-Stevenson Cutis Gyrata Syndrome;
[0609] FGFR3, related to angiogenesis, wound healing,
achondroplasia, Muenke craniosynostosis, Crouzon syndrome,
acanthosis nigricans, thanatophoric dysplasia, bladder carcinomas,
and multiple myeloma; [0610] FGFR4, related to cancer of the
breast, lung, colon, medullary thyroid, pancreas, ovary, prostate,
endometrium, and fallopian tube, head and neck squamous cell
carcinomas and leiomyosarcoma; [0611] Flt1, related to non-small
cell lung carcinoma, prostate carcinoma, and colorectal cancer;
[0612] Flt3, related to acute myeloid leukemia, myelodysplastic
syndrome, acute lymphoblastic leukemia; [0613] Flt4, related to
primary lymphoedema; [0614] Fms, related to immune disorders,
including rheumatoid arthritis, systemic lupus erythematosis (SLE),
Wegener's granulomatosis, and transplant rejection, inflammatory
diseases including Chronic Obstructive Pulmonary Disease (COPD),
emphysema, and atherosclerosis, metabolic disorders, including
insulin resistance, hyperglycemia, and lipolysis, disorders of bone
structure or mineralization, including osteoporosis, increased risk
of fracture, hypercalcemia, and bone metastases, kidney diseases,
including nephritis (e.g. glomerulonephritis, interstitial
nephritis, Lupus nephritis), tubular necrosis, diabetes-associated
renal complications, and hypertrophy and cancers, including
multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia
(CML), breast cancer, and ovarian cancer; [0615] Frk, related to
acute myeloid leukemia and type 1 diabetes; [0616] Fyn, related to
Alzheimer's disease, schizophrenia and prevention of metastases,
e.g. in melanoma and squamous cell carcinoma; [0617] GSK3
(Gsk3.alpha. and/or Gsk3.beta.), related to CNS disorders such as
Alzheimer's disease, Parkinson's disease, amyotrophic lateral
sclerosis, diabetes type II, bipolar disorders, stroke, cancer,
chronic inflammatory disease, leukopenia, schizophrenia, chronic
pain, neuropathic pain, and traumatic head injury; [0618] HCK,
related to chronic myelogenous leukemia and acute lymphocytic
leukemia; [0619] Her2/Erbb2, related to prostate and breast cancer;
[0620] Her4/Erbb4, related to childhood medulloblastoma; [0621]
IGF1R, related to prostate cancer, hepatocellular carcinoma; [0622]
IKK beta, related to leukemia of T-cells, necrosis, insulin
resistance, and malignant neoplasms; [0623] Irak4, related to
bacterial infections, immunodeficiency syndrome, Crohn's disease,
ulcerative colitis, asthma, chronic bronchitis, cardio hypertrophy,
and kidney hypertension; [0624] Itk, related to allergic asthma;
[0625] Jak1, related to Hepatitis C virus infection; [0626] Jak2,
related to myeloproliferative disorders such as polycythaemia vera,
myelofibrosis, essential thrombocythemia, myeloid metaplasia and
leukemias, including acute lymphoblastic leukemia, chronic
neutrophilic leukemia, juvenile myelomonocytic leukemia, CMML,
Philadelphia chromosome-negative CML, megakaryocyte leukemia, and
acute erythroid leukemia; [0627] Jak3, related to X-linked severe
combined immunodeficiency, myeloproliferative disorders, transplant
rejection and autoimmune diseases such as rheumatoid arthritis,
inflammatory bowel syndrome, Crohn's disease, systemic lupus
erythematosis, ulcerative colitis, psoriasis and multiple
sclerosis; [0628] Jnk (Jnk1, Jnk2, Jnk3), related to metabolic
diseases including type 1 diabetes, type 2 diabetes, metabolic
syndrome, obesity, and hepatic steatosis; cardiovascular diseases
such as atherosclerosis, ischemia (e.g. cerebrovascular ischemia,
liver ischemia), reperfusion injury, cardiac hypertrophy; renal
diseases such as chronic renal failure; neoplastic diseases and
associated complications, including chemotherapy-induced hypoxia,
prostate tumors, myeloid leukemia and cancers of the liver, bone,
skin, brain, pancreas, lung breast, colon, prostate and ovary;
transplant rejection; pain of neuropathic or inflammatory origin
including acute and chronic pain; inflammatory and autoimmune
diseases including age-related macular degeneration, rheumatoid
arthritis, inflammatory bowel disease, ulcerative colitis, Crohn's
disease, systemic lupus erythematosis, Sjogren's Syndrome,
psoriasis, scleroderma, chronic thyroiditis, Grave's disease,
myasthenia gravis, and multiple sclerosis, and inflammation in
other organs including CNS inflammation, pancreatitis, nephritis,
atopic dermatitis, and hepatitis; airway inflammatory diseases such
as asthma; allergy, bronchitis, pulmonary fibrosis, chronic
obstructive pulmonary disease; neurologic diseases such as stroke,
cerebrovascular ischemia, neurodegenerative diseases such as
Parkinson's disease, Alzheimer's disease, amyotrophic lateral
sclerosis, dementia, senile chorea, head and spinal cord trauma,
and Huntington's disease. More particularly, Jnk1 is related to
type 1 diabetes, type 2 diabetes, metabolic syndrome, obesity and
hepatic steatosis, Jnk2 is related to atherosclerosis, and Jnk3 is
related to inflammatory diseases including autoimmune diseases such
as rheumatoid arthritis, inflammatory bowel syndrome, Crohn's
disease, systemic lupus erythematosis, Sjogren's Syndrome,
psoriasis and multiple sclerosis, airway inflammatory diseases such
as asthma, allergy, pulmonary fibrosis, and chronic obstructive
pulmonary disease, and inflammation in other organs, such as CNS
inflammation, pancreatitis, nephritis, and hepatitis; neurologic
diseases such as stroke, cerebrovascular ischemia, and
neurodegenerative diseases such as Parkinson's disease, Alzheimer's
disease, and Huntington's disease; and neoplastic diseases such as
prostate tumors and myeloid leukemia; [0629] Kdr, related to
anti-angiogenesis for treating solid tumor growth (e.g. ovarian,
lung, breast, prancreatic, prostate, colon, gastrointestinal
stromal tumor, non small cell lung cancer, and epidermoid cancer),
metastasis, psoriasis, rheumatoid arthritis, diabetic retinopathy
and age related macular degeneration; [0630] Kit, related to
malignancies, including mast cell tumors, small cell lung cancer,
testicular cancer, gastrointestinal stromal tumors (GISTs),
glioblastoma, astrocytoma, neuroblastoma, carcinomas of the female
genital tract, sarcomas of neuroectodermal origin, colorectal
carcinoma, carcinoma in situ, Schwann cell neoplasia associated
with neurofibromatosis, acute myelocytic leukemia, acute
lymphocytic leukemia, chronic myelogenous leukemia, mastocytosis,
melanoma, and canine mast cell tumors, and inflammatory diseases,
including asthma, rheumatoid arthritis, allergic rhinitis, multiple
sclerosis, inflammatory bowel syndrome, transplant rejection, and
hypercosinophilia; [0631] LCK, related to acute lymphoblastic
leukemia, T-cell lymphoma, lymphopenia, renal carcinoma, colon
carcinoma, severe combined immunodeficiency, multiple sclerosis,
inflammatory bowel and type I diabetes; [0632] MAP2K1, related to
acute myeloid leukemia, breast, ovarian and liver cancer; [0633]
MAP2K2, related to cancer and inflammation; [0634] MAP4K4, related
to cancer and tumor metastasis, diabetes and metabolic syndrome;
[0635] MAPKAPK2, cancer (e.g. prostate, breast), stroke,
menengitis, and inflammatory disorders; [0636] Met, related to
kidney, breast, bladder, non-small-cell lung, colorectal, and
bladder cancers, and hepatocellular carcinoma; [0637] Mnk1, related
to conditions associated with heat shock, nutrient deprivation,
oxidative or osmotic stress, and infection of mammalian cells (e.g.
with viruses such as adenovirus (Ad) or influenza virus), and
autoimmune diseases; [0638] MLK1, related to neurodegenerative
diseases such as Alzheimer's disease and Parkinson's disease, and
inflammatory disorders; [0639] p38, related to acute coronary
syndrome, stroke, atherosclerosis, and inflammatory autoimmune
diseases such as rheumatoid arthritis, inflammatory bowel disease,
and Crohn's disease; [0640] PDGFR (PDGFRA, PDGFRB), related to
idiopathic hypereosinophilic syndrome, chronic eosinophilic
leukemia, glioma, gastrointestinal stromal tumors (GISTs), juvenile
myelomonocytic leukemia, metastatic medulloblastoma, atherogenesis,
and restenosis. More particularly, PDGFRA related to idiopathic
hypereosinophilic syndrome, chronic eosinophilic leukemia, glioma,
gastrointestinal stromal tumors (GISTs), juvenile myelomonocytic
leukemia, metastatic medulloblastoma, atherogenesis, and
restenosis, and PDGFRB related to idiopathic hypereosinophilic
syndrome, chronic eosinophilic leukemia, juvenile myelomonocytic
leukemia, and metastatic medulloblastoma; [0641] PDPK1, related to
cancer and diabetes; [0642] Pim1, related to cancers such as
hematopoietic (e.g. acute myeloid and acute lymphoid leukemias) and
prostate cancers, and non-Hodgkin's lymphomas; [0643] Pim2, related
to lymphomas; [0644] Pim3, related to hepatocellular carcinoma;
[0645] PKC alpha, related to pituitary tumors and prefrontal
cortical dysfunction such as distractibility, impaired judgment,
impulsivity, and thought disorder, also may be used to sensitize
chemotherapy in breast, colon, and non small cell lung cancers;
[0646] PKC beta, related to diabetic retinopathy;
[0647] PKC-theta, related to insulin resistance, T-cell lymphoma;
[0648] Plk1, related to cancers (e.g. lymphoma of the thyroid,
non-Hodgkin's lymphomas, colorectal cancers, leukemias and
melanoma), also useful as sensitizer in chemotherapy; [0649] Pyk2,
related to inflammation (e.g. osteoporosis, polycystic kidney
disease, rheumatoid arthiritis and inflammatory bowel disease), CNS
disease (e.g. Parkinson's disease and Alzheimer's disease), stroke
and cancers (e.g. gliomas, breast cancer, and pancreatic cancer);
[0650] Ret, related to cancer of the thyroid, neuroblastoma,
familial medullary thyroid carcinoma (FMTC), multiple endocrine
neoplasia type IIA and IIB (MEN2A, MEN2B), and neurodegenerative
disorders (e.g. Hirschsprung's disease, Parkinson's disease,
Alzheimer's disease, and amyotrophic lateral sclerosis); [0651]
ROCK (ROCK-1, ROCK-2), related to cancers (e.g. ovarian cancer,
hepatocellular carcinoma, pancreatic cancer), ocular disease (e.g.
glaucoma), cardiac hypertrophy, improved renal perfusion,
transplant rejection, and acute respiratory distress syndrome;
[0652] Ron, related to cancer and inflammation; [0653] Src, related
to cancer and osteoporosis; [0654] Stk6, related to gastric,
bladder, breast, lung, CNS, ovarian, kidney, colon, prostate,
pancreas, and cervical cancers, melanoma, leukemia, and
neuroblastoma; [0655] Syk, related to lymphomas (e.g. mantle cell
lymphoma); [0656] TEC, related to sepsis, septic shock,
inflammation, rheumatoid arthritis, Crohn's disease, irritable
bowel disease (IBD), and ulcerative colitis; [0657] Tie2 (TEK),
related to cancer, arthritis (e.g. rheumatoid arthritis), and
atherosclerosis; [0658] TrkA, related to pain (e.g. chronic pain,
neuropathic pain), cancer, arthritis, diabetic retinopathy, macular
degeneration and psoriasis; [0659] Yes, related to various cancers
including esophageal squamous cell carcinoma; and [0660] Zap70,
related to AIDS, systemic lupus erythematosus, myasthenia gravis,
atherosclerosis, rejection of transplanted organs or tissues,
allograft rejection including acute and chronic allograft
rejection, graft versus host disease, rheumathoid arthritis,
psoriasis, systemic sclerosis, atopic dermatitis, eczematous
dermatitis, alopecia, and inflammation of the nasal mucus membrane,
including all forms of rhinitis.
[0661] Additional aspects and embodiments will be apparent from the
following Detailed Description and from the claims.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0662] As used herein the following definitions apply unless
clearly indicated otherwise:
[0663] "Halogen" refer to all halogens, that is, chloro (Cl),
fluoro (F), bromo (Br), or iodo (I).
[0664] "Hydroxyl" or "hydroxy" refer to the group --OH.
[0665] "Thiol" refers to the group --SH.
[0666] "Lower alkyl" alone or in combination means an
alkane-derived radical containing from 1 to 6 carbon atoms (unless
specifically defined) that includes a straight chain alkyl or
branched alkyl. The straight chain or branched alkyl group is
attached at any available point to produce a stable compound. In
many embodiments, a lower alkyl is a straight or branched alkyl
group containing from 1-6, 1-4, or 1-2, carbon atoms, such as
methyl, ethyl, propyl, isopropyl, butyl, t-butyl, and the like. A
"substituted lower alkyl" denotes lower alkyl that is independently
substituted, unless indicated otherwise, with one or more,
preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents, attached
at any available atom to produce a stable compound, wherein the
substituents are selected from the group consisting of --F, --OH,
--NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2,
--C(S)NH.sub.2, --S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2,
--NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2,
--OR.sup.o, --SR.sup.o, --OC(O)R.sup.o, --OC(S)R.sup.o,
--C(O)R.sup.o, --C(S)R.sup.o, --C(O)OR.sup.o, --C(S)OR.sup.o,
--S(O)R.sup.o, --S(O).sub.2R.sup.o, --C(O)NHR.sup.o,
--C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o, --C(S)NR.sup.oR.sup.o,
--S(O).sub.2NHR.sup.o, --S(O).sub.2NR.sup.oR.sup.o,
--C(NH)NHR.sup.o, --C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o,
--NHC(S)R.sup.o, --NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NR.sup.oS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.c, --R.sup.f, and
--R.sup.g. Furthermore, possible substitutions include subsets of
these substitutions, such as are indicated herein, for example, in
the description of compounds of Formula III, attached at any
available atom to produce a stable compound. For example "fluoro
substituted lower alkyl" denotes a lower alkyl group substituted
with one or more fluoro atoms, such as per fluoro alkyl, where
preferably the lower alkyl is substituted with 1, 2, 3, 4 or 5
fluoro atoms, also 1, 2, or 3 fluoro atoms. While it is understood
that substitutions are attached at any available atom to produce a
stable compound, when optionally substituted alkyl is an R group of
a moiety such as --OR (e.g. alkoxy), --SR (e.g. thioalkyl), --NHR
(e.g. alkylamino), --C(O)NHR, and the like, substitution of the
alkyl R group is such that substitution of the alkyl carbon bound
to any O, S, or N of the moiety (except where N is a heteroaryl
ring atom) excludes substituents that would result in any O, S, or
N of the substituent (except where N is a heteroaryl ring atom)
being bound to the alkyl carbon bound to any O, S, or N of the
moiety. "C.sub.2-6 alkyl" denotes lower alkyl containing 2-6 carbon
atoms. A "substituted C.sub.2-6alkyl" denotes optionally
substituted lower alkyl containing 2-6 carbon atoms. A "substituted
methyl" denotes methyl that is independently substituted, unless
indicated otherwise, with 1, 2, or 3 substituents, wherein the
substituents are selected as per optionally substituted lower
alkyl.
[0667] "C.sub.1-3 alkylene" refers to a divalent alkane-derived
radical containing 1-3 carbon atoms, straight chain or branched,
from which two hydrogen atoms are taken from the same carbon atom
or from different carbon atoms. C.sub.1-3alkylene includes
methylene --CH.sub.2--, ethylene --CH.sub.2CH.sub.2--, propylene
--CH.sub.2CH.sub.2CH.sub.2--, and isopropylene
--CH(CH.sub.3)CH.sub.2-- or --CH.sub.2CH(CH.sub.3)--. C.sub.1-3
alkylene substituted with one or more substituents indicates
C.sub.1-3 alkylene that is independently substituted, with one or
more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents as
indicated, attached at any available atom to produce a stable
compound.
[0668] "Lower alkenyl" alone or in combination means a straight or
branched hydrocarbon containing 2-6 carbon atoms (unless
specifically defined) and at least one, preferably 1-3, more
preferably 1-2, most preferably one, carbon to carbon double bond.
Carbon to carbon double bonds may be either contained within a
straight chain or branched portion. Examples of lower alkenyl
groups include ethenyl, propenyl, isopropenyl, butenyl, and the
like. A "substituted lower alkenyl" denotes lower alkenyl that is
independently substituted, unless indicated otherwise, with one or
more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents,
attached at any available atom to produce a stable compound,
wherein the substituents are selected from the group consisting of
--F, --OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH,
--C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, OR.sup.o, --SR.sup.o, --OC(O)R.sup.o,
--OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o, --C(O)OR.sup.o,
--C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.oR.sup.o, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NR.sup.oS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.d, --R.sup.f, and
--R.sup.g. Further, possible substitutions include subsets of these
substitutions, such as are indicated herein, for example, in the
description of compounds of Formula III, attached at any available
atom to produce a stable compound. For example "fluoro substituted
lower alkenyl" denotes a lower alkenyl group substituted with one
or more fluoro atoms, where preferably the lower alkenyl is
substituted with 1, 2, 3, 4 or 5 fluoro atoms, also 1, 2, or 3
fluoro atoms. While it is understood that substitutions are
attached at any available atom to produce a stable compound,
substitution of alkenyl groups are such that --F, --C(O)--,
--C(S)--, --C(NH)--, --S(O)--, --S(O).sub.2--, --O--, --S--, or N
(except where N is a heteroaryl ring atom), are not bound to an
alkene carbon thereof. Further, where alkenyl is a substituent of
another moiety or an R group of a moiety such as --OR, --NHR,
--C(O)R, and the like, substitution of the moiety is such that any
--C(O)--, --C(S)--, --S(O)--, --S(O).sub.2--, --O--, --S--, or N
thereof (except where N is a heteroaryl ring atom) are not bound to
an alkene carbon of the alkenyl substituent or R group. Further,
where alkenyl is a substituent of another moiety or an R group of a
moiety such as --OR, --NHR, --C(O)NHR, and the like, substitution
of the alkenyl R group is such that substitution of the alkenyl
carbon bound to any O, S, or N of the moiety (except where N is a
heteroaryl ring atom) excludes substituents that would result in
any O, S, or N of the substituent (except where N is a heteroaryl
ring atom) being bound to the alkenyl carbon bound to any O, S, or
N of the moiety. An "alkenyl carbon" refers to any carbon within an
alkenyl group, whether saturated or part of the carbon to carbon
double bond. An "alkene carbon" refers to a carbon within an
alkenyl group that is part of a carbon to carbon double bond.
[0669] "Lower alkynyl" alone or in combination means a straight or
branched hydrocarbon containing 2-6 carbon atoms (unless
specifically defined) containing at least one, preferably one,
carbon to carbon triple bond. Examples of alkynyl groups include
ethynyl, propynyl, butynyl, and the like. A "substituted lower
alkynyl" denotes lower alkynyl that is independently substituted,
unless indicated otherwise, with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2, or 3 substituents, attached at any available atom
to produce a stable compound, wherein the substituents are selected
from the group consisting of --F, --OH, --NH.sub.2, --NO.sub.2,
--CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2,
--S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2, --OR.sup.o, --SR.sup.o,
--OC(O)R.sup.o, --OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o,
--C(O)OR.sup.o, --C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.oR.sup.o, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NRS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.d, --R.sup.e, and
--R.sup.g. Further, possible substitutions include subsets of these
substitutions, such as are indicated herein, for example, in the
description of compounds of Formula III, attached at any available
atom to produce a stable compound. For example "fluoro substituted
lower alkynyl" denotes a lower alkynyl group substituted with one
or more fluoro atoms, where preferably the lower alkynyl is
substituted with 1, 2, 3, 4 or 5 fluoro atoms, also 1, 2, or 3
fluoro atoms. While it is understood that substitutions are
attached at any available atom to produce a stable compound,
substitution of alkynyl groups are such that --F, --C(O)--,
--C(S)--, --C(NH)--, --S(O)--, --S(O).sub.2--, --O--, --S--, or N
(except where N is a heteroaryl ring atom) are not bound to an
alkyne carbon thereof. Further, where alkynyl is a substituent of
another moiety or an R group of a moiety such as --OR, --NHR,
--C(O)R, and the like, substitution of the moiety is such that any
--C(O)--, --C(S)--,--S(O)--, --S(O).sub.2--, --O--, --S--, or N
thereof (except where N is a heteroaryl ring atom) are not bound to
an alkyne carbon of the alkynyl substituent or R group. Further,
where alkynyl is a substituent of another moiety or an R group of a
moiety such as --OR, --NHR, --C(O)NHR, and the like, substitution
of the alkynyl R group is such that substitution of the alkynyl
carbon bound to any O, S, or N of the moiety (except where N is a
heteroaryl ring atom) excludes substituents that would result in
any O, S, orN of the substituent (except where N is a heteroaryl
ring atom) being bound to the alkynyl carbon bound to any O, S; or
N of the moiety. An "alkynyl carbon" refers to any carbon within an
alkynyl group, whether saturated or part of the carbon to carbon
triple bond. An "alkyne carbon" refers to a carbon within an
alkynyl group that is part of a carbon to carbon triple bond.
[0670] "Cycloalkyl" refers to saturated or unsaturated,
non-aromatic monocyclic, bicyclic or tricyclic carbon ring systems
of 3-10, also 3-8, more preferably 3-6, ring members per ring, such
as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl, and the like. A
"substituted cycloalkyl" is a cycloalkyl that is independently
substituted, unless indicated otherwise, with one or more,
preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents, attached
at any available atom to produce a stable compound, wherein the
substituents are selected from the group consisting of halogen,
--OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH,
--C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.o, --SR.sup.o, --OC(O)R.sup.o,
--OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o, --C(O)OR.sup.o,
--C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.oR.sup.o, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o, --NR.sup.oC(S)NH
R.sup.o, --NHC(O)NR.sup.oR.sup.o, --NHC(S)NR.sup.oR.sup.o,
--NR.sup.oC(O)NR.sup.oR.sup.o, --NR.sup.oC(S)NR.sup.oR.sup.o,
--NHS(O).sub.2NHR.sup.o, --NR.sup.oS(O).sub.2NH.sub.2,
--NR.sup.oS(O).sub.2NHR.sup.o, --NHS(O).sub.2NR.sup.oR.sup.o,
--NR.sup.oS(O).sub.2NR.sup.oR.sup.o, --NHR.sup.o,
--NR.sup.oR.sup.o, --R.sup.d, --R.sup.e, --R.sup.f, and
--R.sup.g.
[0671] "Heterocycloalkyl" refers to a saturated or unsaturated
non-aromatic cycloalkyl group having from 5 to 10 atoms in which
from 1 to 3 carbon atoms in the ring are replaced by heteroatoms of
O, S or N, and are optionally fused with benzo or heteroaryl of 5-6
ring members. Heterocycloalkyl is also intended to include oxidized
S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring
nitrogen. Heterocycloalkyl is also intended to include compounds in
which a ring carbon may be oxo substituted, i.e. the ring carbon is
a carbonyl group, such as lactones and lactams. The point of
attachment of the heterocycloalkyl ring is at a carbon or nitrogen
atom such that a stable ring is retained. Examples of
heterocycloalkyl groups include, but are not limited to,
morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl,
pyrrolidinyl, pyrrolidonyl, piperazinyl, dihydrobenzofuryl, and
dihydroindolyl. A "substituted heterocycloalkyl" is a
heterocycloalkyl that is independently substituted, unless
indicated otherwise, with one or more, preferably 1, 2, 3, 4 or 5,
also 1, 2, or 3 substituents, attached at any available atom to
produce a stable compound, wherein the substituents are selected
from the group consisting of halogen, --OH, --NH.sub.2, --NO.sub.2,
--CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2,
--S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2, --OR.sup.o, --SR.sup.o,
--OC(O)R.sup.o, --OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o,
--C(O)OR.sup.o, --C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.pR.sup.c, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NR.sup.oS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.d, --R.sup.e, --R.sup.f,
and --R.sup.g.
[0672] "Aryl" alone of in combination refers to a monocyclic or
bicyclic ring system containing aromatic hydrocarbons such as
phenyl or naphthyl, which may be optionally fused with a cycloalkyl
of preferably 5-7, more preferably 5-6, ring members. "Arylene" is
a divalent aryl. A "substituted aryl" is an aryl that is
independently substituted, unless indicated otherwise, with one or
more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents,
attached at any available atom to produce a stable compound,
wherein the substituents are selected from the group consisting of
halogen, --OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH,
--C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.o, --SR.sup.o, --OC(O)R.sup.o,
--OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o, --C(O)OR.sup.o,
--C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.oR.sup.o, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NR.sup.oS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.d, --R.sup.e, --R.sup.f,
and --R.sup.g. A "substituted arylene" is a divalent substituted
aryl.
[0673] "Heteroaryl" alone or in combination refers to a monocyclic
aromatic ring structure containing 5 or 6 ring atoms, or a bicyclic
aromatic group having 8 to 10 atoms, containing one or more,
preferably 1-4, more preferably 1-3, even more preferably 1-2,
heteroatoms independently selected from the group consisting of O,
S, and N. Heteroaryl is also intended to include oxidized S or N,
such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen.
A carbon or nitrogen atom is the point of attachment of the
heteroaryl ring structure such that a stable compound is produced.
Examples of heteroaryl groups include, but are not limited to,
pyridinyl, pyridazinyl, pyrazinyl, quinaoxalyl, indolizinyl,
benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl,
pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl,
isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl,
triazolyl, furanyl, benzofuryl, and indolyl. "Nitrogen containing
heteroaryl" refers to heteroaryl wherein any heteroatoms are N.
"Heteroarylene" is a divalent heteroaryl. A "substituted
heteroaryl" is a heteroaryl that is independently substituted,
unless indicated otherwise, with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2, or 3 substituents, attached at any available atom
to produce a stable compound, wherein the substituents are selected
from the group consisting of halogen, --OH, --NH.sub.2, --NO.sub.2,
--CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2,
--S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2, --OR.sup.o, --SR.sup.o,
--OC(O)R.sup.o, --OC(S)R.sup.o, --C(O)R.sup.o, --C(S)R.sup.o,
--C(O)OR.sup.o, --C(S)OR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--C(O)NHR.sup.o, --C(S)NHR.sup.o, --C(O)NR.sup.oR.sup.o,
--C(S)NR.sup.oR.sup.o, --S(O).sub.2NHR.sup.o,
--S(O).sub.2NR.sup.oR.sup.o, --C(NH)NHR.sup.o,
--C(NH)NR.sup.pR.sup.c, --NHC(O)R.sup.o, --NHC(S)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NR.sup.oC(S)R.sup.o,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o,
--NHC(O)NHR.sup.o, --NHC(S)NHR.sup.o, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(S)NH.sub.2, --NR.sup.oC(O)NHR.sup.o,
--NR.sup.oC(S)NHR.sup.o, --NHC(O)NR.sup.oR.sup.o,
--NHC(S)NR.sup.oR.sup.o, --NR.sup.oC(O)NR.sup.oR.sup.o,
--NR.sup.oC(S)NR.sup.oR.sup.o, --NHS(O).sub.2NHR.sup.o,
--NR.sup.oS(O).sub.2NH.sub.2, --NR.sup.oS(O).sub.2NHR.sup.o,
--NHS(O).sub.2NR.sup.oR.sup.o, --NR.sup.oS(O).sub.2NR.sup.oR.sup.o,
--NHR.sup.o, --NR.sup.oR.sup.o, --R.sup.d, --R.sup.e, --R.sup.f,
and --R.sup.g. "Substituted heteroarylene" is a divalent
substituted heterOaryl.
[0674] The variables R.sup.o, R.sup.p, R.sup.c, R.sup.d, R.sup.e,
R.sup.f and R.sup.g as used in the description of optional
substituents for alkyl, alkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl are defined as follows:
[0675] each R.sup.o, R.sup.p, and R.sup.c are independently
selected from the group consisting of R.sup.d, R.sup.e, R.sup.f,
and R.sup.g, or R.sup.p and R.sup.c combine with the nitrogen to
which they are attached to form a 5-7 membered heterocycloalkyl or
a 5 or 7 membered nitrogen containing heteroaryl, wherein the 5-7
membered heterocycloalkyl or 5 or 7 membered nitrogen containing
heteroaryl are optionally substituted with one or more, preferably
1, 2, 3, 4 or 5, also 1, 2, or 3 substituents selected from the
group consisting of halogen, --NO.sub.2, --CN, --OH, --NH.sub.2,
--OR.sup.u, --SR.sup.u, --NHR.sup.u, --NR.sup.uR.sup.u, --R.sup.x,
and --R.sup.y; [0676] each R.sup.d is independently lower alkyl,
wherein lower alkyl is optionally substituted with one or more,
preferably 1, 2, 3, 4 or 5, also 1, 2 or 3 substituents selected
from the group consisting of fluoro, --OH, --NH.sub.2, --NO.sub.2,
--CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2,
--S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2, --NHC(S)NH.sub.2,
--NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2, --OR.sup.k, --SR.sup.k,
--OC(O)R.sup.k, --OC(S)R.sup.k, --C(O)R.sup.k, --C(S)R.sup.k,
--C(O)OR.sup.k, --C(S)OR.sup.k, --S(O)R.sup.k, --S(O).sub.2R.sup.k,
--C(O)NHR.sup.k, --C(S)NHR.sup.k, --C(O)NR.sup.kR.sup.k,
--C(S)NR.sup.kR.sup.k, --S(O).sub.2NHR.sup.k,
--S(O).sub.2NR.sup.kR.sup.k, --C(NH)NHR.sup.k,
--C(NH)NR.sup.mR.sup.n, --NHC(O)R.sup.k, --NHC(S)R.sup.k,
--NR.sup.kC(O)R.sup.k, --NR.sup.kC(S)R.sup.k,
--NHS(O).sub.2R.sup.k, --NR.sup.kS(O).sub.2R.sup.k,
--NHC(O)NHR.sup.k, --NHC(S)NHR.sup.k, --NR.sup.kC(O)NH.sub.2,
--NR.sup.kC(S)NH.sub.2, --NR.sup.kC(O)NHR.sup.k,
--NR.sup.kC(S)NHR.sup.k, --NHC(O)NR.sup.kR.sup.k,
--NHC(S)NR.sup.kR.sup.k, --NR.sup.kC(O)NR.sup.kR.sup.k,
--NR.sup.kC(S)NR.sup.kR.sup.k, --NHS(O).sub.2NHR.sup.k,
--NR.sup.kS(O).sub.2NH.sub.2, --NR.sup.kS(O).sub.2NHR.sup.k,
--NHS(O).sub.2NR.sup.kR.sup.k, --NR.sup.kS(O).sub.2NR.sup.kR.sup.k,
--NHR.sup.k, --NR.sup.kR.sup.k, --R.sup.i, and --R.sup.j; [0677]
each R.sup.e is independently lower alkenyl, wherein lower alkenyl
is optionally substituted with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2 or 3 substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH,
--C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.k, --SR.sup.k, --OC(O)R.sup.k,
--OC(S)R.sup.k, --C(O)R.sup.k, --C(S)R.sup.k, --C(O)OR.sup.k,
--C(S)OR.sup.k, --S(O)R.sup.k, --S(O).sub.2R.sup.k,
--C(O)NHR.sup.k, --C(S)NHR.sup.k, --C(O)NR.sup.kR.sup.k,
--C(S)NR.sup.kR.sup.k, --S(O).sub.2NHR.sup.k,
--S(O).sub.2NR.sup.kR.sup.k, --C(NH)NHR.sup.k,
--C(NH)NR.sup.mR.sup.n, --NHC(O)R.sup.k, --NHC(S)R.sup.k,
--NR.sup.kC(O)R.sup.k, --NR.sup.kC(S)R.sup.k,
--NHS(O).sub.2R.sup.k, --NR.sup.kS(O).sub.2R.sup.k,
--NHC(O)NHR.sup.k, --NHC(S)NHR.sup.k, --NR.sup.kC(O)NH.sub.2,
--NR.sup.kC(S)NH.sub.2, --NR.sup.kC(O)NHR.sup.k,
--NR.sup.kC(S)NHR.sup.k, --NHC(O)NR.sup.kR.sup.k,
--NHC(S)NR.sup.kR.sup.k, --NR.sup.kC(O)NR.sup.kR.sup.k,
--NR.sup.kC(S)NR.sup.kR.sup.k, --NHS(O).sub.2NHR.sup.k,
--NR.sup.kS(O).sub.2NH.sub.2, --NR.sup.kS(O).sub.2NHR.sup.k,
--NHS(O).sub.2NR.sup.kR.sup.k, --NR.sup.kS(O).sub.2NR.sup.kR.sup.k,
--NHR.sup.k, --NR.sup.kR.sup.k, --R.sup.h, and --R.sup.j; [0678]
each R.sup.f is independently lower alkynyl, wherein lower alkynyl
is optionally substituted with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2 or 3 substituents selected from the group
consisting of fluoro, --OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH,
--C(S)OH, --C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.k, --SR.sup.k, --OC(O)R.sup.k,
--OC(S)R.sup.k, --C(O)R.sup.k, --C(S)R.sup.k, --C(O)OR.sup.k,
--C(S)OR.sup.k, --S(O)R.sup.k, --S(O).sub.2R.sup.k,
--C(O)NHR.sup.k, --C(S)NHR.sup.k, --C(O)NR.sup.kR.sup.k,
--C(S)NR.sup.kR.sup.k, --S(O).sub.2NHR.sup.k,
--S(O).sub.2NR.sup.kR.sup.k, --C(NH)NHR.sup.k,
--C(NH)NR.sup.mR.sup.n, --NHC(O)R.sup.k, --NHC(S)R.sup.k,
--NR.sup.kC(O)R.sup.k, --NR.sup.kC(S)R.sup.k,
--NHS(O).sub.2R.sup.k, --NR.sup.kS(O).sub.2R.sup.k,
--NHC(O)NHR.sup.k, --NHC(S)NHR.sup.k, --NR.sup.kC(O)NH.sub.2,
--NR.sup.kC(S)NH.sub.2, --NR.sup.kC(O)NHR.sup.k, --NR.sup.kC(S)N
HR.sup.k, --NHC(O)NR.sup.kR.sup.k, --NHC(S)NR.sup.kR.sup.k,
--NR.sup.kC(O)NR.sup.kR.sup.k, --NR.sup.kC(S)NR.sup.kR.sup.k,
--NHS(O).sub.2NHR.sup.k, --NR.sup.kS(O).sub.2NH.sub.2,
--NR.sup.kS(O).sub.2NHR.sup.k, --NHS(O).sub.2NR.sup.kR.sup.k,
--NR.sup.kS(O).sub.2NR.sup.kR.sup.k, --NHR.sup.k,
--NR.sup.kR.sup.k, --R.sup.h, and --R.sup.j; [0679] each R.sup.g is
independently selected from the group consisting of cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl, wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2 or 3
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2,
--C(S)NH.sub.2, --S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2,
--NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2,
--OR.sup.k, --SR.sup.k, --OC(O)R.sup.k, --OC(S)R.sup.k,
--C(O)R.sup.k, --C(S)R.sup.k, --C(O)OR.sup.k, --C(S)OR.sup.k,
--S(O)R.sup.k, --S(O).sub.2R.sup.k, --C(O)NHR.sup.k,
--C(S)NHR.sup.k, --C(O)NR.sup.kR.sup.k, --C(S)NR.sup.kR.sup.k,
--S(O).sub.2NHR.sup.k, --S(O).sub.2NR.sup.kR.sup.k,
--C(NH)NHR.sup.k, --C(NH)NR.sup.mR.sup.n, --NHC(O)R.sup.k,
--NHC(S)R.sup.k, --NR.sup.kC(O)R.sup.k, --NR.sup.kC(S)R.sup.k,
--NHS(O).sub.2R.sup.k, --NR.sup.kS(O).sub.2R.sup.k,
--NHC(O)NHR.sup.k, --NHC(S)NHR.sup.k, --NR.sup.kC(O)NH.sub.2,
--NR.sup.kC(S)NH.sub.2, --NR.sup.kC(O)NHR.sup.k,
--NR.sup.kC(S)NHR.sup.k, --NHC(O)NR.sup.kR.sup.k,
--NHC(S)NR.sup.kR.sup.k, --NR.sup.kC(O)NR.sup.kR.sup.k,
--NR.sup.kC(S)NR.sup.kR.sup.k, --NHS(O).sub.2NHR.sup.k,
--NR.sup.kS(O).sub.2NT-I2, --NR.sup.kS(O).sub.2NHR.sup.k,
--NHS(O).sub.2NR.sup.kR.sup.k, --NR.sup.kS(O).sub.2NR.sup.kR.sup.k,
--NHR.sup.k, --NR.sup.kR.sup.k, --R.sup.h, --R.sup.i, and
--R.sub.j; [0680] wherein R.sup.k, R.sup.m, and R.sup.n at each
occurrence are independently selected from the group consisting of
R.sup.h, R.sup.i, and R.sup.j, or R.sup.m and R.sup.n combine with
the nitrogen to which they are attached form a 5-7 membered
heterocycloalkyl or a 5 or 7 membered nitrogen containing
heteroaryl, wherein the 5-7 membered heterocycloalkyl or 5 or 7
membered nitrogen containing heteroaryl are optionally substituted
with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3
substituents selected from the group consisting of halogen,
--NO.sub.2, --CN, --OH, --NH.sub.2, OR.sup.u, --SR.sup.u,
--NHR.sup.u, --NR.sup.uR.sup.u, --R.sup.x, and --R.sup.y; [0681]
wherein each R.sup.h is independently lower alkyl optionally
substituted with one or more, preferably 1, 2, 3, 4 or 5, also 1,
2, or 3 substituents selected from the group consisting of fluoro,
--OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH,
--C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.r, --SR.sup.r, --OC(O)R.sup.r,
--OC(S)R.sup.r, --C(O)R.sup.r,--C(S)R.sup.r, --C(O)OR.sup.r,
--C(S)OR.sup.r, --S(O)R.sup.r, --S(O).sub.2R.sup.r,
--C(O)NHR.sup.r, --C(S)NHR.sup.r, --C(O)NR.sup.rR.sup.r,
--C(S)NR.sup.rR.sup.r, --S(O).sub.2NHR.sup.r,
--S(O).sub.2NR.sup.rR.sup.r, --C(NH)NHR.sup.r,
--C(NH)NR.sup.sR.sup.r, --NHC(O)R.sup.r, --NHC(S)R.sup.r,
--NR.sup.rC(O)R.sup.r, --NR.sup.rC(S)R.sup.r,
--NHS(O).sub.2R.sup.r, --NR.sup.rS(O).sub.2R.sup.r,
--NHC(O)NHR.sup.r, --NHC(S)NHR.sup.r, --NR.sup.rC(O)NH.sub.2,
--NR.sup.rC(S)NH.sub.2, --NR.sup.rC(O)NHR.sup.r,
--NR.sup.rC(S)NHR.sup.r, --NHC(O)NR.sup.rR.sup.r,
--NHC(S)NR.sup.rR.sup.r, --NR.sup.rC(O)NR.sup.rR.sup.r,
--NR.sup.rC(S)NR.sup.rR.sup.r, --NHS(O).sub.2NHR.sup.r,
--NR.sup.rS(O).sub.2NH.sub.2, --NR.sup.rS(O).sub.2NHR.sup.r,
--NHS(O).sub.2NR.sup.rR.sup.r, --NR.sup.rS(O).sub.2NR.sup.rR.sup.r,
--NHR.sup.r, --NR.sup.rR.sup.r, --R.sup.i, and --R.sup.j; [0682]
wherein each R.sup.f is independently selected from the group
consisting of lower alkenyl and lower alkynyl, wherein lower
alkenyl or lower alkynyl are optionally substituted with one or
more, preferably 1, 2, 3, 4 or 5, also 1, 2 or 3 substituents
selected from the group consisting of fluoro, --OH, NH.sub.2,
--NO.sub.2, --CN, --C(O)OH, --C(S)OH, --C(O)NH.sub.2,
--C(S)NH.sub.2, --S(O).sub.2NH.sub.2, --NHC(O)NH.sub.2,
--NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2, --C(NH)NH.sub.2,
--OR.sup.r, --SR.sup.r, --OC(O)R.sup.r, --OC(S)R.sup.r,
--C(O)R.sup.r, --C(S)R.sup.r, --C(O)OR.sup.r, --C(S)OR.sup.r,
--S(O)R.sup.r, --S(O).sub.2R.sup.r,--C(O)NHR.sup.r,
--C(S)NHR.sup.r, --C(O)NR.sup.rR.sup.r, --C(S)NR.sup.rR.sup.r,
--S(O).sub.2NHR.sup.r, --S(O).sub.2NR.sup.rR.sup.r,
--C(NH)NHR.sup.r, --C(NH)NR.sup.sR.sup.r, --NHC(O)R.sup.r,
--NHC(S)R.sup.r, --NR.sup.rC(O)R.sup.r, --NR.sup.rC(S)R.sup.r,
--NHS(O).sub.2R.sup.r, --NR.sup.rS(O).sub.2R.sup.r,
--NHC(O)NHR.sup.r, --NHC(S)NHR.sup.r, --NR.sup.rC(O)NH.sub.2,
--NR.sup.tC(S)NH.sub.2, --NR.sup.rC(O)NHR.sup.r,
--NR.sup.rC(S)NHR.sup.r, --NHC(O)NR.sup.rR.sup.r,
--NHC(S)NR.sup.rR.sup.r, --NR.sup.tC(O)NR.sup.rR.sup.r,
--NR.sup.rC(S)NR.sup.tR.sup.s, --NHS(O).sub.2NHR.sup.r,
--NR.sup.rS(O).sub.2NH.sub.2, --NR.sup.rS(O).sub.2NHR.sup.r,
--NHS(O).sub.2NR.sup.rR.sup.r, --NR.sup.tS(O).sub.2NR.sup.tR.sup.s,
--NHR.sup.r, --NR.sup.rR.sup.r, and --R.sup.j; [0683] wherein each
R.sup.j is independently selected from the group consisting of
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally
substituted with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2
or 3 substituents selected from the group consisting of halogen,
--OH, --NH.sub.2, --NO.sub.2, --CN, --C(O)OH, --C(S)OH,
--C(O)NH.sub.2, --C(S)NH.sub.2, --S(O).sub.2NH.sub.2,
--NHC(O)NH.sub.2, --NHC(S)NH.sub.2, --NHS(O).sub.2NH.sub.2,
--C(NH)NH.sub.2, --OR.sup.r, --SR.sup.r, --OC(O)R.sup.r,
--OC(S)R.sup.r, --C(O)R.sup.r, --C(S)R.sup.r, --C(O)OR.sup.r,
--C(S)OR.sup.r, --S(O)R.sup.r, --S(O).sub.2R.sup.r,
--C(O)NHR.sup.r, --C(S)NHR.sup.r, --C(O)NR.sup.rR.sup.r,
--C(S)NR.sup.rR.sup.r, --S(O).sub.2NHR.sup.r,
--S(O).sub.2NR.sup.rR.sup.r, --C(NH)NHR.sup.r,
--C(NH)NR.sup.sR.sup.t, --NHC(O)R.sup.r, --NHC(S)R.sup.r,
--NR.sup.rC(O)R.sup.r, --NR.sup.rC(S)R.sup.r,
--NHS(O).sub.2R.sup.r, --NR.sup.rS(O).sub.2R.sup.r,
--NHC(O)NHR.sup.r, --NHC(S)NHR.sup.r, --NR.sup.rC(O)NH.sub.2,
--NR.sup.rC(S)NH.sub.2, --NR.sup.rC(O)NHR.sup.r,
--NR.sup.rC(S)NHR.sup.r, --NHC(O)NR.sup.rR.sup.r,
--NHC(S)NR.sup.rR.sup.r, --NR.sup.rC(O)NR.sup.rR.sup.r,
--NR.sup.rC(S)NR.sup.rR.sup.r, --NHS(O).sub.2NHR.sup.r,
--NR.sup.rS(O).sub.2NH.sub.2, --NR.sup.rS(O).sub.2NHR.sup.r,
--NHS(O).sub.2NR.sup.rR.sup.r, --NR.sup.rS(O).sub.2NR.sup.rR.sup.r,
--NHR.sup.r, --NR.sup.rR.sup.r, cycloalkylamino, and --R.sup.x;
[0684] wherein each R.sup.r, R.sup.s, and R.sup.t at each
occurrence are independently selected from the group consisting of
lower alkyl, C.sub.3-6 alkenyl, C.sub.3-6 alkynyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl, wherein lower alkyl is
optionally substituted with one or more, preferably 1, 2, 3, 4 or
5, also 1, 2, or 3 substituents selected from the group consisting
of --R.sup.y, fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino,
provided, however, that any substitution of the lower alkyl carbon
bound to any O, S, or N, of --O.sup.r, --SR.sup.r, --C(O)OR.sup.r,
--C(S)OR.sup.r, --C(O)NHR.sup.r, --C(S)NHR.sup.r,
--C(O)NR.sup.rR.sup.r, --C(S)NR.sup.rR.sup.r,
--S(O).sub.2NHR.sup.r, --S(O).sub.2NR.sup.rR.sup.r,
--C(NH)NHR.sup.r, --NR.sup.rC(O)R.sup.r, --NR.sup.rC(S)R.sup.r,
--NR.sup.rS(O).sub.2R.sup.r, --NHC(O)NHR.sup.r, --NHC(S)NHR.sup.r,
--NR.sup.rC(O)NH.sub.2, --NR.sup.rC(S)NH.sub.2,
--NR.sup.rC(O)NHR.sup.r, --NR.sup.rC(S)NHR.sup.r,
--NHC(O)NR.sup.rR.sup.r, --NHC(S)NR.sup.rR.sup.r,
--NR.sup.rC(O)NR.sup.rR.sup.r, --NR.sup.rC(S)NR.sup.rR.sup.r,
--NHS(O).sub.2NHR.sup.r, --NR.sup.rS(O).sub.2NH.sub.2,
--NR.sup.rS(O).sub.2NHR.sup.r, --NHS(O).sub.2NR.sup.rR.sup.r,
--NR.sup.rS(O).sub.2NR.sup.rR.sup.r, --NHR.sup.r, or
--NR.sup.rR.sup.r is selected from the group consisting of fluoro
and --R.sup.y, and wherein C.sub.3-6 alkenyl or C.sub.3-6 alkynyl
are optionally substituted with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2, or 3 substituents selected from the group
consisting of --R.sup.y, fluoro, lower alkyl, fluoro substituted
lower alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino, provided, however, that any
substitution of the C.sub.3-6 alkenyl or C.sub.3-6 alkynyl carbon
bound to any O, S, or N, of --OR.sup.r, --SR.sup.r, --C(O)OR.sup.r,
--C(S)OR.sup.r, --C(O)NHR.sup.r, --C(S)NHR.sup.r,
--C(O)NR.sup.rR.sup.r, --C(S)NR.sup.rR.sup.r,
--S(O).sub.2NHR.sup.r, --S(O).sub.2NR.sup.rR.sup.r,
--C(NH)NHR.sup.r, --NR.sup.rC(O)R.sup.r, --NR.sup.rC(S)R.sup.r,
--NR.sup.rS(O).sub.2R.sup.r, --NHC(O)NHR.sup.r, --NHC(S)NHR.sup.r,
--NR.sup.rC(O)NH.sub.2, --NR.sup.rC(S)NH.sub.2,
--NR.sup.rC(O)NHR.sup.r, --NR.sup.rC(S)NHR.sup.r,
--NHC(O)NR.sup.rR.sup.r, --NHC(S)NR.sup.rR.sup.r,
--NR.sup.rC(O)NR.sup.rR.sup.r, --NR.sup.rC(S)NR.sup.rR.sup.r,
--NHS(O).sub.2NHR.sup.r, --NR.sup.rS(O).sub.2NH.sub.2,
--NR.sup.rS(O).sub.2NHR.sup.r, --NHS(O).sub.2NR.sup.rR.sup.r,
--NR.sup.rS(O).sub.2NR.sup.rR.sup.r, --NHR.sup.r, or
--NR.sup.rR.sup.r is selected from the group consisting of fluoro,
lower alkyl, fluoro substituted lower alkyl, and --R.sup.y, and
wherein cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are
optionally substituted with one or more, preferably 1, 2, 3, 4 or
5, also 1, 2, or 3 substituents selected from the group consisting
of halogen, --OH, --NH.sub.2, --NO.sub.2, --CN, lower alkyl, fluoro
substituted lower alkyl, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino, or R.sup.s and
R.sup.t combine with the nitrogen to which they are attached form a
5-7 membered heterocycloalkyl or a 5 or 7 membered nitrogen
containing heteroaryl, wherein the 5-7 membered heterocycloalkyl or
5 or 7 membered nitrogen containing heteroaryl are optionally
substituted with one or more, preferably 1, 2, 3, 4 or 5, also 1,
2, or 3 substituents selected from the group consisting of halogen,
--NO.sub.2, --CN, --OH, --NH.sub.2OR.sup.u, --SR.sup.u,
--NHR.sup.u, --NR.sup.uR.sup.u, --R.sup.x, and --R.sup.y; [0685]
wherein each R.sup.u is independently selected from the group
consisting of lower alkyl, C
.sub.3-6 alkenyl, C.sub.3-6 alkynyl, cycloalkyl, heterocycloalkyl,
aryl, and heteroaryl, wherein lower alkyl is optionally substituted
with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3
substituents selected from the group consisting of --R.sup.y,
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino, provided,
however, that any substitution of the lower alkyl carbon bound to
thc O of --OR.sup.u, S of --SR.sup.u, or N of --NHR.sup.u is fluoro
or --R.sup.y, and wherein C.sub.3-6 alkenyl or C.sub.3-6 alkynyl
are optionally substituted with one or more, preferably 1, 2, 3, 4
or 5, also 1, 2, or 3 substituents selected from the group
consisting of --R.sup.y, fluoro, --OH, --NH.sub.2, lower alkyl,
fluoro substituted lower alkyl, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino, provided,
however, that any substitution of the C.sub.3-6 alkenyl or
C.sub.3-6 alkynyl carbon bound to the O of --OR.sup.u, S of
--SR.sup.u, or N of --NHR.sup.u is fluoro, lower alkyl, fluoro
substituted lower alkyl, or --R.sup.y, and wherein cycloalkyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3
substituents selected from the group consisting of halogen, --OH,
--NH.sub.2, --NO.sub.2, --CN, lower alkyl, fluoro substituted lower
alkyl, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino; [0686] wherein each R.sup.x is
selected from the group consisting of lower alkyl, lower alkenyl
and lower alkynyl, wherein lower alkyl is optionally substituted
with one or more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3
substituents selected from the,group consisting of --R.sup.y,
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino, and wherein
lower alkenyl or lower alkynyl are optionally substituted with one
or more, preferably 1, 2, 3, 4 or 5, also 1, 2, or 3 substituents
selected from the group consisting of --R.sup.y, fluoro, --OH,
--NH.sub.2, lower alkyl, fluoro substituted lower alkyl, lower
alkoxy, fluoro substituted lower alkoxy, lower alkylthio, fluoro
substituted lower alkylthio, mono-alkylamino, di-alkylamino, and
cycloalkylamino; [0687] wherein each R.sup.y is selected from the
group consisting of cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl, and
heteroaryl are optionally substituted with one or more, preferably
1, 2, 3, 4 or 5, also 1, 2, or 3 substituents selected from the
group consisting of halogen, --OH, --NH.sub.2, --NO.sub.2, --CN,
lower alkyl, fluoro substituted lower alkyl, lower alkoxy, fluoro
substituted lower alkoxy, lower alkylthio, fluoro substituted lower
alkylthio, mono-alkylamino, di-alkylamino, and cycloalkylamino.
[0688] In some embodiments, all occurrences of optionally
substituted lower alkyl, optionally substituted C.sub.2-6 alkyl,
optionally substituted lower alkenyl, or optionally substituted
lower alkynyl are optionally substituted with one or more, also 1,
2 or 3 groups or substituents selected from the group consisting of
fluoro, --NO.sub.2, --CN, --OR.sup.1a, --SR.sup.1a,
--NR.sup.1aR.sup.1a, --OC(O)R.sup.1a, --OC(S)R.sup.1a,
--C(O)R.sup.1a, --C(S)R.sup.1a, --C(O)OR.sup.1a, --C(S)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --C(NH)NR.sup.1aR.sup.1a,
--NR.sup.1aC(O)R.sup.1a, --NR.sup.1aC(S)R.sup.1a,
--NR.sup.1aS(O).sub.2R.sup.1a, rNR.sup.1aC(O)NR.sup.1aR.sup.1a,
--NR.sup.1aC(S)NR.sup.1aR.sup.1a,
--NR.sup.1aS(O).sub.2NR.sup.1aR.sup.1a, --S(O)R.sup.1a,
--S(O).sub.2R.sup.1a, cycloalkyl, heterocycloalkyl, aryl and
heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with one or more, also 1, 2
or 3 groups or substituents selected from the group consisting of
halogen, --NO.sub.2, --CN, --OR.sup.1a, --SR.sup.1a,
--NR.sup.1aR.sup.1a, --OC(O)R.sup.1a, --OC(S)R.sup.1a,
--C(O)R.sup.1a, --C(S)R.sup.1a, --C(O)OR.sup.1a, --C(S)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --C(NH)NR.sup.1aR.sup.1a,
--NR.sup.1aC(O)R.sup.1a, --NR.sup.1aC(S)R.sup.1a,
--NR.sup.1aS(O).sub.2R.sup.1a, --NR.sup.1aC(O)NR.sup.1aR.sup.1a,
--NR.sup.1aC(S)NR.sup.1aR.sup.1a,
--NR.sup.1aS(O).sub.2NR.sup.1aR.sup.1a, --S(O)R.sup.1a,
--S(O).sub.2R1;1, --R.sup.1b, and lower alkyl optionally
substituted with one or more, also 1, 2 or 3 groups or substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and --R.sup.1b, and all occurrences of optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted 5-7 membered heterocycloalkyl, optionally substituted
aryl, optionally substituted arylene, optionally substituted
heteroaryl, optionally substituted heteroarylene, or optionally
substituted 5 or 7 membered nitrogen containing heteroaryl are
optionally substituted with one or more, also 1, 2, or 3 groups or
substituents selected from the group consisting of halogen,
--NO.sub.2, --CN, --OR.sup.1a, --SR.sup.1a, --NR.sup.1aR.sup.1a,
--OC(O)R.sup.1a, --OC(S)R.sup.1a, --C(O)R.sup.1a, --C(S)R.sup.1a,
--C(O)OR.sup.1a, --C(S)OR.sup.1a, --C(O)NR.sup.1aR.sup.1a,
--C(S)NR.sup.1aR.sup.1a, --S(O).sub.2NR.sup.1aR.sup.1a,
--C(NH)NR.sup.1aR.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, --NR.sup.1aS(O).sub.2R.sup.1a,
--NR.sup.1aC(O)NR.sup.1aR.sup.1a, --NR.sup.1aC(S)NR.sup.1aR.sup.1a,
--NR.sup.1aS(O).sub.2NR.sup.1aR.sup.1a, --S(O)R.sup.1a,
--S(O).sub.2R.sup.1a, --R.sup.1b, and lower alkyl optionally
substituted with one or more, also 1, 2 or 3 groups or substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and --R.sup.1b, wherein R.sup.1a is selected from the group
consisting of hydrogen, provided, however, that hydrogen is not
bound to any of C(S), C(O), S(O), or S(O).sub.2of --OC(O)R.sup.1a,
--OC(S)R.sup.1a, --C(O)R.sup.1a, --C(S)R.sup.1a,
--NR.sup.1aC(O)R.sup.1a, --NR.sup.1aC(S)R.sup.1a,
--NR.sup.1aS(O).sub.2R.sup.1a, --S(O)R.sup.1a, or
--S(O).sub.2R.sup.1a, --R.sup.1b, and lower alkyl optionally
substituted with one or more, also 1, 2 or 3 groups or substituents
selected from the group consisting of fluoro, --OH, --NH.sub.2,
lower alkoxy, fluoro substituted lower alkoxy, lower alkylthio,
fluoro substituted lower alkylthio, mono-alkylamino, di-alkylamino,
and --R.sup.1b, provided, however, that any substitution of the
alkyl carbon bound to O, S, or N of --OR.sup.1a, --SR.sup.1a,
--NR.sup.1aR.sup.1a, --C(O)OR.sup.1a, --C(S)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --C(NH)NR.sup.1aR.sup.1a,
--NR.sup.1aC(O)R.sup.1a, --NR.sup.1aC(S)R.sup.1a,
--NR.sup.1aS(O).sub.2R.sup.1a, --NR.sup.1aC(O)NR.sup.1aR.sup.1a,
--NR.sup.1aC(S)NR.sup.1aR.sup.1a, or
--NR.sup.1aS(O).sub.2NRuR.sup.1a, is fluoro or --R.sup.1b, and
wherein --R.sup.1b is selected from the group consisting of
cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein
cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more, also 1, 2 or 3 groups or substituents
selected from the group consisting of halogen, --CN, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and cycloalkylamino.
[0689] In some embodiments, all occurrences of optionally
substituted lower alkyl, optionally substituted C.sub.2-6 alkyl,
optionally substituted lower alkenyl, or optionally substituted
lower alkynyl are optionally substituted with one or more, also 1,
2 or 3 groups or substituents selected from the group consisting of
fluoro, --CN, --OR.sup.1a, --SR.sup.1a, --NR.sup.1aR.sup.1a,
--C(O)R.sup.1a, --C(S)R.sup.1a, --C(Q)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, --NR.sup.1aS(O).sub.2R.sup.1a,
--S(O)R.sup.1a, --S(O).sub.2R.sup.b, cycloalkyl, heterocycloalkyl,
aryl and heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with one or more, also 1, 2
or 3 groups or substituents selected from the group consisting of
halogen, --CN, --OR.sup.1a, --SR.sup.1a, --NR.sup.1aR.sup.1a,
--C(O)R.sup.1a, --C(S)R.sup.1a, --C(O)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, --NR.sup.1aS(O).sub.2R.sup.1a,
--S(O)R.sup.1a, --S(O).sub.2R.sup.1a, --R.sup.1b, and lower alkyl
optionally substituted with one or more, also 1, 2 or 3 groups or
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and --R.sup.1b, and all occurrences of optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted 5-7 membered heterocycloalkyl, optionally
substituted aryl, optionally substituted arylene, optionally
substituted heteroaryl, optionally substituted heteroarylene, or
optionally substituted 5 or 7 membered nitrogen containing
heteroaryl are optionally substituted with one or more, also 1, 2,
or 3 groups or substituents selected from the group consisting of
halogen, --CN, --OR.sup.1a, --SR.sup.1a, --NR.sup.1aR.sup.1a,
--C(O)R.sup.1a, --C(S)R.sup.1a, --C(O)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, --NR.sup.1aS(O).sub.2R.sup.1a,
--S(O)R.sup.1a, --S(O).sub.2R.sup.1a, --R.sup.1b, and lower alkyl
optionally substituted with one or more, also 1, 2 or 3 groups or
substituents selected from the group consisting of fluoro, --OH,
--NH.sub.2, lower alkoxy, fluoro substituted lower alkoxy, lower
alkylthio, fluoro substituted lower alkylthio, mono-alkylamino,
di-alkylamino, and --R.sup.1b, wherein R.sup.1a is selected from
the group consisting of hydrogen, provided, however, that hydrogen
is not bound to any of C(S), C(O), S(O), or S(O).sub.2of
--C(O)R.sup.1a, --C(S)R.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, --NR.sup.1aS(O).sub.2R.sup.1a,
--S(O)R.sup.1a, or --S(O).sub.2R.sup.1a, --R.sup.1b, and lower
alkyl optionally substituted with one or more, also 1, 2 or 3
groups or substituents selected from the group consisting of
fluoro, --OH, --NH.sub.2, lower alkoxy, fluoro substituted lower
alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and --R.sup.1b, provided, however,
that any substitution of the alkyl carbon bound to O, S, or N of
--OR.sup.1a, --SR.sup.1a, --NR.sup.1aR.sup.1a, --C(O)OR.sup.1a,
--C(O)NR.sup.1aR.sup.1a, --C(S)NR.sup.1aR.sup.1a,
--S(O).sub.2NR.sup.1aR.sup.1a, --NR.sup.1aC(O)R.sup.1a,
--NR.sup.1aC(S)R.sup.1a, or --NR.sup.1aS(O).sub.2R.sup.1a, is
fluoro or --R.sup.1b, and wherein --R.sup.1b is selected from the
group consisting of cycloalkyl, heterocycloalkyl, aryl and
heteroaryl, wherein cycloalkyl, heterocycloalkyl, aryl and
heteroaryl are optionally substituted with one or more, also 1, 2
or 3 groups or substituents selected from the group consisting of
halogen, --CN, --OH, --NH.sub.2, lower alkoxy, fluoro substituted
lower alkoxy, lower alkylthio, fluoro substituted lower alkylthio,
mono-alkylamino, di-alkylamino, and cycloalkylamino.
[0690] "Lower alkoxy" denotes the group --OR.sup.z, where R.sup.z
is lower alkyl. "Substituted lower alkoxy" denotes lower alkoxy in
which R.sup.z is lower alkyl substituted with one or more
substituents as indicated herein, for example, in the description
of compounds of Formula III, including descriptions of substituted
cycloalkyl, cycloheteroalkyl, aryl and heteroaryl, attached at any
available atom to produce a stable compound. Preferably,
substitution of lower alkoxy is with 1, 2, 3, 4, or 5 substituents,
also 1, 2, or 3 substituents. For example "fluoro substituted lower
alkoxy" denotes lower alkoxy in which the lower alkyl is
substituted with one or more fluoro atoms, where preferably the
lower alkoxy is substituted with 1, 2, 3, 4 or 5 fluoro atoms, also
1, 2, or 3 fluoro atonis. While it is understood that substitutions
on alkoxy are attached at any available atom to produce a stable
compound, substitution of alkoxy is such that O, S, or N (except
where N is a heteroaryl ring atom), are not bound to the alkyl
carbon bound to the alkoxy O. Further, where alkoxy is described as
a substituent of another moiety, the alkoxy oxygen is not bound to
a carbon atom that is bound to an O, S, or N of the other moiety
(except where N is a heteroaryl ring atom), or to an alkene or
alkyne carbon of the other moiety.
[0691] "Lower alkylthio" denotes the group --SR.sup.aa, where
R.sup.aa is lower alkyl. "Substituted lower alkylthio" denotes
lower alkylthio in which R.sup.aa is lower alkyl substituted with
one or more substituents as indicated herein, for example, in the
description of compounds of Formula III, including descriptions of
substituted cycloalkyl, cycloheteroalkyl, aryl and heteroaryl,
attached at any available atom to produce a stable compound.
Preferably, substitution of lower alkylthio is with 1, 2, 3, 4, or
5 substituents, also 1, 2, or 3 substituents. For example "fluoro
substituted lower alkylthio" denotes lower alkylthio in which the
lower alkyl is substituted with one or more fluoro atoms, where
preferably the lower alkylthio is substituted with 1, 2, 3, 4 or 5
fluoro atoms, also 1, 2, or 3 fluoro atoms. While it is understood
that substitutions on alkylthio are attached at any available atom
to produce a stable compound, substitution of alkylthio is such
that O, S, or N (except where N is a heteroaryl ring atom), are not
bound to the alkyl carbon bound to the alkylthio S. Further, where
alkylthio is described as a substituent of another moiety, the
alkylthio sulfur is not bound to a carbon atom that is bound to an
O, S, or N of the other moiety (except where N is a heteroaryl ring
atom), or to an alkene or alkyne carbon of the other moiety.
[0692] "Amino" or "amine" denotes the group --NH.sub.2.
"Mono-alkylamino" denotes the group --NHR.sup.bb where R.sup.bb is
lower alkyl. "Di-alkylamino" denotes the group --NR.sup.bbR.sup.cc,
where R.sup.bb and R.sup.cc are independently lower alkyl.
"Cycloalkylamino" denotes the group --NR.sup.ddR.sup.ee, where
R.sup.dd and R.sup.ee combine with the nitrogen to form a 5-7
membered heterocycloalkyl, where the heterocycloalkyl may contain
an additional heteroatom within the ring, such as O, N, or S, and
may also be further substituted with lower alkyl. Examples of 5-7
membered heterocycloalkyl include, but are not limited to,
piperidine, piperazine, 4-methylpiperazine, morpholine, and
thiomorpholine. While it is understood that when mono-alkylamino,
di-alkylamino, or cycloalkylamino are substituents on other
moieties that are attached at any available atom to produce a
stable compound, the nitrogen of mono-alkylamino, di-alkylamino, or
cycloalkylamino as substituents is not bound to a carbon atom that
is bound to an O, S, or N of the other moiety.
[0693] A "nitrogen protecting group" is a chemical group covalently
bound to a nitrogen atom of a compound that is used to protect the
nitrogen from reaction during a synthetic step. The nitrogen
protecting group may be added to a compound and removed in a
subsequent step by methods known to those of skill in the art.
Nitrogen protecting groups include, without limitation, carbamates,
amides, N-sulfonyl derivatives, groups of formula --C(O)OR, wherein
R is, for example, methyl, ethyl, t-butyl, benzyl, phenylethyl,
CH.sub.2.dbd.CHCH.sub.2--, and the like, groups of the formula
--C(O)R' wherein R' is, for example, methyl, phenyl,
trifluoromethyl, and the like, groups of the formula --SO.sub.2R'',
wherein R'' is, for example, tolyl, phenyl, trifluoromethyl,
2,2,5,7,8-pentamethylchroman-6-yl, 2,3,6-trimethyl-4-methoxyphenyl,
and the like, and silanyl containing groups, such as
2-trimethylsilylethoxymethyl, t-butyldimethylsilyl,
triisopropylsilyl, and the like. Other suitable nitrogen protecting
groups may be found in texts such as T. W. Greene & P. G. M.
Wuts, Protective Groups in Organic Synthesis, John Wiley &
Sons, 1991.
[0694] As used herein, the term "composition" refers to a
formulation suitable for administration to an intended animal
subject for therapeutic purposes that contains at least one
pharmaceutically active compound and at least one pharmaceutically
acceptable carrier or excipient.
[0695] The term "pharmaceutically acceptable" indicates that the
indicated material does not have properties that would cause a
reasonably prudent medical practitioner to avoid administration of
the material to a patient, taking into consideration the disease or
conditions to be treated and the respective route of
administration. For example, it is commonly required that such a
material be essentially sterile, e.g., for injectibles.
[0696] In the present context, the term "therapeutically effective"
or "effective amount" indicates that the materials or amount of
material is effective to prevent, alleviate, or ameliorate one or
more symptoms of a disease or medical condition, and/or to prolong
the survival of the subject being treated.
[0697] In the present context, the terms "synergistically
effective" or "synergistic effect" indicate that two or more
compounds that are therapeutically effective, when used in
combination, provide improved therapeutic effects greater than the
additive effect that would be expected based on the effect of each
compound used by itself.
[0698] As used herein, the terms "ligand" and "modulator" are used
equivalently to refer to a compound that changes (i.e., increases
or decreases) the activity of a target biomolecule, e.g., an enzyme
such as a kinase. Generally a ligand or modulator will be a small
molecule, where "small molecule refers to a compound with a
molecular weight of 1500 daltons or less, or preferably 1000
daltons or less, 800 daltons or less, or 600 daltons or less. Thus,
an "improved ligand" is one that possesses better pharmacological
and/or pharmacokinetic properties than a reference compound, where
"better" can be defined by one skilled in the relevant art for a
particular biological system or therapeutic use.
[0699] In the context of binding compounds and ligands, the term
"derivative" or "derivative compound" refers to a compound having a
chemical structure that contains a common core chemical structure
as a parent or reference compound, but differs by having at least
one structural difference, e.g., by having one or more substituents
added and/or removed and/or replaced, and/or by having one or more
atoms replaced with different atoms. Unless clearly indicated to
the contrary, the term "derivative" does not mean that the
derivative is synthesized using the parent compound as a starting
material or as an intermediate, although in some cases, the
derivative may be synthesized from the parent.
[0700] Thus, the term "parent compound" refers to a reference
compound having structural features also found in the derivative
compound. Often but not always, a parent compound has a simpler
chemical structure than the derivative.
[0701] By "chemical structure" or "chemical substructure" is meant
any definable atom or group of atoms that constitute an
individually identifiable portion of a molecule, such as a
substituent moiety, a core which is optionally substituted, and the
like. Normally, chemical substructures of a ligand can have a role
in binding of the ligand to a target molecule, or can influence the
three-dimensional shape, electrostatic charge, and/or
conformational properties of the ligand.
[0702] The term "binds" in connection with the interaction between
a target and a potential binding compound indicates that the
potential binding compound associates with the target to a
statistically significant degree as compared to association with
proteins generally (i.e., non-specific binding). Thus, the term
"binding compound" refers to a compound that has a statistically
significant association with a target molecule. Preferably a
binding compound interacts with a specified target with a
dissociation constant (K.sub.D) of 1 mM or less, 1 .mu.M or less,
100 nM or less, 10 nM or less, or 1 nM or less.
[0703] In the context of compounds binding to a target, the terms
"greater affinity" and "selective" indicates that the compound
binds more tightly than a reference compound, or than the same
compound in a reference condition, i.e., with a lower dissociation
constant. In some embodiments, the greater affinity is at least 2,
3, 4, 5, 8, 10, 50, 100, 200, 400, 500, 1000, or 10,000-fold
greater affinity.
[0704] As used herein in connection with compounds of the
invention, the term "synthesizing" and like terms means chemical
synthesis from one or more precursor materials. Further, by
"assaying" is meant the creation of experimental conditions and the
gathering of data regarding a particular result of the experimental
conditions. For example, enzymes can be assayed based on their
ability to act upon a detectable substrate. A compound or ligand
can be assayed based on its ability to bind to a particular target
molecule or molecules.
[0705] As used herein, the term "modulating" or "modulate" refers
to an effect of altering a biological activity, especially a
biological activity associated with a particular biomolecule such
as a protein kinase. For example, an agonist or antagonist of a
particular biomolecule modulates the activity of that biomolecule,
e.g., an enzyme, by either increasing (e.g. agonist, activator), or
decreasing (e.g. antagonist, inhibitor) the activity of the
biomolecule, such as an enzyme. Such activity is typically
indicated in terms of an inhibitory concentration (IC.sub.50) or
excitation concentration (EC.sub.50) of the compound for an
inhibitor or activator, respectively, with respect to, for example,
an enzyme.
[0706] In the context of the use, testing, or screening of
compounds that are or may be modulators, the term "contacting"
means that the compound(s) are caused to be in sufficient proximity
to a particular molecule, complex, cell, tissue, organism, or other
specified material that potential binding interactions and/or
chemical reaction between the compound and other specified material
can occur.
[0707] As used herein in connection with amino acid or nucleic acid
sequence, the term "isolate" indicates that the sequence is
separated from at least a portion of the amino acid and/or nucleic
acid sequences with which it would normally be associated.
[0708] In connection with amino acid or nucleic sequences, the term
"purified" indicates that the subject molecule constitutes a
significantly greater proportion of the biomolecules in a
composition than the proportion observed in a prior composition,
e.g., in a cell culture. The greater proportion can be 2-fold,
5-fold, 10-fold, or more than 10-fold, with respect to the
proportion found in the prior composition.
I. General
[0709] The present invention concerns compounds of Formula I and
all sub-generic formulae, compounds of Formula II and all
sub-generic formulae, and compounds of Formula III and all
sub-generic formulae that are modulators of protein kinases, for
example without limitation, the compounds are modulators of at
least one of the kinases selected from the group consisting of Abl,
Akt1, Akt2, Akt3, ALK, Alk5, B-Raf, Brk, Btk, Cdk2, CDK4, CDK5,
CDK6, CHK1, c-Raf-1, Csk, EGFR, EphA1, EphA2, EphB2, EphB4, Erk2,
Fak, FGFR1, FGFR2, FGFR3, FGFR4, Flt1, Flt3, Flt4, Fms, Frk, Fyn,
Gsk3.alpha., Gsk3.beta., HCK, Her2/Erbb2, Her4/Erbb4, IGF1R, IKK
beta, Irak4, Itk, Jak1, Jak2, Jak3, Jnk1, Jnk2, Jnk3, Kdr, Kit,
LCK, MAP2K1, MAP2K2, MAP4K4, MAPKAPK2, Met, Mnk1, MLK1, p38,
PDGFRA, PDGFRB, PDPK1, Pim1, Pim2, Pim3, PKC alpha, PKC beta, PKC
theta, Plk1, Pyk2, Ret, ROCK1, ROCK2, Ron, Src, Stk6, Syk, TEC,
Tie2, TrkA, Yes, and Zap70, and the use of such compounds in the
treatment of diseases or conditions.
II. Kinase Targets and Indications of the Invention
[0710] Protein kinases play key roles in propagating biochemical
signals in diverse biological pathways. More than 500 kinases have
been described, and specific kinases have been implicated in a wide
range of diseases or conditions (i.e., indications), including for
example without limitation, cancer, cardiovascular disease,
inflammatory disease, neurological disease, and other diseases. As
such, kinases represent important control points for small molecule
therapeutic intervention. Description of specific target protein
kinases contemplated by the present invention follow:
[0711] Abl: Target Abl (i.e., Abelson Murine Leukemia Viral
Oncogene Homolog 1) is a 122.9 kDa tyrosine kinase encoded by
chromosome 9q34.1 (symbol: ABL1.) The mature protein comprises SH3
(i.e., Src homology region 3) and SH.sub.2(i.e., Src homology
region 2) domains and the TK (i.e., tyrosine kinase) domain.
[0712] OMIM indicates Abl is expressed ubiquitously and can be
localized to the nucleus where it binds DNA. Accordingly, Abl has
been implicated in processes of cell differentiation, cell
division, cell adhesion, and stress response. Alterations of the
gene ABL1 by a t(9;22)(q34;q11) chromosomal rearrangement or viral
transduction lead to malignant transformation, as in chronic
myeloid leukemia (CML). The kinase activity of Abl is negatively
regulated by the constituent SH3 domain, and deletion of the SH3
domain turns ABL1 into an oncogene. The t(9;22) translocation
occurs in greater than 90% of chronic myelogeneous leukemia, 25 to
30% of adult and 2 to 10% of childhood acute lymphoblastic leukemia
(ALL), and rare cases of acute myelogenous leukemia. The
translocation results in the head-to-tail fusion of the BCR and ABL
genes (Chissoe et al., Genomics 1995, 27: 67-82). The DNA-binding
activity of Abl is regulated by CDC2-mediated phosphorylation
suggesting a cell cycle function for ABL. Welch & Wang (Cell
1993, 75: 779-790) showed that the tyrosine kinase activity of
nuclear Abl is regulated in the cell cycle through a specific
interaction with the retinoblastoma protein RB1. A domain in the
C-terminus of RB1 binds to the ATP-binding lobe of Abl, resulting
in kinase inhibition. The RB-ABL interaction is not affected by the
viral oncoproteins that bind to RB. Hyperphosphorylation of RB
correlates with release of Abl and activation of Abl in S phase
cells. In the nucleus Abl can enhance transcription, and this
activity is inhibited by RB. Thus, nuclear Abl is an S
phase-activated tyrosine kinase that might participate directly in
the regulation of transcription. (Online Mendelian Inheritance in
Man, OMIM (TM). Johns Hopkins University, Baltimore, Md., MIM
Number: 189980: Dec. 20, 2005: World Wide Web URL:
http://www3.ncbi.nlm.nih.gov/omim/). Abl inhibitors may be useful
in treating leukemia, including chronic myelogeneous leukemia,
acute lymphoblastic leukemia, and acute myelogenous leukemia,
[0713] Akt1: Target kinase Akt1 (i.e., v-akt murine thymoma viral
oncogene homolog 1) is a 55.7 kDa STK encoded by chromosome
14q32.32 (symbol: AKT1). Akt1 is also known as protein kinase
B-alpha, or PKB-alpha. OMIM indicates phosphoinositide 3-kinases
(i.e., PI3Ks) generate specific inositol lipids implicated in the
regulation of cell growth, proliferation, survival,
differentiation, and cytoskeletal changes. One of the best
characterized targets of PI3K lipid products is the protein kinase
AKT, or protein kinase B (PKB). In quiescent cells, PKB resides in
the cytosol in a low-activity conformation. Upon cellular
stimulation, PKB is activated through recruitment to cellular
membranes by PI3K lipid products and by phosphorylation by 3-prime
phosphoinositide-dependent kinase-1. Most proliferating cells are
programmed to undergo apoptosis unless specific survival signals
are provided. PDGF promotes cellular proliferation, inhibits
apoptosis, and activates the RAS/PIK3/AKT1/IKBKA/NFKB1 pathway
(Romashkova and Makarov, Nature 1999, 401: 86-90). In this pathway,
NFKB1 does not induce c-myc and apoptosis, but instead induces
putative antiapoptotic genes. In response to PDGF, Akt1 transiently
associates with IKBK and induces IKBK activation (OMIM MIM Number:
164730: Oct. 26, 2005) Aberrant Akt1 activity is correlated with)
including gastric and prostate tumors, colorectal, ovarian,
pancreatic, and breast cancer, glioblastoma and leukemia. Sun et
al., Am J Pathol 2001, 159(2):431-7, report that significantly
increased AKT1 kinase activity was detected in primary carcinomas
of prostate (16 of 30), breast (19 of 50), and ovary (11 of 28).
Tanno et al., Cancer Res 2001, 61 (2):589-93, provide evidence for
a link between AKT signaling and the regulation of IGF-IR
expression and demonstrate that active AKT promotes the
invasiveness of pancreatic cancer cells through the up-regulation
of IGF-IR expression. Neri et al., Mol Cancer Res 2003,
1(3):234-46, suggest that an up-regulation of the PI3K/AKT1 pathway
might be one of the survival mechanisms responsible for the onset
of resistance to chemotherapeutic and differentiating therapy in
patients with acute leukemia. Akt1 activity is also important in
schizophrenia and bipolar disorders, Emamian et al., Nat Genet
2004, 36(2):115-6, present convergent evidence for a decrease in
AKT1 protein levels and levels of phosphorylation of GSK3beta at
Ser9 in the peripheral lymphocytes and brains of individuals with
schizophrenia; a significant association between schizophrenia and
an AKT1 haplotype associated with lower AKT1 protein levels; and a
greater sensitivity to the sensorimotor gating-disruptive effect of
amphetamine, conferred by AKT1 deficiency. Akt1 inhibitors may be
useful in treating cancer, including gastric, prostate, colorectal,
ovarian, pancreatic and breast cancer, glioblastoma and leukemia,
and also for use in combination with other chemotherapeutic
drugs.
[0714] Akt2: Target kinase Akt2 (i.e., v-akt murine thymoma viral
oncogene homolog 2) is a 55.8 kDa STK encoded by chromosome
19q13.1-13.2 (symbol: AKT2). Akt2 is also known as protein kinase B
beta, PKB-beta. OMIM indicates that the Akt2 isoform of Akt (see
e.g., Akt1 and Akt3) is enriched in insulin-responsive tissues and
has been implicated in the metabolic actions of insulin. Glucose
homeostasis depends on insulin responsiveness in target tissues,
most importantly, muscle and liver. The critical initial steps in
insulin action include phosphorylation of scaffolding proteins and
activation of phosphatidylinositol 3-kinase. These early events
lead to activation of the serine-threonine protein kinase Akt, also
known as protein kinase B. Cho et al., Science 2001, 292:1728-1731,
showed that mice deficient in Akt2 are impaired in the ability of
insulin to lower blood glucose because of defects in the action of
the hormone on liver and skeletal muscle. Ablation of Akt2 in mice
results in a mild but statistically significant fasting
hyperglycemia due to peripheral insulin resistance and
nonsuppressible hepatic glucose production accompanied by
inadequate compensatory hyperinsulinemia (OMIM MIM Number: 164731:
Oct. 26, 2005). Arboleda et al., Cancer Res 2003, 63(1):196-206
showed that AKT2 transfected breast and ovarian cancer cells
demonstrated increased adhesion and invasion through collagen IV
because of up-regulation of beta1 integrins and that AKT2 cells
were more metastatic than control cells in vivo. Yamamoto et al.,
Clin Cancer Res 2004, 10(8):2846-50 studied the prognostic
significance of Akt2 and activated Akt expression in pancreatic
ductal adenocarcinoma (PDAC), concluding that p-Akt expression is a
significant prognostic indicator for PDAC and inhibition of Akt is
a possible molecular approach for treatment of PDAC. Yuan et al.,
Oncogene 2000, 19(19):2324-30, demonstrate that activation of AKT2
is a common occurrence in human ovarian cancer and that the PI
3-kinase/Akt pathway may be an important target for ovarian cancer
intervention. Yuan et al., J Biol Chem 2003, 278(26):23432-40,
demonstrate that constitutively active AKT2 renders
cisplatin-sensitive A2780S ovarian cancer cells resistant to
cisplatin, whereas phosphatidylinositol 3-kinase inhibitor or
dominant negative AKT2 sensitizes A2780S and cisplatin-resistant
A2780CP cells to cisplatin-induced apoptosis through regulation of
the ASK1/JNK/p38 pathway. Akt2 inhibitors may be useful in treating
cancer, including ovarian and pancreatic cancers, pancreatic ductal
adenocarcinoma, and metastases of breast and ovarian cancer, and
also for use in combination with other chemotherapeutic drugs,
where such use sensitizes the tumor cells to the effects of the
other chemotherapeutics.
[0715] Akt3: Target kinase Akt3 (i.e., v-akt murine thymoma viral
oncogene homolog 3) is a 55.8 kDa STK encoded by chromosome
Iq43-q44 (symbol:AKT3); Akt3 is also known as PKB gamma. Akt3 was
identified as a protein kinase with high homology with the protein
kinases A and C, hence, the name PKB. Akt3 comprises a PH domain
that preferentially binds PtdIns(3,4,5)P.sub.3 and
PtdIns(3,4)P.sub.2 over other phosphatidyl inositols (PIs). In
quiescent cells, PKB resides in the cytosol in a low-activity
conformation.
[0716] Upon cellular stimulation, PKB is activated through
recruitment to cellular membranes by PI3K lipid products and
phosphorylation by 3'-phosphoinositide-dependent kinase-1 (PDK1).
Active PKB then appears to detach from the plasma membrane and to
translocate through the cytosol to the nucleus.
[0717] Stahl et al. have found that selective activation of the
Akt3 protein promotes cell survival and tumor development in 43 to
60% of nonfamilial melanomas. The predominant Akt isoform active in
melanomas was identified by showing that small interfering RNA
(siRNA) against only Akt3, and not Akt1 or Akt2, lowered the amount
of phosphorylated (active) Akt in melanoma cells. The amount of
active Akt3 increased progressively during melanoma tumor
progression with highest levels present in advanced-stage
metastatic melanomas. Mechanisms of Akt3 deregulation occurred
through a combination of overexpression of Akt3 accompanying copy
number increases of the gene and decreased PTEN protein function
occurring through loss or haploinsufficiency of the PTEN gene.
Targeted reduction of Akt3 activity with siRNA or by expressing
active PTEN protein stimulated apoptotic signaling, which reduced
cell survival by increasing apoptosis rates thereby inhibiting
melanoma tumor development. Therefore, Akt3 is a selective target
in melanoma cells which provides therapeutic opportunities for
subjects in advanced stages of the disease (Stahl et al., Cancer
Res. 2004, 64:7002-10). Nakatani et al., J Biol Chem 1999,
274(31):21528-32 showed that in estrogen receptor-deficient breast
cancer cells and androgen-insensitive prostate cells, the amount of
Akt3 enzymatic activity was approximately 20-60-fold higher than in
cells that were estrogen- or androgen-responsive. These and other
results indicate that Akt3 may contribute to the more aggressive
clinical phenotype of the estrogen receptor-negative breast cancers
and androgen-insensitive prostate carcinomas and inhibitors may
provide therapeutic benefits in treating these cancers. Akt3
inhibitors may be useful in treating cancer, including estrogen
receptor-negative breast cancers, androgen-insensitive prostate
carcinomas, and melanomas.
[0718] ALK: Target kinase ALK (i.e., anaplastic lymphoma kinase) is
a 176.4 kDa receptor tyrosine kinase encoded by chromosome 2p23
(symbol: ALK). ALK appears to play a role in the development of the
central nervous system. Perkins, et al., show that systemic ALCL is
highly associated with anaplastic lymphoma kinase (ALK) gene
translocations with over-expression of ALK protein. Anaplastic
large cell lymphoma (ALCL) comprises 10-15% of childhood
non-Hodgkin lymphomas (NHL) (Perkins et al., Br J Haematol 2005,
131(5):624-7). Marzec et al., states aberrant expression of the ALK
tyrosine kinase as a chimeric protein with nucleophosmin (NPM) and
other partners plays a key role in malignant cell transformation of
T-lymphocytes and other cells. Further, studies with inhibitors of
NPM/ALK enzyme activity suggest ALK as an attractive therapeutic
target in T-cell lymphomas and other malignancies that express the
kinase in an active form (Marzec et al., Lab Invest 2005,
85(12):1544-54). ALK inhibitors may be useful in treating cancer,
including anaplastic large cell lymphoma and other T cell
lymphomas.
[0719] Alk5: Target kinase Alk5 (i.e., Activin receptor-like kinase
5) is a 56.0 kDa STK encoded by chromosome 9q22 (symbol: TGFBR1).
Alk5, the gene for which was isolated by Franzen et al (Cell 1993,
75: 681-692), is also known as transforming growth factor-beta
receptor, type I, from which term the gene symbol derives. Among
other activities, Alk5 mediates the induction of multiple genes
involved in cell-matrix interactions. Alk5 inhibitors may be useful
in treating pancreatic and biliary cancers and cutaneous T-cell
lymphoma.
[0720] B-Raf: Target kinase B-Raf (i.e., v-raf murine sarcoma viral
oncogene homolog B1) is a 84.4 kDa serine/threonine kinase encoded
by chromosome 7q34 (symbol: BRAF). The mature protein comprises RBD
(i.e., Ras binding domain), C1 (i.e., protein kinase C conserved
region 1) and STK (i.e., serine/threonine kinase) domains.
[0721] Target kinase B-Raf is involved in the transduction of
mitogenic signals from the cell membrane to the nucleus and may
play a role in the postsynaptic responses of hippocampal neurons.
As such, genes of the RAF family encode kinases that are regulated
by Ras and mediate cellular responses to growth signals. Indeed,
B-Raf kinase is a key component of the
RAS.fwdarw.Raf.fwdarw.MEK.fwdarw.ERK/MAP kinase signaling pathway,
which plays a fundamental role in the regulation of cell growth,
division and proliferation, and, when constitutively activated,
causes tumorigenesis. Among several isoforms of Raf kinase, the
B-type, or B-Raf, is the strongest activator of the downstream MAP
kinase signaling.
[0722] The BRAF gene is frequently mutated in a variety of human
tumors, especially in malignant melanoma and colon carcinoma. The
most common reported mutation was a missense thymine (T) to adenine
(A) transversion at nucleotide 1796 (T1796A; amino acid change in
the B-Raf protein is Val<600> to Glu<600>) observed in
80% of malignant melanoma tumors. Functional analysis reveals that
this transversion is the only detected mutation that causes
constitutive activation of B-Raf kinase activity, independent of
RAS activation, by converting B-Raf into a dominant transforming
protein.
[0723] Niihori et al., report that in 43 individuals with
cardio-facio-cutaneous (CFC) syndrome, they identified two
heterozygous KRAS mutations in three individuals and eight BRAF
mutations in 16 individuals, suggesting that dysregulation of the
RAS-RAF-ERK pathway is a common molecular basis for the three
related disorders (Niihori et al., Nat Genet. 2006,
38(3):294-6).
[0724] B-Raf inhibitors may be useful in treating neurologic
diseases such as ischemic stroke, multi-infarct dementia, head
injury, spinal cord injury, Alzheimer's disease (AD), Parkinson's
disease; neoplastic diseases including, but not limited to,
melanoma, glioma, sarcoma, carcinoma (e.g. lung, breast,
pancreatic, renal), lymphoma (e.g. histiocytic lymphoma) and cancer
of the thyroid, lung (e.g. small cell lung cancer), liver, breast,
ovary and colon, neurofibromatosis, myelodysplastic syndrome,
leukemia, tumor angiogenesis; pain of neuropathic or inflammatory
origin, including acute pain, chronic pain, and migraine;
cardiovascular diseases including heart failure, cardiac
hypertrophy, thrombosis (e.g. thrombotic microangiopathy
syndromes), atherosclerosis, reperfusion injury; inflammation
including, but not limited to, psoriasis, polycystic kidney disease
(PKD), arthritis and autoimmune diseases and conditions,
osteoarthritis, endometriosis, scarring, vascular restenosis,
fibrotic disorders, rheumatoid arthritis, inflammatory bowel
disease (IBD); immunodeficiency diseases, organ transplant
rejection, graft versus host disease; renal or prostatic diseases
including diabetic nephropathy, nephrosclerosis,
glomerulonephritis, prostate hyperplasia; metabolic disorders,
obesity; infection, including, but not limited to Helicobacter
pylori and Influenza virus, fever, sepsis; pulmonary diseases
including chronic obstructive pulmonary disease (COPD) and acute
respiratory distress syndrome (ARDS); genetic developmental
diseases such as Noonan's syndrome, Costello syndrome,
(faciocutaneoskeletal syndrome), leopard syndrome,
cardio-faciocutaneous syndrome (CFC), and neural crest syndrome
abnormalities causing cardiovascular, skeletal, intestinal, skin,
hair and endocrine diseases.
[0725] c-Raf-1: Target kinase c-Raf-1 (i.e., v-raf murine sarcoma
viral oncogene homolog 1) is a 73.0 kDa STK encoded by chromosome
3p25 (symbol: RAF1). c-Raf-1 can be targeted to to the mitochondria
by BCL2 (i.e., oncogene B-cell leukemia 2) which is a regulator of
apoptotic cell death. Active c-Raf-1 improves BCL2-mediated
resistance to apoptosis, and c-Raf-1 phosphorylates BAD (i.e.,
BCL2-binding protein). c-Raf-1 is implicated in carcinomas,
including colorectal, ovarian, lung and renal cell carcinoma.
C-Raf-1 is also implicated as an important mediator of tumor
angiogenesis (Hood, J. D. et al., 2002, Science 296, 2404). C-Raf-1
inhibitors may also be useful for the treatment of acute myeloid
leukemia and myelodysplastic syndromes (Crump, Curr Pharm Des 2002,
8(25):2243-8). Raf-1 activators may be useful as treatment for
neuroendocrine tumors, such as medullary thyroid cancer, carcinoid,
small cell lung cancer and pheochromocytoma (Kunnimalaiyaan et al.,
Anticancer Drugs 2006, 17(2):139-42). C-Raf-1 inhibitors may be
useful in treating colorectal, ovarian, lung and renal cell
carcinoma, acute myeloid leukemia, myelodysplastic syndromes, tumor
angiogenesis, and neuroendocrine tumors such as medullary thyroid
cancer, carcinoid, small cell lung cancer and pheochromocytoma.
[0726] Brk: Target kinase Brk (i.e. breast tumor kinase, also known
as PTK6) is a 51.8 kDa non-receptor tyrosine kinase encoded by
human chromosome 20q13.3 (symbol: BRK). The kinase has an SH3
domain, an SH.sub.2domain, and a catalytic domain. In normal
tissues, the expression of Brk (breast tumor kinase) is restricted
to differentiating epithelial cells of the skin and
gastrointestinal tract. According to Harvey and Crompton, Brk is
expressed in over 60% of breast carcinoma tissue samples and breast
tumour cell lines, but not normal mammary tissue or benign lesions.
They used RNA interference to efficiently and specifically
downregulate Brk protein levels in breast carcinoma cells, and
determined that this results in a significant suppression of their
proliferation (Harvey and Crompton, Oncogene, 2003, 22(32):
5006-5010). Lin et al. identified protein-tyrosine kinases that may
be involved in the development and progression of head and neck
squamous cell carcinoma (HNSCC), and their findings suggest that
the signaling pathways, mediated through EphA1, Brk, and Ron may be
involved in the development and progression of HNSCC (Lin et al.,
Arch Otolaryngol Head Neck Surg. 2004, 130(3):311-6). Llor et al.
examined BRK expression in the normal gastrointestinal tract, colon
tumor cell lines, and primary colon tumor samples and showed BRK is
expressed in normal epithelial cells of the gastrointestinal tract
that are undergoing terminal differentiation. BRK expression also
increased during differentiation of the Caco-2 colon adenocarcinoma
cell line. Modest increases in BRK expression were detected in
primary colon tumors by RNase protection, in situ hybridization,
and immunohistochemical assays (Llor et al., Clin Cancer Res. 1999,
5(7):1767-77). Brk inhibitors may be useful in treating cancer,
such as breast and colon cancer, and head and neck squamous cell
carcinoma.
[0727] Btk: Target kinase Btk (i.e., Bruton's tyrosine kinase) is a
76.3 kDa tyrosine kinase encoded by chromosome Xq21.33-q22 (symbol:
BTK). The mature kinase comprises a PH (i.e., Pleckstrin homology)
domain, a BTK (i.e., Bruton's tyrosine kinase motif) motif, two SH3
domains, and a TK domain. Mao et al. determined the X-ray crystal
structure of the Btk kinase domain in its unphosphorylated state to
2.1-angstrom resolution (Mao et al., J. Biol. Chem., 2001,
276:41435).
[0728] As a member of the BTK/Tcc family of protein tyrosine
kinases (i.e., PTKs), cytoplasmic Btk is involved in signal
transduction pathways regulating growth and differentiation of
B-lineage lymphoid cells (Rawlings, D. J., and Witte, O. N., 1994.
Immunol. Rev. 138:105-119; Kurosaki, T., 1997, Curr Opin. Immunol
9:309-318; Uckun, F. M., 1998, Biochemical Pharmacology
56:683-691). As such, Btk participates in signal transduction
pathways initiated by the binding of a variety of extracellular
ligands to their cell surface receptors. For example, following
ligation of B cell antigen receptors (BCR), Btk activation by the
concerted actions of the PTKs Lyn and Syk (Kurosaki, T. (1997) Curr
Opin. Immunol. 9, 309-318) is required for induction of
phospholipase C-[gamma]2 mediated calcium mobilization (Kurosaki,
T., 1997, Curr Opin. Immunol. 9:309-318). Furthermore, Btk
regulates B cell antigen receptor-mediated JNK1 response through
Rac1 and phospholipase C-gamma2 activation.
[0729] Mutations in the human BTK gene are the cause of X-linked
agammaglobulinemia (XLA), a male immune deficiency disorder
characterized by a lack of mature, immunoglobulin producing,
peripheral B cells (Tsukada, S., et al. (1993) Cell 72, 279-290;
and Vetrie, D., et al. (1993) Nature 361, 226-233) and associated
with a failure of Ig heavy chain rearrangement. Patients are
unusually prone to bacterial infection but not to viral infection.
A clinical picture resembling rheumatoid arthritis develops in
many. Before antibiotics, death occurred in the first decade. In
the more usual X-linked form of the disease, plasma cells are
lacking. A rarer form of agammaglobulinemia (Hitzig, W. H et al.
1961, Med. Wschr., 91:1625), which is inherited as an autosomal
recessive, shows marked depression of the circulating lymphocytes,
and lymphocytes are absent from the lymphoid tissue. Mensink et al.
(Clin. Genet., 1987, 31:91) mapped XLA to Xq21.3-q22. Schwaber
(Clin. Invest., 1992, 89:2053) presented direct evidence that of a
failure of V(D)J recombination which causes arrest in the
transition from pre-B cell to B lymphocyte. XLA patients have been
classified in 2 general groups: those presenting at an early age
with particularly severe infections and those with less severe
disease in which production of immunoglobulin is sustained at
low-to-normal levels well into the first decade of life. In the
latter cases, an oncogenetic change may occur in which the
defective tyrosine kinase no longer can sustain the B-cell
population, and a progressive reduction in immunoglobulin
production occurs (Ohta, Y et al., Proc. Nat. Acad. Sci., 1994,
91:9062)
[0730] Btk is an inhibitor of the Fas/APO-1 death inducing
signaling complex (DISC) in B-lineage lymphoid cells (Vassilev, A.,
et al., 1998, J. Biol. Chem., 274:1646-1656). Additionally, Btk
prevents ceramide- and vincristine-induced apoptosis. The fate of
leukemia/lymphoma cells may reside in the balance between the
opposing proapoptotic effects of caspases activated by DISC and an
upstream anti-apoptotic regulatory mechanism involving Btk and/or
its substrates (Vassilev, A., et al., 1998, J. Biol. Chem.
274:1646-1656).
[0731] Accordingly, inhibitors of Btk are likely to enhance the
drug sensitivity of B-lineage (e.g. leukemia/lymphoma such as acute
lymphocytic leukemia) cells. Thus, pharmacological agents with
Btk-modulatory activity can be used as chemosensitizing agents for
treating Btk-expressing malignancies or diseases caused by
proliferation and antibody production of BTK-expressing B-cells,
and as B-cell reconstituting agents in humoral immunodeficiencies
with decreased numbers or absence of B-cells. Furthermore, Btk
modulating agents are useful as immunosuppressive agents for
prevention of hyperacute rejection of organs in transplantation,
which is directed by B-cells, autoimmune diseases, and conversion
of immunity to drugs (e.g. antibodies or biologicals) or blood
products (e.g. coagulation factors such as Factor VIII) in patients
who develop antibodies to such agents.
[0732] Significant additional research has defined the role of Btk
in the cell. For example, Cheng et al. (Proc. Nat. Acad. Sci.,
1994, 91:8152) showed that Btk interacts with the SH3 domains of
Fyn, Lyn, and Hck, all of which are activated upon stimulation of
B- and T-cell receptors. These findings extended the range of
interactions mediated by SH3 domains and provide indication of a
link between Btk and previously established signaling pathways in B
lymphocytes. Further, linkage studies involving 1,114 progeny
backcross revealed colocalization of X-linked immunodeficiency
(xid) mutation in mice with the BTK gene for (Thomas, J. D. et al.,
Science 1993, 261:355). And further, Uckun et al. (Uckun, F. M et
al. 1996, Science 273: 1096) reported that DT-40 lymphoma B cells
rendered Btk deficient through targeted disruption of the BTK gene
did not undergo radiation-induced apoptosis. Finally, Btk plays a
key role in endotoxin-induced TNF.alpha. release from monocytes
(Horwood, N. J. et al., J. Exp. Med. 2003, 197:1603).
[0733] Btk inhibitors may be useful in treating multiple sclerosis,
systemic lupus erythematosis, rheumatoid arthritis, and Graves'
disease, in addition to the diseases discussed above.
[0734] Cyclin dependent kinases: The cyclin dependent kinases (Cdk)
play a major role in the signaling of cell cycles. The Cdk binds
cyclin protein to form complex involved in the various stages of
the cell cycle. Aberrant cell cycle progression occurs in cancers,
often involving the activity of Cdk, such that inhibitors of Cdk
are potential anti-cancer therapeutics.
[0735] Cdk2: Target kinase Cdk2 (i.e., Cyclin dependent kinase 2)
is a 33.9 kDa serine/threonine kinase (STK) encoded by chromosome
12q13 (symbol: CDK2). Cdk2 is also known as p33 protein kinase, and
cell division protein kinase 2. De Bondt et al. reported the X-ray
crystallographic structure of Cdk2 (De Bondt et al., Nature 1993,
363: 595-602). Cdk2 is involved in control of the human cell cycle,
Cdk2 activation requires association with cyclins and leads to cell
proliferation. Further, inhibition of cellular proliferation occurs
upon association of inhibitors (e.g., cyclin-dependent kinase
inhibitor 1B) with the cyclin-Cdk2 complex. Cyclin E/Cdk2 complexes
play a role in carcinogenesis, for example, Cdk2 amplication is
associated with concurrent Cyclin E amplification in several
cancers, including colorectal and ovarian cancer (Kitahara et al.,
Int. J. Cancer, 1995, 53: 1986-1989; Marone et al., Int. J. Cancer,
1998, 75: 31-39). Teixeira et al. show that retinoic acid inhibits
cell cycle progression of MCF-7 human breast cancer cells by
inhibiting cdk2 mRNA and protein production and by decreasing cdk2
activity (Teixeira et al., Mol Endocrinol 1997, 11(9):1191-202).
Cipriano and Chen studied the TGF-beta1 effect on normal human
prostate and carcinoma cells, and showed that normal cells were
sensitive to growth inhibition, whereas tumor cells were not or
only minimally inhibited regardless of the concentration of
TGF-beta1, and correlated these results to Cdk2 activity. Their
results indicate that a lack of inhibition of the Cdk2 activity
correlates with insensitivity to TGF-beta1 in prostate tumor cells
(Cipriano and Chen, Oncogene 1998, 17(12):1549-56). Cdk2 inhibitors
may be useful in treating cancer, including prostate, breast,
colorectal and ovarian cancer.
[0736] Cdk4: Target kinase Cdk4 (i.e., Cyclin dependent kinase 4)
is a 33 kDa STK encoded by chromosome 12q14 (symbol: CDK4). Lam et
al. reported expression of CDK4 and CDK6 was elevated relative to
matched normal brain tissue in eight of 18 glioblastoma multiforme
(GBM) tumours (44%). Their data attests to the functional
importance of both CDK4 and CDK6 in astrocytic tumourigenesis,
particularly during the later stages of tumour progression (Lam et
al, Br J Neurosurg 2000, 14(1):28-32). Backlund et al. found that
loss of both wild-type copies of any of the three tumour suppressor
genes CDKN2A, CDKN2B and RB1 or gene amplification of CDK4,
disrupting the Rb1 pathway, were associated with shorter survival
in anaplastic astrocytoma patients (Backlund et al., Br J Cancer
2005, 93(1):124-30). Yu et al. report that the ability of cyclin D1
to activate cyclin-dependent kinase CDK4 underlies the critical
role for cyclin D1 in breast cancer formation. They also found that
the continued presence of CDK4-associated kinase activity is
required to maintain breast tumorigenesis (Yu et al., Cancer Cell
2006, 9(1):23-32). Cdk4 inhibitors may be useful in treating
cancer, including glioblastoma (e.g. glioblastoma multiforme),
anaplastic astrocytoma, and breast cancer.
[0737] Cdk5: Target kinase Cdk5 (i.e., Cyclin dependent kinase 5)
is a 33.3 kDa STK encoded by chromosome 7q36 (symbol: GDK5). Cruz
et al. state that proteolytic cleavage of p35 generates p25,
leading to aberrant Cdk5 activation. The accumulation of p25 is
implicated in several neurodegenerative diseases. Their findings
provide compelling evidence that in vivo deregulation of Cdk5 by
p25 plays a causative role in neurodegeneration and the development
of neurofibrillary pathology (Cruz et al., Neuron 2003,
40(3):471-83). Takahashi et al. investigated the Cdk5 distribution
pattern in diffuse Lewy body disease brains using
immunohistochemistry. Their data suggest that Cdk5 may be
associated with Lewy body formation (Takahashi et al., Brain Res
2000, 862(1-2):253-6). Cdk5 inhibitors may be useful in treating
neurodegenerative disorders, including Alzheimer's disease,
amyotrophic lateral sclerosis and Lewy body disease.
[0738] Cdk6: Target kinase Cdk6 (i.e., Cyclin dependent kinase 6)
is a 36.9 kDa STK encoded by chromosome 7q21-q22 (symbol: CDK6).
Lam et al. reported expression of CDK4 and CDK6 was elevated
relative to matched normal brain tissue in eight of 18 glioblastoma
multiforme (GBM) tumours (44%). Their data attests to the
functional importance of both CDK4 and CDK6 in astrocytic
tumourigenesis, particularly during the later stages of tumour
progression (Lam et al., Br J Neurosurg 2000, 14(1):28-32).
Costello et al., applied restriction landmark genomic scanning to
matched samples of glioma and normal brain DNA and found
tumor-specific amplification of the gene encoding cyclin-dependent
kinase 6 (CDK6). They also corroborated this finding by identifying
both amplification-associated and amplification-independent
increases in CDK6 protein levels in gliomas relative to matched
normal brain samples (Costello et al., Cancer Res 1997,
57(7):1250-4). Corcoran et al. found in two samples from patients
with splenic lymphoma with villous lymphocytes (SLVL), the CDK6
protein was markedly over expressed. They suggest that
dysregulation of CDK6 gene expression contributes to the
pathogenesis of SLVL and splenic marginal zone lymphoma (SMZL)
(Oncogene 1999, 18(46):6271-7). Chilosi et al. provide evidence
that CDK6 is abnormally expressed in T-cell lymphoblastic
lymphoma/leukemia (T-LBL/ALL) and may be involved in the
pathogenesis of this malignancy (Chilosi et al., Am J Pathol 1998,
152(1):209-17). Cram et al. show that Indole-3-carbinol (I3C) can
induce a G(1) cell cycle arrest of human MCF-7 breast cancer cells
that is accompanied by the selective inhibition of cyclin-dependent
kinase 6 (CDK6) expression (Cram et al., J Biol Chem 2001,
276(25):22332-40). Cdk6 inhibitors may be useful in treating
cancer, including glioblastoma multiforme, non-Hodgkin's lymphoma,
splenic marginal zone lymphoma, T-cell lymphoblastic lymphoma
(T-LBL) and T-cell acute lymphoblastic leukemia (T-ALL).
[0739] CHK1: Target kinase Chk1 (i.e., Checkpoint kinase) is a 54.4
kDa STK encoded by chromosome 11q24 (symbol: CHEK1, CHK1). CHK1 is
involved in DNA damage checkpoint. Carassa et al., to understand
the role of Chk1 and Chk2 in the cellular response to different
anticancer agents, knocked down the expression of each protein or
simultaneously of both proteins by using the small interfering RNA
technique in a HCT-116 colon carcinoma cell line and in its
isogenic systems in which p53 and p21 had been inactivated by
targeted homologous recombination. They show that inhibition of
Chk1 but not of Chk2 in p21(-/-) and p53(-/-) cells caused a
greater, abrogation of G(2) block induced by ionizing radiation and
cis-diamine-dichloroplatinum treatments and a greater sensitization
to the same treatments than in the parental cell line with p53 and
p21 wild type proteins. Their data further emphasise the role of
Chk1 as a molecular target to inhibit in tumors with a defect in
the G(1) checkpoint with the aim of increasing the selectivity and
specificity of anticancer drug treatments (Carrassa et al., Cell
Cycle 2004, 3(9):1177-81). Similarly, studies by Hirose et al.
focused on the mechanism by which Temozolomide (TMZ) induces G(2)-M
arrest and on whether inhibition of such G(2)-M arrest might
sensitize glioma cells to TMZ-induced toxicity. U87MG glioma cells
treated with TMZ underwent G(2)-M arrest associated with Chk1
activation and phosphorylation of both cdc25C and cdc2. These
TMZ-induced effects were inhibited by the Chk1 kinase inhibitor
UCN-01. Although not in itself toxic, UCN-01 increased the
cytotoxicity of TMZ 5-fold, primarily by inhibiting cellular
senescence and increasing the percentage of cells bypassing G(2)-M
arrest and undergoing mitotic catastrophe. In addition to enhancing
TMZ-induced cytotoxicity in p53-proficient cells, UCN-01 also
blocked TMZ-induced Chk1 activation and transient G(2)-M arrest in
p53-deficient U87MG-E6 cells and similarly enhanced TMZ-induced
mitotic catastrophe and cell death. Taken together, their results
indicate that Chk1 links TMZ-induced DNA mismatch repair to G(2)-M
arrest. Furthermore, inhibition of the cytoprotective G(2) arrest
pathway sensitizes cells to TMZ-induced cytotoxicity and may
represent a novel, mechanism-based means of increasing TMZ efficacy
in both p53 wild-type and p53 mutant glioma cells. (Hirose et al.,
Cancer Res 2001, 61(15):5843-9). As such, CHK1 inhibitors may be
used in combination therapy to improve the therapeutic efficacy of
chemotherapeutic drugs. CHK1 inhibitors may be useful in
combination with chemotherapeutic drugs in treating cancer.
[0740] Csk: Target kinase Csk (i.e., c Src kinase) is a 50.7 kDa
tyrosine kinase encoded by chromosome 15q23-q25 (symbol: CSK). Csk,
cloned by Partanen et al. (Oncogene 1991, 6: 2013-2018), is a
cytoplasmic tyrosine kinase that downregulates the tyrosine kinase
activity of the Src oncoprotein through tyrosine phosphorylation of
the Src carboxy terminus. Activation of Csk may be therapeutic for
cancers in which Src is activated, such as in colon and pancreatic
carcinomas (Lutz et al., Biochem Biophys Res Commun 1998,
243(2):503-8; Cam et al., Cancer 2001, 92(1):61-70). Zheng and She
state that the lymphoid-specific phosphatase (LYP) encoded by
PTPN22 is involved in preventing spontaneous T-cell activation by
dephosphorylating and inactivating T-cell receptor-associated Csk
kinase. They genotyped 396 type 1 diabetic patients and 1,178
control subjects of Caucasian descent from north central Florida
and report a strong association between type 1 diabetes and a
polymorphism (R.sup.620W) in the PTPN22 gene. In vitro experiments
have shown that the mutant 620W LYP protein (1858T) does not bind
Csk. Together with previous reports of the association between
PTPN22 and type 1 diabetes, as well as rheumatoid arthritis and
systemic lupus erythematosus, these results provide compelling
evidence that LYP is a critical player in multiple autoimmune
disorders (Zheng and She, Diabetes 2005, 54(3):906-8). Csk
modulators, including inhibitors, may be useful in treating
autoimmune diseases, including type 1 diabetes, rheumatoid
arthritis and systemic lupus erythematosus.
[0741] EGFR: Target kinase EGFR (i.e., Epidermal Growth Factor
Receptor) is a 134.3 kDa transmembrane tyrosine kinase coded by
chromosome 7p12.3-p12.1 (symbol: EGFR). OMIM indicates that EGF
enhances phosphorylation of several endogenous membrane proteins,
including EGFR. EGFR has 2 components of different molecular
weight; both contain phosphotyrosine and phosphothreonine but only
the higher molecular weight form contains phosphoserine (Carlin and
Knowles, Proc. Nat. Acad. Sci. 1982, 79: 5026-5030.). Carlin et al.
(Cytogenet. Cell Genet. 1982, 32: 256) showed that the specific
cell surface antigen previously called SA7 (Aden and Knowles,
Immunogenetics 1976, 3: 209-211) is identical to EGFR. EGFR
signaling involves small GTPases of the Rho family, and EGFR
trafficking involves small GTPases of the Rab family. Lanzetti et
al. (Nature 2000, 408: 374-377) reported that the EPS8 protein
connects these signaling pathways. EPS8 is a substrate of EGFR that
is held in a complex with SOS1 by the adaptor protein E3B1, thereby
mediating activation of RAC. Through its SH3 domain, EPS8 interacts
with RNTRE. Further, Lanzetti et al. (ibid) showed that RNTRE is a
RAB5 GTPase-activating protein whose activity is regulated by EGFR.
By entering in a complex with EPS8, RNTRE acts on RAB5 and inhibits
internalization of EGFR. Furthermore, RNTRE diverts EPS8 from its
RAC-activating function, resulting in the attenuation of RAC
signaling. Thus, depending on its state of association with E3B1 or
RNTRE, EPS8 participates in both EGFR signaling through RAC and
EGFR trafficking through RAB5 (OMIM MIM Number: 131550: Dec. 16,
2005).
[0742] EGFR is implicated in breast cancer, colorectal, and bladder
cancer, and modulation of EGFR activity is a therapeutic route to
amelioration of these pathologic states (Xu et al., Mol Cancer Ther
2005, 4(3):435-42). An important unmet need has emerged in non
small cell lung cancer patients who initially respond to treatment
with EGFR inhibitors but then develop resistance to the initial
drug (Koboyashi, S. et al. N Engl J Med. 2005, 352:786-92). EGFR is
also a possible target for treating glioblastoma multiforme
(Raizer, J Neurooncol 2005, 74(1):77-86), and squamous cell
carcinomas, for example in the esophagus (Hanawa et al., Int J
Cancer 2006, 118(5):1173-80), head and neck (Hambek et al.,
Anticancer Res 2004, 24(6):3881-6), and oral cavity and tongue
(Ekberg et al., Int J Oncol 2005, 26(5):1177-85). Unlu and Leake
studied the effect of epidermal growth factor (EGF) and a specific
inhibitor of EGFR, on the growth and invasiveness of the prostate
cancer cell lines PC3 and DU145. Their results indicate that EGF is
a potent stimulative agent for both growth and invasion in prostate
cancer cells, and that targeting the EGFR function inhibits not
only tumor growth but also invasiveness (Unlu and Leake, Int J Biol
Markers 2003, 18(2):139-46). EGFR inhibitors may be useful in
treating cancer, including breast, colon, bladder, prostate and non
small cell lung cancer, squamous cell carcinomas of the head and
neck, oral cavity, and esophagus, and glioblastoma multiforme.
[0743] EphA1: Target kinase EphA1 (i.e., Ephrin Receptor A1) is a
108.1 kDa tyrosine kinase encoded by chromosome 7q32-q34 (symbol:
EPHA1). OMIM indicates that the EPH and EPH-related receptors
comprise the largest subfamily of receptor protein-tyrosine
kinases. They have been implicated in mediating developmental
events, particularly in the nervous system. Receptors in the Eph
subfamily typically have a single kinase domain and an
extracellular region containing a Cys-rich domain and 2 fibronectin
type III repeats. The ligands for Eph receptors have been named
"ephrins" by the Eph Nomenclature Committee (Cell 1997, 90:
403-404). Based on their structures and sequence relationships,
ephrins are divided into the ephrin-A (EFNA) class, which are
anchored to the membrane by a glycosylphosphatidylinositol linkage,
and the ephrin-B (EFNB) class, which are transmembrane proteins.
The Eph family of receptors are divided into 2 groups based on the
similarity of their extracellular domain sequences and their
affinities for binding ephrin-A and ephrin-B ligands. The Eph
Nomenclature Committee (ibid.) proposed that Eph receptors
interacting preferentially with ephrin-A proteins be called EphA
and Eph receptors interacting preferentially with ephrin-B proteins
be called EphB. Maru et al. (1988) reported characterization of the
novel receptor tyrosine kinase gene, EPH. The splicing points of
kinase domain-encoding exons were completely distinct from those of
other protein tyrosine kinase genes, suggesting that this is the
earliest evolutionary split within this family. In Northern blot
analysis, EPH gene mRNA was detected in liver, lung, kidney, and
testes of rat; screening of 25 human cancers of various cell types
showed preferential expression in cells of epithelial origin.
Overexpression of EPH mRNA was found in a hepatoma and a lung
cancer without gene amplification. Southern blot analysis of DNAs
from human-mouse hybrid clones with an EPH probe showed that this
gene is present on human chromosome 7. Two other receptor tyrosine
kinase genes, MET and EGFR, are on the same chromosome. By in situ
hybridization, Yoshida et al. (1989) assigned the EPH locus to
7q32-q36. Although ephrins form a high-affinity multivalent complex
with their receptors present on axons, axons can be rapidly
repelled rather than being bound (OMIM MIM Number: 179610: Sep. 5,
2000). Lin et al. identified 5 PTKs that were overexpressed in head
and neck squamous cell carcinoma (HNSCC) using a reverse
transcriptase-PCR technique and confirmed the overexpression of 3
known PTKs in some of the 8 archival HNSCC specimens studied. Their
finding suggests that the signaling pathways mediated through
EphA1, Brk, and Ron may be involved in the development and
progression of HNSCC (Lin et al., Arch Otolaryngol Head Neck Surg
2004, 130(3):311-6). EphA1 inhibitors may be useful in treating
cancer, including liver and lung cancer, and head and neck squamous
cell carcinoma.
[0744] EphA2: Target kinase EphA2 (i.e., Ephrin Receptor A2) is a
108.3 kDa tyrosine kinase encoded by chromosome 1p36.1 (symbol:
EPHA2). EphA2, similar to other ephrin receptors, is found in
epithelial, lymphoid, and especially neuron tissue where EphA2 is
critically involved in short-range contact-mediated axonal
guidance. Further, EphA2 is highly expressed in metastatic melanoma
cells. Ephrin A1, a ligand for EphA2, was shown to be up regulated
during melanoma progression (Easty et al., Int J Cancer 1999,
84(5):494-501). Hattori et al. showed that ephrin-A2 forms a stable
complex with the metalloproteinase Kuzbanian, involving
interactions outside the cleavage region and the protease domain.
Eph receptor binding triggered ephrin-A2 cleavage in a localized
reaction specific to the cognate ligand. The cleavage-inhibiting
mutation in ephrin-A2 delayed axon withdrawal. Hattori et al.
(ibid) concluded that their studies reveal mechanisms for protease
recognition and control of cell surface proteins, and, for
ephrin-A2, they may provide a means for efficient axon detachment
and termination of signaling (Hattori et al., Science 2000, 289:
1360-1365). Ireton and Chen review EphA2 as a promising target for
cancer therapeutics, indicating EphA2 is overexpressed in breast,
prostate, lung, and colon cancers (Ireton and Chen, Curr Cancer
Drug Targets 2005, 5(3):149-57). Landen et al. state that EphA2 is
involved in many processes crucial to malignant progression, such
as migration, invasion, metastasis, proliferation, survival and
angiogenesis. Inducing EphA2 downregulation by any one of several
mechanisms (antibody-mediated inhibition of signalling,
antibody-mediated downregulation of total EphA2 expression and
siRNA-mediated inhibition of expression) has been shown to decrease
tumour growth, prolong survival and inhibit angiogenesis in
multiple preclinical models of ovarian, breast and pancreatic
cancer. Targeting EphA2 is especially attractive in ovarian cancer,
in which overexpression is present in >75% of cases (Landen et
al., Expert Opin Ther Targets 2005, 9(6):1179-87). Abraham et al.
state that the clinical significance of the expression of EphA2 was
observed in breast, prostate, colon, skin, cervical, ovarian, and
lung cancers. They studied EphA2 to determine the expression of
EphA2 and its ligand, Ephrin A-1, and E-cadherin in carcinoma of
the urinary bladder, and determine EphA2 as a new target for
therapy in bladder cancer. They conclude EphA2 may serve as a novel
target for bladder cancer therapy, (Abraham et al., Clin Cancer Res
2006, 12(2):353-60;). EphA2 inhibitors may be useful in treating
cancer, including bladder, breast, prostate, colon, skin, cervical,
ovarian, pancreatic, and lung cancer and melanoma.
[0745] EphB2: Target kinase EphB2 (i.e., Ephrin Receptor B2) is a
117.5 kDa transmembrane tyrosine kinase encoded by chromosome
1p36.1-p35 (symbol: EPHB2). Mann et al. state that forward and
reverse signaling mediated by EphB tyrosine kinase receptors and
their transmembrane ephrin-B ligands play important roles in axon
pathfinding. In their investigations of growth cones from the
ventral (EphB receptor-bearing) and dorsal (ephrin-B-bearing)
embryonic Xenopus retina designed to investigate the signaling
mechanisms in both forward and reverse directions, it is reported
that unclustered, but not clustered, EphB2 ectodomains trigger fast
(5-10 min) transient collapse responses in growth cones. This
collapse response is mediated by low levels of intracellular cyclic
GMP and requires proteasome function. In contrast, clustered, but
not unclustered, ephrin-B1 ectodomains cause slow (30-60 min)
growth cone collapse that depends on high cGMP levels and is
insensitive to inhibition of the proteasomal pathway. Upon
receptor-ligand binding, endocytosis occurs in the reverse
direction (EphB2-Fc into dorsal retinal growth cones), but not the
forward direction, and is also sensitive to proteasomal inhibition.
Endocytosis is functionally important because blocking of EphB2
internalization inhibits growth cone collapse. They state their
data reveals that distinct signaling mechanisms exist for B-type
Eph/ephrin-mediated growth cone guidance and suggest that
endocytosis provides a fast mechanism for switching off signaling
in the reverse direction (Mann et al., J Neurobiol 2003,
57(3):323-36). Nakada et al. demonstrate that migrating
glioblastoma cells overexpress EphB2 in vitro and in vivo; glioma
migration and invasion are promoted by activation of EphB2 of
inhibited by blocking EphB2. Dysregulation of EphB2 expression or
function may underlie glioma invasion (Nakada et al., Cancer Res
2004, 64(9):3179-85). Wu et al., investigated the expression of
EphB2 and EphB4 in breast carcinomas. Clinicopathological and
survival correlations were statistically analyzed in a series of 94
breast carcinomas, 9 normal specimens and 4 breast carcinoma cell
lines. Both EphB2 and EphB4 RTPCR products could be detected in all
specimens. Increased EphB2 protein expression was negatively
associated with overall survival (Wu et al., Pathol Oncol Res 2004,
10(1):26-33). Hafner et al. studied expression profiles of 12
different Eph receptors and 8 ephrins in human lung, colorectal,
kidney, liver, and brain cancers. They report EphB2 was
up-regulated 9-fold in hepatocellular carcinoma (Hafner et al.,
Clinical Chemistry 2004, 50:490-99). Umeda et al. studied the
expression of ephrinB2 and EphB receptors within fibroproliferative
membranes in patients with ocular angiogenic diseases collected
during vitrectomy. EphB2 and EphB3 expression was observed on
fibroproliferative membranes that were harvested from patients with
proliferative diabetic retinopathy (EphB2, 90.0%; EphB3, 70.0%) and
retinopathy of prematurity (EphB2, 35.0%; EphB3, 45.0%). Their data
suggest that the ephrinB2-EphB2/B3 system may play an important
role in ocular angiogenesis (Umeda et al., Am J Ophthalmol 2004,
138(2):270-9). EphB2 inhibitors may be useful in treating cancer,
including breast cancer, hepatocellular carcinoma and glioblastoma,
and for use in treating ocular angiogenesis diseases, including
retinopathy (e.g. retinopathy of prematurity and proliferative
diabetic retinopathy).
[0746] EphB4: Target kinase EphB4 (i.e., Ephrin Receptor B4) is a
108.3 kDa transmembrane tyrosine kinase encoded by chromosome 7q22
(symbol: EPHB4). EphB4 belongs to the Eph family of receptor
tyrosine kinases. Developmental studies have shown that the Eph
receptors, by regulating cell adhesion and cell movement in the
embryo, are important for the proper organization and integrity of
tissues. Because tissue disorganization and abnormal cell adhesion,
movement, and survival characterize the more advanced stages of
cancer, the inappropriate functioning of an Eph receptor in breast
tumor cells enhances malignancy. Xia et al. studied the biological
function of the receptor tyrosine kinase EphB4 in bladder cancer.
All of nine bladder cancer cell lines examined expressed EphB4.
Further, they showed EphB4 knockdown using specific siRNA and
antisense oligodeoxynucleotides molecules led to a profound
inhibition in cell viability associated with apoptosis via
activation of caspase-8 pathway and downregulation of antiapoptotic
factor, bc1-x1. Furthermore, EphB4 knockdown significantly
inhibited tumor cell migration and invasion. EphB4 knockdown in an
in vivo murine tumor xenograft model led to a nearly 80% reduction
in tumor volume associated with reduced tumor proliferation,
increased apoptosis and reduced tumor micro vasculature (Xia et
al., Oncogene 2006, 25(5):769-80). Xia et al. also studied the
expression and biological role of EphB4 in prostate cancer. They
found EphB4 mRNA is expressed in 64 of 72 (89%) prostate tumor
tissues assessed and EphB4 protein expression is found in the
majority (41 of 62, 66%) of tumors, and 3 of 20 (15%) normal
prostate tissues. They also showed knockdown of the EphB4 protein
using EphB4 short interfering RNA or antisense oligodeoxynucleotide
significantly inhibits cell growth/viability, migration, and
invasion, and induces apoptosis in prostate cancer cell lines (Xia
et al., Cancer Res 2005, 65(11):4623-32). Lee et al. used RT-PCR,
western blotting and immunohistochemical techniques to examine
EphB4 expression and protein levels in human prostate cancer cell
lines LNCaP, DU145 and PC3. Immunohistochemistry was also used to
examine localisation of EphB4 in tissue samples from 15 patients
with prostate carcinomas. All three prostate cancer cell lines
expressed the EphB4 gene and protein. EphB4 immunoreactivity in
vivo was significantly greater in human prostate cancers as
compared with matched normal prostate epithelium and there appeared
to be a trend towards increased expression with higher grade
disease (Lee et al., BMC Cancer 2005, 5:119). Stephenson et al.
used commercially available cDNA arrays to identify EphB4 as a gene
that is up-regulated in colon cancer tissue when compared with
matched normal tissue from the same patient (Stephenson et al., BMC
Mol Biol 2001, 2:15). Takai et al. used fluorescent
immunohistochemistry to analyze serial frozen sections of 20
endometrial carcinomas and 20 normal endometria for EphB4 and
ephrin-B2 protein expression. Further, they analyzed the
relationship between the patient's characteristics and the
percentages of EphB4- and ephrin-B2-stained cells. They indicate
the results demonstrate that increased EphB4 and ephrin-B2
expression may reflect or induce in endometrial carcinomas
increased potential for growth and tumorigenicity (Takai et al.,
Oncol Rep 2001, 8(3):567-73). Sinha et al., studied expression of
EphB4 in six men with primary squamous cell carcinoma of the head
and neck (HNSCC) that had metastasized to the cervical lymph nodes.
They obtained specimens of the primary tumor, the nodal metastasis,
and the adjacent normal mucosa, and performed immunocytochemistry
on each. They observed EphB4 expression in all primary and
metastatic tumors and no expression in the normal tissue. In each
of the six patients, expression was greater in the metastatic tumor
than in the primary tumor (Sinha et al., Ear Nose Throat J 2003,
82(11):866, 869-70, 887). EphB4 inhibitors may be useful in
treating cancer, including breast, bladder, prostate, colon, and
endometrial cancers and head and neck squamous cell carcinoma.
[0747] Erk2: Target kinase Erk2 (i.e., extracellular
signal-regulated kinase 2) is a 41.4 kDa dual function
serine/threonine-tyrosine kinase encoded by chromosome 22q11.2
(symbol: MAPK1). Erk2 is a member of the mitogen-activated protein
(MAP) kinase family and is alternatively known as mitogen-activated
protein kinase 1 (i.e., MAPK1). MAP kinases act as an integration
point for multiple biochemical signals, and are involved in a wide
variety of cellular processes such as proliferation,
differentiation, transcription regulation and development.
[0748] The activation of Erk2 requires phosphorylation by upstream
kinases. Upon activation, Erk2 translocates to the nucleus of the
stimulated cells, where it phosphorylates nuclear targets, in
addition to other targets including microtubule associated protein
2, myelin basic protein and ELK1. MacKenzie et al. state that the
cAMP-specific phosphodiesterase family 4, subfamily D, isoform 3
(i.e., PDE4D3) is shown to have FQF (i.e., Phe-Gln-Phe) and KIM
(i.e., Kinase Interaction Motif) docking sites for Erk2. These
sites straddle the Ser(579) target residue for Erk2 phosphorylation
of PDE4D3. Mutation of either or both of these docking sites
prevent Erk2 from being co-immunoprecipitated with PDE4D3, ablate
the ability of epidermal growth factor (EGF) to inhibit PDE4D3
through Erk2 action in transfected COS cells, and attenuate the
ability of Erk2 to phosphorylate PDE4D3 in vitro. The two conserved
NH(2)-terminal blocks of sequence, called upstream conserved
regions 1 and 2 (i.e., UCR1 and UCR.sup.2), that characterize PDE4
long isoforms, are proposed to amplify the small, inherent
inhibitory effect that Erk2 phosphorylation exerts on the PDE4D
catalytic unit. In contrast to this, the lone intact UCR.sup.2
region found in PDE4D1 directs COOH-terminal Erk2 phosphorylation
to cause the activation of this short isoform. From the analysis of
PDE4D3 truncates, it is suggested that UCR1 and UCR.sup.2 provide a
regulatory signal integration module that serves to orchestrate the
functional consequences of Erk2 phosphorylation. The PDE4D gene
thus encodes a series of isoenzymes that are either inhibited or
activated by Erk2 phosphorylation and thereby offers the potential
for ERK2 activation either to increase or decrease cAMP levels in
cellular compartments (MacKenzie et al., J Biol Chem 2000,
275(22):16609-17).
[0749] According to OMIM, Pleschka et al. (Nature Cell Biol., 2001,
3: 301-305) proposed that Erk2 regulates a cellular factor involved
in the viral nuclear export protein function. They suggested that
local application of MEK inhibitors may have only minor toxic
effects on the host while inhibiting viral replication without
giving rise to drug-resistant virus variants (OMIM MIM Number:
176948: Oct. 27, 2005). Erk2 is involved in cytokine signaling and
is a target for treating inflammation. Ramesh and Philipp state
that lipoproteins are the key inflammatory molecule type of
Borrelia burgdorferi, the spirochete that causes Lyme disease. They
investigated whether specific inhibition of p38 and Erk1/2 MAPK
would inhibit TNF-alpha and IL-6 production and thus astrocyte
apoptosis, and proliferation, respectively. Lipoprotein-stimulated
IL-6 production was unaffected by the MAPK inhibitors. In contrast,
inhibition of both p38 and Erk1/2 significantly diminished
TNF-alpha production, and totally abrogated production of this
cytokine when both MAPK pathways were inhibited simultaneously.
MAPK inhibition thus may be considered as a strategy to control
inflammation and apoptosis in Lyme neuroborreliosis (Ramesh and
Philipp, Neurosci Lett 2005, 384(1-2):112-6). The role of Erk2 in
signaling of cell differentiation, proliferation and survival
suggests that inhibition of Erk2 may be therapeutic for several
types of cancer. Husain et al. studied the effect of NSAIDs on MAPK
activity and phosphorylation in gastric cancer. They conclude that
NS-398 (a selective COX-2 inhibitor) and indomethacin (a
non-selective NSAID) significantly inhibit proliferation and growth
of human gastric cancer cell line MKN28. This effect is mediated by
NSAID-induced inhibition of MAPK (ERK2) kinase signaling pathway,
essential for cell proliferation (Husain et al., Life Sci 2001,
69(25-6):3045-54). Erk2 inhibitors may be useful in treating
cancer, including gastric cancer, and in treating inflammation,
including control of inflammation and apoptosis in Lyme
neuroborreliosis.
[0750] Fak: Target kinase Fak (i.e., Focal adhesion kinase 1, aka
protein tyrosine kinase 2, PTK2) is a 119.2 kDa tyrosine kinase
encoded by chromosome 8q24.3 (symbol: PTK2). The structure of Fak
comprises a B41 (i.e., Band 4.1 homology) domain in addition to the
TK domain. Fak and a related protein Pyk2/CAK-beta are cytoplasmic
non-receptor protein tyrosine kinases. Localization of Fak via its
C-terminal Focal Adhesion Targeting domain to focal
complexes/adhesions (sites of integrin receptor clustering) is a
prerequisite for Fak activation. Auto- or trans-phosphorylation of
Fak on Tyr397 allows for docking of the Src family kinases, among
other molecules. Src kinases phosphorylate Fak not only on the Tyr
residues in the activation loop of the FAK kinase, but also create
binding sites for downstream signaling components. Moreover, FAK
can be activated by other receptors, linking them to the integrin
signaling pathway. Activation of FAK can promote cell spreading,
locomotion, survival and anchorage-dependent growth.
[0751] FAK-related non-kinase (FRNK), the C-terminal portion of FAK
is expressed in some cell types by activating transcription from an
alternative promoter. FRNK is proposed to function as an endogenous
inhibitor of Fak signaling.
[0752] The role of Fak in signaling of cell proliferation,
migration, adhesion and survival may be targeted in therapeutics
for several types of cancer. Lightfoot et al. used
immunohistochemical techniques to assess FAK expression in patients
with fibrocystic disease (FCD), atypical ductal hyperplasia (ADH),
ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma
(IDC). The pattern of FAK expression in DCIS was significantly
higher than ADH (p<0.0001) and IDC (p=0.02). They conclude that
FAK overexpression in preinvasive, DCIS tumors precedes tumor cell
invasion or metastasis, suggesting that FAK may function as a
survival signal and be an early event in breast tumorigenesis
(Lightfoot et al., Breast Cancer Res Treat 2004, 88(2):109-16).
Miyazaki et al. performed an immunohistochemical analysis of FAK
protein expression to determine the relationship between FAK
overexpression and clinicopathological factors in oesophageal
squamous cell carcinoma (ESCC). They concluded that FAK
overexpression of ESCC was related to cell differentiation, tumour
invasiveness, and lymph node metastasis. Consequently, patients
with ESCC who had FAK overexpression had a poor prognosis (Miyazaki
et al., Br J Cancer 2003, 89(1):140-5). Smith et al. tested
antisense oligonucleotide inhibitors of FAK, in combination with
5-fluorouracil (5-FU), to increase its sensitivity in human
melanoma cell lines. They conclude their data show that the
downregulation of FAK by antisense oligonucleotide combined with
5-FU chemotherapy results in a greater loss of adhesion and greater
apoptosis in melanoma cells than treatment with either agent alone,
suggesting that the combination may be a potential therapeutic
agent for human melanoma in vivo (Smith et al., Melanoma Res 2005,
15(5):357-62). In a review, Natarajan et al. summarize data that
has demonstrated 1) elevated FAK expression in anaplastic
astrocytoma and glioblastoma tumor biopsy samples, 2) a role for
FAK in the promotion of glioblastoma cell proliferation, survival
and migration in vitro, and 3) a role for FAK in the promotion of
glioblastoma cell proliferation in vivo in an animal model
(Natarajan et al., Cancer J 2003, 9(2):126-33). Rovin et al.
investigated FAK expression in human prostate specimens by using
immunohistochemistry. In their conclusion, they suggest that the
sustained elevated levels of FAK expression during prostate tumor
cell progression is consistent with a role for FAK in the
development and maintenance of prostate carcinoma (Rovin et al.,
Prostate 2002, 53(2):124-32). Itoh et al. investigated whether
focal adhesion kinase (FAK) is involved in the progression of human
hepatocellular carcinoma (HCC). They conclude their data suggests
that FAK plays an important role in promoting tumor progression,
especially vascular invasion, in HCC (Itoh et al., Clin Cancer Res
2004, 10(8):2812-7). von Sengbusch et al. state that organ-specific
tumor cell adhesion to extracellular matrix (ECM) components and
cell migration into host organs often involve integrin-mediated
cellular processes that can be modified by environmental conditions
acting on metastasizing tumor cells, such as shear forces within
the blood circulation. Since the focal adhesion kinase (FAK)
appears to be essential for the regulation of the integrin-mediated
adhesive and migratory properties of tumor cells, they investigated
its role in early steps of the metastatic cascade using in vitro
and in vivo approaches. They summarize that FAK appears to be
involved in early events of integrin-mediated adhesion of
circulating carcinoma cells under fluid flow in vitro and in vivo.
This kinase may take part in the establishment of definitive
adhesive interactions that enable adherent tumor cells to resist
fluid shear forces, resulting in an organ-specific formation of
distant metastases (von Sengbusch et al., Am J Pathol 2005,
166(2):585-96).
[0753] Westhoff et al., note that elevated expression of the
nonreceptor tyrosine kinases Src and Fak correlates with malignancy
potential and poor clinical prognosis in colon and breast tumors.
They also state that recent studies monitoring focal adhesion
dynamics in cells deficient for Fak and Src implicate Src and Fak
as critical mediators of integrin adhesion turnover that promote
cell migration. Cells devoid of FAK exhibit impaired migration and
have large peripheral focal adhesion structures, while cells
lacking the three ubiquitous Src family members Src, Fyn, and Yes
also demonstrate altered distribution of focal adhesions and
impaired cell migration. Src kinase activity is clearly necessary
for focal adhesion turnover and cell motility, presumably by
tyrosine phosphorylation of key focal adhesion substrates, such as
FAK. The extracellular regulated kinase (ERK)/mitogen-activated
protein kinase (MAPK), pathway is also important in regulating
focal/adhesion dynamics during cell motility and it is likely that
ERK/MAPK contributes to Src-induced focal adhesion turnover.
Further, Westhoff et al. have recently reported that ERK/MAPK,
which is recruited to focal adhesions following v-Src activation,
is required for maximal activity of the protease calpain 2
promoting focal adhesion turnover and migration of
v-Src-transformed cells. ERK/MAPK-induced activation of calpain 2
is also required for epidermal growth factor-induced substrate
deadhesion and cell motility. Six major tyrosine phospho-acceptor
sites have been identified on Fak, at positions 397, 407, 576, 577,
861, and 925. Tyr 397 becomes phosphorylated (presumed by
auto-phosphorylation) upon integrin engagement. This leads to the
formation of a consensus binding site for the Src SH.sub.2 domain,
promoting association between Src and FAK. Phosphorylation of the
remaining tyrosine residues on Fak is considered to be
Src-dependent. Westhoff et al., generated a FAK mutant (4-9F-Fak)
in which each of the putative Src-induced tyrosine phosphorylation
sites (Tyr 407, 576, 577, 861, and 925) has been mutated to a
phenylalanine (Phe). They found that v-Src-induced phosphorylation
of FAK on tyrosine residues is necessary to enhance the adaptor
function of Fak with regard to assembly of the calpain 2/FAK/p42ERK
complex. Src-induced phosphorylation of Fak is also required for
Fak to undergo proteolytic cleavage by calpain in v-Src-transformed
cells and is necessary for calpain-mediated focal adhesion turnover
during transformation and cell migration. In addition, they show
that Src-induced phosphorylation of Fak also regulates F-actin
assembly and cell spreading. We further demonstrate a role for
Src-induced tyrosine phosphorylation of Fak in survival and
anchorage-independent growth of transformed cells (Westhoff et al.,
Molecular and Cellular Biology, 2004, 24: 8113-8133).
[0754] Fak inhibitors may be useful in treating cancer, including
colon and breast cancers, melanoma, ductal carcinoma in situ,
oesophageal squamous cell carcinoma, anaplastic astrocytoma and
glioblastoma, and human hepatocellular and prostate carcinomas, as
well as in reducing tumor metastasis. They may also be used in
combination with other chemotherapeutic drugs to provide
synergistic effects in treating cancers such as melanoma.
[0755] FGFR kinase family: The FGFRs (i.e., Fibroblast Growth
Factor Receptors) comprise a family of related but individually
distinct tyrosine kinase receptors. They have a similar protein
structure, with 3 immunoglobulin-like domains in the extracellular
region, a single membrane spanning segment, and a cytoplasmic
tyrosine kinase domain. The fibroblast growth factor receptors that
have been identified are FGFR1, FGFR2, FGFR3, which is mutant in
achondroplasia; and FGFR4. Sequence analysis of the 4.5-kb human
FGFR2 gene shows an open reading frame encoding the typical
membrane-spanning, tyrosine kinase receptor structure of the FGFR
gene family. A discussion of FGFR1, FGFR2, FGFR3, and FGFR4
follows.
[0756] FGFR
[0757] FGFR1: Target FGFR1 (i.e., Fibroblast Growth Factor Receptor
1) is a 91.9 kDa transmembrane tyrosine kinase encoded by
chromosome 8p11.2p11.1 (symbol: FGFR1). FGFR1 is also known as
FMS-like tyrosine kinase 2 (i.e., Flt2). FGFR1 is implicated in a
variety of cancers (e.g. 8p11 syndrome, Braun & Shannon, Cancer
Cell 2004, 5:203). Additionally, FGFR1 is an important mediator of
tumor angiogenesis (Compagni et al., Cancer Res. 2000, 60:7163).
The X-ray crystallographic structure of FGFR1 bound to fibroblast
growth factor 2 has been reported by Plotnikov et al (Cell 1999,
98: 641-650). Rossi et al report an FGFR1 P252R mutation in
families affected by Pfeiffer syndrome (Rossi et al., Clin
Dysmorphol. 2003, 12(4):269-74). FGFR1 inhibitors may be useful in
treating 8p11 myeloproliferative syndrome.
[0758] FGFR2: Target FGFR2 (i.e., Fibroblast Growth Factor Receptor
2) is a 92 kDa transmembrane tyrosine kinase encoded by chromosome
10q26 (symbol: FGFR2). According to OMIM, from a human tumor cDNA
library, Houssaint et al. (Proc. Nat. Acad. Sci. 1990, 87:
8180-8184) isolated a gene encoding a putative receptor-like
protein-tyrosine kinase that they called TK14. The amino acid
sequence was closely related to that of the mouse protein bck
(bacterially expressed kinase), and more distantly related to the
sequences of a chicken basic fibroblast growth factor receptor (73%
sequence homology) and its presumed human equivalent, the FLG
protein. Overexpression of the TK14 protein by transfection of
COS-1 cells led to the appearance of new cell-surface binding sites
for both acidic and basic fibroblast growth factors (OMIM MIM
Number: 176943: Apr. 6, 2006).
[0759] Sequence analysis of the 4.5-kbp human FGFR2 gene shows an
open reading frame encoding the typical membrane-spanning, tyrosine
kinase receptor structure of the FGFR gene family. Two alternative
gene products have been characterized: KGFR and BEK. These two
isoforms are identical except for a 49-amino acid sequence spanning
the second half of the third Ig loop in the extracellular region.
This local diversity is due to the presence of alternative exons
within FGFR2, exon B being expressed in the BEK product and exon
K26 in KGFR. Control of these alternative splice sites is thought
to involve transacting factors (Gilbert et al., Molec. Cell. Biol.
1993, 13: 5461-5468). The variation in expressed gene product is
highly significant because the ligand-binding characteristics of
KGFR and BEK are quite distinct. Furthermore, they have different
patterns of expression in murine embryogenesis. Whereas KGFR
appears to have a role in skin development, BEK is preferentially
expressed in osteogenesis. BEK transcripts are concentrated in the
frontal bones, maxilla, mandibula, and ossicles of the middle
ear.
[0760] To elucidate the structural determinants governing
specificity in FGF signaling, Plotnikov et al. (Cell 2000, 101:
413-424) determined the crystal structures of FGF1 and FGF2
complexed with the immunoglobulin-like ligand-binding domains 2 and
3 (D.sub.2 and D3) of FGFR1 and FGFR2, respectively. They found
that highly conserved FGF-D.sub.2 and FGF-linker (between D.sub.2
and D3) interfaces define a general binding site for all FGF-FGFR
complexes. Specificity is achieved through interactions between the
N-terminal and central regions of FGFs and 2 loop regions in D3
that are subject to alternative splicing. These structures provide
a molecular basis for FGF1 as a universal FGFR ligand and for
modulation of FGF-FGFR specificity through primary sequence
variations and alternative splicing (OMIM MIM Number: 176943: Apr.
6, 2006).
[0761] Defects in FGFR2 are a cause of Crouzon Syndrome (CS); also
called craniofacial dysostosis type I (CFD1). CS is an autosomal
dominant syndrome characterized by craniosynostosis (premature
fusion of the skull sutures), hypertelorism, exophthalmos and
external strabismus, parrot-beaked nose, short upper lip,
hypoplastic maxilla, and a relative mandibular prognathism (OMIM
MIM Number: #123500: May 4, 2000).
[0762] Further, defects in FGFR2 are a cause of Jackson-Weiss
syndrome (JWS). JWS is an autosomal dominant craniosynostosis
syndrome characterized by craniofacial abnormalities and
abnormality of the feet including broad great toes with medial
deviation and tarsal-metatarsal coalescence (OMIM MIM Number:
176943: Apr. 6, 2006).
[0763] Further, defects in FGFR2 are a cause of Apert Syndrome,
also known as acrocephalo-syndactyly type I (ACS1), which is
characterized by craniosynostosis (premature fusion of the skull
sutures) and severe syndactyly (cutaneous and bony fusion of the
digits), and is autosomal dominant.
[0764] Further, defects in FGFR2 are a cause of Pfeiffer Syndrome
(PS), also known as acrocephalosyndactyly type V (ACS5), which is
characterized by craniosynostosis with deviation and enlargement of
the thumbs and great toes, brachymesophalangy, with phalangeal
ankylosis and a varying degree of soft tissue syndactyly. Three
subtypes of PS have been described: mild autosomal dominant form
(type 1); cloverleaf skull, elbow ankylosis, early death, sporadic
(type 2); craniosynostosis, early demise, sporadic (type 3).
[0765] Further, defects in FGFR2 are the cause of beare-stevenson
cutis gyrata syndrome (BSCGS) which is an autosomal dominant
condition characterized by the furrowed skin disorder of cutis
gyrata, acanthosis nigricans, craniosynostosis, craniofacial
dysmorphism, digital anomalies, umbilical and anogenital
abnormalities and early death (OMIM MIM Number: 176943: Apr. 6,
2006).
[0766] Further, defects, in FGFR2 are the cause of Antley-Bixler
syndrome which is characterized by trapezoidocephaly, midface
hypoplasia, humeroradial synostosis, bowing of femora, fractures
and other abnormalities (OMIM MIM Number: #207410: Nov. 29, 2005).
FGFR2 inhibitors may be useful in treating Crouzon Syndrome,
Jackson-Weiss Syndrome, Apert Syndrome, craniosynostosis, Pfeiffer
Syndrome, acrocephalo syndactyly type V, and Beare-Stevenson Cutis
Gyrata Syndrome.
[0767] FGFR3: Target kinase FGFR3 (i.e., Fibroblast Growth Factor
Receptor 3) is a 87.7 kDa transmembrane tyrosine kinase encoded by
chromosome 4p16.3 (symbol: FGFR3). As a member of the fibroblast
growth factor family, FGFR3 is involved in a variety of activities,
including mitogenesis, angiogenesis, and wound healing.
Furthermore, FGFR3 plays a role in the development and maintenance
of bone and brain tissue. FGFR3 regulates bone growth by limiting
the formation of bone from cartilage, particularly in the long
bones. In addition, FGFR3 is activated by translocation in
approximately 15% of multiple myeloma (Trudel, S., Blood 2004,
103:3521). FGFR3 inhibitors may be useful in treating angiogenesis
disorders, wounds, achondroplasia, Muenke craniosynostosis, Crouzon
syndrome, acanthosis nigricans, thanatophoric dysplasia, bladder
carcinomas, and multiple myeloma.
[0768] FGFR4: Target kinase FGFR4 (i.e., Fibroblast Growth Factor
Receptor 4) is a 88.0 kDa transmembrane tyrosine kinase encoded by
chromosome 5q35.3 (symbol: FGFR4). According to OMIM, Partanen et
al. (EMBO J. 1991, 10: 1347-1354) reported the cDNA cloning and
analysis of a novel member of the fibroblast growth factor receptor
(FGFR) gene family expressed in K562 erythroleukemia cells. Its
deduced amino acid sequence was 55% identical with the previously
characterized FGFRs, FLG (FGFR1) and BEK, and had the structural
characteristics of an FGFR family member including 3
immunoglobulin-like domains in its extracellular part. The
expression pattern of FGFR4 was found to be distinct from that of
FLG and BEK and also distinct from that of FGFR3, which had also
cloned from K562 crythroleukemia cells. To elucidate further the
physiologic relevance of protein-tyrosine kinases and to search for
additional members of the gene family as possible factors in
carcinogenesis, Holtrich et al. (Proc. Nat. Acad. Sci. 1991, 88:
10411-10415) amplified mRNA from lung tissue by the polymerase
chain reaction (PCR) using PTK-specific primers followed by
sequencing of the clones. They identified a novel protein-tyrosine
kinase, which they called TKF (tyrosine kinase related to
fibroblast growth factor receptor). Among a wide variety of cells
and tissues tested, including human lymphocytes and macrophages,
TKF was found to be expressed only in lung and in some tumors of
lung origin as well as in malignancies not derived from lung
tissues. Sequence comparison has demonstrated that TKF is identical
to FGFR4 (OMIM MIM Number: 134935: May 3, 2002).
[0769] The FGFR4 protein interacts with specific growth factors to
conduct signals from the environment outside the cell to the
nucleus. Animal studies indicate that this FGFR4 gene is involved
in muscle development and the maturation of bone cells in the
skull. FGFR4 may also play a role in the development and
maintenance of specialized cells (called foveal cones) in the
light-sensitive layer (retina) at the back of the eye. Aberrant
expression of FGFR4 is correlated with cancer of the breast, ovary,
endometrium, and fallopian tube, and with leiomyosarcoma. FGFR4
inhibitors may be useful in treating cancer of the breast, lung,
colon, medullary thyroid, pancreas, ovary, prostate, endometrium,
and fallopian tube, head and neck squamous cell carcinomas and
leiomyosarcoma.
[0770] Flt1: Target kinase Flt1 (i.e., Fms like tyrosine kinase 1)
is a 150.7 kDa transmembrane tyrosine kinase encoded by chromosome
13q12 (symbol: FLT1), also known as VEGFR1 (i.e., Vascular
Endothelial Growth Factor Receptor 1). According to OMIM, oncogene
FLT belongs to the SRC gene family and is related to oncogene ROS.
Like other members of this family, it shows tyrosine protein kinase
activity that is important for the control of cell proliferation
and differentiation via interaction with PLC-gammas, PTPN11, GRB2,
CRK, NCK1 and other proteins. The name is due to the resemblance of
the sequence structure of the FLT gene to that of the FMS gene.
VEGF and its high-affinity binding receptors, the tyrosine kinases
Flk1 and Flt1, are thought to be important for the development of
embryonic vasculature. Studying transgenic mice in whom the Flk1
gene was disrupted, Shalaby et al. (Nature 1995, 376: 62-65)
demonstrated a total failure of embryonic mice to develop blood
vessels and failure of blood island formation in the yolk sac. Fong
et al. (Nature 1995, 376: 65-69) reported that in mice Flt1 is
essential for the organization of embryonic vasculature, but is not
essential for endothelial cell differentiation. Transgenic mouse
embryos homozygous for a targeted mutation in the Flt1 locus formed
endothelial cells in both embryonic and extraembryonic regions, but
assembled these cells into abnormal vascular channels and died in
utero at mid-somite stages. At earlier stages, the blood islands of
homozygous mice were abnormal, with angioblasts in the interior as
well as on the periphery. Fong et al. (ibid.) suggested that the
Flt1 signaling pathway may regulate normal endothelial cell-cell or
cell-matrix interactions during vascular development (OMIM MIM
Number: 165070: Mar. 27, 2006). Flt1 inhibitors may be useful in
treating non-small cell lung carcinoma, prostate carcinoma, and
colorectal cancer.
[0771] Flt3: Target kinase Flt3 (i.e., Fms-like tyrosine kinase 3)
is a transmembrane tyrosine kinase of 112.8 kDa encoded by
chromosome 13q12 (symbol: FLT3). According to OMIM, Rosnet et al.
(Genomics 1991, 9: 380-385) isolated a novel member of the class 3
receptors discussed above. They demonstrated that this gene of the
tyrosine kinase family, called FLT3, has strong sequence
similarities with other members of the group. Lymphohematopoietic
stem cells serve as a reservoir for virtually all blood cells but
make up only approximately 0.01% of human or murine marrow cells.
The ability to isolate and expand this population has clinical
applications in bone marrow transplantations for cancer and genetic
diseases. Small et al. (Proc. Nat. Acad. Sci. 1994, 91: 459-463)
cloned the cDNA for stem cell tyrosine kinase 1, the human homolog
of murine Flk2/Flt3, from a CD34+ hematopoietic stem cell-enriched
library. The cDNA encoded a protein of 993 amino acids with 85%
identity and 92% similarity to the murine homolog. STK1, which is
identical to FLT3, is a member of the type III receptor tyrosine
kinase family that includes KIT, FMS, and platelet-derived growth
factor receptor. STK1 expression in human blood and marrow is
restricted to CD34+ cells, a population greatly enriched by
stem/progenitor cells. Antisense oligonucleotides directed against
STK1 sequences inhibited hematopoietic colony formation, most
strongly in long-term bone marrow cultures. The data suggested that
STK1 may function as a growth factor receptor on hematopoietic stem
and/or progenitor cells (OMIM MIM Number: 136351: Mar. 3,
2005).
[0772] Levis et al., state that Internal tandem duplication (ITD)
mutations of the receptor tyrosine kinase FLT3 have been found in
20% to 30% of patients with acute myeloid leukemia (AML). These
mutations constitutively activate the receptor and appear to be
associated with a poor prognosis. In their study, dose-response
cytotoxic assays were performed with AG1295, a tyrosine kinase
inhibitor active against FLT3, on primary blasts from patients with
AML, and they found that AG1295 was specifically cytotoxic to AML
blasts harboring FLT3/ITD mutations. They suggest that these
mutations contribute to the leukemic process and that the FLT3
receptor represents a therapeutic target in AML (Levis et al.,
Blood 2001, 98:885-887). Flt3 inhibitors may be useful in treating
acute myeloid leukemia, myelodysplastic syndrome, acute
lymphoblastic leukemia.
[0773] Flt4: Target kinase Flt4 (i.e., Fms-like tyrosine kinase 4)
is a transmembrane tyrosine kinase of 145.6 kDa encoded by
chromosome 5q35.3 (symbol: FLT4). Flt4 is also known as VEGFR.sup.3
(i.e., Vascular Endothelial Growth Factor Reccptor 3). According to
OMIM, by screening a placenta cDNA library with a mouse Flt3 probe,
Galland et al. (Genomics 1992, 13: 475-478) isolated a human gene
encoding a putative receptor-type tyrosine kinase, FLT4. The
deduced amino acid sequence of the intracellular portion of the
molecule showed that it was strongly related to FLT1 and KDR and to
a lesser degree to members of the class 3 receptor-type tyrosine
kinases: FMS, PDGFR, KIT, and FLT3. Primary lymphoedema, a rare,
autosomal dominant disorder that leads to a disabling and
disfiguring swelling of the extremities and, when untreated, tends
to worsen with time, has been linked to the FLT4 locus (Karkkainen
et al., Nat. Genet. 2000, 25: 153-9). All disease-associated
alleles, analyzed had missense mutations and encoded proteins with
an inactive tyrosine kinase, preventing downstream gene activation
(OMIM MIM Number: 136352: Nov. 19, 2003). Flt4 inhibitors may be
useful in treating primary lymphoedema.
[0774] Fms: Target kinase Fms (i.e., feline McDonough sarcoma) is a
member of the family of genes originally isolated from the Susan
McDonough strain of feline sarcoma viruses. Fms is a transmembrane
tyrosine kinase of 108.0 kDa coded by chromosome 5q33.2-q33.3
(symbol: CSF1R). The structure of the transmembrane receptor Fms
comprises two Ig-like domains, a IgC2-like domain, two additional
Ig-like domains, a TM domain, and the TK domain.
[0775] Fms is the receptor for the macrophage colony-stimulating
factor (M-CSF), and is crucial for the growth and differentiation
of the monocyte-macrophage lineage. Upon binding of M-CSF to the
extracellular domain of Fms, the receptor dimerizes and
trans-autophosphorylates cytoplasmic tyrosine residues.
[0776] M-CSF, first described by Robinson and co-workers (Blood,
1969, 33:396-9), is a cytokine that controls the production,
differentiation, and function of macrophages. M-CSF stimulates
differentiation of progenitor cells to mature monocytes, and
prolongs the survival of monocytes. Furthermore, M-CSF enhances
cytotoxicity, superoxide production, phagocytosis, chemotaxis, and
secondary cytokine production of additional factors in monocytes
and macrophages. Examples of such additional factors include
granulocyte colony stimulating factor (G-CSF), interleukin-6
(IL-6), and iriterleukin-8 (IL-8). M-CSF stimulates hematopoiesis,
promotes differentiation and proliferation of osteoclast progenitor
cells, and has profound effects on lipid metabolism. Furthermore,
M-CSF is important in pregnancy. Physiologically, large amounts of
M-CSF are produced in the placenta, and M-CSF is believed to play
an essential role in trophoblast differentiation (Motoyoshi. Int J
Hematol. 1998, 67:109-22). The elevated serum M-CSF levels of early
pregnancy may participate in the immunologic mechanisms responsible
for the maintenance of the pregnancy (Flanagan & Lader, Curr
Opin Hematol. 1998, 5:181-5).
[0777] Aberrant expression and/or activation of Fms has been
implicated in acute myeloid leukemia, AML (Ridge et al., Proc. Nat.
Acad. Sci., 1990, 87:1377-1380). Mutations at codon 301 are
believed to lead to neoplastic transformation by ligand
independence and constitutive tyrosine kinase activity of the
receptor. The tyrosine residue at codon 969 has been shown to be
involved in a negative regulatory activity, which is disrupted by
amino acid substitutions. Accordingly, Fms mutations are most
prevalent (20%) in chronic myelomonocytic leukemia and AML type M4
(23%), both of which are characterized by monocytic
differentiation.
[0778] A condition related to AML is chronic myeloid leukemia
(CML). During the myeloid blast crisis (BC) of CML, non-random
additional chromosome abnormalities occur in over 80% of patients.
However, these cytogenetic changes have been reported to precede
the clinical signs of CML-BC by several months to years suggesting
that other biological events may participate in the multistep
process of acute transformation of CML. The autocrine production of
growth factors has been shown to occur in several hematological
malignancies and particularly in AML. Specchia et al [Br J
Haematol. 1992 March; 80(3):310-6] have demonstrated that IL-1 beta
gene is expressed in almost all cases of CML in myeloid blast
crisis, and that a high proportion of cases showed constitutive
expression of the M-CSF gene. Many of the same patients in the
Specchia et al study demonstrated simultaneous co-expression of
Fms. After exposure of leukemic cells to phorbol myristate acetate
(PMA), release of M-CSF protein was documented in three of five
patients studied; however, no significant interleukin-3 (IL-3),
granulocyte-macrophage colony-stimulating factor (GM-CSF) or
granulocyte colony-stimulating factor (G-CSF), was detected in
these patients. This demonstrates that different patterns of growth
factors secretion exist in AML and CML, and that distinct molecular
events are likely involved in the control of leukemic
proliferation.
[0779] The observation that production of M-CSF, the major
macrophage growth factor, is increased in tissues during
inflammation (Le Meur et al, J. Leukocyte Biology. 2002;
72:530-537) provides a role for Fms in certain diseases. For
example, COPD is characterized by airflow limitation that is not
fully reversible. The airflow limitation is usually progressive and
associated with an abnormal inflammatory response of the lungs to
noxious particles or gases. The chronic inflammation of COPD is
observed through the airways, parenchyma, and pulmonary
vasculature. The inflammatory cell population consists of
neutrophils, macrophages, and T lymphocytes, along with eosinophils
in some patients. Macrophages are postulated to play an
orchestrating role in COPD inflammation by releasing mediators such
as TNF-a, IL-8 and LTB4, which are capable of damaging lung
structures and/or sustaining neutrophilic inflammation.
[0780] Further, M-CSF/fms signaling is critical to osteoclast
formation and survival of osteoclast precursors. For example,
estrogen loss in menopause results in increased M-CSF and thus
increased osteoclast number and bone resorption which leads to
increased risk of fracture and osteoporosis. Accordingly, blockage
of this signal is a target for the inhibition of bone resorption
(Teitelbaum, Science. 2000; 289:1504; Rohan, Science. 2000;
289:1508).
[0781] Atherosclerosis, an inflammatory disease of the vessel
walls, is associated with significant morbidity and mortality. A
effect for Fms inhibition in the treatment and prevention of
atherosclerosis depends on several observations (Libby, Nature.
2002; 420:868-874). First, monocytes resident in the arterial
intima increase expression of scavenger receptors and internalize
modified lipoproteins. The resulting lipid-laden macrophages
develop into foam cells characteristic of the atherosclerotic
lesion. Macrophages in atheroma secrete cytokines and growth
factors involved in lesion progression. Additionally, macrophages
replicate within the intima. Through Fms, M-CSF activates the
transition from monocyte to lipid-laden macrophage and augments
expression of scavenger receptor A. Indeed, atherosclerotic plaques
over-express M-CSF which is critical for atherosclerotic
progression. Mice deficient in M-CSF have been found to experience
less severe atherosclerosis than mice with normal M-CSF
(Rajavashisth, et. al., J. Clin. Invest. 1998; 101:2702-2710; Qiao,
et. al., Am. J. Path. 1997; 150:1687-1699). Accordingly, inhibitors
of Fms disrupt M-CSF signaling, compromising monocyte to macrophage
foam cell progression, macrophage survival and replication, and
cytokine signaling that participates in lesion progression.
[0782] Wegener's granulomatosis, also known as vasculitis, is
characterized by granulomatous inflammation of the blood vessels
with necrosis. This inflammation limits blood flow to organs with
consequent damage. Although the disease can involve any organ
system, Wegener's granulomatosis mainly affects the respiratory
tract (i.e., sinuses, nose, trachea, and lungs) and the kidneys.
The endothelium plays a central role in the immunopathology of
several vascular disorders in many inflammatory conditions such as
Wegener's granulomatosis in which use of intravenous immunoglobulin
(IV Ig) has been shown to be beneficial (see e.g., Basta et al, J
Clin Invest 1994, 94:1729-1735). It has been reported (Xu et al,
Am. J. Path., 1998; 153:1257-1266) that IV Ig inhibits endothelial
cell proliferation in a dose- and time-dependent manner and
down-regulates the expression of adhesion molecule mRNA (ICAM-1 and
VCAM-1), chemokine mRNA (MCP-1, M-CSF, and GM-CSF), and
proinflammatory cytokine mRNA (TNF-.alpha., IL-1 , and IL-6)
induced by TNF-.alpha. or IL-1 . These results may explain, at
least in part, the therapeutic effect of IV Ig in vascular and
inflammatory disorders. Additionally, these results suggest that
inhibition of M-CSF activity at the level of interaction with Fms
is an efficacious treatment strategy.
[0783] The role of M-CSF and Fms in emphysema appears to involve
the regulation of elastin metabolism through control of matrix
metalloproteins. M-CSF has a role in the modulation of the
accumulation and function of alveolar macrophages (AMs) in vivo
(Shibata et al, Blood 2001, 98: pp. 2845-2852). Osteopetrotic
(Op/Op) mice have no detectable M-CSF and show variable
tissue-specific reductions in macrophage numbers. Accordingly, it
was hypothesized that AMs would be decreased in number and have
altered function in Op/Op mice because of the absence of M-CSF.
Shibata et al found that lung macrophages identified in lung
sections were decreased in number in 20-day-old Op/Op mice but not
Op/Gp mice older than 4 months compared with findings in
age-matched littermate controls. The numbers of AMs recovered by
bronchoalveolar lavage (BAL) were also reduced in young but not
adult Op/Op mice compared with controls. Importantly, AMs of Op/Op
mice spontaneously release higher levels of matrix
metalloproteinases (MMPs) than AMs of controls. Consistent with an
increased release of MMP, Op/Op mice have abnormal elastin
deposition and spontaneously develop emphysema in the absence of
molecular or cellular evidence of lung inflammation. Accordingly,
the modulation of metalloelastase activity in macrophages by M-CSF
may control the degradation of elastin fibers in lungs or blood
vessels.
[0784] Metastatic cancer cells cause bone destruction, with
associated fracture, pain, deformation, and hypercalcaemia, due to
production of osteoclasticogenic factors including M-CSF by tumor
cells (Clohisy et al, Clin. Orthop. 2000, 373: 104-14). Binding of
M-CSF to the Fms product stimulates formation of osteoclasts and
osteolytic activity (Kodama et al, J. Exp. Med. 1991, 173: 269-72;
Feng et al, Endocrinology 2002, 143:4868-74). Accordingly,
inhibition of osteoclast activity at the level of Fms offers a
compelling target for amelioration of bone metastasis.
[0785] Nephritis is inflammation of the kidneys. It may be caused
for example by a bacterial infection of the kidneys or exposure to
a toxin. However, nephritis more commonly develops from an abnormal
immune reaction, which can occur, for example, when an antibody
attacks either the kidney itself or an antigen attached to kidney
cells, or when an antigen-antibody complex formed elsewhere in the
body attaches to cells in the kidney. Some types of nephritis
involve infiltration of kidney tissues by white blood cells and
deposits of antibodies. In other types of nephritis, inflammation
may consist of tissue swelling or scarring without white bloodcells
or antibodies. Furthermore, nephritis can occur anywhere in the
kidneys. With respect to the glomeruli, progressive damage to
glomeruli causes urine production to fall and metabolic waste
products to build up in the blood. When damage to glomeruli is
severe, inflammatory cells and injured glomerular cells accumulate,
compressing the capillaries within the glomerulus and interfering
with filtration. Scarring may develop, impairing kidney function
and reducing urine production. In some cases, microthrombi may form
in the small blood vessels, further decreasing kidney function.
Less commonly, nephritis involves the tubulointerstitial tissues;
such inflammation is called tubulointerstitial nephritis. When
inflammation damages the tubules and the tubulointerstitial
tissues, the kidneys may become unable to concentrate urine,
eliminate (excrete) metabolic waste products from the body, or
balance the excretion of sodium and other electrolytes, such as
potassium. When the tubules and tubulointerstitial tissues are
damaged, kidney failure often develops. Accordingly, inhibition of
Fms offers a target for therapeutic intervention in nephritis due
to the modulation of the inflammatory response comprising the
etiology of the disease.
[0786] Lupus nephritis, i.e., renal involvement in systemic lupus
erythematosus (SLE), is a common disease manifestation with a poor
prognosis. At least three potentially overlapping, immunopathogenic
mechanisms for lupus nephritis are supported by experimental data.
First, circulating immune complexes consisting chiefly of DNA and
anti-DNA are deposited in the kidney. Resulting complement
activation and chemotaxis of neutrophils leads to a local
inflammatory proccss. Second, in situ formation of antigen and
antibody complexes may similarly lead to complement activation and
leucocyte mediated injury. Third, antibodies against specific
cellular targets may produce renal injury. An additional mechanism
is observed in SLE patients with the antiphospholipid antibody
syndrome. Glomerular thrombosis can result from the
hypercoagulability that accompanies antibodies directed against
negatively charged phospholipid-protein complexes (e.g. biologic
false positive VDRL, anticardiolipin antibodies, and lupus
anticoagulant). Mesangial lupus nephritis is accompanied by normal
diagnostic findings or with a mild degree of proteinuria but
typically absence of hypertension or abnormal urinary sediment.
Focal and diffuse proliferative lupus glomerulonephritis are often
associated with the worst prognosis for renal survival and can be
accompanied by nephrotic syndrome, significant hypertension and
abnormal urine sediment. Membranous lupus nephritis often presents
with proteinuria, moderate to high grade, but usually normal
urinary sediment in the absence of hypertension. Mesangial lupus
nephropathy is generally associated with an excellent prognosis,
whereas proliferative lupus nephropathy, especially diffuse
variant, is often characterized by hypertension, red cell casts and
significant deterioration of renal function. Nephrotic syndrome in
the absence of hypertension, active urinary sediment or significant
hypocomplementemia suggest the membranous variant of lupus
nephropathy. Membranous nephropathy generally is associated with a
good prognosis and relative preservation of renal function.
However, in the presence of persistent nephrotic range proteinuria,
membranous lupus nephropathy can, in fact, lead to loss of renal
function and end stage renal disease (ESRD). Accordingly,
inhibition of Fms offers a target for therapeutic intervention in
lupus due to the modulation of the inflammatory response comprising
the etiology of the disease.
[0787] Macrophage accumulation is a prominent feature in many forms
of glomerulonephritis. Local proliferation of macrophages within
the kidney has been described in human and experimental
glomerulonephritis and may have an important role in augmenting the
inflammatory response. Isbel et al (Nephrol Dial Transplant 2001,
16: 1638-1647) examined the relationship between local macrophage
proliferation and renal expression of M-CSF. Glomerular and
tubulointerstitial M-CSF expression was found to be up-regulated in
human glomerulonephritis, being most prominent in proliferative
forms of disease. Because this correlates with local macrophage
proliferation, it suggests that increased renal M-CSF production
plays an important role in regulating local macrophage
proliferation in human glomerulonephritis. In a model of renal
inflammation (UUO--unilateral ureteric obstruction) anti-Fms
antibody treatment reduced macrophage accumulation (Le Meur et.al.,
J Leukocyte Biology, 2002, 72: 530-537). Accordingly, inhibition of
Fms offers a target for therapeutic intervention in
glomerulonephritis.
[0788] Insulin resistance and obesity are hallmark of type II
diabetes and there is a strong correlation exists between insulin
resistance and abdominal visceral fact accumulation (Bjorntrop,
Diabetes Metab. Res. Rev., 1999, 15: 427-441). Current evidence
indicates that macrophages accumulating in adipose tissue release
TNF-a and other factors that cause adipocyte changes (hypertrophy,
lipolysis, reduced insulin sensitivity) and also promote insulin
resistance in surrounding tissues. Therefore, macrophage
accumulation in type 2 diabetes is important for disease
progression. Accordingly, inhibition of Fms has potential in
preventing the development of insulin resistance and
hyperglycemia.
[0789] Similarly, the observation that production of M-CSF, the
major macrophage growth factor, is increased in tissues during
inflammation points out a role for Fms in diseases, such as for
example inflammatory diseases. More particularly, because elevated
levels of M-CSF are found in the disease state, modulation of the
activity of Fms can ameliorate disease associated with increased
levels of M-CSF.
[0790] Fms inhibitors may be useful in treating to immune
disorders, including rheumatoid arthritis, systemic lupus
erythematosis (SLE), Wegener's granulomatosis, and transplant
rejection, inflammatory diseases including Chronic Obstructive
Pulmonary Disease (COPD), emphysema, and atherosclerosis, metabolic
disorders, including insulin resistance, hyperglycemia, and
lipolysis, disorders of bone structure or mineralization, including
osteoporosis, increased risk of fracture, hypercalcemia, and bone
metastases, kidney diseases, including nephritis (e.g.
glomerulonephritis, interstitial nephritis, Lupus nephritis),
tubular necrosis, diabetes-associated renal complications, and
hypertrophy and cancers, including multiple myeloma, acute myeloid
leukemia, chronic myeloid leukemia (CML), breast cancer, and
ovarian cancer.
[0791] Frk: Target kinase Frk (Fyn-related kinase) is a 58.5 kDa
tyrosine kinase encoded by chromosome 6q21-q22.3 (symbol: FRK). The
structure comprises an SH.sub.2, an SH3 and a tyrosine kinase
domain. Hosoya et al., report the the identification of a SRC-like
tyrosine kinase gene, FRK (Fyn-related kinase), fused with ETV6 in
a patient with acute myelogenous leukemia carrying
t(6;12)(q21;p13). The ETV6/FRK protein was shown to be
constitutively autophosphorylated on its tyrosine residues.
ETV6/FRK phosphorylated histones H.sub.2B and H4 in vitro to a
greater extent than did FRK, suggesting it had elevated kinase
activity. ETV6/FRK could transform both Ba/F3 cells and NIH3T3
cells, which depended on its kinase activity (Hosoya et al., Genes
Chromosomes Cancer 2005, 42(3):269-79). Welsh et al. concluded that
FRK/RAK contributes to cytokine-induced beta-cell death, and
inhibition of this kinase could provide means to suppress beta-cell
destruction in Type I diabetes (Welsh et al., Biochem J 2004,
382(1):261-8). Frk inhibitors may be useful in treating acute
myeloid leukemia and type I diabetes.
[0792] Fyn: Target kinase Fyn (i.e., Fyn oncogene related to Src,
Fgr, Yes) is a 60.6 kDa non-receptor tyrosine kinase encoded by
chromosome 6q21 (symbol: FYN). Fyn is involved in regulation of
mast cell degranulation in a synergistic confluence of Fyn and Lyn
(i.e., v-Yes-1 Yamaguchi sarcoma viral related oncogene homolog)
pathways at the level of protein kinase C and calcium regulation.
Fyn inhibitors may be useful in treating Alzheimer's disease,
schizophrenia and in prevention of metastases, e.g. in melanoma and
squamous cell carcinoma.
[0793] Gsk3.alpha., Gsk3 : Target kinase Gsk3 (i.e., Glycogen
synthase kinase 3 beta) is a 46.8 kDa STK encoded by chromosome
3q13.3 (symbol: GSK3B). Target kinase Gsk3.alpha. (i.e., Glycogen
synthase kinase 3 alpha) is a 51.0 kDa STK encoded by chromosome
19q13.2 (symbol: GSK3A). Gsk3 is a pro line-directed
serine-threonine kinase that was initially identified as a
phosphorylating and inactivating glycogen synthase. Two isoforms,
alpha and beta, show a high degree of amino acid homology
(Stambolic & Woodgett, Biochem. J. 1994, 303: 701-704,). GSK3B
is involved in energy metabolism, neuronal cell development, and
body pattern formation (Plyte et al., Biochim, Biophys. Acta 1992,
1114: 147-162,). The X-ray crystallographic structure of Gsk3 has
been reported by Dajani et al. (Cell 2001, 105: 721-732). Klein
& Melton (Proc. Nat. Acad. Sci. 1996, 93: 8455-8459) proposed
that Gsk3 is the endogenous target of lithium in diverse systems.
For example, lithium potently and specifically inhibits Gsk3
activity in vitro. This suggests a mechanism whereby lithium can
mimic insulin action, and lithium inhibition of the Gsk3 pathway in
the brain could explain the actions of lithium action in
manic-depressive illness in addition to its effects on development
and its insulin like activity.
[0794] Phiel et al., show that therapeutic concentrations of
lithium, a GSK-3 inhibitor, block the production of Abeta peptides
by interfering with APP cleavage at the gamma-secretase step, but
do not inhibit Notch processing. Importantly, lithium also blocks
the accumulation of Abeta peptides in the brains of mice that
overproduce APP. The target of lithium in this setting is GSK-3
alpha, which is required for maximal processing of APP. Since GSK-3
also phosphorylates tau protein, the principal component of
neurofibrillary tangles, inhibition of GSK-3alpha offers a new
approach to reduce the formation of both amyloid plaques and
neurofibrillary tangles, two pathological hallmarks of Alzheimer's
disease (Phiel et al., Nature 2003, 423:435-439). Eldar-Finkelman
states that GSK-3 inhibitors might prove useful as therapeutic
compounds in the treatment of conditions associated with elevated
levels of enzyme activity, such as type 2 diabetes and Alzheimer's
disease. The pro-apoptotic feature of GSK-3 activity suggests a
potential role for its inhibitors in protection against neuronal
cell death, and in the treatment of traumatic head injury and
stroke. Finally, selective inhibitors of GSK-3 could mimic the
action of mood stabilizers such as lithium and valproic acid and be
used in the treatment of bipolar mood disorders (Eldar-Finkelman,
Trends Mol Med 2002, 8:126-132). Martinez et al. state that
glycogen synthase kinase 3 (GSK-3) was initially described as a key
enzyme involved in glycogen metabolism, but is now known to
regulate a diverse array of cell functions. Two forms of the
enzyme, GSK-3alpha and GSK-3beta, have been previously identified.
Small molecules inhibitors of GSK-3 may, therefore, have several
therapeutic uses, including the treatment of neurodegenerative
diseases, diabetes type II, bipolar disorders, stroke, cancer, and
chronic inflammatory disease (Martinez et al., Med Res Rev 2002,
22(4):373-84). GSK inhibitors may be useful in treating CNS
disorders such as Alzheimer's disease, Parkinson's disease,
amyotrophic lateral sclerosis, diabetes type II, bipolar disorders,
stroke, cancer, chronic inflammatory disease, leucopenia,
schizophrenia, chronic pain, neuropathic pain, and traumatic head
injury.
[0795] HCK: Target kinase HCK (hemopoietic cell kinase) is a 59.5
kDa tyrosine kinase encoded by chromosome 20q11.21 (symbol: HCK).
The protein structure comprises an SH3, and SH.sub.2 and a
bipartite kinase domain. HCK inhibitors may be useful in treating
chronic myelogenous leukemia and acute lymphocytic leukemia.
[0796] Her2/Erbb2: Target kinase Her2/Erbb2 (i.e., Human EGF
receptor 2) is a 137.9 kDa transmembrane tyrosine kinase encoded by
chromosome 17q11.2-q12/17q21.2 (symbol: ERBB2). According to OMIM,
the ERBB2 locus 17q21 is the chromosome 17 breakpoint in acute
promyelocyte leukemia (APL). Amplification of ERBB2 is observed in
human salivary gland adenocarcinoma (Semba et al., Proc. Nat. Acad.
Sci. 1985, 82:6497-6501) and in a gastric cancer cell line
(Fukushige et al., Molec. Cell. Biol. 1986, 6:955-958).
Overexpression of ERBB2 has been implicated in the neoplastic
transformation of prostate cancer. Interleukin-6 (IL6) is a
cytokine that was initially recognized as a regulator of immune and
inflammatory responses, but also regulates the growth of many tumor
cells, including prostate cancer. Qui et al. showed that treatment
of a prostate cancer cell line with IL6 induces tyrosine
phosphorylation of ERBB2 and ERBB3, but not ERBB1/EGFR. ERBB2 also
forms a complex with the gp130 subunit of the IL6 receptor (IL6R)
in an IL6-dependent manner. This association is important because
the inhibition of ERBB2 activity results in abrogation of
IL6-induced MAPK activation (Qui et al., Nature 1998, 393:83-85).
Thus, ERBB2 is a critical component of IL6 signaling through the
MAP kinase pathway. Additionally, overexpression of ERBB2 confers
Taxol resistance in breast cancers by inhibiting p34 (CDC2)
activation (Yu et al., Molec. Cell 1998, 2:581-91) (OMIM MIM
Number: 164870: Jan. 30, 2006). Her2/Erbb2 inhibitors may be useful
in treating prostate and breast cancer.
[0797] Her4/Erbb4: Target kinase Her4/Erbb4 (i.e., Human EGF
receptor 4) is a 146.8 kDa transmembrane tyrosine kinase encoded by
chromosome 2q33.3-q34 (symbol: ERBB4). According to OMIM, the
HER.sup.4/ERBB4 gene is a member of the type I receptor tyrosine
kinase subfamily that includes EGFR, ERBB2, and ERBB3. The gene
product of ERBB4 is a receptor for NDF/heregulin, which are
essential for neuronal development, Her4/Erbb4-/- mouse embryos
exhibit axonal misprojections which correlate with aberrant
migration of a subpopulation of hindbrain-derived cranial neural
crest cells. Accordingly, Her4/Erbb4 signaling provides patterning
information essential for the proper migration of neural crest
cells (OMIM MIM Number: 600543: Jul. 27, 2005). Her4/Erbb4
inhibitors may be useful in treating childhood medulloblastoma.
[0798] IGF1R: Target kinase IGF1R (insulin-like growth factor 1
receptor) is a 154.8 kDa receptor tyrosine kinase encoded by
chromosome 15q26.1 (symbol: IGF1R). Overexpressed in breast and
prostate cancer, acting to enhance tumor cell survival. IGF1R
inhibitors may be useful in treating prostate cancer and
hepatocellular carcinoma.
[0799] IKK beta: Target kinase IKK beta (i.e., inhibitor of nuclear
factor kappa B kinase beta) is a 86.6 kDa STK encoded by chromosome
8p11.2 (symbol: IKBKB). According to OMIM, IKK beta phosphorylates
serine residues of I-kappa-B proteins which marks them for
destruction via the ubiquitination pathway, thereby allowing
activation of the NF-kappa-B complex. Activated NF-.kappa.B complex
translocates into the nucleus and binds DNA at kappa-B-binding
motifs. Yin et al (Nature 396: 77-80, 1998) have shown that the
antiinflammatory properties of aspirin and salicylate are mediated
in part by their specific inhibition of IKK-beta, thereby
preventing activation by NF-kappa-B of genes involved in the
pathogenesis of the inflammatory response. Rossi et al (403:
103-108, 2000) demonstrated a novel mechanism of antiinflammatory
activity that was based on the direct inhibition and modification
of the IKK-beta subunit of IKK. Since IKK-beta is responsible for
the activation of NF-kappa-B by proinflammatory stimuli, Rossi et
al. (ibid/) suggested that their findings explained how
cyclopentenone prostaglandins function and can be used to improve
the utility of COX2 inhibitors (OMIM MIM Number: 603258: Nov. 16,
2005).
[0800] IKKbeta inhibitors may be useful in treating leukemia of
T-cells, necrosis, neoplasms, insulin resistance, and malignant
neoplasms.
[0801] Irak4: Target kinase Irak4 (i.e., Interleukin 1 receptor
associated kinase 4) is a 51.5 kDa serine/threonine kinase encoded
by chromosome 12q12 (symbol: IRAK4). Interleukin-1 receptor
associated kinases (e.g., IRAK1) are important mediators in the
signal transduction of Toll-like receptor (TLR, e.g., TLR.sup.4)
and IL1R family members are collectively referred to as TIRs. Irak4
functions in this signal transduction pathway. The structure of
Irak4 comprises a DEATH domain adjacent a STK domain. The DEATH
domain is a protein-protein interaction motif found in certain
proteins of the apoptotic pathway.
[0802] Irak4 was originally identified as NY-REN-64, one of 65
human tumor antigens recognized by autologous antibodies from
patients with renal cell carcinoma using serological analysis of
recombinant cDNA expression libraries (SEREX). Sequence analysis of
the NY-REN-64 cDNA clone identified in the SEREX screen revealed a
novel gene encoding a transcript of 2.8 kilobases and a predicted
protein of 460 amino acids (Genbank Accession AF155118) noted to
bear a protein kinase motif (Scanlan et al., Int. J. Cancer, 1999,
83,456-464). Based on its homology to the other IL-1
receptor-associated kinases, this gene has more recently been
placed in the IRAK family and given the name IL-1
receptor-associated kinase-4.
[0803] Irak4 is required for the efficient recruitment of IRAK1 to
the IL-1 receptor complex following IL-1 engagement, triggering
intracellular signaling cascadcs leading to transcriptional
up-regulation and mRNA stabilization. Irak4 Phosphorylates Irak1.
Effective Irak4 functioning is crucial for protective immunity
against specific bacteria, including pyogenic bacterial, but is
redundant against other microorganism. Irak4 inhibitors may be
useful in treating immunodeficiency syndrome, Crohn's disease,
ulcerative colitis, asthma, chronic bronchitis, cardio hypertrophy,
and kidney hypertension.
[0804] Itk: Target kinase Itk (i.e., IL-2 inducible T-cell kinase)
is a tyrosine kinase of 71.8 kDa encoded by chromosome 5q31-q32
(symbol: ITK). Itk is a T-cell specific homology of kinase Btk. The
EMT Tec family kinases are non-receptor type protein-tyrosine
kinase that are highly expressed in many hematopoietic cell lines.
The TEC-family protein tyrosine kinases ITK, RLK(TXK) and TEC have
been identified as key components of T-cell-receptor signalling
that contribute to the regulation of phospholipase C-gamma, the
mobilization of Ca2+ and the activation of mitogen-activated
protein kinases. Recent data also show that TEC kinases contribute
to T-cell-receptor-driven actin reorganization and cell
polarization, which are required for productive T-cell activation.
Functional studies have implicated TEC kinases as important
mediators of pathways that control the differentiation of CD4+ T
helper cells (Schwartzberg et al., 2005, Nature immunology,
5:284).
[0805] T cells express three TEC kinases, ITK, RLK and TEC, all of
which are activated downstream of the T-cell receptor (TCR) ( Berg,
L. J et al., Annu. Rev. Immunol., 2005, 23:549) and have been shown
to be involved in signaling through the TCR (Schaeffer, E. M. et
al., 1999, Science, 284:638). Although ITK, RLK and TEC are all
found in T cells, they are expressed at different levels and by
different subpopulations. (Lucas, J. A et al., 2003, Immunol. Rev.,
191;119. Colgan, J. et al. 2004, Immunity, 21:189). High expression
of TEC was seen in each of 3 patients examined with myelodysplastic
syndrome (Sato K et al., 1994, Leukemia, 8:1663). Although no human
disease has been associated with mutations of the TEC kinases that
are expressed by T cells, ITK-deficient mice have specific defects
in T Helper 2 (TH2)-cell responses and reduced pathology in models
of allergic asthma (Fowell et al. 1999, Immunity, 11:399). Specific
ITK inhibitors reduce disease in a mouse model of allergic asthma
19, TEC kinases are activated through phosphorylation by SRC-family
kinases, such as LCK, and recruitment to the plasma membrane
through binding of PtdIns(3,4,5)P3, where they are brought into
TCRsignalling complexes through interactions with SLP76, LAT and
other molecules (Bunnell, S. C. et al. 2000, J. Biol. Chem.,
275:2219,).
[0806] Consistent with a role for ITK in allergic responses,
increased ITK expression has been seen in peripheral blood T cells
from humans with atopic dermatitis (Matsumoto, Y. et al., 2002,
Int. Arch. Allergy Immunol. 129:327). Importantly, Itk-/- mice
cannot mount effective TH2-cell responses to infection with many
pathogens that are used to evaluate TH2-cell differentiation,
including Nippostrongylus brasiliensis, Schistosoma mansoni and
Leishmania major (Fowell, D. J. et al. 1999, Immunity, 11:399.
Schaeffer, E. et al. 2001, Nature Immunol., 2:1183).
[0807] TH2-cell responses have been implicated in the pathology of
allergic asthma, which is characterized by an increased number of
TH2 cells in the lungs, increased TH2-cytokine production,
increased mucus production in the lungs and inflammation of the
airways (Cohn, L et al. 2004, Annu. Rev. Immunol., 22:789). For
several reasons, ITK, however, might be an ideal therapeutic target
for TH2-cell-mediated diseases, provided that the inhibitor has a
high degree of specificity. Itk inhibitors may be useful in
treating allergic asthma.
[0808] Jak1: Target kinase Jak1 (i.e., Janus kinase 1) is a 132 kDa
tyrosine kinase, encoded by chromosome 1p31.3 (symbol: JAK1). Jak1
inhibitors may be useful in treating Hepatitis C virus
infection.
[0809] Jak2: Target kinase Jak2 (i.e., Janus kinase 2) is a 130.7
kDa tyrosine kinase encoded by chromosome 9p24 (symbol: JAK2). Jak2
inhibitors may be useful in treating myeloproliferative disorders
such as polycythaemia vera, myelofibrosis, essential
thrombocythemia, myeloid metaplasia and leukemias, including acute
lymphoblastic leukemia, chronic neutrophilic leukemia, juvenile
myelomonocytic leukemia, CMML, Philadelphia chromosome-negative
CML, megakaryocytic leukemia, and acute erythroid leukemia
[0810] Jak3: Target kinase Jak3 (i.e., Janus kinase 3) is a 125.1
kDa tyrosine kinase encoded by chromosome 19p13.1 (symbol: JAK3).
According to OMIM, JAK3 is a member of the Janus kinase (JAK)
family of tyrosine kinases involved in cytokine receptor-mediated
intracellular signal transduction. Interleukin-2 (IL2) signaling
requires the dimerization of IL2 receptor-beta (IL2RB) with the
common gamma chain (gamma-c; IL2RG). Mutations in the IL2RG gene
cause X-linked severe combined immunodeficiency. Interleukins IL2,
IL4, IL7, IL9, and IL15, whose receptors are known to contain the
common gamma chain, induce the tyrosine phosphorylation and
activation of Jak3. Truncations of gamma-c and a point mutation of
gamma-c, causing moderate X-linked combined immunodeficiency,
decrease the association between the common gamma chain and Jak3.
Since mutations in the IL2RG gene in at least some XSCID and XCID
patients prevent normal Jak3 activation, mutations in Jak3 may
result in an XSCID-like phenotype (OMIM MIM Number: 600173: Apr. 4,
2006). A related kinase, Jak2, is activated through mutation in
patients with a variety of myeloproliferative disorders (Kralovics
R. et al. N Engl J Med. 2005 352:1779-90). The role of Jak3 in B
and T lymphocyte maturation and T cell function makes Jak3 a target
for treating transplant rejection and autoimmune diseases. Jak3
inhibitors may be useful in treating X-linked severe combined
immunodeficiency, myeloproliferative disorders, transplant
rejection and autoimmune diseases such as rheumatoid arthritis,
inflammatory bowel syndrome, Crohn's disease, systemic lupus
erythematosis, ulcerative colitis, psoriasis and multiple
sclerosis.
[0811] Jnk1: Target Jnk1 (i.e., c-Jun kinase 1) is a 48.3 kDa
serine/threonine kinase encoded by chromosome 10q11.22 (symbol:
MAPK8), also known as mitogen-activated protein kinase 8. Jnk1 is a
mitogen-activated protein kinase (i.e., MAPK) which form a family
of serine-threonine protein kinases that participate in a major
signaling system by which cells transduce extracellular stimuli
into intracellular responses. MAPKs comprise the extracellular
regulated kinases, or ERKs, for example Erk2, and the
stress-activated protein kinases (SAPKs). MAPKs respond to
activation by environmental stress and pro-inflammatory cytokines
by phosphorylating a number of transcription factors, primarily
components of AP-1 such as c-Jun and ATF2 and thus regulates AP-1
transcriptional activity. In T-cells, Jnk1 and Jnk2 are required
for polarized differentiation of T-helper cells into Th1 cells.
Jnk1 is activated by threonine and tyrosine phosphorylation by
either of two dual specificity kinases, MAP2K4 and MAP2K7 and
inhibited by dual specificity phosphatases, such as DUSP1. Jnk1
inhibitors may be useful in treating type 1 diabetes, type 2
diabetes, metabolic syndrome, obesity and hepatic steatosis.
[0812] Jnk2: Target kinase Jnk2 (i.e., c-Jun kinase 2) is 48.1 kDa
serine/threonine kinase encoded by chromosome 5q35 (symbol: MAPK9).
According to OMIM, the transcriptional activity of the c-Jun
protooncoprotein is augmented through phosphorylation at two sites
by c-Jun kinases (JNKs). Using in-gel kinase assays, Hibi et al.
(1993) identified 2 JNKs, 46 and 55 kD in size. The 46-kD protein
Jnk1 was shown to be a member of the mitogen-activated protein
kinase (MAPK) family. Using a JNK1 cDNA as a probe, Kallunki et al.
(1994) and Sluss et al. (1994) isolated cDNAs encoding the 55-kD
protein, which both designated Jnk2. Kallunki et al. (1994)
reported that the sequence of the predicted 424-amino acid JNK2
protein is 83% identical to that of JNK1. Both JNKs contain a
thr-pro-tyr phosphorylation motif. Northern blot analysis revealed
that JNK2 is expressed as multiple transcripts in many cell types
(OMIM MIM Number: 602896: Jul. 7, 2005).
[0813] Jnk2 responds to activation by environmental stress and
pro-inflammatory cytokines by phosphorylating a number of
transcription factors, primarily components of AP-1 such as c-Jun
and ATF2 and thus regulates AP-1 transcriptional activity. In
T-cells, JNK1 and JNK2 are required for polarized differentiation
of T-helper cells into Th1 cells. Jnk2 isoforms display different
binding patterns: alpha-1 and alpha-2 preferentially bind to c-Jun,
whereas beta-1 and beta-2 bind to ATF2. However, there is no
correlation between bindirig and phosphorylation, which is achieved
at about the same efficiency by all isoforms. Jnk2 is activated by
threonine and tyrosine phosphorylation by either of two dual
specificity kinases, MAP2K4 and MAP2K7. Further, Jnk2 is inhibited
by dual specificity phosphatases, such as DUSP1. Jnk2 inhibitors
may be useful in treating atherosclerosis.
[0814] Jnk3: Target kinase Jnk3 (i.e., c-Jun kinase 3) is 52.6 kDa
serine/threonine kinase encoded by chromosome 4q21-q22 (symbol:
MAPK 10). According to OMIM, the c-Jun kinases (JNKs) are members
of the mitogen-activated protein kinase (MAPK) family that activate
the Jun transcription factor. Gupta et al. (EMBO J. 1996, 15:
2760-2770) isolated brain cDNAs encoding 10 different JNK isoforms,
8 of which were derived from either JNK1 or JNK2. The other 2 cDNAs
were from a gene that the authors designated JNK3. JNK3 contains an
extended N-terminal region not found in JNK1 or JNK2. The 2 JNK3
isoforms, called JNK3-alpha-1 and JNK3-alpha-2, have different C
termini. By SDS-PAGE of in vitro transcription/translation
products, Gupta et al. (ibid.) determined that JNK3-alpha-1
migrates as a 45-to-48-kD doublet and JNK3-alpha-2 migrates as a
54-to-57-kD doublet. They stated that the lower band probably
represents translation from a second in-frame start codon that
corresponds to the first codon in JNK1 and JNK2. All the JNKs were
activated by treatment of cells with the inflammatory cytokine IL1.
Multiple JNK isoforms were shown to be inactivated by MKP1.
Comparison of the binding activity of the JNK isoforms demonstrated
that they differ in their interactions with the ATF2 (CREB2), ELK1,
and Jun transcription factors. Gupta et al. (ibid.) suggested that
individual JNKs selectively target specific transcription factors
in vivo, providing a mechanism for the generation of
tissue-specific responses to the activation of the JNK signal
transduction pathway. Mohit et al. (Neuron 1995, 14: 67-78)
identified JNK3, or p49-3F12 kinase, as the gene encoding a 49-kD
antigen found in the hippocampus and neocortex. The distribution of
JNK3-expressing neurons closely matches that of Alzheimer disease
targeted neurons in those areas of the brain. Northern blot
analysis revealed that JNK3 is expressed as a 2.7-kb mRNA
exclusively in the nervous system. Mice defective for Jnk3 are
resistant to excitotoxicity induced apoptosis (Yang D. D. et al.,
Nature 1997, 389:865) (OMIM MIM Number: 602897: Mar. 13, 2006).
[0815] Jnk3 responds to activation by environmental stress and
pro-inflammatory cytokines by phosphorylating a number of
transcription factors, primarily components of AP-1 such as c-Jun
and ATF2 and thus regulates AP-1 transcriptional activity. Jnk3 is
required for stress-induced neuronal apoptosis and the pathogenesis
of glutamate excitotoxicity, Jnk3 is activated by threonine and
tyrosine phosphorylation by two dual specificity kinases, MAP2K4
and MAP2K7. MAP2K7 phosphorylates MAPK10 on Thr-221 causing a
conformational change and a large increase in Vmax. MAP2K4 then
phosphorylates Tyr-223 resulting in a further increase in Vmax.
Jnk3 is inhibited by dual specificity phosphatases, such as
DUSP1.
[0816] Jnk3 inhibitors may be useful in treating inflammatory
diseases including autoimmune diseases such as rheumatoid
arthritis, inflammatory bowel syndrome, Crohn's disease, systemic
lupus erythematosis, Sjogren's Syndrome, psoriasis and multiple
sclerosis, airway inflammatory diseases such as asthma, allergy,
pulmonary fibrosis, and chronic obstructive pulmonary disease, and
inflammation in other organs, such as CNS inflammation,
pancreatitis, nephritis, and hepatitis; neurologic diseases such as
stroke, cerebrovascular ischemia, and neurodegenerative diseases
such as Parkinson's disease, Alzheimer's disease, and Huntington's
disease; and neoplastic diseases such as prostate tumors and
myeloid leukemia.
[0817] Kdr: Target kinase Kdr (i.e., Kinase Insert Domain Receptor)
is a transmembrane tyrosine kinase of 151.5 kDa encoded by
chromosome 4q12 (symbol: KDR). Kdr has a complex secondary
structure comprising three Ig-like (i.e., immunoglobulin-like)
domains, one IGC2 (i.e., immunoglobulin-like C2-type) domain, two
additional Ig-like domains, one additional IGC2 domain, one TM
(i.e., transmembrane) domain, and a split TK domain. Kdr, also
known as VEGFR2 (i.e., Vascular Endothelial Growth Factor Receptor
2), Flt2, and Flk1 (i.e., fetal liver kinase 1), is the receptor
for Vegf and VegfC (i.e., Vascular endothelial growth factor C) and
plays a key role in vascular development and regulation of vascular
permeability. Walter et al. (Genes Chromosomes Cancer, 2002,
33:295-303) has proposed based on an observed mutation in the
kinase domain of KDR that a potential mechanism involved in
hemangioma formation is the alteration of the VEGF signaling
pathway in endothelial and/or pericytic cells.
[0818] Due to the role of angiogenesis, and aberrant control
thereof in pathologic states, Kdr is a target for therapeutic
intervention. Angiogenesis is the process by which new blood vessel
growth occurs from pre-existing vasculature and is mediated through
multiple pro-angiogenic factors, including for example Kdr. Under
normal adult physiological conditions, angiogenesis occurs during
wound healing, organ regeneration, and in some aspects of female
reproductive function. Angiogenesis is also important for the
progression of many pathological disorders such as solid tumor
growth (ovarian, lung, breast, prancreatic, prostate, colon,
gastrointestinal stromal tumor, non small cell lung cancer, and
epidermoid cancer), metastasis, psoriasis, rheumatoid arthritis,
diabetic retinopathy and age related macular degeneration, (Hoeben
et al., 2004, Pharmacol. Rev. 56:549-580).
[0819] When the dimeric cytokine VEGF binds to the receptor
tyrosine kinases Flt-1 and/or KDR, receptor dimerization occurs,
followed by autophosphorylation which leads to kinase activation
and phosphorylation of intracellular substrates. This receptor
tyrosine kinase activity initiates a cellular signaling pathway
which leads to endothelial cell proliferation and migration that is
necessary for the process of angiogenesis.
[0820] Tumors that grow beyond 1-2 mm in size require the process
of angiogenesis in order to receive the appropriate nutrients and
oxygen that is required for tumor progression. Accordingly,
inhibition of this process by small molecules that bind to the
surface of receptor tyrosine kinases such as, for example, Kdr,
inhibits tumor growth in both animal and human models. Direct
evidence of the role of VEGF as a tumor angiogenesis factor in vivo
is shown in studies in which VEGF expression or VEGF activity was
inhibited. This, has been achieved with anti-VEGF antibodies, with
dominant-negative VEGFR-2 mutants which inhibited signal
transduction, and with anti-sense VEGF RNA techniques. All
approaches led to the reduction in tumor cell lines in vivo as a
result of inhibited tumor angiogenesis. (Scappaticci, 2002, J.
Clin. Oncology 20(18):3906-3927 and references within).
[0821] Kdr inhibitors may be useful in treating solid tumor growth
(e.g. ovarian, lung, breast, prancreatic, prostate, colon,
gastrointestinal stromal tumor, non small cell lung cancer, and
epidermoid cancer), metastasis, psoriasis, rheumatoid arthritis,
diabetic retinopathy and age related macular degeneration.
[0822] Kit: Target kinase Kit (i.e., feline Hardy-Zuckerman 4
sarcoma viral oncogene) is a 109.9 kDa transmembrane tyrosine
kinase encoded by chromosome 4q12 (symbol: KIT). Receptor protein
tyrosine kinases (RPTKs) regulate key signal transduction cascades
that control cellular growth and proliferation. The Stem Cell
Factor (SCF) receptor Kit is a type III transmembrane RPTK that
includes five extracellular immunoglobulin (IG) domains, a single
transmembrane domain, and a split cytoplasmic kinase domain
separated by a kinase insert segment. Kit plays an important role
in the development of melanocytes, mast, germ, and hematopoietic
cells.
[0823] Stem Cell Factor (SCF) is a protein encoded by the S1 locus,
and has also been called kit ligand (KL) and mast cell growth
factor (MGF), based on the biological properties used to identify
it (reviewed in Tsujimura, Pathol Int 1996, 46:933-938; Loveland,
et al., J. Endocrinol 1997, 153:337-344; Vliagoftis, et al., Clin
Immunol 1997, 100:435-440; Broudy, Blood 1997, 90:1345-1364;
Pignon, Hermatol Cell Ther 1997, 39:114-116; and Lyman, et al.,
Blood 1998, 91:1101-1134,). Herein the abbreviation SCF refers to
the ligand for Kit.
[0824] SCF is synthesized as a transmembrane protein with a
molecular weight of 220 or 248 Dalton, depending on alternative
splicing of the mRNA to encode exon 6. The larger protein can be
proteolytically cleaved to form a soluble, glycosylated protein
which noncovalently dimerizes. Both the soluble and membrane-bound
forms of SCF can bind to and activate Kit. For example, in the
skin, SCF is predominantly expressed by fibroblasts, keratinocytes,
and endothelial cells, which modulate the activity of melanocytes
and mast cells expressing Kit. In bone, marrow stromal cells
express SCF and regulate hematopoiesis of Kit expressing stem
cells. In the gastrointestinal tract, intestinal epithelial cells
express SCF and affect the interstitial cells of Cajal and
intraepithelial lymphocytes. In the testis, sertoli cells and
granulosa cells express SCF which regulates spermatogenesis by
interaction with Kit on germ cells.
[0825] According to OMIM, Signaling from Kit is essential for
primordial germ cell growth both in vivo and in vitro. Many
downstream effectors of the KIT signaling pathway have been
identified in other cell types, but how these molecules control
primordial germ cell survival and proliferation are unknown.
Determination of the KIT effectors acting in primordial germ cells
has been hampered by the lack of effective methods to manipulate
easily gene expression in these cells. De Miguel et al. (2002)
overcame this problem by testing the efficacy of
retroviral-mediated gene transfer for manipulating gene expression
in mammalian germ cells. They found that primordial germ cells can
successfully be infected with a variety of types of retroviruses.
They used this method to demonstrate an important role of the AKT1
in regulating primordial germ cell growth (OMIM MIM Number: 164920:
Apr. 17, 2006).
[0826] Aberrant expression and/or activation of Kit has been
implicated in a variety of pathologic states. For example, evidence
for a contribution of Kit to neoplastic pathology includes its
association with leukemias and mast cell tumors, small cell lung
cancer, testicular cancer, and some cancers of the gastrointestinal
tract and central nervous system. In addition, Kit has been
implicated in playing a role in carcinogenesis of the female
genital tract sarcomas of neuroectodermal origin, and Schwann cell
neoplasia associated with neurofibromatosis. It was found that mast
cells are involved in modifying the tumor micro environment and
enhancing tumor growth (Yang et al., J Clin Invest. 2003,
112:1851-1861; Viskochil, J Clin Invest. 2003, 112:1791-1793).
[0827] Kit inhibitors may be useful in treating malignancies,
including mast cell tumors, small cell lung cancer, testicular
cancer, gastrointestinal stromal tumors (GISTs), glioblastoma,
astrocytoma, neuroblastoma, carcinomas of the female genital tract,
sarcomas of neuroectodermal origin, colorectal carcinoma, carcinoma
in situ, Schwann cell neoplasia associated with neurofibromatosis,
acute myelocytic leukemia, acute lymphocytic leukemia, chronic
myelogenous leukemia, mastocytosis, melanoma, and canine mast cell
tumors, and inflammatory diseases, including asthma, rheumatoid
arthritis, allergic rhinitis, multiple sclerosis, inflammatory
bowel syndrome, transplant rejection, and hypereosinophilia.
[0828] LCK: Target kinase MCK (i.e., lymphocyte-specific protein
tyrosine kinase) is a 57.9 kDa membrane associated non receptor
tyrosine kinase encoded by chromosome 1p34.3 (symbol: LCK). The
protein structure comprises an SH3 and SH.sub.2 domain. LCK
inhibitors may be useful in treating acute lymphoblastic leukemia,
T-cell lymphoma, lymphopenia, renal carcinoma, colon carcinoma,
severe combined immunodeficiency, multiple sclerosis, inflammatory
bowel and type 1 diabetes.
[0829] MAP2K1: Target kinase MAP2K1 (i.e., Mitogen-activated
protein kinase kinase 1) is a threonine/tyrosine kinase of 43.3 kDa
encoded by chromosome 15q22.1-q22.33 (symbol: MAP2K1). According to
OMIM, MAP2K1 is also known as MEK1 (i.e., MAPK/ERK Kinase 1).
Mitogen-activated protein (MAP) kinases, also known as
extracellular signal-regulated kinases (ERKs) are thought to act as
an integration point for multiple biochemical signals because they
are activated by a wide variety of extracellular signals, are
rapidly phosphorylated on threonine and tyrosine residues, and are
highly conserved in evolution (Crews et al., Science 1992 258:
478-480). MAP2K1 is a critical protein kinase lying upstream of MAP
kinase which stimulates the enzymatic activity of MAP kinase Crews
et al. (ibid.) found that Mek1 (i.e., MAP2K1) expressed in bacteria
phosphorylates the Erk gene product in vitro. They showed that the
Mek1 gene is highly expressed in murine brain. Seger et al. (J.
Biol. Chem., 1992, 267:25628-25631) cloned a cDNA encoding the
human homolog of Mek1, symbolized MKK1 by them, from a human T-cell
cDNA library. When overexpressed in COS cells, the predicted
43,439-Da protein led to increased phorbol ester-stimulated MAP
kinase kinase activity. They also isolated a related cDNA, called
MKK1b, that appears to be an alternatively spliced form of MKK1.
Seger et al. (ibid.) detected a 2.6-kb MKK1 transcript by Northern
blot analysis in all tissues examined. Zheng and Guan (J. Biol.
Chem., 1993, 268: 11435-11439) also cloned a human cDNA
corresponding to MEK1. They noted that the 393-amino acid protein
shares 99% amino acid identity with murine Mek1 and 80% homology
with human MEK2. The authors characterized biochemically the human
MEK1 and MEK2 gene products. The gene is also symbolized MAP2K1, or
PRKMK1. MAP2K1 catalyzes the concomitant phosphorylation of a
threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located
in MAP kinases and activates ERK1 and ERK2 MAP kinases. Certain
inhibitors of MEK1 are potent anti-cancer agents (Sebolt-Leopold,
J. S., et a)., Nat. Med. 1999, 5:810) (OMIM MIM Number: 176872:
Jun. 6, 2005). MAP2K1 inhibitors may be useful in treating acute
myeloid leukemia, breast, ovarian and liver cancer.
[0830] MAP2K2: Target kinase MAP2K2 (i.e., Mitogen-activated
protein kinase kinase 2) is a threonine/tyrosine kinase of 44.4 kDa
encoded by chromosome 7q32 (symbol: MAP2K2); MAP2K2 is also known
as Mek2; see MAP2K1 above. According to OMIM, Zheng and Guan
(ibid.) isolated and sequenced 2 human cDNAs encoding members of
the MAP kinase kinase (MAP2K) family, designated MEK1 and MEK2 by
them. The MEK2 cDNA encodes a predicted 400-amino acid protein that
shares 80% sequence identity with human MEK1. Zheng and Guan
(ibid.) showed that recombinant MEK2 and MEK1 both could activate
human Erk1 in vitro. They further characterized biochemically both
MAP2Ks (OMIM MIM Number: 601263: Oct. 23, 2003).
[0831] The mitogen-activated protein kinase (MAPK) pathway is a
major pathway in the cellular signal transduction cascade from
growth factors to the cell nucleus. The pathway involves kinases at
two, levels: MAP kinase kinases (MAPKK), and their substrates MAP
(mitogen activated protein) kinases (MAPK). There are different
isoforms in the MAP kinase family. [For review, see Seger, R.;
Krebs, E. G. FASEB, 9, 726, (1995)]. The compounds of this
invention can inhibit the action of one or both of these kinases:
MEK, a MAP kinase kinase, and its substrate ERK, a MAP kinase. ERK
(extracellular regulated kinases), a p42 MAPK, is found to be
essential for cell proliferation and differentiation.
Overexpression and/or over activation of MEK or ERK has been found
to be associated with various human cancers [For example,
Sivaraman, V. S. et al., C. C. J. Clin. Invest., 99, 1478 (1997)].
It has been demonstrated that inhibition of MEK prevents activation
of ERK and subsequent activation of ERK substrates in cells,
resulting in inhibition of cell growth stimulation and reversal of
the phenotype of ras-transformed cells [Dudley, D. T. et al., Proc.
Nat. Acad. Sci., 92, 7686 (1995)]. MAP2K2 inhibitors may be useful
in treating cancer and inflammation.
[0832] MAP4K4: Target kinase MAP4K4 (i.e., Mitogen-activated
protein kinase kinase 4) is a serine threonine kinase of 152.1 kDa
encoded by chromosome 2q11.2 (symbol: MAP4K4) and is also known as
HGK. MAP4K4 inhibitors may be useful in treating cancer, tumor
metastasis, diabetes and metabolic syndrome.
[0833] MAPKAPK2: Target kinase MAPKAPK2 (i.e., Mitogen activated
protein kinase activated protein kinase 2) is 45.6 kDa
serine/threonine kinase encoded by chromosome 1q32 (symbol:
MAPKAPK2). According to OMIM, Stokoe et al. (Biochem. J. 1993,
296:843-849) described a protein kinase, which they designated
MAPKAP kinase-2, that was active only after phosphorylation by
mitogen-activated protein kinase (MAP kinase). They identified
several features that distinguish MAPKAP kinase-2 from the MAPKAP
kinase-1 family. Stokoe et al. (ibid.) stated that MAPKAP kinase-2
was identified based on its in vitro phosphorylation of glycogen
synthase; however, its phosphorylation of glycogen synthase had not
been shown in vivo. Stokoe et al. (ibid.) cloned a partial human
MAPKAP kinase-2 cDNA from a teratocarcinoma cell line cDNA library.
The cDNA sequence revealed the following features (in 5-prime to
3-prime order): a proline-rich region containing 2 putative
SH3-binding sites, a kinase catalytic domain, a threonine residue
phosphorylated by MAP kinase, and a nuclear localization signal. By
Northern analysis, Stokoe et al. (ibid.) demonstrated that the gene
is expressed as a 3.3-kb transcript in all of the 6 human tissues
tested. The physiological substrate of MAPKAP kinase 2 appears to
be the small heat shock protein (HSP27/HSP25) (OMIM MIM Number:
602006: Mar. 3, 2005).
[0834] In vitro, MAPKAP kinase 2 can phosphorylate glycogen
synthase at Ser-7 and tyrosine hydroxylase (on Ser-19 and Ser-40).
This kinase phosphorylates Ser in the peptide sequence,
Hyd-X--R--X(2)-S, where Hyd is a large hydrophobic residue. MAPKAP
kinase 2 is activated by two distinct pathways: the first involves
the stimulation of p42/p44 MAPK by growth factors, and the second,
triggered by stress and heat shock, depends on the activation of
MPK2 and upstream MAPKK/MAPKKK. MAPKAPK2, inhibitors may be useful
in treating cancer (e.g. prostate, breast), stroke, menengitis, and
inflammatory disorders.
[0835] Met: Target kinase Met (i.e., Hepatocyte growth factor
receptor) is 155.5 kDa transmembrane tyrosine kinase encoded by
chromosome 7q31 (symbol: MET). According to OMIM, Cooper et al.
(Nature 1984, 311: 29-33) cloned a transforming gene from a
chemically transformed human osteosarcoma-derived cell line and
mapped it to 7p11.4-qter. Identity to all previously known
oncogenes except ERBB was ruled out by the fact that they are
encoded by other chromosomes; identity to ERBB is probably excluded
by failure of direct hybridizations of the 2 probes. MET was the
designation suggested by Cooper et al. (ibid.). Dean et al. (Nature
1985, 318: 385-388) showed that MET is in the tyrosine kinase
family of oncogenes. It appeared to be most closely related in
sequence to the human insulin receptor and ABL oncogene. From the
sequence of MET cDNA, Park et al. (Proc. Nat. Acad. Sci. 1987, 84:
6379-6383) concluded that this oncogene is a cell-surface receptor
for a then unknown ligand. The cellular MET protooncogene product
is a receptor-like tyrosine kinase comprised of disulfide-linked
subunits of 50 kD (alpha) and 145 kD (beta). In the fully processed
Met product, the alpha subunit is extracellular, and the beta
subunit has extracellular, transmembrane, and tyrosine kinase
domains as well as sites of tyrosine phosphorylation (OMIM MIM
Number: 164860: Oct. 18, 2005). Met inhibitors may be useful in
treating a variety of neoplasms including kidney, breast, bladder,
non-small-cell lung, colorectal, and bladder cancers, and in
hepatocellular carcinoma.
[0836] MLK1: Target kinase MLK1 (i.e., mixed-lineage kinase 1, aka
mitogen-activated protein kinase kinase kinase 9) is a 121.9 kDa
serine/threonine kinase encoded by chromosome 14q24.3-q31 (symbol:
MAP3K9). MLK1 is expressed in epithelial tumor cell lines of colon,
breast and esophageal origin. Silva et al. review the mixed lineage
kinase (MLK)-c-jun N-terminal kinase (JNK) signaling cascade, which
leads to the phosphorylation and activation of the transcription
factor c-jun. There is much evidence, from in vitro and in vivo
studies, that this cascade can mediate cell death. In addition,
there is evidence that it is operative upstream in the death
process. It is possible that abrogation of this pathway may
forestall death before irreversible cellular injury. They review
the evidence that inhibition of the MLKs can prevent dopamine
neuron cell death and the degeneration of their axons (Silva et
al., Mov Disord 2005, 20(6):653-64). Lund et al. state that MLK
inhibitor CEP-1347 blocks the activation of the c-Jun/JNK apoptotic
pathway in neurons exposed to various stressors and attenuates
neurodegeneration in animal models of Parkinson's disease (PD).
Microglial activation may involve kinase pathways controlled by
MLKs and might contribute to the pathology of neurodegenerative
diseases. They explored the possibility that CEP-1347 modulates the
microglial inflammatory response [tumour necrosis factor-alpha
(TNF-alpha), interleukin-6 (IL-6), and monocyte chemotactic
protein-1 (MCP-1)] and report that the MLK inhibitor CEP-1347
reduced cytokine production in primary cultures of human and murine
microglia, and in monocyte/macrophage-derived cell lines,
stimulated with various endotoxins or the plaque forming peptide
Abeta1-40. Moreover, CEP-1347 inhibited brain TNF production
induced by intracerebroventricular injection of lipopolysaccharide
in mice. As expected from a MLK inhibitor, CEP-1347 acted upstream
of p38 and c-Jun activation in microglia by dampening the activity
of both pathways. These data imply MLKs as important, yet
unrecognized, modulators of microglial inflammation, and
demonstrate a novel anti-inflammatory potential of CEP-1347 (Lund
et al., J Neurochem 2005, 92(6): 1439-51). MLK1 inhibitors may be
useful in treating neurodegenerative disorders such as Alzheimer's
and Parkinson's disease and inflammatory disorders.
[0837] Mnk1: Target kinase Mnk1 (i.e., MAP kinase interacting
serine/threonine kinase 1) is a 51.3 kDa STK encoded by chromosome
1p34.1 (symbol: MKNK1). According to OMIM, Fukunaga and Hunter
(EMBO J. 1997, 16:1921-1933) observed that the C-terminal region of
Mnk1 was phosphorylated and activated vivo and in vitro by Erk1 and
p38 MAP kinases, but not by JNK/SAPK. Waskiewicz et al., (EMBO J.
1997, 16:1909-1920) reported that in vitro, Mnk1 rapidly
phosphorylates eIF4E at the physiologically relevant site, ser209.
In cells, they observed that Mnk1 is posttranslationally modified
and enzymatically activated in response to mitogenic and stress
stimuli. This activation could be blocked by inhibitors of MAP
kinase kinase-1 and p38, and Waskiewicz et al. (ibid.) concluded
that Mnk1 is downstream of multiple MAP kinases (OMIM MIM Number;
606724: Feb. 27, 2002).
[0838] Accordingly, dephosphorylation of eIF4E strongly correlates
with inhibition or impairment of cap-dependent mRNA translation
under certain stress conditions such as heat shock, nutrient
deprivation, oxidative or osmotic stress, and infection of
mammalian cells with certain viruses such as adenovirus (Ad) or
influenza virus, among others. Mnk1 inhibitors may be useful in
treating conditions associated with heat shock, nutrient
deprivation, oxidative or osmotic stress, infection of mammalian
cells (e.g. with viruses such as adenovirus (Ad) or influenza
virus), and autoimmune diseases.
[0839] p38: Target kinase p38 (i.e., Mitogen-activated Protein
Kinase 14) is a 41.5 kDa STK encoded by chromosome 6p21.3-p21.2
(symbol: MAPK14). According to OMIM, production of interleukin-1
and tumor necrosis factor (TNF) from stimulated human monocytes is
inhibited by a series of pyridinyl-imidazole compounds called
CSAIDs (cytokine-suppressive antiinflammatory drugs). These agents
have shown activity in a variety of animal models of acute and
chronic inflammation. Using radiolabeled chemical probes for
radioligand binding assays and photoaffinity labeling experiments,
Lee et al. (Nature 1994, 372:739-746) identified, purified,
cDNA-cloned, and biochemically characterized 2 CSBPs (CSAID-binding
proteins) as molecular targets of pyridinyl-imidazole cytokine
inhibitors. They designated the 2 closely related mitogen-activated
protein kinases (MAPKs) CSBP1 and CSBP2. Binding of
pyridinyl-imidazole compounds inhibited CSBP kinase activity and
was directly correlated with their ability to inhibit cytokine
production, suggesting that the CSBPs are critical for cytokine
production. Lee et al. (ibid.) considered the 2 to be products of
alternative splicing. The 4.2-kb CSBP mRNA encodes a predicted
360-amino acid protein and was expressed in all tissues tested.
CSBP1 and CSBP2 are identical except for a 75-nucleotide stretch
within the coding region. Han et al. (Science 1994, 265: 808-811)
cloned the mouse homolog as a protein that is tyrosine
phosphorylated as part of the protein kinase cascades induced by
endotoxic lipopolysaccharide. They named this 38-kD protein p38. As
p38 is a member of the stress-activated protein kinase (SAPK) class
of MAPKs, Goedert et al. (Genomics 1997, 41:501-502) referred to
this protein as SAPK2A. Zervos et al. (Proc. Nat. Acad. Sci. 1995,
92:10531-10534) identified p38 as a human protein that interacts
with MAX protein and designated it MXI2. The MXI2 gene encodes a
297-residue protein whose sequence indicates that it is related to
the extracellular signal-regulated kinases (ERK protein kinases).
MXI2 in yeast interacts with Max and with the G terminus of c-Myc.
MXI2 phosphorylates MAX both in vitro and in vivo. The authors
speculated that phosphorylation by MXI2 may effect the ability of
MAX to oligomerize with itself and its partners, bind DNA, or
regulate gene expression (OMIM MIM Number: 600289: Feb. 13,
2006).
[0840] There are four known isoforms of p38, i.e., p38-.alpha.,
p38.beta., p38.gamma., and p38.delta.. The .alpha. and .beta.
isoforms are expressed in inflammatory cells and are key mediators
of TNF-.alpha. production. Inhibiting the p38.alpha. and .beta.
enzymes incells results in reduced levels of TNF-.alpha.
expression. Also, administering p38.alpha. and .beta. inhibitors in
animal models of inflammatory disease has proven that such
inhibitors are effective in treating those diseases. Accordingly,
the p38 enzymes serve an important role in inflammatory processes
mediated by IL-1 and TNF-.alpha.. Compounds that reportedly inhibit
p38 kinase and cytokines such as IL-1 and TNF-.alpha. for use in
treating inflammatory diseases are disclosed in the following
published international patent applications: WO 00/12497
(quinazoline derivatives as p38 kinase inhibitors); WO 00/56738
(pyridine and pyrimidine derivatives for the same purpose); WO
00/12497 (discusses the relationship between p38 kinase
inhibitors); and WO 00/12074 (piperazine and piperidine compounds
useful as p38 inhibitors).
[0841] p38 responds to activation by environmental stress,
pro-inflammatory cytokines and lipopolysaccharide (LPS) by
phosphorylating a number of transcription factors, such as ELK1 and
ATF2 and several downstream kinases, such as MAPKAPK2 and MAPKAPK5.
Additionally, p38 plays a critical role in the production of some
cytokines, for example IL-6. p38 phosphorylates ELK1 and ATF2.
[0842] p38 is activated by threonine and tyrosine phosphorylation
by either of two dual specificity kinases, MAP2K3 or MAP2K6, and
potentially also MAP2K4 and it is inhibited by dual specificity
phosphatases, such as DUSP1. p38 is specifically inhibited by the
binding of pyridinyl-imidazole compounds, which are
cytokine-suppressive anti-inflammatory drugs (CSAID).
[0843] p38 inhibitors have the potential to treat a number of
diseases, including, but not limited to acute coronary syndrome,
stroke, atherosclerosis, and inflammatory diseases such as
rheumatoid arthritis, inflammatory bowel disease, and Crohn's
disease.
[0844] PDGFR kinase family: Related to Fms and Kit are two
platelet-derived growth factor receptors, alpha (i.e., PDGFRA) and
beta (PDGFRB). The gene coding for PDGFRA is located on chromosome
4q12 in the same region of chromosome 4 as the oncogene coding for
Kit. Most gastrointestinal stromal tumors (GIST) have activating
mutations in Kit, and most patients with GISTs respond well to
Gleevec, which inhibits Kit. Heinrich et al. (Science 2003,
299:708-10.) have shown that approximately 35% of GISTs lacking Kit
mutations have intragenic activation mutations in the gene encoding
pdgfra, and that tumors expressing Kit or PDGFRA were
indistinguishable with respect to activation of downstream
signaling intermediates and cytogenetic changes associated with
tumor progression. Thus, Kit and PDGFRA mutations appear to be
alternative and mutually exclusive oncogenic mechanisms in GISTs.
PDGF is a potent growth factor and chemoattractant for smooth
muscle cells (SMCs), and the renarrowing of coronary arteries
following angioplasty is due in part to the enhanced proliferation
of SMCs in response to increased levels of PDGF. Therefore,
compounds that inhibit the kinase activity of PDGFr may be useful
in the treatment of restenosis. In addition, since PDGF and PDGFr
are overexpressed in several types of human gliomas, small
molecules capable of suppressing PDGFr activity have potential
utility as anticancer therapeutics [Nister, M., J. Biol, Chem.,
266, 16755 (1991); Strawn, L. M., J. Biol. Chem. 269, 21215
(1994)].
[0845] PDGFR.alpha.: Target PDGFR.alpha. (i.e., Plate Derived
Growth Factor Receptor, alpha) is a 122.7 kDa transmembrane
tyrosine kinate encoded by chromosome 4q12 (symbol: PDGFRA).
According to OMIM, The KIT oncogene, another member of the PDGF
growth factor receptor subfamily, is located in the same region of
chromosome 4 (Stenman et al., Genes Chromosomes Cancer 1989,
1:155-158). PDGFR 1 (i.e. PDGFRB) and CSF1R (i.e. FMS) are also
membrane-spanning growth factor receptors with tyrosine kinase
activity. The PDGFR 1 and CSF1R genes appear to have evolved from a
common ancestral gene by gene duplication, in as much as these 2
genes are tandemly linked on chromosome 5 (Roberts et al., Cell
1988, 55: 655-661). They are oriented head-to-tail with the 5-prime
exon of FMS located only 500 bp from the last 3-prime exon of
PDGFRB. An analogous situation may exist for the PDGFR.sup.2 (i.e.
PDGFRA) and KIT genes on chromosome 4. From an evolutionary point
of view, it is possible that the distribution of these 4 loci,
PDGFR.sup.2, KIT, PDGFR1, and FMS, on chromosomes 4 and 5 is a
result of gene duplication and chromosome doubling
(tetraploidization) (OMIM MIM Number: 173490: Mar. 21, 2005).
PDGFRA inhibitors may be useful in treating idiopathic
hypereosinophilic syndrome, chronic eosinophilic leukemia, glioma,
gastrointestinal stromal tumors (GISTs), juvenile myelomonocytic
leukemia, metastatic medulloblastoma, atherogenesis, and
restenosis.
[0846] PDGFR.beta.: Target PDGFR.beta. (i.e., Plate Derived Growth
Factor Receptor, beta) is a 124.0 kDa transmembrane tyrosine kinate
encoded by chromosome 5q31-q32 (symbol: PDGFRB). According to OMIM,
stimulation of cell proliferation of the receptor for PDGF has been
implicated in atherogenesis and in cell transformation by the SIS
oncogene. Escobedo et al. (1986) sequenced the receptor and cloned
its gene. Gronwald et al. (Proc. Nat. Acad. Sci. 1988,
85:3435-3439) cloned a cDNA coding for human PDGFR and studied its
expression. The cDNA contained an open reading frame that coded for
a protein of 1,106 amino acids. In transfectants, Gronwald et al.
(ibid.) found that the PDGFR clone expressed a high affinity
receptor specific for the BB isoform of PDGF, i.e., PDGF dimers
composed of 2 B chains. There may be a separate class of PDGF
receptor that binds both the homodimers and the heterodimer.
Claesson-Welsh et al. (Molec. Cell. Biol. 1988, 8:3476-3486)
determined the structure of the human PDGF receptor as deduced from
a full-length cDNA clone. The receptor expressed in Chinese hamster
ovary cells was found to bind specifically to B-chain-containing
PDGF molecules. With the description of a second PDGF receptor, it
is necessary to use the symbol PDGFR1. Matsui et al. (1989)
designated the second type of PDGFR as type alpha because PDGF
binding was blocked by AA as well as BB isoforms of the ligand; the
product of the earlier cloned PDGF receptor was termed type beta
(OMIM MIM Number: 173410: Nov. 19, 2003).
[0847] PDGFRP has been implicated in hematological cancers, such as
chronic myelomonocytic leukemia (CMML), in which a significant
number of patients have a t(5;12)(q33;p13) translocation resulting
in TEL-PDGFR.beta. fusion protein. Golub et al. report that the
consequence of the t(5;12) translocation is expression of a fusion
transcript in which the tyrosine kinase domain of the
platelet-derived growth factor receptor beta (PDGFR.beta.) on
chromosome 5 is coupled to a novel ets-like gene, tel, on
chromosome 12. The tel-PDGFR beta fusion demonstrates the oncogenic
potential of PDGFR.beta. and may provide a paradigm for early
events in the pathogenesis of AML (Golub et al., Cell 1994,
77:307-316). PDGFRB inhibitors may be useful in treating idiopathic
hypereosinophilic syndrome, chronic eosinophilic leukemia, juvenile
myelomonocytic leukemia, and metastatic medulloblastoma.
[0848] PDPK1: Target PDPK1 (3-phosphoinositide dependent protein
kinase-1) is a 63.2 kDa serine/threonine kinase encoded by
chromosome 16p13.3 (symbol: PDPK1). PDPK1 inhibitors may be useful
in treating cancers and diabetes.
[0849] Pim1: Target Pim1 (i.e., Proviral Integration Site 1) is a
35.7 kDa serine/threonine kinase encoded by chromosome 6p21.2
(symbol: PIM1) found in the cytoplasm and nucleus. The structure of
Pim1 comprises a STK (i.e., serine/threonine kinase) domain. X-ray
crystal structures of Pim1 bound to various small molecules have
recently been solved (Jacobs, et al., J Biol Chem 2005 280:
13728-34; Qian, et al., J Biol Chem 2005 280: 6130-7; Kumar, et
al., J Mol Biol 2005 348: 183-93).
[0850] Pim1 is the first described member of a unique family of
serine/threonine kinases, which includes at least two other kinases
(Pim2 and PIM3) with significant sequence homology to Pim1 (van der
Lugt, et al., Embo J 1995 14: 2536-44; Feldman, et al., J Biol Chem
1998 273: 16535-43). The Pim1 protooncogene was originally
identified as a genetic locus frequently activated by the proviral
insertion of Moloney murine leukemia virus into mouse T cell
lymphomas (Cuypers et al., Cell 1984, 37:141-150). Several
substrates of Pim1 phosphorylation have been identified, including
c-Myb (Winn, et al., Cell Cycle 2003 2: 258-62), BAD (Yan, et al.,
J Biol Chem 2003 278: 45358-67; Aho, et al., FEBS Lett 2004 571:
43-9), SOCS-1 (Chen, et al., Proc Natl Acad Sci USA 2002 99:
2175-80), Cdc25A (Mochizuki, et al., J Biol Chem 1999 274:
18659-66), HP1 (Koike, et al., FEBS Lett 2000 467: 17-21), PAP-1
(Maita, et al., Eur J Biochem 2000 267: 5168-78), p21.sup.cipt/waf)
(Wang, et al., Biochim Biophys Acta 2002 1593: 45-55), PTP-U2S
(Wang, et al., Arch Biochem Biophys 2001 390: 9-18), and NFATc1
(Rainio, et al., J Immunol 2002 168:1524-7). Pim1 has been shown to
have diverse biological roles in cell survival, proliferation,
differentiation, and immune response. (Wang, et al., J Vet Sci 2001
2: 167-79; Bachmann, et al., Int J Biochem Cell Biol 2005 37:
726-30). However, mice lacking all three Pim genes have recently
been shown to be viable and demonstrate that the PIM kinases are
important for growth factor signaling, but are not essential for
development (Mikkers, et al., Mol Cell Biol 2004 24: 6104-15).
During embryonal development PIM genes are expressed in partially
overlapping fashion in cells in both immune and central nervous
system as well as in epithelia (Eichmann A, Yuan L, Breant C,
Alitalo K, and Koskinen P J. (2000) Developmental expression of PIM
kinases suggests functions also outside of the hematopoietic
system. Oncogene 19: 1215-1224). PIM-1, the prototypical member of
the PIM family is located both in the cytoplasm and nucleus, but
its precise role in these two locations has not been fully
elucidated.
[0851] Dysfunction of Pim1 has been implicated in the progression
of multiple cancers, including several hematopoietic and prostate
cancers. Although the exact mechanisms by which Pim1 participates
in cell transformation have not been completely elucidated, several
reports point to the ability of Pim1 to pirolongcell survival
(Lilly, et al., Cancer Res 1997 57: 5348-55); Lilly, et al.,
Oncogene 1999 18: 4022-31; Moroy, et al., Proc Natl Acad Sci USA
1993 90: 10734-8). Overexpression of Pim1 has been observed in
myeloid and lymphoid acute leukemia and Pim1 is constitutively
expressed in some myeloid leukemia cell lines (Lilly, et al.,
Oncogene 1992 7: 727-32; Amson, et al., Proc Natl Acad Sci USA 1989
86: 8857-61). Increased Pim1 expression has also been identified in
neoplastic prostate cancer specimens from patients by cDNA
microarray analysis and by anti-Pim1 antibody staining
(Dhanasekaran, et al., Nature 2001 412: 822-6). In a transgenic
murine model of prostate cancer in which human c-myc is expressed,
the gene expression profile is consistent with that seen in human
prostate cancer, including upregulation of Pim1 (Ellwood-Yen, et
al., Cancer Cell 2003 4: 223-38). In addition, Pim1 may participate
in deregulated cell growth in prostate cancer through the hormone
independent activation of the androgen receptor, a typical
characteristic of advanced prostate cancer that offers poor patient
prognosis. (Kim, et al., Oncogene 2004 23: 1838-44). The PIM-1
proto-oncogene has also been implicated in human hematopoietic
malignancies with its overexpression frequently detected in human
hematopoietic cell lines as well as in fresh tumor cells from
patients with leukemia (Nagarajan et al. Proc. Natl. Acad. Sci.
USA, 1986, 83:2556-2560; Meeker et al., Oncogene Res. 1987, 1:
87-101; Amson et al., Proc. Natl. Acad. Sci. USA, 1989, 86:
8857-8861).
[0852] In diffuse large cell lymphoma (DLCL), the most common form
of non-Hodgkin's lymphoma, Pim1 has been shown to undergo
chromosomal translocations, resulting in its overexpression
(Akasaka, et al., Cancer Res 2000 60: 2335-41). A recent study
showed that Pim1 was also the target of an aberrant somatic
hypermutation in DLCL (Pasqualucci, et al., Nature 2001 412:
341-6). Hypermutation sites are distributed in both 5' UTR and
coding sequence, and independent of the chromosomal translocations.
Notably, there are seven missense mutations introduced into the
coding exons of the gene. These missense mutations may affect the
three-dimensional structure and, in some Cases, the kinase activity
of the Pim1 protein. Hypermutations are also detected in Pim1 found
in primary central nervous system lymphomas (Montesinos-Rongen, et
al., Blood 2004 103: 1869-75) and multiple subtypes of AIDS-induced
non-Hodgkin's lymphomas (Gaidano, et al., Blood 2003 102: 1833-41).
Inhibition of Pim1 kinase activity by small molecules has the
potential to offer a therapeutic benefit in these diseases.
[0853] Transgenic mice with PIM-1 driven by Emu enhancer sequence's
demonstrated that PIM-1 function as a weak oncogene because by
itself it does not lead to tumor formation but does so after a
second oncogenic gene become overexpressed. In 75% of the tumors
over-expressing PIM-1, the second gene found to be over-expressed
is c-myc (van der Houven van Oordt C W, Schouten T G, van Krieken J
H, van Dierendonck J H, van der Eb A J, Breuer M L. (1998)
X-ray-induced lymphomagenesis in Emu-PIM-1 transgenic mice: an
investigation of the co-operating molecular events. Carcinogenesis
19:847-853). In fact when crosses were made between Emu-PIM
transgenic mice and E mu-myc transgenic mice, the combination of
genes is so oncogenic that the offsprings die in utero due to pre B
cell lymphomas (Verbeek S, van Lohuizen M, van der Valk M, Domen J,
Kraal G, and Berns A. (1991) Mice bearing the Emu-myc and Emu-PIM-1
transgenes develop pre-B-cell leukemia prenatally. Mol. Cell.
Biol., 11: 1176-1179).
[0854] Mice deficient for PIM-1 show normal synaptic transmission
and short-term plasticity but failed to consolidate enduring LTP
(i.e., long term potentiation) even though PIM-2 and PIM-3 are
expressed in the hippocampus (Konietzko U, Kauselmann G, Scafidi J,
Staubli U, Mikkers H, Berns A, Schweizer M, Waltereit R, and Kuhl
D. (1999) PIM kinase expression is induced by LTP stimulation and
required for the consolidation of enduring LTP. EMBO J. 18:
3359-3369).
[0855] Various factors are known to enhance the transcription of
PIM-1 kinase in mouse and human. PIM-1 closely cooperates with
another oncoprotein, c-myc, in triggering intracellular signals
leading to both transformation and apoptosis and the selective
inhibition of apoptotic signaling pathways leading to Bcl-2 (van
Lohuizen M, Verbeek S, Krimpenfort P, Domen J, Saris C,
Radaszkiewicz T, and Berns A. (1989) Predisposition to
lymphomagenesis in PIM-1 transgenic mice: cooperation with c-myc
and N-myc in murine leukemia virus-induced tumors. Cell 56:673-682;
Breuer M L, Cuypers H T, Berns A. (1989). Evidence for the
involvement of PIM-2, a new common proviral insertion site, in
progression of lymphomas. EMBO J. 8:743-748; Verbeek S, van
Lohuizen M, van der Valk M, Domen J, Kraal G, and Berns A. (1991)
Mice bearing the E mu-myc and E mu-PIM-1 transgenes develop
pre-B-cell leukemia prenatally. Mol. Cell. Biol. 11: 1176-1179;
Shirogane T, Fukada T, Muller J M, Shima D T, Hibi M, and Hirano T.
(1999) Synergistic roles for PIM-1 and c-Myc in STAT3-mediated cell
cycle progression and antiapoptosis. Immunity, 11: 709-719). PIM-1
kinase is induced by T cell antigen receptor cross linking by
cytokines and growth factors and by mitogens including IL2, IL3,
IL6, IL9, IL12, IL15, GM-CSF, G-CSF, IFNa, INFg, prolactin, ConA,
PMA and anti-CD3 antibodies (Zhu N, Ramirez L M, Lee R L, Magnuson
N S, Bishop G A, and Gold M R. (2002) CD40 signaling in B cells
regulates the expression of the PIM-1 kinase via the NF-kappa B
pathway. J Immunol. 168: 744-754). PIM-1 expression is rapidly
induced after cytokine stimulation and the proliferative response
to cytokines is impaired in cells from PIM-1 deficient mice (Domen
J, van der Lugt N M, Acton D, Laird P W, Linders K, Berns A. (1993)
PIM-1 levels determine the size of early B lymphoid compartments in
bone marrow. J. Exp. Med. 178: 1665-1673).
[0856] Members of the PIM family of kinases interact with Socs-1
protein, a potent inhibitor of JAK activation thereby playing a
major role in signaling down stream of cytokine receptors. The
phosphorylation of Socs-1 by PIM family of kinases prolongs the
half-life of Socs-1 protein, thus potentiating the inhibitory
effect, of Socs-1 on JAK-STAT activation (Chen X P, Losman J A,
Cowan S, Donahue E, Fay S, Vuong B Q, Nawijn M C, Capece D, Cohan V
L, Rothman P. (2002) PIM serine/threonine kinases, regulate the
stability of Socs-1 protein. Proc. Natl. Acad. Sci. USA
99:2175-2180.). PIM-1 is expressed during G1/S phase of the cell
cycle suggesting that it is involved in cell cyclc regulation
(Liang H, Hittelman W, Nagarajan L., Ubiquitous expression and cell
cycle regulation of the protein kinase PIM-1. (1996) Arch Biochem
Biophys. 330:259-265).). PIM-1 kinase activity and the protein
level is increased in CD 40 mediated B cell signaling and this
increase in PIM-1 level is mediated through the activation of
NF-.kappa.B (Zhu et al. 2002. supra). PIM-1 can physically interact
with NFATc transcription factors enhancing NFATc dependant
transactivation and IL2 production in Jurkat cells (Rainio E M,
Sandholm J, Koskinen P J. (2002) Cutting edge: Transcriptional
activity of NFATc1 is enhanced by the PIM-1 kinase. J. Immunol.
168:1524-1527). This indicates a novel phosphorylation dependant
regulatory mechanism targeting NFATc1 through which PIM-1 acts as
down stream effector of ras to facilitate IL2 dependant
proliferation and survival of lymphoid cells (ibid.).
[0857] Pim1 is shown to interact with many other targets.
Phosphorylation of Cdc25A phosphatase, a direct transcriptional
target of c-myc, increase its phosphatase activity both in-vivo and
in-vitro indicating that Cdc25A link PIM-1 and c-myc in cell
transformation and apoptosis (Mochizuki T, Kitanaka C, Noguchi K,
Muramatsu T, Asai A, and Kuchino Y. (1999) Physical and functional
interactions between PIM-1 kinase and Cdc25A phosphatase.
Implications for the PIM-1-mediated activation of the c-Myc
signaling pathway; J. Biol. Chem. 274:18659-18666). PIM-1 also
phosphorylate PTP-U2S, atyrosine phosphatase associated with
differentiation and apoptosis in myeloid cells, decreasing its
phosphatase activity and hence preventing premature onset of
apoptosis following PMA-induced differentiation (Wang et al. (2001)
Pim-1 negatively regulates the activity of PTP-U2S phosphatase and
influences terminal differentiation and apoptosis of monoblastoid
leukemia cells. Arch. Biochem. Biophys. 390:9-18). The
phosphorylation of another PIM-1 target, heterochromatin protein 1
(HP1) has been shown to be involved in transcription repression
(Koike et al., FEBS Lett. 2000, 467:17-21).
[0858] Pim1 inhibitors may be useful in treating cancers such as
hematopoietic (e.g. acute myeloid and acute lymphoid leukemias) and
prostate cancers, and non-Hodgkin's lymphomas.
[0859] Pim2: Target kinase Pim2 (i.e., Serine/threonine-protein
kinase Proviral Integration Site 2) is a 34.2 kDa STK encoded by
chromosome Xp11.23 (symbol: PIM2). Pim2 has also been shown to play
a role in cell survival and the control of apoptosis and its value
as an inhibitor target has been considered (Yan, et al., J Biol
Chem 2003 278: 45358-67; Giles, Blood 2005 105:4158-4159; Fox, et
al., Genes Dev 2003 17: 1841-54). Pim2 has been found to be
overexpressed in some lymphomas (Cohen, et al., Leuk Lymphoma 2004
45: 951-5). Additionally, Pim2 is required for the rapamycin
resistant T-cell survival and the rapamycin-resistant growth of
nontransformcd hematopoietic cells (Hammerman, et al., Blood 2005
105: 4477-83; Fox, et al., J Exp Med 2005 201: 259-66). Pim2
inhibitors may be useful in treating lymphomas.
[0860] Pim3: Target kinase Pim3 (i.e., Serine/threonine-protein
kinase Proviral Integration Site 3) is a 35.8 kDa STK encoded by
chromosome 22q13 (symbol: PIM3). Pim3 has recently been shown to be
overexpressed in human hepatocellular carcinoma cells and its
ablation resulted in attenuated cell proliferation and enhanced
apoptosis, suggesting that Pim3 can also participate in abnormal
cell growth and inhibition of apoptosis (Fujii, et al., Int J
Cancer 2005 114: 209-18). Pim3 inhibitors may be useful in treating
hepatocellular carcinoma.
[0861] PKC alpha: Target kinase PKC alpha (i.e., Protein kinase C
alpha) is a 76.8 kDa STK encoded by chromosome 17q22-q23.2 (symbol:
PRKCA). Protein kinase C (PKC) is the major phorbol ester receptor.
Nine mammalian members of the PKC family have been identified and
designated alpha, beta, gamma, delta, epsilon, zeta, eta, theta,
and lambda. According to OMIM, Parker et al. (Science 1986,
233:853-859) purified PKC from bovine brain and through the use of
oligonucleotide probes based on partial amino acid sequence,
derived cDNA clones from bovine cDNA libraries. Activation of PKC
by calcium ions and the second messenger diacylglycerol is thought
to play a central role in the induction of cellular responses to a
variety of ligand-rcceptor systems and in the regulation of
cellular responsiveness to external stimuli. Birnbaum et al.
(Science 2004, 306:882-884) showed that high levels of PKC activity
in prefrontal cortex, as seen for example during stress exposure,
markedly impaired behavioral and electrophysiologic measures of
working memory. Birnbaum (ibid.) concluded that excessive PKC
activation can disrupt prefrontal cortical regulation of behavior
and thought, possibly contributing to signs of prefrontal cortical
dysfunction such as distractibility, impaired judgment,
impulsivity, and thought disorder (OMIM MIM Number: 176960: Apr.
17, 2006). A mutation in PKC-alpha (D294G) correlates with
pituitary tumor. PKC alpha inhibitors may be useful in treating
pituitary tumors and prefrontal cortical dysfunction such as
distractibility, impaired judgment, impulsivity, and thought
disorder, also may be used to sensitize chemotherapy in breast,
colon, and non small cell lung cancers.
[0862] PKC beta: Target kinase PKC beta (i.e., Protein kinase C,
beta I) is a 76.7 kDa STK encoded by chromosome 16p11.2 (symbol:
PRKCB1). According to OMIM, Leitges et al. (Science 1996,
273:788-791) found that mice homozygous for a targeted disruption
of the PRKCB1 gene develop an immunodeficiency characterized by
impaired humoral immune responses and reduced cellular responses of
B cells similar to X-linked immunodeficiency (Xid) in mice. Thus,
they concluded that the 2 isoforms, PKC-beta-I (PRKCB1) and
PKC-beta-II (PRKCB2), play an important role in B-cell activation
and may be functionally linked to Bruton tyrosine kinase in antigen
receptor-mediated signal transduction (OMIM MIM Number: 176970:
Mar. 3, 2006). In general, inhibitors PKC beta and PKC isoforms may
be effective in treating disorders characterized by dysregulated
NFKB survival signaling. PKC beta inhibitors may be useful in
treating diabetic retinopathy.
[0863] PKC theta: Target kinase PKC-theta (i.e., Protein kinase,
theta) is a 81.9 kDa STK encoded by chromosome 10p15 (symbol:
PRKCQ). According to OMIM, in an attempt to find PKC isoforms that
are involved in growth control and/or activation of T lymphocytes,
Baier et al., (J. Biol. Chem. 1993, 268:4997-5004) used a human
peripheral blood lymphocyte-derived cDNA library was employed to
identify a novel PKC isoform, termed PKC-theta. The gene encodes a
protein of approximately 80 kD, expressed predominantly in lymphoid
tissues and hematopoietic cell lines, particularly T cells. The
alpha form (PRKCA) has been mapped to chromosome 17, the beta form
(PRKGB1) to chromosome 16, the gamma form (PRKCG) to chromosome 19,
and the delta form (PRKCD) to chromosome 3. By fluorescence in situ
hybridization, Erdel et al. (Genomics 1995, 25:595-597) assigned
the PRKCQ gene to 10p15. Blanco and Brown (Mammalian Genome 1997,
8:70-71) mapped the homolog Pkcq to mouse chromosome 2 by analysis
of an interspecific backcross. Sun et al. (Nature 2000,
404:402-407) demonstrated that PKC-theta is essential for T-cell
antigen receptor (TCR)-mediated T-cell activation but dispensable
during TCR-dependent thymocyte development. They generated mice
deficient in Pkc-theta by homologous recombination. Mutant mice
were normal and fertile. TCR-initiated NF-kappa-B activation was
absent from PKC-theta-/- mature T lymphocytes, but was intact in
thymocytes. Activation of NF-kappa-B by tumor necrosis factor-alpha
and interleukin-1 was unaffected in the mutant mice. Induction of
JNK was normal in T cells from mutant mice. Sun et al. (ibid.)
concluded that PKC-theta functions in a unique pathway that links
the TCR signaling complex to the activation of NF-kappa-B in mature
T lymphocytes. Using hyperinsulinemic-euglycemic clamps, Kim et al.
(J. Clin. Invest. 2004, 114:823-827) demonstrated that skeletal
muscle and hepatic insulin action did not differ between wildtype
and Pkc-theta null mice. A 5-hour lipid infusion decreased
insulin-stimulated skeletal muscle glucose uptake in the wildtype
mice that was associated with 40 to 50% decreases in
insulin-stimulated tyrosine phosphorylation of insulin receptor
substrate-1 (IRS1) and IRS1-associated PI3K activity. In contrast,
Pkc-theta inactivation prevented fat-induced defects in insulin
signaling and glucose transport in skeletal muscle. Kim et al.
(ibid.) concluded that PKC-theta is a crucial component mediating
fat-induced insulin resistance in skeletal muscle (OMIM MIM Number:
600448: Oct. 15, 2004).
[0864] Li et al. report that PKC plays a critical role in
competitive activity-dependent synapse modification at the
neuromuscular synapse in vitro and in vivo. This action involves a
reduction of the strength of inactive inputs to muscle cells that
are activated by other inputs. A decrease of postsynaptic
responsiveness and a loss of postsynaptic acetyl choline receptors
account for the heterosynaptic loss in vitro. The loss is not seen
in preparations in which PKC has been blocked pharmacologically.
Here, they show that the loss does not occur in in vitro
preparations made from animals genetically modified to lack the
theta isoform of PKC. Synapse elimination in the newborn period in
vivo is delayed but is eventually expressed in knock-out animals.
PKC-dependent synapse reduction is suppressed in heterologous
cultures combining normal nerve and PKC-theta-deficient muscle, as
might be expected from the postsynaptic locus of the changes that
underlie the activity-dependent plasticity. Preparations in which
PKC-theta-deficient neurons innervated normal muscle also exhibited
a marked deficit in PKC-deficient synapse reduction. The
presynaptic action of PKC-theta implied by this observation is
blocked by TTX, and the authors propose that the activity-related
synapse strengthening is decreased by presynaptic PKC-theta. Thus,
PKC-theta in both presynaptic and postsynaptic elements plays a
critical role in activity-dependent synapse modulation and loss (Li
et al., Journal of Neuroscience 2004, 24(15):3762-3769). PKC theta
inhibitors may be useful in treating insulin resistance, T-cell
lymphoma.
[0865] Plk1: Target kinase Plk1 (i.e., Polo like kinase 1) is a
68.3 kDa STK encoded by chromosome 16p12.3 (symbol: PLK1). Plk1 is
a regulator required for multiple mitotic processes, including
bipolar mitotic spindle formation, actin ring formation, and
chromosomal segregation. According to OMIM, Holtrich et al. (Proc.
Nat. Acad. Sci. 1994, 91:1736-1740) observed that PLK1 transcripts
are present at high level in tumors of various origins. In
vertebrate cells, the nuclear entry of mitosis-promoting factor
(MPF) during prophase is thought to be essential for the induction
and coordination of M-phase events. Phosphorylation of cyclin B1 is
central to its nuclear translocation. Toyoshima-Morimoto et al.
(Nature 2001, 410:215-220) purified a protein kinase from Xenopus
M-phase extracts that phosphorylates a crucial serine residue
(S147) in the middle of the nuclear export signal sequence of
cyclin B1. They identified this kinase as Plx1, a Xenopus homolog
of PLK1. During cell cycle progression in HeLa cells, a change in
the kinase activity of endogenous Pls1 toward S147 and/or S133
correlates with a kinase activity in the cell extracts. An
anti-PLK1 antibody depleted the M-phase extracts of the kinase
activity toward S147 and/or S133. An anti-phospho-S147 antibody
reacted specifically with cyclin B1 only during G2/M phase. A
mutant cyclin B1 in which S133 and S147 were replaced by alanines
remained in the cytoplasm, whereas wildtype cyclin B1 accumulated
in the nucleus during prophase. Further, coexpression of
constitutively active Plk1 stimulates nuclear entry of cyclin B1.
Toyoshima-Morimoto et al. (ibid.) concluded that Plk1 may be
involved in targeting MPF to the nucleus during prophase (OMIM MIM
Number: 602098: Aug. 5, 2005). Plk1 inhibitors may be useful in
treating cancers (e.g. lymphoma of the thyroid, non-Hodgkin's
lymphomas, colorectal cancers, leukemias and melanoma), also useful
as sensitizer in chemotherapy.
[0866] Pvk2: Target kinase Pyk2 (i.e., Protein-Tyrosine Kinase 2)
is a 115.9 kDa tyrosine kinase of the FAK family (see e.g., target
Fak) encoded by chromosome 8p21.1 (symbol: PTK2B). As with target
kinase Fak, Pyk2 comprises B41 and TK domains, and Pyk2 is also
known as Fak2 (i.e., Focal Adhesion Kinase 2). Because Pyk2 is
calcium dependent, it is also known as CADAK (i.e.,
calcium-dependent tyrosine kinase).
[0867] As mentioned, another member of the FAK subfamily is kinase
Fak. Pyk2 and Fak shares 65% sequence identity in the kinase domain
and have similar domain structure: an N-terminus domain for
integrin binding, and a C-terminus domain for Paxillin binding. Fak
is ubiquitously expressed while Pyk2 exhibits a more restricted
tissue expression pattern primarily in neuronal and hematopoetic
tissues.
[0868] According to OMIM, Focal adhesion kinases (i.e., FAKs) are
cytoplasmic protein-tyrosine kinases associated with focal
adhesions and whose activity is induced by ligand binding to
various receptors including those of, for example, integrin and
growth factors. FAKs are known to target paxillin and are
substrates for Src family kinases (Calalb et al., Molec. Cell.
Biol. 1995, 15:954-963). Herzog et al. (Genomics 1996, 32:484-486)
identified a gene for another focal adhesion kinase by
low-stringency screening of a hippocampus cDNA library. They
symbolized the gene FAK2. The FAK2 cDNA encodes a predicted
1,009-amino acid protein with 42% identity to FAK1. Northern blot
analysis detected a 4.5-kb mRNA in brain, kidney, spleen, and
lymphocytes. Protein-tyrosine kinases in the central nervous system
are activated in response to a variety of neurotrophic factors that
control neuronal differentiation and survival via cell surface
receptors. Also, protein phosphorylation is involved in membrane
excitability and the function of ion channels. Lev et al. (Nature
1995, 376:737-745) discovered a nonreceptor type protein kinase
that is highly expressed in adult rat brain. The kinase, which they
symbolized PYK2 (proline-rich tyrosine kinase-2), was cloned from a
rat spinal cord cDNA library using degenerate PGR primers
corresponding to conserved tyrosine kinase motifs of PYK1 (see
Manser et al., Nature 1993, 363:364-367). Lev et al. (ibid.) cloned
the human homolog from a human fetal brain cDNA library using the
rat sequence as a probe. The predicted protein of 1,009 amino acids
has 61% sequence identity to the FAK1 protein (Ptk2) (OMIM MIM
Number: 601212: Jan. 19, 2005). PKC-theta may represent an
important signaling intermediate between neuropeptide activated
receptors or neurotransmitters that increase calcium flux and the
downstream signals that regulate neuronal activity. PKC-theta
interacts with the SH.sub.2 domain of Grb2, and may phosphorylate
the voltage-gated potassium channel protein Kv1.2. Its activation
is highly correlated with the stimulation of c-Jun N-terminal
kinase activity.
[0869] Pyk2 regulates multiple signaling events crucial for
macrophage morphology and migration. It mediates the Jak-dependent
activation of MAPK and Stat1. By rapidly translocating to the
vicinity of the immune synapse after T cell receptor stimulation,
Pyk2 plays an essential role in T cell activation and polarized
secretion of cytokines. The morphology and behavior of macrophages
in Pyk2-/- mice were impaired. Macrophages isolated from mutant
mice failed to become polarized, to undergo membrane ruffling, and
to migrate in response to chemokine stimulation. Moreover, the
contractile activity in the lamellipodia of Pyk2-/- macrophages was
impaired, as revealed by measuring the rearward movement toward the
nucleus of fibronectin-coated beads on the lamellipodia in
opposition to an immobilizing force generated by optical
tweezers.
[0870] Pyk2 is implicated in several therapeutic areas including
inflammation (e.g. osteoporosis, Polycystic Kidney Disease,
rheumatoid arthritis and some bowel diseases) and CNS disease like
Parkinson's disease and Alzheimer's disease). Pyk2 in osteoclasts
is an adhesion kinase, localized in the sealing zone, activated by
ligation of v3 integrin, and phosphorylated by Src kinase. Methods
for preventing cell death in a subject and their application in the
treatment of neurodegenerative diseases and conditions, such as
Alzheimer's disease, stroke, Parkinson's disease have been
patterned by Griswold-Prenner Irene and Powell Kyle.
[0871] Pyk2 is also a potential therapeutic target for tumors. Pyk2
is a novel effector of fibroblast growth factor receptor 3
activation. Pyk2 facilitates EGFR- and c-Src-mediated Stat3
activation and has a role in triggering Stat3-induced oncogenesis.
HER.sup.3, but not HER.sup.2, mediates the phosphorylation of the
C-terminal region of PYK2 to promote a mitogenic response through
activation of the MAPK pathway. Furthermore, PYK2 phosphorylation
by HER.sup.3 induces tumor invasion. A central role of PYK2 in
signaling downstream of HER.sup.3 is substantiated by the
demonstration that expression of a dominant-negative PYK2-KM
construct abrogates the Heregulin-induced MAPK activity and
inhibits the invasive potential of glioma cells.
[0872] Overexpression of wild-type RAFTK significantly enhanced
breast cancer cell invasion, while overexpression of the mutants
Tyr402 or Tyr881 of RAFTK inhibited this migration. Therefore, Pyk2
may serve as a mediator and an integration point between focal
adhesion molecules in HRG-mediated signaling in T47D breast cancer
cells.
[0873] A murine pancreatic cancer cell line overexpressing Pyk2,
mPanc02, was treated with a Pyk2-dominant negative adenovirus
(Ad-Pyk2DN), or GFP (ad-GFP) adenovirus. The dominant negative Pyk2
adenovirus is able to decrease tumor growth and increase survival
in several in vivo tumor models.
[0874] Although no point mutation of Pyk2 has been reported to be
significant in any disease, human umbilical vein endothelial cells
express mRNA transcripts for both the full length isoform Pyk2 and
the truncated isoform Pyk2-H containing the C-terminal
deletion.
[0875] Pyk 2 inhibitors may be useful in treating inflammation
(e.g. osteoporosis, polycystic kidney disease, rheumatoid arthritis
and inflammatory bowel disease), CNS disease (e.g. Parkinson's
disease and Alzheimer's disease), stroke and cancers (e.g. gliomas,
breast cancer, and pancreatic cancer).
[0876] Ret: Target Ret (i.e., Rearranged during Transfection) is a
124.3 kDa tyrosine kinase encoded by chromosome 10q11.2 (symbol:
RET). Ret is also known as c-ret (i.e., cellular ret). The domain
structure of Ret comprises cadherin, transmembrane, and TK domains.
Cadherins are glycoproteins involved in Ca2+-mediated cell-cell
adhesion; see e.g., Yap et al., Annu Rev Cell Dev Biol. 1997;
13:119-46).
[0877] According to OMIM, the RET protooncogene is one of the
receptor tyrosine kinases, cell-surface molecules that transduce
signals for cell growth and differentiation. The RET gene was
defined as an oncogene by a classical transfection assay. RET can
undergo oncogenic activation in vivo and in vitro by cytogenetic
rearrangement (Gricco et al., Cell 1990, 60:557-563). Mutations in
the RET gene are associated with multiple endocrine neoplasia, type
IIA (MEN2A), multiple endocrine neoplasia, type IIB (MEN2B),
Hirschsprung disease (HSCR; aganglionic megacolon), and medullary
thyroid carcinoma (MTC) (OMIM MIM Number: 164761: Jan. 27,
2006).
[0878] Ret (Rearranged during Transformation) was identified as a
rearranged human oncogene in the classic NIH3T3 transformation
assay (Takahashi et al., 1985, Cell 42(2):581-8) and subsequently
characterized as a Receptor Tyrosine kinase (Takahashi et al.,
1988, Oncogene 3(5):571-8).
[0879] Ret and NTRK1 (i.e., Neurotrophic tyrosine receptor kinase
1) are receptor tyrosine kinase (RTK) proteins which play a role in
the development and maturation of specific components of the
nervous system. Their alterations have been associated to several
human diseases, including some forms of cancer and developmental
abnormalities. These features have contributed to the concept that
one gene can be responsible for more than one disease. Moreover,
both genes (encoding for the two RTKs show genetic alterations that
belong to either "gain of function" or "loss of function" class of
mutations. In fact, receptor rearrangements or point mutations
convert Ret and NTRK1 into dominantly acting transforming genes
leading to thyroid tumors, whereas inactivating mutations,
associated with Hirschsprung's disease (HSCR) and congenital
insensitivity to pain with anhidrosis (CIPA), impair Ret and NTRK1
functions, respectively.
[0880] Implication of Ret in human tumorigenesis was indicated by
the frequent identification of rearranged Ret sequences that
transformed NIH3T3 cells in the DNA isolated from Papillary Thyroid
Carcinoma DNAs. In these cases, the Ret gene was fused to as yet
unknown PTC DNA sequences in the tumor DNA but not the normal
patient DNA (Grieco et al., 1990, Cell 60(4):557-63). In addition,
the chromosomal mapping of Ret to chromosome 10q11.2 co-localized
with genetic mapping data that implicated a gene involved in
patients with MEN2A (Multiple Endocrine Neoplasia 2A) (Ishizaka et
al. 1989 Oncogene 4(12): 1519-21). Expression analysis of the RET
oncogene in a number of human tumors consistently detected
expression of normal-sized transcripts of the RET proto-oncogene in
human pheochromocytomas and in human medullary thyroid carcinomas,
both of familial and sporadic type (Santoro et al., 1990, Oncogene
5:1595-8).
[0881] Further analysis of the tumor DNA of patients with Multiple
endocrine neoplasia type 2A (MEN 2A) and familial medullary thyroid
carcinoma (FMTC) identified mutations in the RET sequence resulting
in amino acid changes in the encoded Ret protein (Donis-Keller
1993, Hum Mol Genet. 2(7):851-6). Likewise, mutations in the RET
gene were correlated with Hirschsprung disease, a developmental
disorder with genetic deletions and mutations in the chromosomal
location of the RET gene (Luo et al., 1993, Hum Mol Genet.
2(11):1803-8).
[0882] By early 1994, multiple papers describe the inactivation of
the RET gene in patients with Hirschsprung disease and similar
phenotype in knock out mice. In addition, activating mutations in
Ret are now identified in patients with MEN2A, MEN2B, and FMTC
(reviewed by van Heyningen V., 1994, Nature 367(6461):319-20).
[0883] It was determined that Ret regulates cell survival. Signal
transduction molecules that form a complex with Ret as a result of
these phosphoryl moieties, such as GRB2, SOS, ras, and raf,
propagate a signal in the cell that promotes neural survival. Thus,
compounds that promote the interactions of the se stimulatory
molecules of Ret would enhance the activity of c-Ret.
Alternatively, protein phosphatases can remove the phosphoryl
moieties placed on the intracellular region of Ret in response to
GDNF, and thus inhibit the signaling capability c-Ret. Thus,
compounds that inhibit phosphatases of Ret will probably enhance
the signaling capacity of c-Ret.
[0884] Ret is implicated in the development and survival of
enteric, synaptic, and sensory neurons and neurons of the renal
system upon stimulation by GDNF (Jing, et al., 1996, Cell
85:1113-1124; Trupp, et al., 1996, Nature 381:785-789; Durbec, et
al., 1996, Nature 381:789-793). Lack of function mutations in Ret
can lead to Hirschsprung's disease, for example, which manifests
itself as a decrease in intestinal tract innervation in mammals.
Thus, compounds that activate Ret are potential therapeutic agents
for the treatment of neurodegenerative disorders, including, but
not limited to, Hirschsprung's disease, Parkinson's disease,
Alzheimer's disease, and amyotrophic lateral sclerosis. Compounds
that inhibit Ret function can also be anti-cancer agents as
over-expression of Ret in cells is implicated in cancers, such as
cancer of the thyroid.
[0885] Modulation of Ret activity may also be useful in treating
cancers of the nerve tissue, such as neuroblastoma, even if an
abnormality is not found the signaling pathway.
[0886] As stated above, RET gene is responsible for MEN2 syndromes,
which are inherited in an autosomal dominant fashion with high
penetrance and diverse clinical manifestations. The predominant RET
mutation is missense mutation which is restricted to 9 codons
(codons 609, 611, 618, 620, 630, 634, 768, 804 and 918). The MEN2
syndromes have 3 subtypes: multiple endocrine neoplasia type 2A
(MEN2A), MEN2B, and familial medullary thyroid carcinoma (FMTC).
Missense mutations at exon 10 (codons 609, 611, 618, and 620) and
exon 11 (codons 630 and 634) have been identified in 98% of MEN2A
families and in 85% of FMTC families. Missense mutations at codons
768 and 804 have been known to be responsible for 5.about.10% of
FMTC cases. In addition, missense mutations at exon 16 (codon 918)
have been found in 95% of MEN2B cases.
[0887] Ret inhibitors may be useful in treating cancer of the
thyroid, neuroblastoma, familial medullary thyroid carcinoma
(FMTC), multiple endocrine neoplasia type IIA and IIB (MEN2A,
MEN2B), and neurodegenerative disorders (e.g. Hirschsprung's
disease, Parkinson's disease, Alzheimer's disease, and amyotrophic
lateral sclerosis)
[0888] Ron: Target kinase Ron (i.e., Ron protein tyrosine kinase)
is a 152.2 kDa transmembrane tyrosine kinase encoded by chromosome
3p21.3 (symbol: MST1R), also known as macrophage stimulating
protein receptor (i.e., MSP receptor). Ron, a member of the Met
hepatocyte growth factor receptor family, was originally cloned
from a human foreskin keratinocyte cDNA library by Ronsin et al
(Oncogene 1993, 8: 1195-1202). RON is expressed in various cell
types including macrophages, epithelial and hematopoietic cells.
Ron activation results in a variety of cellular responses in vitro,
such as activation of macrophages, proliferation, migration, and
invasion, which suggest a broad biologic role in vivo. Hemizygous
mice (Ron+/-) grow to adulthood; however, these mice are highly
susceptible to endotoxic shock and appeared to be compromised in
their ability to downregulate nitric oxide production. Accordingly,
Ron plays a role in early mouse development and may play a limited
role in the inflammatory response. Further, Ron may be involved in
cancer development and progression (OMIM MIM Number: 600168: Jan.
20, 2006). Ron inhibitors may be useful in treating cancer and
inflammation.
[0889] ROCK (ROCK1 and ROCK2): Target kinase ROCK1 (i.e.,
Rho-associated, coiled-coil containing protein kinase 1) is a 158.2
kDa serine/threonine kinase encoded by chromosome 18q11.1 (symbol:
ROCK1). Target kinase ROCK2 (i.e., Rho-associated, coiled-coil
containing protein kinase 2) is a 160.9 kDa serine/threonine kinase
encoded by chromosome 2p24 (symbol: ROCK2). ROCK inhibitors may be
useful in treating related to cancers (e.g. ovarian cancer,
hepatocellular carcinoma, pancreatic cancer), ocular disease (e.g.
glaucoma), cardiac hypertrophy, improved renal perfusion,
transplant rejection, and acute respiratory distress syndrome.
[0890] Src: Target kinase Src (i.e., v-Src Avian Sarcoma
Schmidt-Ruppin A-2 viral oncogene) is a 59.8 kDa non-receptor
tyrosine kinase encoded by chromosome 20q12-q13 (symbol; SRC). The
structure of Src comprises SH3 and SH.sub.2 domains adjacent to the
TK domain. According to OMIM, Azarnia et al. (Science 1988,
239:398-401) found that overexpression of the SRC gene in NIH 3T3
cells caused reduction of cell-to-cell transmission of molecules in
the 400- to 700-dalton range. Downregulation was enhanced by point
mutation of tyrosine-527, whereas mutation of tyrosine-416
suppressed both the downregulation of communication by the tyr-527
mutation and that by gene overexpression. The regulation of
communication by SRC may be important in the control of embryonic
development and cellular growth. Luttrell et al. (Science 1999,
283:655-661) demonstrated that c-src binds to the amino terminus of
beta-arrestin-1 in a complex resulting from the stimulation of
beta-2 adrenergic receptors. Activated beta-2-adrenergic receptor
bound beta-arrestin-1, which then bound c-src. This interaction
targeted the complex to clathrin-coated pits and allowed for
beta-2-adrenergic activation of the MAP kinases Erk1 and Erk2.
TRANCE, a TNF family member, and its receptor, RANK, are critical
regulators of dendritic cell and osteoclast function. Wong et al.
(Molec. Cell 1999, 4:1041-1049) demonstrated that TRANCE activates
the antiapoptotic serine/threonine kinase PKB (AKT1) through a
signaling complex involving SRC and TRAF6. A deficiency in SRC or
addition of SRC family kinase inhibitors blocked TRANCE-mediated
PKB activation in osteoclasts. SRC and TRAF6 interacted with each
other and with RANK upon receptor engagement. TRAF6, in turn,
enhanced the kinase activity of SRC, leading to tyrosine
phosphorylation of downstream signaling molecules such as CBL.
These results defined a mechanism by which TRANCE activates SRC
family kinases and PKB, and provided evidence of cross-talk between
TRAF proteins and SRC family kinases. Using a colon cancer cell
line, Avizienyte et al. (Nature Cell Biol. 2002, 4:632-638) studied
the role of SRC in cell adhesion and metastasis. Transfection and
overexpression of a constitutively active SRC mutant reduced
cell-cell contacts and caused redistribution of adherens junction
components to discrete adhesion-like structures at the tips of
membrane protrusions. Expression of active SRC also impaired the
movement of E-cadherin from the cell interior to the plasma
membrane following exposure to high calcium. Avizienyte et al.
(ibid.) provided evidence that the alpha-V and beta-1 integrins and
FAK were required for the adhesion changes induced by SRC.
Sandilands et al. (Dev. Cell 2004, 7:855-869) found that RhoB
colocalized with active Src in the cytoplasm of mouse embryonic
fibroblasts, and they presented evidence that RhoB is a component
of `outside-in` signaling pathways that coordinate Src activation
with translocation to transmembrane receptors (OMIM MIM Number:
190090: Jan. 7, 2005).
[0891] The Src family of cytoplasmic protein tyrosine kinases
consists of at least eight members (Src, Fyn, Lyn, Yes, Lck, Fgr,
Hck and Blk) that participate in a variety of signaling pathways
[Schwartzberg, P. L., Oncogene, 17, 1463 (1998)]. The prototypical
member of this tyrosine kinase family is p60src (Src). Src is
involved in proliferation and migration responses in many cell
types. In limited studies, Src activity has been shown to be
elevated in breast, colon (.sup..about.90%), pancreatic (>90%)
and liver (>90%) tumors. Greatly increased Src activity is also
associated with metastasis (>90%) and poor prognosis. Antisense
Src message impedes growth of colon tumor cells in nude mice
[Staley et al., Cell Growth & Differentation, 8, 269 (1997)],
suggesting that Src inhibitors should slow tumor growth. In
addition to its role in cell proliferation, Src also acts in stress
response pathways, including the hypoxia response. Previous studies
have shown that colonic tumor cells genetically engineered to
express antisense Src message form tumors demonstrating reduced
vascularization in nude mouse models [Ellis, et al., J. Biol.
Chem., 273, 1052 (1998)], suggesting that Src inhibitors would be
anti-angiogenic as well as antiproliferative.
[0892] Apart from its role in cancer, Src also appears to play a
role in osteoporosis. Mice genetically engineered to be deficient
in src production were found to exhibit osteopetrosis, the failure
to resorb bone [Soriano, P., Cell, 64, 693 (1991); Boyce, B. F., J.
Clin. Invest., 90, 1622 (1992)]. This defect was characterized by a
lack of osteoclast activity. Since osteoclasts normally express
high levels of Src, inhibition of Src kinase activity may be useful
in the treatment of osteoporosis [Missbach, M., Bone, 24, 437
(1999)]. Src inhibitors may be useful in treating cancer and
osteoporosis.
[0893] Stk6: Target Stk6 (i.e., Serine/Threonine protein kinase 6)
is a 45.8 kDa serine/threonine kinase encoded by chromosome
20q13.2-q13.3 (symbol: STK6). According to OMIM, Kimura et al. (J.
Biol. Chem. 1997, 272:13766-13771) cloned a cDNA encoding a novel
human serine/threonine kinase, STK6, that has high homology with
the Aurora and Ip11 kinases. Mutations in these yeast kinases are
known to cause abnormal spindle formation and missegregation of
chromosomes. Northern and Western blot analyses revealed a high
level of STK6 expression product in testis and proliferating
culture cells such as HeLa cells. The endogenous levels of STK6
protein and protein kinase activity were tightly regulated during
cell cycle progression in HeLa cells. The protein was upregulated
during G2/M and rapidly reduced after mitosis. Immunofluorescence
studies revealed specific localization of STK6 protein to the
spindle pole region during mitosis. The results suggested that
STK6, like Aurora and Ip11, is involved in cell growth and/or
chromosome segregation (OMIM MIM Number: 602687: Apr. 1, 2003).
[0894] Aurora A belongs to the family of STKs that are involved in
mitotic events such as centrosome separation and chromosome
segregation and are therefore essential for cell proliferation
(Bischoff & Plowman, Trends Cell Biol. 1999, 9:454-459); Giet
& Prigent, 1999, Cell Science 112: 3591-3601). Inhibitors of
the Aurora kinase family therefore have the potential to block
growth of all tumors.
[0895] The three identified Aurora kinases of this family are known
under various names: Aurora-A (Aurora 1), Aurora B (Aurora 2) and
Aurora C (Aurora 3). Alternate names for Aurora A, described here
further are serine/threonine protein kinase 15 (STK 15), BTAK
Aurora-related kinase 1 (ARK1). The mouse homolog of Aurora A, STK6
also referred to as AIK, is highly homologous to STK15. All Aurora
kinase family members are highly homologous proteins responsible
for mitotic events such as centrosome maturation and segregation,
chromosome segregation, mitotic spindle function and cytokinesis.
Peak expression of Aurora occurs during the G2 and mitotic phase in
cycling cells and then decreases and remains low or undetectable in
resting cells (Shindo et al., 1998, Biochem. Biophys. Res. Commun.
244: 285-292). In mammalian cells proposed substrates for Aurora
include histone H3, a protein involved in chromosome condensation,
and CENP-A, myosin II regulatory light chain, protein phosphatase
1, TPX2, all of which are required for cell devision.
[0896] Zhou et al.; (Zhou et al., 1998, Nature Genet. 20: 189-193)
found that Aurora is involved in the induction of centrosome
duplication-distribution abnormalities and aneuploidy in mammalian
cells. Centrosomes appear to maintain genomic stability through the
establishment of bipolar spindles during cell division, ensuring
equal segregation of replicated chromosomes to 2 daughter cells.
Deregulated duplication and distribution of centrosomes are
implicated in chromosome segregation abnormalities, leading to
aneuploidy seen in many cancer cell types. Zhou et al., (Zhou et
al., 1998, Nature Genet. 20: 189-193) found amplification of Aurora
A in approximately 12% of primary breast tumors, as well as in
breast, ovarian, colon, prostate, neuroblastoma, and cervical
cancer cell lines. Additionally, high expression of Aurora A mRNA
was detected in tumor cell lines without evidence of gene
amplification. Ectopic expression of Aurora A in mouse NIH 3T3
cells led to the appearance of abnormal centrosome number
(amplification) and transformation in vitro. Finally,
over-expression of Aurora A in near-diploid human breast epithelial
cells revealed similar centrosome abnormality, as well as induction
of aneuploidy. These findings suggested that Aurora A is a critical
kinase-encoding gene, whose over-expression leads to centrosome
amplification, chromosomal instability, and transformation in
mammalian cells.
[0897] The AURKA gene is over expressed in many human cancers.
Ectopic over-expression of aurora kinase A in mammalian cells
induces centrosome amplification, chromosome instability, and
oncogenic transformation, a phenotype characteristic of
loss-of-function mutations of p53. Katayama et al. (Katayama et
al., 2004, Nature Genet 36: 55-62) showed that Aurora A kinase,
phosphorylates p53 at ser315, leading to its ubiquitination by MDM2
and proteolysis. P53 is not degraded in the presence of inactive
Aurora A or ubiquitination-defective MDM2. Silencing of Aurora
kinase A results in less phosphorylation of p53 at ser315, greater
stability of p53, and cell-cycle arrest at G2-M. Cells depleted of
Aurora kinase A are more sensitive to cisplatin-induced apoptosis,
and elevated expression of aurora kinase A abolishes this response.
In a sample of bladder tumors with wildtype p53, (Katayama et al.,
2004, Nature Genet. 36: 55-62) found a correlation between elevated
expression of aurora kinase A and low p53 concentration. They
concluded that Aurora A kinase is a key regulatory component of the
p53 pathway and that over-expression of Aurora A leads to increased
degradation of p53, causing down-regulation of checkpoint-response
pathways and facilitating oncogenic transformation of cells.
[0898] By immunoprecipitation of epitope-tagged proteins from
transfected HEK293 cells, Kunitoku et al. (Kunitoku et al., 2003,
Dev. Cell 5: 853-854) demonstrated direct interaction between the
genes CENPA and AURKA. In vitro, AURKA phosphorylated CENPA on
ser7, a residue that is also phosphorylated by AURKB. Examination
of the role of both kinases in the phosphorylation of CENPA
revealed that the reaction is mediated sequentially by AURKA and
AURKB in early mitosis. Mitotic cells in which phosphorylation of
CENPA on ser7 was prevented exhibited a substantial proportion of
misaligned chromosomes resulting from a defect in the ability of
kinetochores to attach to microtubules.
[0899] By yeast 2-hybrid analysis of HeLa cells, Hirota et al.
(Hirota et al., 2003, Cell 114: 585-598) determined that AURKA
interacts with Ajuba (JUB). The two proteins interacted in mitotic
cells and became phosphorylated as they did so. In vitro analysis
revealed that Ajuba induced the autophosphorylation and consequent
activation of AURKA. Depiction of Ajuba prevented activation of
AURKA at centrosomes in late G2 phase and inhibited mitotic entry.
Hirota et al. (Hirota et al., 2003, Cell 114: 585-598) concluded
that Ajuba is an essential activator of AURKA in mitotic
commitment.
[0900] The mammalian Aurora kinase family has been implicated in
tumorigenesis of a variety of different cancers. The main role of
Aurora A in tumor development is in controlling chromosome
segregation during mitosis (Bischoff & Plowman, Trends Cell
Biol. 1999, 9:454-459). Overexpression of Aurora A transforms
rodent fibroblasts (Bischoff et al., 1998, EMBO J. 17:3052-3065).
The elevated levels of Aurora A induce misregulation of chromosome
segregation that results in cells containing multiple centrosomes
and multipolar spindles leading to aneuploidy, a distinctive
feature of most cancers. (Zhou et al., 1998, Nature Genet.
20:189-193). Ewart-Toland et al. (Ewart-Toland et al., 2003, Nature
Genet. 34:403-412) found that tumors from individuals carrying the
91A allele showed more evidence of aneuploidy than those from
individuals who were homozygous for the common 91T allele. They
concluded that individuals with even one copy of the Aurora A 91A
allele develop tumors that have on average a higher degree of
aneuploidy than those from individuals homozygous for 91T. The
oncogenic activity of Aurora kinases is likely to be linked to the
generation of such genetic instability. Miyoshi et al. (Miyoshi et
al., 2001, Int. J. Cancer 92: 370-373) and Sakakura et al.
(Sakakura, et al., 2001, Br. J. Cancer 84: 824-831) report a
correlation between amplification of the Aurora A locus and
chromosomal instability in mammary and gastric tumors.
[0901] Over-expression of Aurora kinases have been reported in a
wide range of human tumors. Aurora A expression is elevated in
tumor cell lines derived from colon, breast, lung, melanoma,
kidney, ovary, pancreas, CNS, gastric tract and leukemia cells
(Tatsuka et al., 1998, Cancer Res. 58(21): 4811-6.) Elevated
expression of Aurora A has been detected in over 50% of colorectal,
ovarian and gastric tumors and in 94% of invasive duct
adenocarcinomas of the breast (Colorectal tumors: Bischoff et al.,
1998, EMBO J. 17:3052-3065, Takahashi et al., 2000, Jpn. J. Cancer
Res. 91:1007-1014, Ovarian tumors: Gritsko et al., 2003, Clin.
Cancer Res. 9:1420-1426; gastric tumors: Sakakura, et al., 2001,
Br. J. Cancer 84: 824-831; breast tumors: Tanaka et al., 1999,
Cancer Res. 59:2041-2044). High levels of Aurora A have also been
reported in renal, cervical, neuroblastoma, melanoma, lymphoma,
pancreatic and prostate tumor cell lines (Bischoff et al., 1998,
EMBO J. 17:3052-3065; Zhou et al., 1998, Nature Genetics
20:189-193; Li et al., 2003, Clin. Cancer Res. 9(3): 991-997).
Amplification and over-expression of Aurora A is further observed
in human bladder cancers and where it is associated with aneuploidy
and aggressive clinical behavior (Sen et al., 2002, J. Natl. Cancer
Inst. 94(17): 1320-1329). Isola et al. (Isola et al., 1995, Am. J.
Pathology 147: 905-911) further found that amplification of the
Aurora A locus correlates with poor prognosis for patients with
node-negative breast cancer.
[0902] Based on the known function of the Aurora kinases,
inhibition of their activity should disrupt mitosis leading to cell
cycle arrest. In vivo, an Aurora inhibitor therefore slows tumor
growth and induces regression. Stk6 inhibitors may be useful in
treating gastric, bladder, breast, lung, CNS, ovarian, kidney,
colon, prostate, pancreas, and cervical, cancers, melanoma,
leukemia, and neuroblastoma.
[0903] Syk: Target kinase Syk (i.e., spleen tyrosine kinase) is a
72.1 kDa tyrosine kinase encoded by chromosome 9q22.2 (symbol:
SYK). Syk inhibitors may be useful in treating lymphomas, such as
mantle cell lymphoma.
[0904] TEC: Target kinase TEC (i.e., tec protein tyrosine kinase)
is a 73.6 kDa non receptor tyrosine kinase encoded by chromosome
4p12 (symbol: TEC). TEC inhibitors may be useful in treating
sepsis, septic shock, inflammation, rheumatoid arthritis, Crohn's
disease, irritable bowel disease (IBD), and ulcerative colitis.
[0905] Tie2: Target kinase Tie2 (i.e., tyrosine kinase,
endothelial) is a 125.8 kDa receptor tyrosine kinase encoded by
chromosome 9p21 (symbol: TEK). Tie2 inhibitors may be useful in
treating cancer, arthritis (e.g. rheumatoid arthritis), and
atherosclerosis.
[0906] TrkA: Target kinase TrkA (i.e., neurotrophic tyrosine
kinase, receptor, type IV is a 87.5 kDa tyrosine kinase encoded by
chromosome Iq21-q22 (symbol: NTRK1). TrkA inhibitors may be useful
in treating pain (e.g. chronic pain, neuropathic pain), cancer,
arthritis, diabetic retinopathy, macular degeneration and
psoriasis.
[0907] Yes: Target kinase Yes (i.e., Yamaguchi Sarcoma Oncogene
homolog 1) is a 60.8 kDa tyrosine kinase encoded by chromosome
18p11.31-p11.21 (symbol: YES1). The structure of Yes comprises SH3
and SH.sub.2 domains followed by a TK domain. The YES oncogene is
homologous to the Yamaguchi sarcoma-virus gene, and the amino acid
sequence of Yes shows a high degree of homology with that of the
SRC gene product of Rous sarcoma virus. The Yes kinase is highly
expressed in multiple mammalian cell types, including neurons,
spermatozoa, platelets, and epithelial cells. The target kinase Yes
is amplified and overexpressed in various cancers including
esophageal squamous cell carcinoma. Yes inhibitors may be useful in
treating cancers including esophageal squamous cell carcinoma.
[0908] Zap70: Target kinase Zap70 (i.e., Zeta-chain associated
protein kinase, 70 kDa) is a 69.9 kDa tyrosine kinase encoded by
chromosome 2q11-13 (symbol: ZAP70). Zap70 was first reported by
Chan et al. (Cell 1992, 71: 649-662). The mature protein comprises
two SH.sub.2 domains and a TK domain.
[0909] Zap70 is crucial to the transduction of the T-cell receptor
(TCR) signalling pathway, which leads ultimately to cellular
differentiation and proliferation. (Weiss & Imboden 1987, Adv.
Immunol. 41: 1-38). On stimulation of the T-cell antigen receptor
tyrosine phosphorylation takes place in a number of intracellular
substrates, mediated by sequential activation of two distinct
families of cytoplasmic PTKs. One substrate is the TCR-zeta chain,
which can mediate the transduction of extracellular stimuli into
cellular effector functions. The Src kinases Lck and Fyn
phosphorylate tyrosine residues on the TCR zeta-chain, contained
within conserved sequences known as immunoreceptor tyrosine-based
activation motifs (ITAMs). The Zap70 protein associates with the
phosphorylated ITAMs in the zeta chain of the activated TCR complex
(Chan et al., 1991, PNAS 88: 9166-9170. Recruitment of Zap70 to the
TCR and its subsequent phosphorylation and activation triggers all
downstream signaling events (Irving & Weiss, 1991, Cell 64:
891-902). This interaction is believed to be critical for TCR
signaling, since zeta-phosphopeptides that block the interaction of
Zap70 with the zeta-chain also inhibit TCR signaling events (Wange
et al., 1995, J. Biol. Chem. 270:944-948).
[0910] The essential role of Zap70 in T-cell function has been
demonstrated in human patients, human T-cell lines and mice. Elder
et al. (Elder et al., 1997, J. of Pediatric Hematology/Oncology
19(6): 546-550) reported studies of human patient suffering from a
rare form of severe combined immune deficiency syndrome (SCID). The
patient was found to be homozygous for a 13-bp deletion involving
nucleotides 1719-1731 of the ZAP70 gene, resulting in premature
termination 35 codons downstream and yielding a mutant protein 82
residues shorter than wildtype Zap70 . This kind of patients have
profound immunodeficiency, lack CD8+ T-cells and have CD4+ T-Cells
that are unresponsive to T-cell receptor (TCR)-mediated
stimulation. Following TCR activation the CD4+ cells show severe
defects in Ca2+ mobilization, tyrosine phosphorylation of
down-stream substrates, proliferation and IL-2 production (Elder et
al., Pediatric Research 39: 743-748).
[0911] ZAP70 deficient human Jurkat cells also demonstrate the
important function of ZAP70 in T-cell receptor signaling. A Jurkat
clone (p116) lacking the Zap70 protein was shown to have defects in
TCR signaling which was correctable by re-introduction of wild type
ZAP70 (Williams et al., 1998, Molecular and Cellular Biology 18
(3): 1388-1399.)
[0912] Studies of ZAP70 deficient mice also underline the crucial
role of the PTK in T-cell signal transduction. ZAP70 deficient mice
had neither CD4 nor CD8 single-positive T cells, but human Zap70
reconstitutes both CD4 and CD8 single-positive populations. Besides
the defects in T-cell development the TCR signalling in thymocytes
was found to be profoundly impaired, suggesting that Zap70 is a
central signalling molecule during thymic selection for CD4 and CD8
lineage. (Negishi et al., 1995, Nature 376: 435-438, Sakaguchi et
al. (Sakaguchi et al. 2003, Nature 426: 454-460) reported that the
mouse strain SKG, which is derived from a closed breeding colony of
BALB/c mice, spontaneously develops chronic arthritis. This
autosomal recessive trait was found to be caused by a mutation
(W163C) in the second SH.sub.2domain of Zap70. The phenotype showed
altered signal transduction from TCRs and a change in the threshold
of T-cells to thymic selection, leading to the positive selection
of otherwise negatively selected autoimmune T-cells.
[0913] The importance of the Zap70 kinase domain function has been
demonstrated by Elder et al. (Elder et al., 2001, J. Immunology
166(1): 656-661). In studies of humans patients and mice showed
that missense mutations within the highly conserved DLAARN motif
within the kinase domain of Zap70 result in SCID. This mutation
caused the loss of catalytic function of Zap70, resulting in
defective T-cell receptor signaling. The requirement of the
catalytic function of Zap70 was further illustrated by Williams et
al., who found an inactive Zap70 mutant (Lys369Arg) was unable to
restore TCR signaling in a ZAP70 deficient Jurkat cell clone (p116)
(Williams et al., 1998, Mol. Cell Biology 18 (3): 1388-1399).
[0914] Zap70 further participates in early B-cell differentiation
and is a prognostic factor in chronic lymphocytic leukaemia (CLL).
The course of CLL is variable. Crespo and coworkers (Crespo et al.,
2003, New Eng. J. Med. 348: 1764-1775) found that Zap70 expression
by cells in CLL is a simple and reliable surrogate for the
identification of immunoglobulin heavy chain variable region
mutations. In the aggressive progression of the disease, Zap70 is
associated with CLL cells expressing an unmutated configuration of
the immunoglobulin heavy chain variable region gene (IgVH)
(Carreras et al., 2005, Journal of Pathology 205 (4): 507-513).
Whereas in indolent disease, the CLL cells usually express a
mutated immunoglobulin heavy chain variable region gene but lack
expression of ZAP70. Rassenti et al. (Rassenti et al., 2004, New
Eng. J. Med. 351: 893-901) found that although the presence of an
unmutated immunoglobulin heavy chain variable region gene in CLL
patients was strongly associated with expression of Zap70, Zap70
was a stronger predictor of the need for treatment in B-cell CLL.
T-cell Zap70 overexpression in CLL patients was found to not only
correlate with Zap70 levels in CLL cells, but also with clinical
stage and disease progression. (Herishanu et al., 2005, Leukemia
advance online publication).
[0915] The protein tyrosine kinase Zap70 functions in the signaling
pathway that plays an essential role in T-cell activation and
development. After TCR stimulation, Zap70 is associated with the
receptor complex through the interaction of its two SH.sub.2
domains with the doubly phosphorylated ITAMs. The association of
Zap70 with the TCR ITAMs facilitates its autophosphorylation and
the tyrosine phosphorylation of Zap70 mediated by Src family PTKs,
e.g. Lck and Fyn. (Iwashima et al., 1994, Science 263:1136-1139).
The recruitment of adaptor molecules like Lat and Slp76 to Zap70 in
turn couple to more distal signaling pathways including Ras and
PLC-gamma (Chu et al., 2003, Journal of Biology 2 (3): 2-21).
[0916] Mutations in the ZAP70 gene are associated with the
selective T-cell defect (STD) and severe combined immunodeficiency
(SCID) in human patients. The variety of point mutations, missense
mutations, deletions and chromosomal rearrangements are identified
in Zap70 all result in the same phenotype (OMIM database with
genetic mutations).
[0917] Zap70 inhibitors may be useful in treating autoimmune,
inflammatory, proliferative and hyperproliferative diseases,
immunologically mediated diseases, AIDS, systemic lupus
erythematosus, myasthenia gravis, atherosclerosis, rejection of
transplanted organs or tissues, allograft rejection including acute
and chronic allograft rejection, graft versus host disease,
rheumathoid arthritis, psoriasis, systemic sclerosis, atopic
dermatitis, eczematous dermatitis, alopecia, and inflammation of
the nasal mucus membrane, including all forms of rhinitis.
III. Binding Assays
[0918] The methods of the present invention can involve assays that
are able to detect the binding of compounds to a target molecule.
Such binding is at a statistically significant level, preferably
with a confidence level of at least 90%, more preferably at least
95, 97, 98, 99% or greater confidence level that the assay signal
represents binding to the target molecule, i.e., is distinguished
from background. Preferably controls are used to distinguish target
binding from non-specific binding. A large variety of binding
assays are known for different target types and can be used for
this invention.
[0919] Binding compounds can also be characterized by their effect
on the activity of the target molecule. Compounds of the present
invention (i.e., compounds of Formula I, including Formulae Ia-Iz,
and all sub-embodiments disclosed herein, or Formula II, including
Formulae IIa-IIo, and all sub-embodiments disclosed herein) may be
assayed with respect to a particular kinase to assess inhibitory
concentration (IG.sub.50) or excitation concentration (EC.sub.50)
of the compound with respect to that kinase. The IC.sub.50 (or
EC.sub.50) is defined as the concentration of compound at which 50%
of the activity of the target kinase activity being measured is
lost (or gained) relative to activity when no compound is present.
Activity can be measured using methods known to those of ordinary
skill in the art, e.g., by measuring any detectable product or
signal produced by occurrence of an enzymatic reaction, or other
activity by a protein being measured. Compounds will have an
IG.sub.50 or EC.sub.50 of less than 10 .mu.M, also less than 1
.mu.M, also less than 100 nM, also less than 10 nM or less than 1
nM.
[0920] By "background signal" in reference to a binding assay is
meant the signal that is recorded under standard conditions for the
particular assay in the absence of a test compound, molecular
scaffold, or ligand that binds to the target molecule. Persons of
ordinary skill in the art will realize that accepted methods exist
and are widely available for determining background signal.
[0921] By "standard deviation" is meant the square root of the
variance. The variance is a measure of how spread out a
distribution is. It is computed as the average squared deviation of
each number from its mean. For example, for the numbers 1, 2, and
3, the mean is 2 and the variance is:
.sigma. 2 = ( 1 - 2 ) 2 + ( 2 - 2 ) 2 + ( 3 - 2 ) 2 3 = 0.667 .
##EQU00001##
[0922] Surface Plasmon Resonance
[0923] Binding parameters can be measured using surface plasmon
resonance, for example, with a BIAcore.RTM. chip (Biacore, Japan)
coated with immobilized binding components. Surface plasmon
resonance is used to characterize the microscopic association and
dissociation constants of reaction between an sFv or other ligand
directed against target molecules. Such methods are generally
described in the following references which are incorporated herein
by reference. Vely F. et al., (2000) BIAcore.RTM. analysis to test
phosphopeptide-SH.sub.2 domain interactions, Methods in Molecular
Biology. 121:313-21; Liparoto et al., (1999) Biosensor analysis of
the interleukin-2 receptor complex, Journal of Molecular
Recognition. 12:316-21; Lipschultz et al., (2000) Experimental
design for analysis of complex kinetics using surface plasmon
resonance, Methods. 20(3):310-8; Malmqvist., (1999) BIACORE: an
affinity biosensor system for characterization of biomolecular
interactions, Biochemical Society Transactions 27:335-40; Alfthan,
(1998) Surface plasmon resonance biosensors as a tool in antibody
engineering, Biosensors & Bioelectronics. 13:653-63; Fivash et
al., (1998) BIAcore for macromolecular interaction, Current Opinion
in Biotechnology. 9:97-101; Price et al.; (1998) Summary report on
the ISOBM TD-4 Workshop: analysis of 56 monoclonal antibodies
against the MUC1 mucin. Tumour Biology 19 Suppl 1:1-20; Malmqvist
et al, (1997) Biomolecular interaction analysis: affinity biosensor
technologies for functional analysis of proteins, Current Opinion
in Chemical Biology. 1:378-83; O'Shannessy et al., (1996)
Interpretation of deviations from pseudo-first-order kinetic
behavior in the characterization of ligand binding by biosensor
technology, Analytical Biochemistry. 236:275-83; Malmborg et al.,
(1995) BIAcore as a tool in antibody engineering. Journal of
Immunological Methods. 183:7-13; Van Regenmortel, (1994) Use of
biosensors to characterize recombinant proteins; Developments in
Biological Standardization. 83:143-51; and O'Shannessy, (1994)
Determination of kinetic rate and equilibrium binding constants for
macromolecular interactions: a critique of the surface plasmon
resonance literature, Current Opinions in Biotechnology.
5:65-71.
[0924] BIAcore.RTM. uses the optical properties of surface plasmon
resonance (SPR) to detect alterations in protein concentration
bound to a dextran matrix lying on the surface of a gold/glass
sensor chip interface, a dextran biosensor matrix. In brief,
proteins are covalently bound to the dextran matrix at a known
concentration and a ligand for the protein is injected through the
dextran matrix. Near infrared light, directed onto the opposite
side of the sensor chip surface is reflected and also induces an
evanescent wave in the gold film, which in turn, causes an
intensity dip in the reflected light at a particular angle known as
the resonance angle. If the refractive index of the sensor chip
surface is altered (e.g., by ligand binding to the bound protein) a
shift occurs in the resonance angle. This angle shift can be
measured and is expressed as resonance units (RUs) such that 1000
RUs is equivalent to a change in surface protein concentration of 1
ng/mm.sup.2. These changes are displayed with respect to time along
the y-axis of a sensorgram, which depicts the association and
dissociation of any biological reaction.
[0925] High Throughput Screening (HTS) Assays
[0926] HTS typically uses automated assays to search through large
numbers of compounds for a desired activity. Typically HTS assays
are used to find new drugs by screening for chemicals that act on a
particular enzyme or molecule. For example, if a chemical
inactivates an enzyme it might prove to be effective in preventing
a process in a cell which causes a disease. High throughput methods
enable researchers to assay thousands of different chemicals
against each target molecule very quickly using robotic handling
systems and automated analysis of results.
[0927] As used herein, "high throughput screening" or "HTS" refers
to the rapid in vitro screening of large numbers of compounds
(libraries); generally tens to hundreds of thousands of compounds,
using robotic screening assays. Ultra high-throughput Screening
(uHTS) generally refers to the high-throughput screening
accelerated to greater than 100,000 tests per day.
[0928] To achieve high-throughput screening, it is advantageous to
house samples on a multicontainer carrier or platform. A
multicontainer carrier facilitates measuring reactions of a
plurality of candidate compounds simultaneously. Multi-well
microplates may be used as the carrier. Such multi-well
microplates, and methods for their use in numerous assays, are both
known in the art and commercially available.
[0929] Screening assays may include controls for purposes of
calibration and confirmation of proper manipulation of the
components of the assay. Blank wells that contain all of the
reactants but no member of the chemical library are usually
included. As another example, a known inhibitor (or activator) of
an enzyme for which modulators are sought, can be incubated with
one sample of the assay, and the resulting decrease (or increase)
in the enzyme activity used as a comparator or control. It will be
appreciated that modulators can also be combined with the enzyme
activators or inhibitors to find modulators which inhibit the
enzyme activation or repression that is otherwise caused by the
presence of the known the enzyme modulator. Similarly, when ligands
to a sphingolipid target are sought, known ligands of the target
can be present in control/calibration assay wells.
[0930] Measuring Enzymatic and Binding Reactions During Screening
Assays
[0931] Techniques for measuring the progression of enzymatic and
binding reactions, e.g., in multicontainer carriers, are known in
the art and include, but are not limited to, the following.
[0932] Spectrophotometric and spectrofluorometric assays are well
known in the art. Examples of such assays include the use of
colorimetric assays for the detection of peroxides, as described in
Gordon, A. J. and Ford, R. A., (1972) The Chemist's Companion: A
Handbook Of Practical Data, Techniques, And References. John Wiley
and Sons, N.Y., Page 437.
[0933] Fluorescence spectrometry may be used to monitor the
generation of reaction products. Fluorescence methodology is
generally more sensitive than the absorption methodology. The use
of fluorescent probes is well known to those skilled in the art.
For reviews, see Bashford et al., (1987) Spectrophotometry and
Spectrofluorometry: A Practical Approach; pp. 91-114, IRL Press
Ltd.; and Bell, (1981) Spectroscopy In Biochemistry. Vol. I, pp.
155-194, CRC Press.
[0934] In spectrofluorometric methods, enzymes are exposed to
substrates that change their intrinsic fluorescence when processed
by the target enzyme. Typically, the substrate is nonfluorescent
and is converted to a fluorophore through one or more reactions. As
a non-limiting example, SMase activity can be detected using the
Amplex.RTM. Red reagent (Molecular Probes, Eugene, Oreg.). In order
to measure sphingomyelinase activity using Amplex.RTM. Red, the
following reactions occur. First, SMase hydrolyzes sphingomyelin to
yield ceramide and phosphorylcholine. Second, alkaline phosphatase
hydrolyzes phosphorylcholine to yield choline. Third, choline is
oxidized by choline oxidase to betaine. Finally, H.sub.2O.sub.2, in
the presence of horseradish peroxidase, reacts with Amplex.RTM. Red
to produce the fluorescent product, Resorufin, and the signal
therefrom is detected using spectrofluorometry.
[0935] Fluorescence polarization (FP) is based on a decrease in the
speed of molecular rotation of a fluorophore that occurs upon
binding to a larger molecule, such as a receptor protein, allowing
for polarized fluorescent emission by the bound ligand. FP is
empirically determined by measuring the vertical and horizontal
components of fluorophore emission following excitation with plane
polarized light. Polarized emission is increased when the molecular
rotation of a fluorophore is reduced. A fluorophore produces a
larger polarized signal when it is bound to a larger molecule (i.e.
a receptor), slowing molecular rotation of the fluorophore. The
magnitude of the polarized signal relates quantitatively to the
extent of fluorescent ligand binding. Accordingly, polarization of
the "bound" signal depends on maintenance of high affinity
binding.
[0936] FP is a homogeneous technology and reactions are very rapid,
taking seconds to minutes to reach equilibrium. The reagents are
stable, and large batches may be prepared, resulting in high
reproducibility. Because of these properties, FP has proven to be
highly automatable, often performed with a single incubation with a
single, premixed, tracer-receptor reagent. For a review, see Owicki
et al., (1997), Application of Fluorescence Polarization Assays in
High-Throughput Screening, Genetic Engineering News, 17:27.
[0937] FP is particularly desirable since its readout is
independent of the emission intensity (Checovich, W. J., et al.,
(1995) Nature 375:254-256; Dandliker, W. B., et al., (1981) Methods
in Enzymology 74:3-28) and is thus insensitive to the presence of
colored compounds that quench fluorescence emission. FP and FRET
(see below) are well-suited for identifying compounds that block
interactions between sphingolipid receptors and their ligands. See,
for example, Parker et al., (2000) Development of high throughput
screening assays using fluorescence polarization: nuclear
receptor-ligand-binding and kinase/phosphatase assays, J Biomol
Screen 5:77-88.
[0938] Fluorophores derived from sphingolipids that may be used in
FP assays are commercially available. For example, Molecular Probes
(Eugene, Oreg.) currently sells sphingomyelin and one ceramide
flurophores. These are, respectively,
N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)sp-
hingosyl phosphocholine (BODIPY.RTM. FL C5-sphingomyelin);
N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoyl)s-
phingosyl phosphocholine (BODIPY.RTM. FL C12-sphingomyelin); and
N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)sp-
hingosine (BODIPY.RTM. FL C5-ceramide). U.S. Pat. No. 4,150,949,
(Immunoassay for gentamicin), discloses fluorescein-labelled
gentamicins, including fluoresceinthiocarbanyl gentamicin.
Additional fluorophores may be prepared using methods well known to
the skilled artisan.
[0939] Exemplary normal-and-polarized fluorescence readers include
the POLARION.RTM. fluorescence polarization system (Tecan AG,
Hombrechtikon, Switzerland). General multiwell plate readers for
other assays are available, such as the VERSAMAX.RTM. reader and
the SPECTRAMAX.RTM. multiwell plate spectrophotometer (both from
Molecular Devices).
[0940] Fluorescence resonance energy transfer (FRET) is another
useful assay for detecting interaction and has been described. See,
e.g., Heim et al., (1996) Curr. Biol. 6:178-182; Mitra et al.,
(1996) Gene 173:13-17; and Selvin et al., (1995) Meth. Enzymol
246:300-345. FRET detects the transfer of energy between two
fluorescent substances in close proximity, having known excitation
and emission wavelengths. As an example, a protein can be expressed
as a fusion protein with green fluorescent protein (GFP). When two
fluorescent proteins are in proximity, such as when a protein
specifically interacts with a target molecule, the resonance energy
can be transferred from one excited molecule to the other. As a
result, the emission spectrum of the sample shifts, which can be
measured by a fluorometer, such as a fMAX multiwell fluorometer
(Molecular Devices, Sunnyvale Calif.).
[0941] Scintillation proximity assay (SPA) is a particularly useful
assay for detecting an interaction with the target molecule. SPA is
widely used in the pharmaceutical industry and has been described
(Hanselman et al., (1997) J. Lipid Res. 38:2365-2373; Kahl et al.,
(1996) Anal. Biochem. 243:282-283; Undenfriend et al., (1987) Anal.
Biochem. 161:494-500). See also U.S. Pat. Nos. 4,626,513 and
4,568,649, and European Patent No. 0,154,734. One commercially
available system uses FLASHPLATE.RTM. scintillant-coated plates
(NEN Life Science Products, Boston, Mass.).
[0942] The target molecule can be bound to the scintillator plates
by a variety of well known means. Scintillant plates are available
that are derivatized to bind to fusion proteins such as GST, His6
or Flag fusion proteins. Where the target molecule is a protein
complex or a multimer, one protein or subunit can be attached to
the plate first, then the other components of the complex added
later under binding conditions, resulting in a bound complex.
[0943] In a typical SPA assay, the gene products in the expression
pool will have been radiolabeled and added to the wells, and
allowed to interact with the solid phase, which is the immobilized
target molecule and scintillant coating in the wells. The assay can
be measured immediately or allowed to reach equilibrium. Either
way, when a radiolabel becomes sufficiently close to the
scintillant coating, it produces a signal detectable by a device
such as a TOPCOUNT NXT.RTM. microplate scintillation counter
(Packard BioScience Co., Meriden Conn.). If a radiolabeled
expression product binds to the target molecule, the radiolabel
remains in proximity to the scintillant long enough to produce a
detectable signal.
[0944] In contrast, the labeled proteins that do not bind to the
target molecule, or bind only briefly, will not remain near the
scintillant long enough to produce a signal above background. Any
time spent near the scintillant caused by random Brownian motion
will also not result in a significant amount of signal. Likewise,
residual unincorporated radiolabel used during the expression step
may be present, but will not generate significant signal because it
will be in solution rather than interacting with the target
molecule. These non-binding interactions will therefore cause a
certain level of background signal that can be mathematically
removed. If too many signals are obtained, salt or other modifiers
can be added directly to the assay plates until the desired
specificity is obtained (Nichols et al., (1998) Anal. Biochem.
257:112-119).
IV. Kinase Activity Assays
[0945] A number of different assays for kinase activity can be
utilized for assaying for active modulators and/or determining
specificity of a modulator for a particular kinase or group or
kinases. In addition to the assay mentioned in the Examples below,
one of ordinary skill in the art will know of other assays that can
be utilized and can modify an assay for a particular application.
For example, numerous papers concerning kinases describe assays
that can be used.
[0946] Additional alternative assays can employ binding
determinations. For example, this sort of assay can be formatted
either in a fluorescence resonance energy transfer (FRET) format,
or using an AlphaScreen (amplified luminescent proximity
homogeneous assay) format by varying the donor and acceptor
reagents that are attached to streptavidin or the phosphor-specific
antibody.
V. Organic Synthetic Techniques
[0947] The versatility of computer-based modulator design and
identification lies in the diversity of structures screened by the
computer programs. The computer programs can search databases that
contain very large numbers of molecules and can modify modulators
already complexed with the enzyme with a wide variety of chemical
functional groups. A consequence of this chemical diversity is that
a potential modulator of kinase function may take a chemical form
that is not predictable. A wide array of organic synthetic
techniques exist in the art to meet the challenge of constructing
these potential modulators. Many of these organic synthetic methods
are described in detail in standard reference sources utilized by
those skilled in the art. One example of such a reference is March,
1994, Advanced Organic Chemistry: Reactions, Mechanisms and
Structure, New York, McGraw Hill. Thus, the techniques useful to
synthesize a potential modulator of kinase function identified by
computer-based methods are readily available to those skilled in
the art of organic chemical synthesis.
[0948] Regarding the synthetic examples described herein, solvents
include polar and non-polar solvents known to those of skill in the
art, including polar aprotic and polar protic solvents. Polar
solvents include, without limitation, protic solvents such as
methanol, ethanol, isopropyl alcohol, t-butanol, n-butanol, acetic
acid, formic acid or water, or aprotic solvents such as
tetrahydrofuran (THF), acetonitrile, dioxane, methylene chloride,
dimethylsulfoxide (DMSO), acetone, N,N-dimethylformamide (DMF),
N,N-dimethylacetamide (DMA), ethyl acetate, 1,2-dimethoxyethane,
1,2-dichloroethane, chloroform, 1,2-dichloroethane, or pyridine.
Polar solvents include a mixture of water with any of the above, or
a mixture of any two or more of the above. Apolar solvents include,
without limitation, toluene, benzene, chlorobenzene, xylenes and
hexanes.
[0949] Regarding the synthetic examples described herein, reducing
agent includes, without limitation, a reducing agent such as
catalytic reducing agents using hydrogen and transition metal
catalysts such as palladium, platinum, rhodium, etc. (e.g.
Pt/acetic acid/H.sub.2); a mixture of trifluoroacetic acid and
triethylsilane, borane tetrahydrofuran complex, diborane, borane
dimethylsulfide complex, and a combination of sodium borohydride
and boron trifluoride; metals such as reduced, iron, zinc powder,
magnesium etc.; metal hydrogen complex compounds such as alkali
metal borohydrides (for example, potassium borohydride, sodium
borohydride, lithium borohydride, zinc borohydride, sodium
triacetoxyborohydride, etc.), aluminum lithium hydride, etc.; metal
hydrides such as sodium hydride, etc.; organic tin compounds
(triphenyltin hydride, etc.); and metal salts such as nickel
compounds, zinc compounds, tin compounds (for example tin(II)
chloride), and samarium iodide/pivalic acid/hexamethylphorphoric
triamide.
[0950] Regarding the synthetic examples described herein, oxidizing
agent includes, without limitation, an oxidizing agent such as
Dess-Martin reagent, TEMPO (2,2,6,6-tetramethylpiperidine-N-oxide),
DDQ (2,3-Dichloro-5,6-dicyano-1,4-benzoquinone), PDC (pyridinium
dichromate), PCC (pyridinium chlorochromate), Pyridine.SO3,
Chromium trioxide, p-nitroperbenzoic acid, magnesium
monoperoxyphthalate, sodium periodate, potassium periodate,
hydrogen peroxide, urea peroxide, alkali metal bromates, cumene
hydroperoxide, tert-butyl peroxide, peracids such as performic
acid, peracetic acid, pertrifluoroacetic acid, perbenzoic acid,
m-chloroperbenzoic acid, o-carboxyperbenzoic acid and the like;
sodium metaperiodate, bichromic acid; bichromates such as sodium
bichromate, potassium bichromate; permanganic acid; permanganates
such as potassium permanganate, sodium permanganate; and lead salts
such as lead tetraacetate.
VI. Alternative Compound Forms or Derivatives
[0951] Compounds contemplated herein are described with reference
to both generic formulae and specific compounds. In addition,
invention compounds may exist in a number of different forms or
derivatives, all within the scope of the present invention. These
include, for example, tautomers, stereoisomers, racemic mixtures,
regioisomers, salts, prodrugs (e.g., carboxylic acid esters),
solvated forms, different crystal forms or polymorphs, and active
metabolites.
(a) Tautomers, Stereoisomers, Regioisomers, and Solvated Forms
[0952] It is understood that some compounds may exhibit
tautomerism. In such cases, the formulae provided herein expressly
depict only one of the possible tautomeric forms. It is therefore
to be understood that the formulae provided herein are intended to
represent any tautomeric form of the depleted compounds and are not
to be limited merely to the specific tautomeric form depicted by
the drawings of the formulae.
[0953] Likewise, some of the compounds according to the present
invention may exist as stereoisomers, i.e. having the same atomic
connectivity of covalently bonded atoms yet differing in the
spatial orientation of the atoms. For example, compounds may be
optical stereoisomers, which contain one or more chiral centers,
and therefore, may exist in two or more stereoisomeric forms (e.g.
enantiomers or diastereomers). Thus, such compounds may be present
as single stereoisomers (i.e., essentially free of other
stereoisomers), racemates, and/or mixtures of enantiomers and/or
diastereomers. As another example, stereoisomers include geometric
isomers, such as cis- or trans-orientation of substituents on
adjacent carbons of a double bond. All such single stereoisomers,
racemates and mixtures thereof are intended to be within the scope
of the present invention. Unless specified to the contrary, all
such steroisomeric forms are included within the formulae provided
herein.
[0954] In some embodiments, a chiral compound of the present
invention is in a form that contains at least 80% of a single
isomer (60% enantiomeric excess ("e.e.") or diastereomeric excess
("d.c.")), or at least 85% (70% e.e. or d.e.), 90% (80% e.e. or
d.e.), 95% (90% e.e. or d.e.), 97.5% (95% e.e. or d.e.), or 99%
(98% e.e. or d.e.). As generally understood by those skilled in the
art, an optically pure compound having one chiral center is one
that consists essentially of one of the two possible enantiomers
(i.e., is enantiomerically pure), and an optically pure compound
having more than one chiral center is one that is both
diastereomerically pure and enantiomerically pure. In some
embodiments, the compound is present in optically pure form.
[0955] For compounds in which synthesis involves addition of a
single group at a double bond, particularly a carbon-carbon double
bond, the addition may occur at either of the double bond-linked
atoms. For such compounds, the present invention includes both such
regioisomers.
[0956] Additionally, the formulae are intended to cover solvated as
well as unsolvated forms of the identified structures. For example,
the indicated structures include both hydrated and noh-hydrated
forms. Other examples of solvates include the structures in
combination with a suitable solvent such as isopropanol, ethanol,
methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine.
(b) Prodrugs and Metabolites
[0957] In addition to the present formulae and compounds described
herein, the invention also includes prodrugs (generally
pharmaceutically acceptable prodrugs), active metabolic derivatives
(active metabolites), and their pharmaceutically acceptable
salts.
[0958] Prodrugs are compounds or pharmaceutically acceptable salts
thereof which, when metabolized under physiological conditions or
when converted by solvolysis, yield the desired active compound.
Prodrugs include, without limitation, esters, amides, carbamates,
carbonates, ureides, solvates, or hydrates of the active compound.
Typically, the prodrug is inactive, or less active than the active
compound, but may provide one or more advantageous handling,
administration, and/or metabolic properties. For example, some
prodrugs are esters of the active compound; during metabolysis, the
ester group is cleaved to yield the active drug. Also, some
prodrugs are activated enzymatically to yield the active compound,
or a compound which, upon further chemical reaction, yields the
active compound.
[0959] In this context, a common example or a prodrug is an alkyl
ester of a carboxylic acid. Relative to compounds of Formula I,
further examples include, without limitation, an amide or carbamate
derivative at the 1-position nitrogen of the azaindole core.
[0960] As described in The Practice of Medicinal Chemistry. Ch.
31-32 (Ed. Wermuth, Academic Press, San Diego, Calif., 2001),
prodrugs can be conceptually divided into two non-exclusive
categories, bioprecursor prodrugs and carrier prodrugs. Generally,
bioprecursor prodrugs are compounds that are inactive or have low
activity compared to the corresponding active drug compound, that
contain one or more protective groups and are converted to an
active form by metabolism or solvolysis. Both the active drug form
and any released metabolic products should have acceptably low
toxicity. Typically, the formation of active drug compound involves
a metabolic process or reaction that is one of the follow
types:
[0961] Oxidative reactions: Oxidative reactions are exemplified
without limitation to reactions such as oxidation of alcohol,
carbonyl, and acid functionalities, hydroxylation of aliphatic
carbons, hydroxylation of alicyclic carbon atoms, oxidation of
aromatic carbon atoms, oxidation of carbon-carbon double bonds,
oxidation of nitrogen-containing functional groups, oxidation of
silicon, phosphorus, arsenic, and sulfur, oxidative N-dealkylation,
oxidative O- and S-dealkylation, oxidative deamination, as well as
other oxidative reactions.
[0962] Reductive reactions: Reductive reactions are exemplified
without limitation to reactions such as reduction of carbonyl
functionalities, reduction of alcohol functionalities and
carbon-carbon double bonds, reduction of nitrogen-containing
function groups, and other reduction reactions.
[0963] Reactions without change in the oxidation state: Reactions
without change in the state of oxidation are exemplified without
limitation to reactions such as hydrolysis of esters and ethers,
hydrolytic cleavage of carbon-nitrogen single bonds, hydrolytic
cleavage of non-aromatic heterocycles, hydration and dehydration at
multiple bonds, new atomic linkages resulting from dehydration
reactions, hydrolytic dehalogenation, removal of hydrogen halide
molecule, and other such reactions.
[0964] Carrier prodrugs are drug compounds that contain a transport
moiety, e.g., that improves uptake and/or localized delivery to a
site(s) of action. Desirably for such a carrier prodrug, the
linkage between the drug moiety and the transport moiety is a
covalent bond, the prodrug is inactive or less active than the drug
compound, the prodrug and any release transport moiety are
acceptably non-toxic. For prodrugs where the transport moiety is
intended to enhance uptake, typically the release of the transport
moiety should be rapid. In other cases, it is desirable to utilize
a moiety that provides slow release, e.g., certain polymers or
other moieties, such as cyclodextrins. (See, e.g., Cheng et al.,
U.S. Patent Publ. No. 20040077595, application Ser. No. 10/656,838,
incorporated herein by reference). Such carrier prodrugs are often
advantageous for orally administered drugs. Carrier prodrugs can,
for example, be used to improve one or more of the following
properties: increased lipophilicity, increased duration of
pharmacological effects, increased site-specificity, decreased
toxicity and adverse reactions, and/or improvement in drug
formulation (e.g., stability, water solubility, suppression of an
undesirable organoleptic or physiochemical property). For example,
lipophilicity can be increased by esterification of hydroxyl groups
with lipophilic carboxylic acids, or of carboxylic acid groups with
alcohols, e.g., aliphatic alcohols. Wermuth, supra.
[0965] Prodrugs may proceed from prodrug form to active form in a
single step or may have one or more intermediate forms which may
themselves have activity or may be inactive.
[0966] Metabolites, e.g., active metabolites overlap with prodrugs
as described above, e.g., bioprecursor prodrugs. Thus, such
metabolites are pharmacologically active compounds or compounds
that further metabolize to pharmacologically active compounds that
are derivatives resulting from metabolic process in the body of a
subject. Of these, active metabolites are such pharmacologically
active derivative compounds. For prodrugs, the prodrug compound is
generally inactive or of lower activity than the metabolic product.
For active metabolites, the parent compound may be either an active
compound or may be an inactive prodrug.
[0967] Prodrugs, and active metabolites may be identified using
routine techniques know in the art. See, e.g., Bertolini et al,
1997, J Med Chem 40:2011-2016; Shan et al., J Pharm Sci 86:756-757;
Bagshawe, 1995, Drug Dev Res 34:220-230; Wermuth, supra.
(c) Pharmaceutically Acceptable Salts
[0968] Compounds can be formulated as or be in the form of
pharmaceutically acceptable salts. Contemplated pharmaceutically
acceptable salt forms include, without limitation, mono, bis, tris,
tetrakis, and so on. Pharmaceutically acceptable salts are
non-toxic in the amounts and concentrations at which they are
administered. The preparation of such salts can facilitate the
pharmacological use by altering the physical characteristics of a
compound without preventing it from exerting its physiological
effect. Useful alterations in physical properties include lowering
the melting point to facilitate transmucosal administration and
increasing the solubility to facilitate administering higher
concentrations of the drug.
[0969] Pharmaceutically acceptable salts include acid addition
salts such as those containing sulfate, chloride, hydrochloride,
fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate,
tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate,
p-toluenesulfonate, cyclohexylsulfamate and quinate.
Pharmaceutically acceptable salts can be obtained from acids such
as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid,
sulfamic acid, acetic acid, citric acid, lactic acid, tartaric
acid, malonic acid, methanesulfonic acid, ethanesulfonic acid,
benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic
acid, fumaric acid, and quinic acid.
[0970] Pharmaceutically acceptable salts also include basic
addition salts such as those containing benzathine, chloroprocaine,
choline, diethanolamine, ethanolamine, t-butylamine,
ethylenediamine, meglumine, procaine, aluminum, calcium, lithium,
magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when
acidic functional groups, such as carboxylic acid or phenol are
present. For example, see Remington's Pharmaceutical Sciences,
19.sup.th ed., Mack Publishing Co., Easton, Pa., Vol. 2, p. 1457,
1995. Such salts can be prepared using the appropriate
corresponding bases.
[0971] Pharmaceutically acceptable salts can be prepared by
standard techniques. For example, the free-base form of a compound
can be dissolved in a suitable solvent, such as an aqueous or
aqueous-alcohol solution containing the appropriate acid and then
isolated by evaporating the solution. In another example, a salt
can be prepared by reacting the free base and acid in an organic
solvent.
[0972] Thus, for example, if the particular compound is a base, the
desired pharmaceutically acceptable salt may be prepared by any
suitable method available in the art, for example, treatment of the
free base with an inorganic acid, such as hydrochloric acid,
hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and
the like, or with an organic acid, such as acetic acid, maleic
acid, succinic acid, mandelic acid, fumaric acid, malonic acid,
pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a
pyranosidyl acid, such as glucuronic acid or galacturonic acid, an
alpha-hydroxy acid, such as citric acid or tartaric acid, an amino
acid, such as aspartic acid or glutamic acid, an aromatic acid,
such as benzoic acid or cinnamic acid, a sulfonic acid, such as
p-toluenesulfonic acid or ethanesulfonic acid, or the like.
[0973] Similarly, if the particular compound is an acid, the
desired pharmaceutically acceptable salt may be prepared by any
suitable method, for example, treatment of the free acid with an
inorganic or organic base, such as an amine (primary, secondary or
tertiary), an alkali metal hydroxide or alkaline earth metal
hydroxide, or the like. Illustrative examples of suitable salts
include organic salts derived from amino acids, such as L-glycine,
L-lysine, and L-arginine, ammonia, primary, secondary, and tertiary
amines, and cyclic amines, such as hydroxyethylpyrrolidine,
piperidine, morpholine or piperazine, and inorganic salts derived
from sodium, calcium, potassium, magnesium, manganese, iron,
copper, zinc, aluminum and lithium. Further examples of
pharmaceutically acceptable salts of compounds of Formulae I-III
include, without limitation, the mono-sodium and bis-potassium
salts thereof.
[0974] The pharmaceutically acceptable salt of the different
compounds may be present as a complex. Examples of complexes
include 8-chlorotheophylline complex (analogous to, e.g.,
dimenhydrinate: diphenhydramine 8-chlorotheophylline (1:1) complex;
Dramamine) and various cyclodextrin inclusion complexes.
[0975] Unless specified to the contrary, specification of a
compound herein includes pharmaceutically acceptable salts of such
compound.
(d) Polymorphic Forms
[0976] In the case of agents that are solids, it is understood by
those skilled in the art that the compounds and salts may exist in
different crystal or polymorphic forms, all of which are intended
to be within the scope of the present invention and specified
formulae.
VII. Administration
[0977] The methods and compounds will typically be used in therapy
for human subjects. However, they may also be used to treat similar
or identical indications in other animal subjects. In this context,
the terms "subject," "animal subject," and the like refer to human
and non-human vertebrates, e.g. mammals, such as non-human
primates, sports and commercial animals, e.g., equines, bovines,
porcines, ovines, rodents, and pets, e.g., canines and felines.
[0978] Suitable dosage forms, in part, depend upon the use or the
route of administration, for example, oral, transdermal,
transmucosal, inhalant or by injection (parenteral). Such dosage
forms should allow the compound to reach target cells. Other
factors are well known in the art, and include considerations such
as toxicity and dosage forms that retard the compound or
composition from exerting its effects. Techniques and formulations
generally may be found in The Science and Practice of Pharmacy,
21.sup.st edition, Lippincott, Williams and Wilkins, Philadelphia,
Pa., 2005 (hereby incorporated by reference herein).
[0979] Compounds of the present invention, i.e. Formula I, can be
formulated as pharmaceutically acceptable salts.
[0980] Carriers or excipients can be used to produce compositions.
The carriers or excipients can be chosen to facilitate
administration of the compound. Examples of carriers include
calcium carbonate, calcium phosphate, various sugars such as
lactose, glucose, or sucrose, or types of starch, cellulose
derivatives, gelatin, vegetable oils, polyethylene glycols and
physiologically compatible solvents. Examples of physiologically
compatible solvents include sterile solutions of water for
injection (WFI), saline solution, and dextrose.
[0981] The compounds can be administered by different routes
including intravenous, intraperitoneal, subcutaneous,
intramuscular, oral, transmucosal, rectal, transdermal, or
inhalant. Oral administration is preferred. For oral
administration, for example, the compounds can be formulated into
conventional oral dosage forms such as capsules, tablets, and
liquid preparations such as syrups, elixirs, and concentrated
drops.
[0982] For inhalants, compounds of the invention may be formulated
as dry powder or a suitable solution, suspension, or aerosol.
Powders and solutions may be formulated with suitable additives
known in the art. For example, powders may include a suitable
powder base such as lactose or starch, and solutions may comprise
propylene glycol, sterile water, ethanol, sodium chloride and other
additives, such as acid, alkali and buffer salts. Such solutions or
suspensions may be administered by inhaling via spray, pump,
atomizer, or nebulizer, and the like. The compounds of the
invention may also be used in combination with other inhaled
therapies, for example corticosteroids such as fluticasone
proprionate, beclomethasone dipropionate, triamcinolone acetonide,
budesonide, and mometasone furoate; beta agonists such as
albuterol, salmeterol, and formoterol; anticholinergic agents such
as ipratroprium bromide or tiotropium; vasodilators such as
treprostinal and iloprost; enzymes such as DNAase; therapeutic
proteins; immunoglobulin antibodies; an oligonucleotide, such as
single or double stranded DNA or RNA, siRNA; antibiotics such as
tobramycin; muscarinic receptor antagonists; leukotriene
antagonists; cytokine antagonists; protease inhibitors; cromolyn
sodium; nedocril sodium; and sodium cromoglycate.
[0983] Pharmaceutical preparations for oral use can be obtained,
for example, by combining the active compounds with solid
excipients, optionally grinding a resulting mixture, and processing
the mixture of granules, after adding suitable auxiliaries, if
desired, to obtain tablets or dragee cores. Suitable excipients
are, in particular, fillers such as sugars, including lactose,
sucrose, mannitol, or sorbitol; cellulose preparations, for
example, maize starch, wheat starch, rice starch, potato starch,
gelatin, gum tragacanth, methyl cellulose,
hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose (CMC),
and/or polyvinylpyrrolidone (PVP: povidone). If desired,
disintegrating agents may be added, such as the cross-linked
polyvinylpyrrolidone, agar, or alginic acid, or a salt thereof such
as sodium alginate.
[0984] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain, for example, gum arabic, talc,
polyvinylpyrrolidone, carbopol gel, polyethylene glycol (PEG),
and/or titanium dioxide, lacquer solutions, and suitable organic
solvents or solvent mixtures. Dye-stuffs or pigments may be added
to the tablets or dragee coatings for identification or to
characterize different combinations of active compound doses.
[0985] Pharmaceutical preparations that can be used orally include
push-fit capsules made of gelatin ("gelcaps"), as well as soft,
sealed capsules made of gelatin, and a plasticizer, such as
glycerol or sorbitol. The push-fit capsules can contain the active
ingredients in admixture with filler such as lactose, binders such
as starches, and/or lubricants such as talc or magnesium stearate
and, optionally, stabilizers. In soft capsules, the active
compounds may be dissolved or suspended in suitable liquids, such
as fatty oils, liquid paraffin, or liquid polyethylene glycols
(PEGs). In addition, stabilizers may be added.
[0986] Alternatively, injection (parenteral administration) may be
used, e.g., intramuscular, intravenous, intraperitoneal, and/or
subcutaneous. For injection, the compounds of the invention are
formulated in sterile liquid solutions, preferably in
physiologically compatible buffers or solutions, such as saline
solution, Hank's solution, or Ringer's solution. In addition, the
compounds may be formulated in solid form and redissolved or
suspended immediately prior to use. Lyophilized forms can also be
produced.
[0987] Administration can also be by transmucosal, topical, or
transdermal means. For transmucosal, topical or transdermal
administration, penetrants appropriate to the barrier to be
permeated are used in the formulation. Such penetrants are
generally known in the art, and include, for example, for
transmucosal administration, bile salts and fusidic acid
derivatives. In addition, detergents may be used to facilitate
permeation. Transmucosal administration, for example, may be
through nasal sprays or suppositories (rectal or vaginal). The
topical compositions of this invention are formulated preferably as
oils, creams, lotions, ointments and the like by choice of
appropriate carriers known in the art. Suitable carriers include
vegetable or mineral oils, white petrolatum (white soft paraffin),
branched chain fats or oils, animal fats and high molecular weight
alcohol (greater than C.sub.12). The preferred carriers are those
in which the active ingredient is soluble. Emulsifiers,
stabilizers, humectants and antioxidants may also be included as
well as agents imparting color or fragrance, if desired. Creams for
topical application are preferably formulated from a mixture of
mineral oil, self-emulsifying beeswax and water in which mixture
the active ingredient, dissolved in a small amount solvent (e.g.,
an oil), is admixed. Additionally, administration by transdermal
means may comprise a transdermal patch or dressing such as a
bandage impregnated with an active ingredient and optionally one or
more carriers or diluents known in the art. To be administered in
the form of a transdermal delivery system, the dosage
administration will, of course, be continuous rather than
intermittent throughout the dosage regimen.
[0988] The amounts of various compounds to be administered can be
determined by standard procedures taking into account factors such
as the compound IC.sub.50, the biological half-life of the
compound, the age, size, and weight of the subject, and the
disorder associated with the subject. The importance of these and
other factors are well known to those of ordinary skill in the art.
Generally, a dose will be between about 0.01 and 50 mg/kg,
preferably 0.1 and 20 mg/kg of the subject being treated. Multiple
doses may be used.
[0989] The compounds of the invention may also be used in
combination with other therapies for treating the same disease.
Such combination use includes administration of the compounds and
one or more other therapeutics at different times, or
co-administration of the compound and one or more other therapies.
In some embodiments, dosage may be modified for one or more of the
compounds of the invention or other therapeutics used in
combination, e.g., reduction in the amount dosed relative to a
compound or therapy used alone, by methods well known to those of
ordinary skill in the art.
[0990] It is understood that use in combination includes use with
other therapies, drugs, medical procedures etc., where the other
therapy or procedure may be administered at different times (e.g.
within a short time, such as within hours (e.g. 1, 2, 3, 4-24
hours), or within a longer time (e.g. 1-2 days, 2-4 days, 4-7 days,
1-4 weeks)) than a compound of the present invention, or at the
same time as a compound of the invention. Use in combination also
includes use with a therapy or medical procedure that is
administered once or infrequently, such as surgery, along with a
compound of the invention administered within a short time or
longer time before or after the other therapy or procedure. In some
embodiments, the present invention provides for delivery of
compounds of the invention and one or more other drug therapeutics
delivered by a different route of administration or by the same
route of administration. The use in combination for any route of
administration includes delivery of compounds of the invention and
one or more other drug therapeutics delivered by the same route of
administration together in any formulation, including formulations
where the two compounds are chemically linked in such a way that
they maintain their therapeutic activity when administered. In one
aspect, the other drug therapy may be co-administered with one or
more compounds of the invention. Use in combination by
co-administration includes administration of co-formulations or
formulations of chemically joined compounds, or administration of
two or more compounds in separate formulations within a short time
of each other (e.g. within an hour, 2 hours, 3 hours, up to 24
hours), administered by the same or different routes.
Co-administration of separate formulations includes
co-administration by delivery via one device, for example the same
inhalant device, the same syringe, etc., or administration from
separate devices within a short time of each other. Co-formulations
of compounds of the invention and one or more additional drug
therapies delivered by the same route includes preparation of the
materials together such that they can be administered by one
device, including the separate compounds combined in one
formulation, or compounds that are modified such that they are
chemically joined, yet still maintain their biological activity.
Such chemically joined compounds may have a linkage that is
substantially maintained in vivo, or the linkage may break down in
vivo, separating the two active components.
EXAMPLES
[0991] Examples related to the present invention are described
below. In most cases, alternative techniques can be used. The
examples are intended to be illustrative and are not limiting or
restrictive to the scope of the invention. In some examples, the
mass spectrometry result indicated for a compound may have more
than one value due to the isotope distribution of an atom in the
molecule, such as a compound having a bromo or chloro
substituent.
Example 1
Synthesis of propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide P-0773 and Related Compounds
[0992] Compound P-0773 was synthesized in five steps from
2,4-difluoro-phenylamine 42 as shown in Scheme 13.
##STR00153##
Step 1--Preparation of 3-amino-2,6-difluoro-benzoic acid benzyl
ester (43)
[0993] To 2,4-difluoro-phenylamine (42, 5.11 mL, 50.7 mmol) in
tetrahydrofuran (250 mL), cooled with dry ice/acetone bath under an
atmosphere of nitrogen, was added n-butyllithium (1.60 M in hexane,
34.0 mL, 54.4 mmol) slowly. After 30 minutes,
1,2-Bis-(chloro-dimethyl-silanyl)-ethane (11.5 g, 53.4 mmol)
dissolved in tetrahydrofuran (40.0 mL) was added to the reaction
slowly. After 1 hour, n-butyllithium (1.60 M in hexane, 31.9 mL,
51.0 mmol) was added slowly to the reaction. The reaction was
stirred at -78.degree. C. for 30 minutes and then allowed to warm
to room temperature over 40 minutes. The reaction was cooled to
-78.degree. C., followed by addition of n-butyllithium (1.60 M in
hexane, 35.1 mL, 56.2 mmol) slowly. After 70 minutes, benzyl
chloroformate (7.97 mL, 55.8 mmol) was added to the reaction. The
reaction mixture was stirred at -78.degree. C. overnight followed
by addition of 2N HCl (120 mL). The reaction was allowed to warm to
room temperature for 2 hours. The organic layer was separated. The
aqueous layer was basified with potassium carbonate and extracted
with ethyl acetate. The organic layers were combined and washed
with brine, dried over anhydrous sodium sulfate, filtrated and
concentrated. The desired compound was isolated by silica gel
column chromatography (ethyl acetate/hexane 20%) to give a
colorless oil (43, 10.6 g, 79.7%). MS(ESI)
[M+H.sup.+].sup.+=264.1.
Step 2--Preparation of
2,6-difluoro-3-(propane-1-sulfonylamino)-benzoic acid benzyl ester
(44)
[0994] To 3-amino-2,6-difluoro-benzoic acid benzyl ester (43, 6.00
g, 22.8 mmol) in methylene chloride (150 mL) was added pyridine
(2.76 mL, 34.2 mmol) and propane-1-sulfonyl chloride (3.80 mL, 33.8
mmol). The reaction was stirred at room temperature overnight. Then
the reaction was poured into water, and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate, filtrated and concentrated. The desired
compound was isolated with silica gel column chromatography to give
a colorless oil (44, 7.0 g, 83.1%). MS(ESI)
[M+H.sup.+].sup.+=370.1.
Step 3--Preparation of
2,6-difluoro-3-(propane-1-sulfonylamino)-benzoic acid (45)
[0995] To 2,6-difluoro-3-(propane-1-sulfonylamino)-benzoic acid
benzyl ester (44, 2.0 g, mmol) in methanol (30 mL) was added 20%
palladium hydroxide on carbon (100 mg). The reaction was stirred
under hydrogen at 1 atm for 15 minutes. The reaction was filtrated
and concentrated to give white solid 45 that was used in the next
step.
Step 4--Preparation of
2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl chloride (46)
[0996] To 2,6-difluoro-3-(propane-1-sulfonylamino)-benzoic acid
(45, 1.50 g, 5.4 mmol) was added toluene (7.0 mL) and thionyl
chloride (15.0 mL, 0.21 mmol). The reaction was heated to reflux
for 3 hours. The reaction was concentrated to give crude compound
that was used in the next step.
Step 5--Preparation of propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0773)
[0997] To aluminum trichloride (8.89 g, 66.7 mmol) was added
methylene chloride (150 mL) under an atmosphere of nitrogen below
5.degree. C. Into this, 5-bromo-7-azaindole (67, 1.64 g, 8.34 mmol)
in methylene chloride (20 mL) was added. The reaction was stirred
for 60.0 minutes and
2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl chloride (46, 3.50
g, 11.8 mmol) in methylene chloride (20 mL) was added. The reaction
was stirred for 6 hours and warmed to room temperature overnight.
The reaction mixture was poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate,
filtrated and concentrated. The desired compound was isolated by
silica gel column chromatography (methylene chloride/methanol 5%)
to give a white solid (P-0773, 1.2 g, 31.4%). MS(ESI)
[M+H.sup.+].sup.+=460.0, 462.0.
[0998]
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-chloro-2-fluo-
ro-phenyl]-methanesulfonamide P-0868
##STR00154##
was prepared following the protocol of Scheme 13, substituting
propane-1-sulfonyl chloride with methanesulfonyl chloride in Step 2
and 2,4-difluoro-phenylamine with 4-chloro-2-fluoro-phenylamine in
Step 1. MS (ESI) [M-H.sup.+].sup.-=443.9, 445.9.
[0999]
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-ph-
enyl]-methanesulfonamide P-0162
##STR00155##
was prepared following the protocol of Scheme 13, substituting
propane-1-sulfonyl chloride with methanesulfonyl chloride in Step
2. This was isolated and reacted further (50 mg, 0.10 mmol) in
methanol (20 mL) by adding 20% palladium hydroxide on carbon (53
mg). The reaction was stirred under hydrogen at 40 psi for 12
hours. The reaction mixture was filtered and concentrated to give
product methanesulfonic acid
[2,4-difluoro-3-1H-pyrrolo-[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-0811 (40 mg, 95%).
##STR00156##
MS(ESI)[M+H.sup.+].sup.+=352.3.
[1000]
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-ph-
enyl]-benzenesulfonamide P-0798 and
N-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]--
3-fluoro-benzenesulfonamide
##STR00157##
were prepared following the protocol of Scheme 13, substituting
propane-1-sulfonyl chloride with benzenesulfonyl chloride and
3-fluoro-benzenesulfonyl chloride, respectively, in Step 2. P-0798
MS(ESI) [M-H.sup.+].sup.-=489.9, 491.9.
[1001] Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-chloro-2-fluoro-pheny-
l]-amide P-0805
##STR00158##
was prepared following the protocol of Scheme 13, substituting
2,4-difluoro-phenylamine with 4-chloro-2-fluoro-phenylamine in Step
1. MS(ESI) [M-H.sup.+].sup.-=471.9, 473.9.
[1002] Propane-1-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-0007
##STR00159##
was prepared following the protocol of Scheme 13, substituting
5-bromo-7-azaindole with 7-azaindole in Step 5. MS(ESI)
[M+H.sup.+].sup.+=380.1.
[1003] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
amide P-0806
##STR00160##
was prepared following the protocol of Scheme 13, substituting
5-bromo-7-azaindole with 5-methoxy-7-azaindole 104 (prepared as
described in Example 16) in Step 5. MS(ESI)
[M-H.sup.+].sup.-=410.1.
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phenyl)-methan-
one P-0265
##STR00161##
[1004] was prepared using the protocol of steps 4 and 5 of Scheme
13, substituting 2,6-difluoro-3-(propane-1-sulfonylamino)-benzoic
acid with 2-fluoro-3-methoxy-benzoic acid in Step 4. MS (ESI)
[M-H.sup.+].sup.-=347, 349.
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-metha-
nesulfonamide P-0170
##STR00162##
[1005] was prepared using the protocol of steps 2, 4, and 5 of
Scheme 13, substituting 3-amino-2,6-difluoro-benzoic acid benzyl
ester with 5-amino-2-fluoro-benzoic acid and propane-1-sulfonyl
chloride with methanesulfonyl chloride in Step 2, to provide the
acid that is carried through in Step 4. MS (ESI)
[M-H.sup.+].sup.-=410.0, 412.0.
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-methyl-phenyl]-metha-
nesulfonamide P-0180
##STR00163##
[1006] was prepared using the protocol of steps 2, 4, and 5 of
Scheme 13, substituting 3-amino-2,6-difluoro-benzoic acid benzyl
ester with 3-amino-2-methyl-benzoic acid and propane-1-sulfonyl
chloride with methanesulfonyl chloride in Step 2, to provide the
acid that is carried through in Step 4. MS (ESI)
[M-H.sup.+].sup.-=405.9, 407.9.
(5-Bromo
-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-chloro-2-fluoro-phenyl)-methan-
one P-0299
##STR00164##
[1007] was prepared using the protocol of Step 5 of Scheme 13,
substituting 2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl
chloride with 3-chloro-2-fluoro-benzoyl chloride. MS (ESI)
[M-H.sup.+].sup.--350.9, 352.9.
Example 2
Synthesis of Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]-amide
P-0955 and Related Compounds
[1008] Compound P-0955 was synthesized in six steps from
4-chloro-2-fluoro-phenylamine 47 as shown in Scheme 14.
##STR00165##
Step 1--Preparation of 3-Amino-6-chloro-2-fluoro-benzoic acid
benzyl ester (48)
[1009] To 4-chloro-2-fluoro-phenylamine (47, 6.30 mL, 57.0 mmol) in
tetrahydrofuran (300 mL), cooled with dry ice/acetone bath under an
atmosphere of nitrogen, was added n-butyllithium (2.500 M in
hexane, 24.4 mL) slowly. After 20 minutes,
1,2-Bis-(chloro-dimethyl-silanyl)-ethane (12.9 g, 60.0 mmol)
dissolved in tetrahydrofuran (40.0 mL) was added to the reaction
slowly. After 1 hour, n-butyllithium (2.50 M in hexane, 25.0 mL)
was added slowly to the reaction. The reaction was stirred at
-78.degree. C. for 20 minutes and allowed to warm to room
temperature over 60 minutes. The reaction was cooled to -78.degree.
C., followed by addition of n-butyllithium (2.50 M in hexane, 26.0
mL) slowly. After 80 minutes, benzyl chloroformate (10.0 mL, 70.0
mmol) was added to the reaction. The reaction mixture was stirred
at -78.degree. C. overnight followed by addition of water (80 mL)
and concentrated hydrochloric acid (25 mL). The reaction was
allowed to warm to room temperature for 2 hours. The organic layer
was separated and the aqueous layer was basified with potassium
carbonate and extracted with ethyl acetate. The organic layers were
combined and Washed with brine, dried over anhydrous sodium
sulfate, filtrated and concentrated. The desired compound was
isolated by silica gel column chromatography (ethyl acetate/hexane
20%) to give a colorless oil (48, 12.5 g, 78.3%). MS(ESI)
[M+H.sup.+].sup.+=280.0.
Step 2--Preparation of
6-chloro-2-fluoro-5-(propane-1-sulfonylamino)-benzoic acid benzyl
ester (49)
[1010] To 3-amino-6-chloro-2-fluoro-benzoic acid benzyl ester (48,
1.20 g, 4.3 mmol) in methylene chloride (28 mL) was added pyridine
(0.52 mL, 6.4 mmol) and propane-1-sulfonyl chloride (0.685 g, 4.8
mmol). The reaction was stirred at room temperature overnight. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was washed with brine, dried over anhydrous
sodium sulfate, filtrated and concentrated. The desired compound
was isolated with silica gel column chromatography to give a
colorless oil (49, 960 mg, 58.0%). MS (ESI)
[M-H.sup.+].sup.-=384.1.
Step 3--Preparation of
6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid
(115)
[1011] To 6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid benzyl ester (49, 6.00 g, 15.6 mmol) in tetrahydrofuran (100
mL) was added 1.0 M aqueous potassium hydroxide (100 mL). The
reaction was heated to reflux overnight. The reaction was poured
into water, acidified to pH 2 with 1 N hydrochloric acid and
extracted with ethyl acetate. The organic portion was dried over
anhydrous sodium sulfate, filtrated and concentrated to give a
white solid 115 (3.95 g, 85.8%).
Step 4--Preparation of 2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid (50)
[1012] To 6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid (115, 0.69 g, 2.3 mmol) in methanol (10 mL) was added 20%
palladium hydroxide on carbon (200 mg). The reaction was stirred
under hydrogen at 50 psi for 2 hours. The reaction was filtrated
and concentrated to give white solid 50 that was used in the next
step. MS(ESI) [M-H.sup.+].sup.-=260.1.
Step 5--Preparation of 2-fluoro-3-(propane-1-sulfonylamino)-benzoyl
chloride (51)
[1013] To 2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid (50,
1.17 g, 4.5 mmol) was added thionyl chloride (10.0 mL). The
reaction was heated to reflux for 3 hours. The reaction was
concentrated to give crude compound 51 that was used in the next
step.
Step 6--Preparation of Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]-amide
(P-0955)
[1014] Aluminum trichloride (2.52 g, 18.9 mmol) and methylene
chloride (60.0 mL) were combined under an atmosphere of nitrogen.
Into the reaction mixture, was added 5-bromo-7-azaindole (67, 630.0
mg, 3.2 mmol) in methylene dichloride (20.0 mL). The reaction was
stirred at room temperature for 70 minutes, then
2-fluoro-3-(propane-1-sulfonylamino)-benzoyl chloride (51, 0.749 g,
2.68 mmol) in methylene chloride (20 mL) was added. The reaction
was stirred at room temperature for 2 hours. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 30% ethyl acetate in hexane to give
compound P-0955 (65 mg, 5.5%). MS(ESI) [M+H.sup.+].sup.+=440.2,
442.2.
[1015] Using the protocol of Scheme 14, substituting
5-bromo-azaindole with either 5-chloro-7-azaindole (80, prepared as
described in Example 9), 5-fluoro-7-azaindole (81, prepared as
described in Example 9) or 7-azaindole in Step 6,
propane-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]-amide
P-1013 (MS(ESI) [M-H.sup.+].sup.-=394.1), propane-1-sulfonic acid
[2-fluoro-3-(5-fluoro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-1028 (MS(ESI) [M-H.sup.+].sup.-=378.1), and propane-1-sulfonic
acid
[2-fluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-1056 (MS(ESI) [M+H.sup.+].sup.+=362.2) were prepared,
respectively;
##STR00166##
Example 3
Propane-1-sulfonic acid
[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-amide P-0088 and Related Compounds
[1016] Compound P-0088 was synthesized in one step from
propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-pheny-
l]-amide P-0773 by Suzuki coupling ((Muyaura and Suzuki, Chem. Rev.
1995, 95:2457) as shown in Scheme 15.
##STR00167##
Step 1--Preparation of propane-1-sulfonic acid
[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-amide (P-0088)
[1017] To propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0773, prepared as described in Example 1, 65.0 mg, 0.14
mmol) in acetonitrile (4.0 mL) was added pyridine-3-boronic acid
(609, 25.0 mg, 0.20 mmol), tetrakis
(triphenylphosphine)palladium(0) (11 mg, 1.0% mmol) and aqueous
potassium carbonate (1.0 M, 2.0 mL). The reaction was heated to
160.degree. C. for 10 minutes in a CEM Discover microwave
instrument. The reaction was poured into water, and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate, filtrated and concentrated. The desired
compound was isolated by silica gel column chromatography
(methylene chloride/methanol 5%) to give a white solid (P-0088, 30
mg, 46.9%). MS(ESI) [M+H.sup.+].sup.+=457.2.
[1018] Additional compounds were prepared following the protocol of
Scheme 15, optionally replacing propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide P-0773 with an appropriate 5-bromo azaindole and/or
pyridine-3-boronic acid 609 with an appropriate boronic acid or
boronic acid ester. The 5-bromo azaindole used was synthesized as
described in either Example 1,2, 5, 10, or 73. The following
compounds were made following this procedure: [1019]
N-[2-Methyl-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-met-
hanesulfonamide (P-0082), [1020]
N-[2-Methyl-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-methanesulfonamide (P-0121), [1021]
N-[4-Fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-methanesulfonamide (P-0079), [1022]
N-[4-Fluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-met-
hanesulfonamide (P-0308), [1023]
N-{4-Fluoro-3-[5-(3-methanesulfonylamino-phenyl)-1H-pyrrolo[2,3-b]pyridin-
e-3-carbonyl]-phenyl}-methanesulfonamide (P-0156), [1024]
N-{4-Fluoro-3-[5-(3-methanesulfonyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl]-phenyl}-methanesulfonamide (P-0297), [1025]
3-[3-(2-Fluoro-5-methanesulfonylamino-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-
-yl]-benzamide (P-0228), [1026]
3-[3-(2-Fluoro-5-hydroxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzamid-
e (P-0037), [1027]
(2-Fluoro-5-hydroxy-phenyl)-[5-(3-methanesulfonyl-phenyl)-1H-pyrrolo[2,3--
b]pyridin-3-yl]-methanone (P-0008), [1028]
N-{3-[3-(2-Fluoro-5-hydroxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-pheny-
l}-methanesulfonamide (P-0716), [1029]
N-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-methanesulfonamide (P-0700), [1030]
N-{4-Chloro-2-fluoro-3-[5-(3-methanesulfonyl-phenyl)-1H-pyrrolo[2,3-b]
pyridine-3-carbonyl]-phenyl}-methanesulfonamide (P-0841), [1031]
N-[4-Chloro-2-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbon-
yl)-phenyl]-methanesulfonamide (P-0734), [1032]
N-{4-Chloro-2-fluoro-3-[5-(3-methanesulfonylamino-phenyl)-1H-pyrrolo[2,3--
b]pyridine-3-carbonyl]-phenyl}-methanesulfonamide (P-0745), [1033]
N-{2,4-Difluoro-3-[5-(3-methanesulfonylamino-phenyl)-1H-pyrrolo[2,3-b]
pyridine-3-carbonyl]-phenyl}-methanesulfonamide (P-0746), [1034]
3-[3-(2,6-Difluoro-3-methanesulfonylamino-benzoyl)-1H-pyrrolo[2,3-b]pyrid-
in-5-yl]-benzamide (P-0721), [1035]
N-[2,4-Difluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-methanesulfonamide (P-0184), [1036]
N-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-benzenesulfonamide (P-0685), [1037] Propane-1-sulfonic acid
[4-chloro-2-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
)-phenyl]-amide (P-0753), [1038] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-a-
mide (P-0636), [1039] Propane-1-sulfonic acid
[4-chloro-2-fluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-amide (P-0776) [1040] Propane-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluor-
o-phenyl}-amide (P-0956), [1041] Propane-1-sulfonic acid
{3-[5-(4-dimethylamino-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4--
difluoro-phenyl}-amide (P-0889), [1042] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l]-phenyl}-amide (P-0877), [1043] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-trifluoromethyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-amide (P-0912), [1044] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(3-methoxy-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l]-phenyl}-amide (P-0874), [1045] Propane-1-sulfonic acid
{3-[5-(3-dimethylamino-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4--
difluoro-phenyl}-amide (P-0876), [1046] Propane-1-sulfonic acid
[2-fluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-0897), [1047] Propane-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2-fluoro-ph-
enyl}-amide (P-1009), [1048]
(2-Fluoro-3-hydroxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl-
)-methanone (P-0027), [1049]
(3-Chloro-2-fluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone (P-0165), [1050] 5-phenyl-1H-pyrrolo[2,3-b]pyridine,
[1051] Propane-1-sulfonic acid
[2-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-amide (P-1251), [1052] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(3-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
]-phenyl}-amide (P-1259), [1053] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
]-phenyl}-amide (P-1260), [1054] Propane-1-sulfonic acid
[2-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-amide (P-1261), [1055] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-pyridin-4-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-amide (P-1262), [1056]
3-{3-[2,6-Difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]-
pyridin-5-yl}-benzoic acid (P-1266), [1057] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(3-morpholin-4-yl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3--
carbonyl]-phenyl}-amide (P-1873), [1058] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(3-morpholin-4-ylmethyl-phenyl)-1H-pyrrolo[2,3-b]pyrid-
ine-3-carbonyl]-phenyl}-amide (P-1878), [1059] Propane-1-sulfonic
acid
{2,4-difluoro-3-[5-(6-methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl]-phenyl}-amide (P-1879), [1060] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(6-morpholin-4-yl-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyrid-
ine-3-carbonyl]-phenyl}-amide (P-1881), [1061] Propane-1-sulfonic
acid
(2,4-difluoro-3-{5-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl]-1H-pyrrolo[-
2,3-b]pyridine-3-carbonyl}-phenyl)-amide (P-1882), [1062]
Propane-1-sulfonic acid
{3-[5-(4-cyano-3,5-dimethyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-
-2,4-difluoro-phenyl}-amide (P-1980), [1063]
N-{2,4-Difluoro-3-[5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-
-3-carbonyl]-phenyl}-4-trifluoromethyl-benzenesulfonamide (P-1996),
[1064]
N-{2,4-Difluoro-3-[5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-
-3-carbonyl]-phenyl}-3-fluoro-benzenesulfonamide (P-1997), [1065]
Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-amide (P-1864), [1066] Propane-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-4-fluoro-ph-
enyl}-amide (P-1432), [1067]
4-{3-[2,6-Difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]-
pyridin-5-yl}-benzamide (P-1546), [1068]
4-{3-[2,6-Difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]-
pyridin-5-yl}-N,N-dimethyl-benzamide (P-1547), [1069]
Propane-1-sulfonic acid
(2,4-difluoro-3-{5-[4-(morpholine-4-carbonyl)-phenyl]-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl}-phenyl)-amide (P-1548), [1070]
3-{3-[2,6-Difluoro-3-(propane-1-sulfonylamino)-benzoyl]-3-1H-pyrrolo[2,3--
b]pyridin-5-yl}-benzamide (P-1549), [1071] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-methyl-1H-imidazol-2-yl)-1H-pyrrolo[2,3-b]pyridine--
3-carbonyl]-phenyl}-amide (P-2006), [1072]
N-{2,4-Difluoro-3-[5-(6-methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-3-fluoro-benzenesulfonamide (P-2012),
[1073]
(3-Difluoromethoxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanone (P-1528),
[1074]
N-{3-[3-(3-Difluoromethoxy-2,6-difluoro-benzoyl)-1H-pyrrolo[2,3-b]p-
yridin-5-yl]-phenyl}-methanesulfonamide (P-1527), [1075]
(2,2-Difluoro-benzo[1,3]dioxol-5-yl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-methanone (P-1558), [1076]
N-{3-[3-(2,2-Difluoro-benzo[1,3]dioxole-5-carbonyl)-1H-pyrrolo[2,3-b]pyri-
din-5-yl]-phenyl}-methanesulfonamide (P-1564), [1077]
3-(2,3-Dihydro-benzo[1,4]dioxin-6-ylmethyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridine (P-1529), [1078]
N-{3-[3-(2,3-Dihydro-benzo[1,4]dioxin-6-ylmethyl)-1H-pyrrolo[2,3-b]pyridi-
n-5-yl]-phenyl}-methanesulfonamide (P-1530), [1079]
(3,5-Bis-difluoromethoxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methanone (P-1520), [1080]
N-{3-[3-(3,5-Bis-difluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]--
phenyl}-methanesulfonamide (P-1522), [1081]
(2,2-Difluoro-benzo[1,3]dioxol-4-yl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-methanone (P-1515), [1082]
N-{3-[3-(2,2-Difluoro-benzo[1,3]dioxole-4-carbonyl)-1H-pyrrolo[2,3-b]pyri-
din-5-yl]-phenyl}-methanesulfonamide (P-1516), [1083]
(2,6-Difluoro-3-methoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone (P-1387), [1084]
(2,6-Difluoro-3-methoxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin--
3-yl)-methanone (P-1388), [1085]
(2,6-Difluoro-3-methoxy-phenyl)-(5-pyridin-4-yl-1H-pyrrolo[2,3-b]pyridin--
3-yl)-methanone (P-1389), [1086]
N-{3-[3-(2,6-Difluoro-3-methoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-p-
henyl}-acetamide (P-1390), [1087]
N-{3-[3-(2,6-Difluoro-3-methoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-p-
henyl}-methanesulfonamide (P-1391), [1088]
3-(2-Fluoro-3-methoxy-benzyl)-5-phenyl-1H-pyrrolo[2,3-b]pyridine
(P-1323), [1089]
3-(2-Fluoro-3-methoxy-benzyl)-5-(4-trifluoromethyl-phenyl)-1H-pyrrolo[2,3-
-b]pyridine (P-1324), [1090]
3-(2-Fluoro-3-methoxy-benzyl)-5-(3-trifluoromethyl-phenyl)-1H-pyrrolo[2,3-
-b]pyridine (P-1325), [1091]
5-(3-Chloro-phenyl)-3-(2-fluoro-3-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridi-
ne (P-1326), [1092]
3-(2-Fluoro-3-methoxy-benzyl)-5-(3-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridi-
ne (P-1327), [1093]
3-(2-Fluoro-3-methoxy-benzyl)-5-(3-trifluoromethoxy-phenyl)-1H-pyrrolo[2,-
3-b]pyridine (P-1328), [1094]
N-{3-[3-(2-Fluoro-3-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl-
}-methanesulfonamide (P-1329), [1095]
(2-Fluoro-3-methoxy-phenyl)-[5-(4-trifluoromethyl-phenyl)-1H-pyrrolo[2,3--
b]pyridin-3-yl]- methanone (P-1330), [1096]
(2-Fluoro-3-methoxy-phenyl)-[5-(3-trifluoromethyl-phenyl)-1H-pyrrolo[2,3--
b]pyridin-3-yl]-methanone (P-1331), [1097]
[5-(3-Chloro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(2-fluoro-3-methoxy-p-
henyl)-methanone (P-1332), [1098]
[5-(4-Chloro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(2-fluoro-3-methoxy-p-
henyl)-methanone (P-1333), [1099]
[5-(3,5-Difluoro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(2-fluoro-3-metho-
xy-phenyl)-methanone (P-1334), [1100]
4-[3-(2-Fluoro-3-methoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-benzonit-
rile (P-1335), [1101]
N-{3-[3-(2-Fluoro-3-methoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-pheny-
l}-acetamide (P-1336), [1102]
(2-Fluoro-3-methoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone (P-1338), [1103]
(2-Fluoro-3-methoxy-phenyl)-[5-(4-trifluoromethoxy-phenyl)-1H-pyrrolo[2,3-
-b]pyridin-3-yl]-methanone (P-1339), [1104]
(2-Fluoro-3-methoxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl-
)-methanone (P-1343), [1105]
(2-Fluoro-3-methoxy-phenyl)-[5-(3-trifluoromethoxy-phenyl)-1H-pyrrolo[2,3-
-b]pyridin-3-yl]-methanone (P-1344), [1106]
(5-Phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethoxy-phenyl)-meth-
anone (P-1408), [1107]
(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethoxy-phenyl-
)-methanone (P-1413), [1108]
N-{3-[3-(3-Trifluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-pheny-
l}-methanesulfonamide (P-1428), [1109]
N-{3-[3-(3-Trifluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-pheny-
l}-acetamide (P-1422), [1110]
(E)-3-{3-[3-(3-Trifluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-p-
henyl}-acrylic acid (P-1417), [1111]
(3-Difluoromethoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-metha-
none (P-1409), [1112]
(3-Difluoromethoxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone (P-1415), [1113]
N-{3-[3-(3-Difluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl-
}-methanesulfonamide (P-1430), [1114]
N-{3-[3-(3-Difluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl-
}-acetamide (P-1424), [1115]
(E)-3-{3-[3-(3-Difluoromethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-ph-
enyl}-acrylic acid (P-1418), [1116]
(3,5-Dimethoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
(P-1406), [1117]
(3,5-Dimethoxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-met-
hanone (P-1411), [1118]
N-{3-[3-(3,5-Dimethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-me-
thanesulfonamide (P-1426), [1119]
N-{3-[3-(3,5-Dimethoxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-ac-
etamide (P-1420), [1120]
(3-Fluoro-5-trifluoromethyl-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3--
yl)-methanone (P-1407), [1121]
(3-Fluoro-5-trifluoromethyl-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methanone (P-1412), [1122]
N-{3-[3-(3-Fluoro-5-trifluoromethyl-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-y-
l]-phenyl}-methanesulfonamide (P-1427), [1123]
N-{3-[3-(3-Fluoro-5-trifluoromethyl-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-y-
l]-phenyl}-acetamide (P-1421), [1124]
(3,5-Dichloro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone (P-1414), [1125]
N-{3-[3-(3,5-Dichloro-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-met-
hanesulfonamide (P-1429), [1126]
N-{3-[3-(3,5-Dichloro-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl}-ace-
tamide (P-1423), [1127]
(5-Phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethyl-phenyl)-metha-
none (P-1405), [1128]
(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethyl-phenyl)-
-methanone (P-1410), [1129]
N-{3-[3-(3-Trifluoromethyl-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl-
}-methanesulfonamide (P-1425), [1130]
N-{3-[3-(3-Trifluoromethyl-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-phenyl-
}-acetamide (P-1419), and [1131]
(E)-3-{3-[3-(3-Trifluoromethyl-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-ph-
enyl}-acrylic acid (P-1416). The following table indicates the
5-bromo azaindole (column 2) and boronic acid (column 3) used to
afford the compound (column 4). Column 1 provides the compound
number and column 5 the observed mass.
TABLE-US-00001 [1131] Boronic 5-Br azaindole acid P-0082
##STR00168## ##STR00169## P-0121 ##STR00170## ##STR00171## P-0079
##STR00172## ##STR00173## P-0308 ##STR00174## ##STR00175## P-0156
##STR00176## ##STR00177## P-0297 ##STR00178## ##STR00179## P-0228
##STR00180## ##STR00181## P-0037 ##STR00182## ##STR00183## P-0008
##STR00184## ##STR00185## P-0716 ##STR00186## ##STR00187## P-0700
##STR00188## ##STR00189## P-0841 ##STR00190## ##STR00191## P-0734
##STR00192## ##STR00193## P-0745 ##STR00194## ##STR00195## P-0746
##STR00196## ##STR00197## P-0210 ##STR00198## ##STR00199## P-0721
##STR00200## ##STR00201## P-0184 ##STR00202## ##STR00203## P-0685
##STR00204## ##STR00205## P-0753 ##STR00206## ##STR00207## P-0636
##STR00208## ##STR00209## P-0776 ##STR00210## ##STR00211## P-0956
##STR00212## ##STR00213## P-0889 ##STR00214## ##STR00215## P-0877
##STR00216## ##STR00217## P-0912 ##STR00218## ##STR00219## P-0874
##STR00220## ##STR00221## P-0876 ##STR00222## ##STR00223## P-0897
##STR00224## ##STR00225## P-1009 ##STR00226## ##STR00227## P-0027
##STR00228## ##STR00229## P-0165 ##STR00230## ##STR00231## P-0857
##STR00232## ##STR00233## ##STR00234## ##STR00235## P-1251
##STR00236## ##STR00237## P-1259 ##STR00238## ##STR00239## P-1260
##STR00240## ##STR00241## P-1261 ##STR00242## ##STR00243## P-1262
##STR00244## ##STR00245## P-1266 ##STR00246## ##STR00247## P-1873
##STR00248## ##STR00249## P-1878 ##STR00250## ##STR00251## P-1879
##STR00252## ##STR00253## P-1881 ##STR00254## ##STR00255## P-1882
##STR00256## ##STR00257## P-1980 ##STR00258## ##STR00259## P-1996
##STR00260## ##STR00261## P-1997 ##STR00262## ##STR00263## P-1864
##STR00264## ##STR00265## P-1432 ##STR00266## ##STR00267## P-1546
##STR00268## ##STR00269## P-1547 ##STR00270## ##STR00271## P-1548
##STR00272## ##STR00273## P-1549 ##STR00274## ##STR00275## P-2006
##STR00276## ##STR00277## P-2012 ##STR00278## ##STR00279## P-1528
##STR00280## ##STR00281## P-1527 ##STR00282## ##STR00283## P-1558
##STR00284## ##STR00285## P-1564 ##STR00286## ##STR00287## P-1529
##STR00288## ##STR00289## P-1530 ##STR00290## ##STR00291## P-1520
##STR00292## ##STR00293## P-1522 ##STR00294## ##STR00295## P-1515
##STR00296## ##STR00297## P-1516 ##STR00298## ##STR00299## P-1387
##STR00300## ##STR00301## P-1388 ##STR00302## ##STR00303## P-1389
##STR00304## ##STR00305## P-1390 ##STR00306## ##STR00307## P-1391
##STR00308## ##STR00309## P-1323 ##STR00310## ##STR00311## P-1324
##STR00312## ##STR00313## P-1325 ##STR00314## ##STR00315## P-1326
##STR00316## ##STR00317## P-1327 ##STR00318## ##STR00319## P-1328
##STR00320## ##STR00321## P-1329 ##STR00322## ##STR00323## P-1330
##STR00324## ##STR00325## P-1331 ##STR00326## ##STR00327## P-1332
##STR00328## ##STR00329## P-1333 ##STR00330## ##STR00331## P-1334
##STR00332## ##STR00333## P-1335 ##STR00334## ##STR00335## P-1336
##STR00336## ##STR00337## P-1338 ##STR00338## ##STR00339## P-1339
##STR00340## ##STR00341## P-1343 ##STR00342## ##STR00343## P-1344
##STR00344## ##STR00345## P-1408 ##STR00346## ##STR00347## P-1413
##STR00348## ##STR00349## P-1428 ##STR00350## ##STR00351## P-1422
##STR00352## ##STR00353## P-1417 ##STR00354## ##STR00355## P-1409
##STR00356## ##STR00357## P-1415 ##STR00358## ##STR00359## P-1430
##STR00360## ##STR00361## P-1424 ##STR00362## ##STR00363## P-1418
##STR00364## ##STR00365## P-1406 ##STR00366## ##STR00367## P-1411
##STR00368## ##STR00369## P-1426 ##STR00370## ##STR00371## P-1420
##STR00372## ##STR00373## P-1407 ##STR00374## ##STR00375## P-1412
##STR00376## ##STR00377## P-1427 ##STR00378## ##STR00379## P-1421
##STR00380## ##STR00381## P-1414 ##STR00382## ##STR00383## P-1429
##STR00384## ##STR00385## P-1423 ##STR00386## ##STR00387## P-1405
##STR00388## ##STR00389## P-1410 ##STR00390## ##STR00391## P-1425
##STR00392## ##STR00393## P-1419 ##STR00394## ##STR00395## P-1416
##STR00396## ##STR00397## MS (ESI) [M + H.sup.+].sup.+ Compound
observed P-0082 ##STR00398## 406.2 P-0121 ##STR00399## 407.2 P-0079
##STR00400## 411.1 P-0308 ##STR00401## 410.2 P-0156 ##STR00402##
503.2 P-0297 ##STR00403## 526.1
P-0228 ##STR00404## 453.1 P-0037 ##STR00405## 376.1 P-0008
##STR00406## 411.1 P-0716 ##STR00407## 424.0 [M - H.sup.+].sub.-
P-0700 ##STR00408## 429.1 P-0841 ##STR00409## 522.1 P-0734
##STR00410## 445.0 P-0745 ##STR00411## 535.0 [M - H.sup.+].sub.-
P-0746 ##STR00412## 519.2 [M - H.sup.+].sup.- P-0210 ##STR00413##
504.2 [M - H.sup.+].sup.- P-0721 ##STR00414## 469.0 P-0184
##STR00415## 428.1 P-0685 ##STR00416## 491.1 P-0753 ##STR00417##
473.1 P-0636 ##STR00418## 456.1 P-0776 ##STR00419## 472.1 P-0956
##STR00420## 490.1 P-0889 ##STR00421## 499.2 P-0877 ##STR00422##
486.3 P-0912 ##STR00423## 524.1 P-0874 ##STR00424## 484.3 [M -
H.sup.+].sup.- P-0876 ##STR00425## 499.3 P-0897 ##STR00426## 438.3
P-1009 ##STR00427## 472.2 P-0027 ##STR00428## 334.1 P-0165
##STR00429## 352.1 P-0857 ##STR00430## 455.1 [M - H.sup.+].sup.-
##STR00431## P-1251 ##STR00432## 439.3 P-1259 ##STR00433## 474.2
P-1260 ##STR00434## 474.2 P-1261 ##STR00435## 490.2 P-1262
##STR00436## 457.2 P-1266 ##STR00437## 500.1 P-1873 ##STR00438##
541.2 P-1878 ##STR00439## 555.3 P-1879 ##STR00440## 487.3 P-1881
##STR00441## 542.3 P-1882 ##STR00442## 555.3 P-1980 ##STR00443##
509.2 P-1996 ##STR00444## 562.2 P-1997 ##STR00445## 512.2 P-1864
##STR00446## 460.2 P-1432 ##STR00447## 472.2 P-1546 ##STR00448##
497.2 P-1547 ##STR00449## 527.3 P-1548 ##STR00450## 569.3 P-1549
##STR00451## 499.3 P-2006 ##STR00452## 560.2 P-2012 ##STR00453##
632.1 P-1528 ##STR00454## P-1527 ##STR00455## P-1558 ##STR00456##
P-1564 ##STR00457## P-1529 ##STR00458## P-1530 ##STR00459## P-1520
##STR00460## P-1522 ##STR00461## P-1515 ##STR00462## P-1516
##STR00463## P-1387 ##STR00464## P-1388 ##STR00465## P-1389
##STR00466## P-1390 ##STR00467## P-1391 ##STR00468## P-1323
##STR00469## P-1324 ##STR00470## P-1325 ##STR00471## P-1326
##STR00472## P-1327 ##STR00473## P-1328 ##STR00474## P-1329
##STR00475## P-1330 ##STR00476## P-1331 ##STR00477## P-1332
##STR00478## P-1333 ##STR00479## P-1334 ##STR00480## P-1335
##STR00481## P-1336 ##STR00482## P-1338 ##STR00483## P-1339
##STR00484## P-1343 ##STR00485## P-1344 ##STR00486## P-1408
##STR00487## P-1413 ##STR00488## P-1428 ##STR00489## P-1422
##STR00490## P-1417 ##STR00491## P-1409 ##STR00492## P-1415
##STR00493## P-1430 ##STR00494## P-1424 ##STR00495## P-1418
##STR00496## P-1406 ##STR00497## P-1411 ##STR00498## P-1426
##STR00499## P-1420 ##STR00500## P-1407 ##STR00501## P-1412
##STR00502## P-1427 ##STR00503## P-1421 ##STR00504## P-1414
##STR00505## P-1429 ##STR00506## P-1423 ##STR00507## P-1405
##STR00508## P-1410 ##STR00509## P-1425 ##STR00510## P-1419
##STR00511## P-1416 ##STR00512##
Example 4
Synthesis of
N-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-ethanesulfonamide P-0728
[1132] Compound P-0728 was synthesized in eight steps from
2,4-difluorophenylamine 42 as shown in Scheme 16.
##STR00513## ##STR00514##
Step 1--Preparation of dibenzyl-(2,4-difluoro-phenyl)-amine
(52)
[1133] To 2,4-difluoro-phenylamine (42, 10.0 g, 77.4 mmol) in
N,N-dimethylformamide (130 mL) were added potassium carbonate (32.1
g, 0.23 mol) and benzyl bromide (21.2 mL, 0.18 mol). The reaction
was stirred at room temperature overnight. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtrated. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 10% ethyl acetate in hexane. The
appropriate fractions were combined and concentrated to provide the
compound (52, 12.0 g, 50%). MS(ESI) [M+H.sup.+].sup.+=310.2.
Step 2--Preparation of 3-dibenzylamino-2,6-difluoro-benzaldehyde
(53)
[1134] To dibenzyl-(2,4-difluoro-phenyl)-amine (52, 4.30 g, 13.9
mmol) in tetrahydrofuran (60 mL), under an atmosphere of nitrogen,
cooled in a -78.degree. C. acetone/dry ice bath, was added
n-butyllithium (2.50 M in hexane, 6.1 mL, 15.3 mmol) slowly. The
reaction was stirred for 1 hour, N,N-dimethylformamide (1.2 mL,
15.3 mmol) was added and the reaction was allowed to warm to room
temperature for 1 hour. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 10% ethyl acetate in hexane to provide
the compound (53, 4.0 g, 85%). MS(ESI) [M+H.sup.+].sup.+=337.2.
Step 3--Preparation of
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]py-
ridin-3-yl)-methanol (54)
[1135] To 3-dibenzylamino-2,6-difluoro-benzaldehyde (53, 0.76 g,
2.3 mmol) in methanol (50 mL) were added
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 0.40 g, 2.1 mmol,
prepared as described in Example 17) and potassium hydroxide (0.50
g, 8.9 mmol) under an atmosphere of nitrogen. The reaction was
stirred at room temperature overnight. The reaction was poured into
water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 5% methanol in methylene
chloride to provide the compound (54, 0.60 g, 50%). MS(ESI)
[M+H.sup.+].sup.+=533.2.
Step 4--Preparation of
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]py-
ridin-3-yl)-methanone (55)
[1136] To
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo-
[2,3-b]pyridin-3-yl)-methanol (54, 0.90 g, 1.7 mmol) in methylene
chloride (20 mL) under an atmosphere of nitrogen was added
Dess-Martin periodane (0.97 g, 2.3 mmol). The reaction was stirred
at room temperature for 15 minutes. The reaction was poured into a
solution of sodium bicarbonate and sodium thiosulfate and extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
elating with 5% methanol in methylene chloride to provide the
compound (55, 0.70 g, 78%). MS(ESI) [M+H.sup.+].sup.+=531.2.
Step 5--Preparation of
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopropylsilan-
yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (56)
[1137] To
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo-
[2,3-b]pyridin-3-yl)-methanone (55, 0.84 g, 1.6 mmol) in
tetrahydrofuran (150 mL) was added sodium hydride (210.0 mg, 60% in
mineral oil, 5.3 mmol) under an atmosphere of nitrogen. The
reaction was stirred for 5 minutes. Triisopropylsilyl chloride
(0.80 mL, 3.8 mmol) was added and the reaction was stirred at room
temperature for 3 hours. The reaction mixture was poured into water
and extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 10% ethyl acetate in hexane to provide
the compound (56, 420 mg, 39%). MS(ESI)
[M+H.sup.+].sup.+=687.4.
Step 6--Preparation of
(3-amino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-py-
rrolo[2,3-b]pyridin-3-yl)-methanone (57)
[1138] To
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopr-
opylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (56, 55.0 mg,
0.080 mmol) in methanol (15 mL) was added 20% palladium hydroxide
on carbon (20 mg). The reaction was stirred under an atmosphere of
hydrogen overnight. The reaction was filtered to remove the
catalyst, and then concentrated to give the crude compound that was
used in the next step.
Step 7--Preparation of
N-[2,4-difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]-
pyridine-3-carbonyl)-phenyl]-ethanesulfonamide (58)
[1139] To
(3-amino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopropylsila-
nyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (57, 35.0 mg, 0.069
mmol) in methylene chloride (6 mL) was added methanesulfonyl
chloride (0.30 mL, 3.9 mmol) and triethylamine (0.40 mL, 2.9 mmol).
The reaction was stirred at room temperature overnight. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was dried over anhydrous sodium sulfate and
concentrated to give the crude compound that was used in the next
step.
Step 8--Preparation of
N-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-ethanesulfonamide (P-0728)
[1140] To
N-[2,4-difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrro-
lo[2,3-b]pyridine-3-carbonyl)-phenyl]-ethanesulfonamide (58, 35.0
mg, 0.060 mmol) in tetrahydrofuran (10 mL) was added
tetra-n-butylammonium fluoride (19 mg, 0.072 mmol). The reaction
was stirred at room temperature for 5 minutes. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 5% methanol in methylene
chloride to provide the compound (P-0728, 5.6 mg, 22%). MS(ESI)
[M+H.sup.+].sup.+=443.1.lamda.
[1141] Also,
(3-Benzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methanone P-0807 and
(3-amino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3--
yl)-methanone P-0763
##STR00515##
were synthesized by reacting
(3-dibenzylamino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopropylsilan-
yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (56, 20.0 mg, 0.033
mmol) and
(3-Amino-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-py-
rrolo[2,3-b]pyridin-3-yl)-methanone (57, 20.0 mg, 0.039 mmol),
respectively, in tetrahydrofuran (5.0 mL) with
tetra-n-butylammonium fluoride (13 mg, 0.050 mmol). The reaction
mixtures were stirred at room temperature for 10 minutes, poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
concentrated. The products were isolated by silica gel column
chromatography eluting with 10% ethyl acetate in hexane to give
P-0807 (2.0 mg, MS(ESI) [M+H.sup.+].sup.+=441.1) and P-0763 (1.7
mg, MS(ESI)[M-H.sup.+].sup.-=351.1) as white solids.
Example 5
Preparation of propane-2-sulfonic acid
[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-amide P-0850
[1142] Compound P-0850 was synthesized in four steps from
2,4-difluorophenylamine 42 as shown in Scheme 17.
##STR00516##
Step 1--Preparation of propane-2-sulfonic acid
(2,4-difluoro-phenyl)-amide (59)
[1143] To 2,4-difluoro-phenylamine (42, 4.0 mL, 40.0 mmol) in
methylene chloride (50 mL) were added pyridine (3.37 mL, 42.3
mmol), propane-2-sulfonyl chloride (6.00 g, 42.3 mmol) and
dimethylaminopyridine (0.20 g, 1.64 mmol) under an atmosphere of
nitrogen. The reaction was stirred at 45.degree. C. overnight. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was washed with brine, dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated and
purified by silica gel column chromatography 3% methanol in
methylene chloride to give a white solid (59, 8.0 g, 85%). MS(ESI)
[M-H.sup.+].sup.-=234.0.
Step 2--Preparation of propane-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (60)
[1144] To propane-2-sulfonic acid (2,4-difluoro-phenyl)-amide (59,
2.35 g, 9.95 mmol) in tetrahydrofuran (70 mL) under an atmosphere
of nitrogen cooled with a dry ice/acetone bath was added 1.60 M of
n-butyllithium (1.60 M in hexane, 6.53 mL, 10.45 mmol). The
reaction was stirred for 40 minutes, and then another portion of
n-butyllithium (1.60 M in hexane, 6.84 mL, 10.94 mmol): The
reaction was stirred for 1 hour and N,N-dimethylformamide (0.92 mL,
11.9 mmol) was added. The reaction was allowed to warm to room
temperature overnight. The reaction was poured into water extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous sodium sulfate and filtrated. The filtrate was
concentrated and purified by silica gel column chromatography
(dichloromethane/methanol 5%) to give the compound (60, 1.4 g,
53.4%). MS (ESI) [M-H.sup.+].sup.-=263.4.
Step 3--Preparation of propane-2-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-amide (61)
[1145] To propane-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (60, 220.0 mg, 0.83 mmol) in
methanol (15 mL) was added 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(89, 150.0 mg, 0.77 mmol, prepared as described in Example 17) and
potassium hydroxide (537.0 mg, 9.6 mmol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature overnight.
The reaction was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over sodium
sulfate and filtrated. The filtrate was concentrated and purified
by silica gel column chromatography eluting with 5% methanol in
dichloromethane to give the compound (61, 160 mg, 45.3%). In this
step, minor compound Propane-2-sulfonic acid
{2,4-difluoro-3-[methoxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-amide was also formed and isolated. MS(ESI)
[M+H.sup.+].sup.+=460.1.
Step 4--Preparation of propane-2-sulfonic acid
[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-amide (P-0850)
[1146] To propane-2-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-amide (61, 40.0 mg, 0.087 mmol) in tetrahydrofuran
(10 mL) was added Dess-Martin periodane (48.0 mg, 0.11 mmol). The
reaction was stirred at room temperature for 5 minutes. The
reaction was poured into sodium thiosulfate and potassium carbonate
solution and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtrated. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 5% methanol in methylene
chloride to give the compound (P-0850, 13.4 mg, 33.5%). MS(ESI)
[M+H.sup.+].sup.+=458.1.
[1147]
N-(2,4-Difluoro-3-formyl-phenyl)-3-trifluoromethyl-benzenesulfonami-
de 579,
N-(2,4-difluoro-3-formyl-phenyl)-4-trifluoromethyl-benzenesulfonam-
ide 580, and
N-(2,4-difluoro-3-formyl-phenyl)-4-fluoro-benzenesulfonamide 581,
and (2,4-difluoro-3-formyl-phenyl)-carbamic acid benzyl ester
##STR00517##
were prepared following Steps 1 and 2 of Scheme 17, substituting
propane-2-sulfonyl chloride with 3-trifluoromethyl-benzenesulfonyl
chloride, 4-trifluoromethyl-benzenesulfonyl chloride,
4-fluoro-benzenesulfonyl chloride, and benzyl chloroformate,
respectively, in Step 1.
[1148] Propane-1-sulfonic acid
[4-chloro-3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-
fluoro-phenyl]-amide P-1004
##STR00518##
was prepared using the protocol of Scheme 17, substituting
2,4-difluoro-phenylamine with 4-chloro-2-fluoro-phenylamine and
propane-2-sulfonyl chloride with propane-1-sulfonyl chloride in
Step 1, and 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-chloro-7-azaindole 80 (see Example 9) in Step 3. MS(ESI)
[M+H.sup.+].sup.+=430.1.
[1149] Propane-1-sulfonic acid
[4-chloro-2-fluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-0904
##STR00519##
was prepared using the protocol of Scheme 17, substituting
2,4-difluoro-phenylamine with 4-chloro-2-fluoro-phenylamine and
propane-2-sulfonyl chloride with propane-1-sulfonyl chloride in
Step 1, and 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
7-azaindole 94 in Step 3. MS(ESI) [M+H.sup.+].sup.+=396.2.
[1150]
[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-carb-
amic acid methyl ester P-0974 and
[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-carbamic
acid benzyl ester P-0894
##STR00520##
were prepared using the protocol of Scheme 17, substituting
propane-2-sulfonyl chloride with benzyl chloroformate in Step 1 and
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with 7-azaindole in
Step 3. The products of Step 3 were a mixture of
{2,4-Difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-phenyl}--
carbamic acid methyl ester and
{2,4-Difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-phenyl}--
carbamic acid benzyl ester that were separated by silica gel column
chromatography and carried through Step 4 separately to provide
P-0974 and P-0894, respectively. P-0974 MS (ESI)
[M-H.sup.+].sup.-=330.1. P-0894 MS (ESI)
[M-H.sup.+].sup.-=406.1.
[1151]
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-carbamic acid methyl ester P-0486 and
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-carbamic acid benzyl ester P-0302
##STR00521##
were prepared using the protocol of Scheme 17, substituting
propane-2-sulfonyl chloride with benzyl chloroformate in Step 1.
The products of Step 3 were a mixture of
{2,4-Difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-carbamic acid methyl ester and
{2,4-Difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-carbamic acid benzyl ester that were separated by
silica gel column chromatography and carried through Step 4
separately to provide P-0486 and P-0302, respectively. P-0486 MS
(ESI) [M+H.sup.+].sup.+=409.1. P-0302 MS (ESI)
[M+H.sup.+].sup.+=485.1.
[1152]
[5-(3-Methanesulfonyl-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(2,3,6-
-trifluoro-phenyl)-methanone P-0102
##STR00522##
was prepared using the protocol of Steps 3 and 4 of Scheme 17,
substituting propane-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide with
2,3,6-Trifluoro-benzaldehyde and
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-(3-methanesulfonyl-phenyl)-1H-pyrrolo[2,3-b]pyridine in Step 3.
MS (ESI) [M-H.sup.+].sup.-=429.0.
[1153] Thiophene-2-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide P-1267
##STR00523##
was prepared using the protocol of Steps 3 and 4 of Scheme 17,
substituting 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-chloro-7-azaindole 80 (see Example 9) and propane-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide 60 with thiophene-2-sulfonic
acid (2,4-difluoro-3-formyl-phenyl)-amide 512 (see Example 21) in
Step 3. MS(ESI) [M+H.sup.+].sup.+=451.9.
[1154] Thiophene-3-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide P-1268
##STR00524##
was prepared using the protocol of Steps 3 and 4 of Scheme 17,
substituting 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-chloro-7-azaindole 80 (see Example 9) and propane-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide 60 with thiophene-3-sulfonic
acid (2,4-difluoro-3-formyl-phenyl)-amide 513 (see Example 21) in
Step 3. MS(ESI) [M+H.sup.+].sup.+=454.1.
[1155] Additional compounds were prepared following the protocol of
Scheme 17, optionally replacing propane-2-sulfonyl chloride with an
appropriate acid chloride in Step 1 and/or
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with an appropriate
azaindole in Step 3. Azaindoles were purchased or synthesized as
described in Examples 6, 13, 14, 16 and 17. Some compounds were
isolated after Step 3, as either hydroxy or methoxy derivatives.
The following compounds were made following this procedure: [1156]
Dimethylamine-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluor-
o-phenyl}-amide (P-1257), [1157]
N-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]--
benzenesulfonamide (P-0798), [1158] Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0773), [1159] Dimethylamine-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0898), [1160]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-benzenesulfonamide (P-0885), [1161]
N-[2,4-Difluoro-3-(5-fluoro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-benzenesulfonamide (P-0902), [1162] Propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-isopropenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thyl]-phenyl}-amide (P-1239), [1163] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-isopropenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-amide (P-0991), [1164] Propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-
-phenyl}-amide (P-1240), [1165] Propane-1-sulfonic acid
{2,4-difluoro-3-[methoxy-(5-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-
-phenyl}-amide (P-1241), [1166] Propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-isopropyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
yl]-phenyl}-amide (P-1242), [1167] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-isopropyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-amide (P-0997), [1168] Propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl-
]-phenyl}-amide (P-1243), [1169] Propane-1-sulfonic acid
{2,4-difluoro-3-[methoxy-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl-
]-phenyl}-amide (P-1244), [1170] Propane-1-sulfonic acid
(2,4-difluoro-3-{hydroxy-[5-(4-methyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]py-
ridin-3-yl]-methyl}-phenyl)-amide (P-1245), [1171]
Propane-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-0933), [1172] Propane-1-sulfonic acid
[2,4-difluoro-3(5-fluoro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-am-
ide (P-0907), [1173] Piperidine-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1020), [1174]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-methoxy-benzenesulfonamide (P-0983), [1175]
N-[3-(4-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-benzenesulfonamide (P-0954), [1176]
N-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-acetamide (P-1002), [1177] Dimethylamine-1-sulfonic acid
[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-amide
(P-0950), [1178] Dimethylamine-1-sulfonic acid
[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phe-
nyl]-amide (P-0837), [1179] Dimethylamine-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1258), [1180] Butane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-1263), [1181] Butane-1-sulfonic acid
[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
amide (P-1264), [1182] Butane-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1265), [1183] Propane-1-sulfonic acid
[3-(5-ethoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1252), [1184] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(2-methoxy-ethoxy)-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l]-phenyl}-amide (P-1253), [1185] Propane-1-sulfonic acid
{3-[5-(2-diethylamino-ethoxy)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-d-
ifluoro-phenyl}-amide (P-1254), [1186]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-ethyl-benzenesulfonamide (P-1700), [1187]
N-[3-(5-Ethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]--
4-trifluoromethyl-benzenesulfonamide (P-1783), [1188]
Thiophene-3-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-pheny-
l]-amide (P-1798), [1189] Benzo[b]thiophene-2-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1799), [1190] 5-Pyridin-2-yl-thiophene-2-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1800), [1191]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-cyano-benzenesulfonamide (P-1822), [1192]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-fluoro-4-methyl-benzenesulfonamide (P-1823), [1193]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-isopropyl-benzenesulfonamide (P-1839), [1194]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-fluoro-benzenesulfonamide (P-1840), [1195]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3,5-difluoro-benzenesulfonamide (P-1841), [1196]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-methyl-benzenesulfonamide (P-1842), [1197]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-oxazol-5-yl-benzenesulfonamide (P-1843), [1198]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-fluoro-
-benzenesulfonamide (P-1865), [1199]
N-{2,4-Difluoro-3-[5-(2-methoxy-ethoxy)-1H-pyrrolo[2,3-b]pyridine-3-carbo-
nyl]-phenyl}-3-fluoro-benzenesulfonamide (P-1871), [1200]
N-{2,4-Difluoro-3-[5-(2-methoxy-ethoxy)-1H-pyrrolo[2,3-b]pyridine-3-carbo-
nyl]-phenyl}-4-fluoro-benzenesulfonamide (P-1872), [1201]
Propane-1-sulfonic acid
(2,4-difluoro-3-{5-[4-(2-methoxy-ethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridin-
e-3-carbonyl}-phenyl)-amide (P-1998), [1202]
N-{2,4-Difluoro-3-[5-(2-methoxy-ethoxy)-1H-pyrrolo[2,3-b]pyridine-3-carbo-
nyl]-phenyl}-4-trifluoromethyl-benzenesulfonamide (P-2005), and
[1203]
N-(2,4-Difluoro-3-{5-[4-(2-methoxy-ethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyrid-
ine-3-carbonyl}-phenyl)-4-trifluoromethyl-benzenesulfonamide
(P-2013). The following table indicates the acid chloride (column
2) and the azaindole (column 3) used to afford the target compounds
(column 4). Column 1 provides the compound number and column 5 the
observed mass. Compounds isolated after Step 3 of Scheme 17 are so
noted in column 1.
TABLE-US-00002 [1203] MS (ESI) Sulfonyl [M + H.sup.+].sup.+
chloride Azaindole Compound observed P-1257 ##STR00525##
##STR00526## ##STR00527## 415.1 P-0798 ##STR00528## ##STR00529##
##STR00530## 489.9 491.1 [M - H.sup.+].sup.- P-0773 ##STR00531##
##STR00532## ##STR00533## 455.9 457.9 [M - H.sup.+].sup.- P-0898
##STR00534## ##STR00535## ##STR00536## 497.0 499.1 P-0885
##STR00537## ##STR00538## ##STR00539## 446.1 448.1 [M -
H.sup.+].sup.- P-0902 ##STR00540## ##STR00541## ##STR00542## 430.1
[M - H.sup.+].sup.- P-1239 Step 3 ##STR00543## ##STR00544##
##STR00545## 422.2 [M - H.sup.+].sup.- P-0991 ##STR00546##
##STR00547## ##STR00548## 420.2 P-1240 Step 3 ##STR00549##
##STR00550## ##STR00551## 396.4 [M - H.sup.+].sup.- P-1241 Step 3
##STR00552## ##STR00553## ##STR00554## 410.3 [M - H.sup.+].sup.-
P-1242 Step 3 ##STR00555## ##STR00556## ##STR00557## 424.3 [M -
H.sup.+].sup.- P-0997 ##STR00558## ##STR00559## ##STR00560## 422.3
P-1243 Step 3 ##STR00561## ##STR00562## ##STR00563## 412.3 [M -
H.sup.+].sup.- P-1244 Step 3 ##STR00564## ##STR00565## ##STR00566##
426.4 [M - H.sup.+].sup.- P-1245 Step 3 ##STR00567## ##STR00568##
##STR00569## 480.3 [M - H.sup.+].sup.- P-0933 ##STR00570##
##STR00571## ##STR00572## 414.2 P-0907 ##STR00573## ##STR00574##
##STR00575## 396.1 [M - H.sup.+].sup.- P-1020 ##STR00576##
##STR00577## ##STR00578## 455.2 P-0983 ##STR00579## ##STR00580##
##STR00581## 476.1 [M - H.sup.+].sup.- P-0954 ##STR00582##
##STR00583## ##STR00584## 448.2 P-1002 ##STR00585## ##STR00586##
##STR00587## 393.2 P-0950 ##STR00588## ##STR00589## ##STR00590##
381.2 P-0837 ##STR00591## ##STR00592## ##STR00593## 458.1 P-1258
##STR00594## ##STR00595## ##STR00596## 415.1 P-1263 ##STR00597##
##STR00598## ##STR00599## 472.1 474.1 P-1264 ##STR00600##
##STR00601## ##STR00602## 424.2 P-1265 ##STR00603## ##STR00604##
##STR00605## 426.0 [M - H.sup.+].sup.- P-1252 ##STR00606##
##STR00607## ##STR00608## 424.2 P-1253 ##STR00609## ##STR00610##
##STR00611## 454.2 P-1254 ##STR00612## ##STR00613## ##STR00614##
495.3 P-1700 ##STR00615## ##STR00616## ##STR00617## 476.2 P-1783
##STR00618## ##STR00619## ##STR00620## 510 P-1798 ##STR00621##
##STR00622## ##STR00623## 495.9 497.6 [M - H.sup.+].sup.- P-1799
##STR00624## ##STR00625## ##STR00626## 502.0 (--) P-1800
##STR00627## ##STR00628## ##STR00629## 531.1 P-1822 ##STR00630##
##STR00631## ##STR00632## 473.1 P-1823 ##STR00633## ##STR00634##
##STR00635## 480.1 P-1839 ##STR00636## ##STR00637## ##STR00638##
488 P-1840 ##STR00639## ##STR00640## ##STR00641## 464 P-1841
##STR00642## ##STR00643## ##STR00644## 482 P-1842 ##STR00645##
##STR00646## ##STR00647## 460.0 [M - H.sup.+].sup.- P-1843
##STR00648## ##STR00649## ##STR00650## 513.0 [M - H.sup.+].sup.-
P-1865 ##STR00651## ##STR00652## ##STR00653## 432.1 P-1871
##STR00654## ##STR00655## ##STR00656## 506.2 P-1872 ##STR00657##
##STR00658## ##STR00659## 506.2 ##STR00660## ##STR00661##
##STR00662## P-1998 ##STR00663## ##STR00664## ##STR00665## 530.3
P-2005 ##STR00666## ##STR00667## ##STR00668## 556.0 P-2013
##STR00669## ##STR00670## ##STR00671## 539.2
Example 6
Synthesis of propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenylamino-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-amide P-0848 and Related Compounds
[1204] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenylamino-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-amide P-0848 was synthesized in five steps from
5-bromo-7-azaindole 67 as shown in Scheme 18.
##STR00672##
Step 1--Preparation of
5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (68)
[1205] To 5-bromo-7-azaindole (67, 1.5 g, 7.6 mmol) in
N,N-dimethylformamide (20 mL) were added sodium hydride (60% in
mineral oil, 0.27 g, 11.0 mmol) and trriisopropylsilyl chloride
(2.6 mL, 12.0 mmol), under an atmosphere of nitrogen. The reaction
was stirred for 2 hours at room temperature. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 10% ethyl acetate in hexane
to give the compound (68, 1.6 g, 59%).
MS(ESI)[M+H.sup.+].sup.+=352.3.
Step 2--Preparation of
5-phenyl-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-5-yl)-amine
(69)
[1206] To 5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(68, 0.10 g, 0.3 mmol) in toluene (5 mL) were added aniline (0.04
mL, 0.42 mmol), sodium tert-butoxide (0.15 g, 1.56 mmol),
tris(dibenzyllideneacetone)dipalladium(0) (9.2 mg, 0.01 mmol) and
(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (6.3 mg, 0.01
mmol). The reaction was heated to 160.degree. C. for 10 minutes in
a CEM Discover microwave instrument. The reaction was concentrated
and purified by silica gel column chromatography eluting with 3%
ethyl acetate in hexane to give the compound (69, 40 mg, 40%).
MS(ESI)[M+H.sup.+].sup.+=366.6.
Step 3--Preparation of
phenyl-(-1H-pyrrolo[2,3-b]pyridine-5-yl)-amine (70)
[1207] To
5-phenyl-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-5-yl)--
amine (69, 0.14 g, mmol) in tetrahydrofuran (3.0 mL) was added
tetra-n-butylammonium fluoride (0.197 g, 0.76 mmol). The reaction
was stirred for 1 hour at room temperature. The reaction was
concentrated and purified by silica gel column chromatography
eluting with 3% ethyl acetate in hexane to give the compound (70,
60 mg, 76%). MS(ESI)[M+H.sup.+].sup.+=210.3.
Step 4--Preparation of propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy(5-phenylamino-1H-pyrrolo[2,3-b]pyridin-3-yl)-met-
hyl]-phenyl}-amide (71)
[1208] To phenyl-(-1H-pyrrolo[2,3-b]pyridine-5-yl)-amine (70, 17.0
mg, 0.09 mmol) in methanol (5.0 mL) were added potassium hydroxide
(92.0 mg, 1.6 mmol) and propane-1-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (73, 19.0 mg, 0.072 mmol,
prepared as described in Example 7) under an atmosphere of
nitrogen. The reaction was stirred for 12 hours at room
temperature. The reaction was concentrated and purified by silica
gel column chromatography eluting with 1% methanol in
dichloromethane to give the compound (71, 17 mg, 50%).
MS(ESI)[M+H.sup.+].sup.+=473.5.
Step 5--Preparation of propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenylamino-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-amide (P-0848)
[1209] To propane-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-phenylamino-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thyl]-phenyl}-amide (71, 7.5 mg, 0.016 mmol) in tetrahydrofuran (3
mL) was added Dess-Martin periodane (6.70 mg, 0.0158 mmol) under an
atmosphere of nitrogen. The reaction was stirred for 20 minutes.
The reaction was concentrated and purified by silica gel column
chromatography eluting with 1% methanol in dichloromethane to give
the compound (P-0848, 6.2 mg, 84%).
MS(ESI)[M+H.sup.+].sup.+=471.2.
[1210] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-morpholin-4-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-amide P-0853, propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-methyl-piperidin-1-yl)-1H-pyrrolo[2,3-b]pyridine-3--
carbonyl]-phenyl}-amide P-0860, and propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-methyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]pyridine-3--
carbonyl]-phenyl}-amide P-1246,
##STR00673##
respectively, were prepared using the protocol of Scheme 18,
substituting aniline with morpholine, 4-methyl-piperidine, and
4-methyl-piperazine, respectively, in Step 2. P-0853 MS(ESI)
[M+H.sup.+].sup.+=465.2. P-0860 MS(ESI) [M+H.sup.+].sup.+=477.3.
P-1246 MS(ESI) [M-H.sup.+].sup.-=478.4.
[1211]
4-[5-(3-Chloro-4-methoxy-phenylamino)-1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl]-indole-1-carboxylic acid butylamide P-1859
##STR00674##
was prepared using the protocol of Scheme 18, substituting aniline
with 3-chloro-4-methoxy-phenylamine in Step 2 and
propane-1-sulfonic acid (2,4-difluoro-3-formyl-phenyl)-amide 73
with 4-Formyl-indole-1-carboxylic acid butylamide 519 (see Example
22) in Step 4. MS(ESI) [M+H.sup.+].sup.+=516.2.
[1212]
(1H-Indol-4-yl)-[5-(4-morpholin-4-yl-phenylamino)-1H-pyrrolo[2,3-b]-
pyridin-3-yl]-methanone P-1792,
[5-(4-Chloro-phenylamino)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(1H-indol-4-yl)--
methanone P-1793,
(1H-Indol-4-yl)-[5-(4-piperidin-1-yl-phenylamino)-1H-pyrrolo[2,3-b]pyridi-
n-3-yl]-methanone P-1794, and
[5-(3-Chloro-4-methoxy-phenylamino)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(1H-in-
dol-4-yl)-methanone P-1795,
##STR00675##
respectively, were prepared using the protocol of Steps 1-3 of
Scheme 18, substituting 5-bromo-7-azaindole with
4-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxylic
acid butylamide (P-1687, prepared as described in Example 22,
Scheme 41) in Step 1 and replacing aniline with
4-morpholin-4-yl-phenylamine, 4-chloro-phenylamine,
4-piperidin-1-yl-phenylamine, and 3-chloro-4-methoxy-phenylamine,
respectively, in Step 2. P-1792 MS (ESI) [M+H.sup.+].sup.+=438.3.
P-1793 MS(ESI)[M+H.sup.+].sup.+=387.1. P-1794 MS (ESI)
[M+H.sup.+].sup.+=436.3. P-1795 MS (ESI)
[M+H.sup.+].sup.+=417.1.
Example 7
Synthesis of propane-1-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide 73
[1213] Compound 73 was synthesized in two steps from
2,4-difluorophenylamine 42 as shown in Scheme 19.
##STR00676##
Step 1--Preparation of Propane-1-sulfonic acid
(2,4-difluoro-phenyl)-amide (72)
[1214] To 2,4-difluoro-phenylamine (42, 3.0 mL, 29.8 mmol) in
tetrahydrofuran (50 mL) were added triethylamine (9.13 mL, 65.5
mmol) and propane-1-sulfonyl chloride (2.90 mL, 25.8 mmol) under an
atmosphere of nitrogen. The reaction was stirred at room
temperature overnight. The reaction was poured into 1 M HCl and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated to give the compound (72, 2.0 g, 28%)
that was used in the next step.
Step 2--Preparation of propane-1-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (73)
[1215] To propane-1-sulfonic acid (2,4-difluoro-phenyl)-amide (72,
1.5 g, 6.38 mmol) in tetrahydrofuran (10 mL) under an atmosphere of
nitrogen, cooled in a -78.degree. C. acetone/dry ice bath was added
lithium diisopropylamide (0.80 M in tetrahydrofuran, 24 mL, freshly
prepared from n-butyllithium and diisopropylamine). After 30
minutes, N,N-dimethyl-formamide (542 .mu.L, 7.018 mmol) was added
dropwise to the reaction. The reaction was stirred for 30 minutes
at -78.degree. C. and then allowed to warm to room temperature for
40 minutes. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 5% ethyl acetate in hexane to give a light yellow
solid (73, 300 mg, 18%). MS(ESI)[M-H.sup.+].sup.-=262.3.
Example 8
Synthesis of propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
P-1116.
[1216] Compound P-1116 was synthesized in four steps from
3-amino-benzoic acid ethyl ester 74 as shown in Scheme 20.
##STR00677##
Step 1--Preparation of 3-(propane-1-sulfonylamino)-benzoic add
ethyl ester (75)
[1217] To 3-amino-benzoic acid ethyl ester (74, 5.0 g, 0.030 mol)
in methylene chloride (30.0 mL) were added pyridine (3.67 mL, 0.045
mol) and propane-1-sulfonyl chloride (3.75 mL, 33.0 mmol). The
reaction was stirred at room temperature for 2 hours. The reaction
mixture was poured into water and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and concentrated. The desired compound was isolated by
silica gel column chromatography eluting with 20% ethyl acetate in
hexane to give a white solid (75, 6.0 g, 74.1%).
Step 2--Preparation of 3-(propane-1-sulfonylamino)-benzoic acid
(16)
[1218] To 3-(propane-1-sulfonylamino)-benzoic acid ethyl ester (75,
1.60 g, 5.90 mmol) in water was added lithium hydroxide (1.0 g, 4.2
mmol) and tetrahydrofuran (20 mL). The reaction was stirred at room
temperature overnight. The reaction mixture was acidified with 1 N
HCl, and extracted with ethyl acetate. The organic layer was dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and washed with ethyl acetate to give the compound
(76, 1.2 g, 84.5%) as white solid.
MS(ESI)[M-H.sup.-].sup.-=242.1.
Step 3--Preparation of 3-(propane-1-sulfonylamino)-benzoyl chloride
(77)
[1219] A solution of 3-(propane-1-sulfonylamino)-benzoic acid (76,
1.20 g, 4.93 mmol) in thionyl chloride was heated to reflux for 3.0
hours. Evaporation of the solvent gave compound 77 as white solid
that was used for the next step.
Step 4--Preparation of propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1116)
[1220] To aluminum trichloride (4.8 g, 36.0 mmol) in methylene
chloride (70.0 mL), under an atmosphere of nitrogen, was added
5-bromo-1H-pyrrolo[2,3-b]pyridine (67, 797 mg, 4.04 mmol) dissolved
in methylene chloride (5.0 mL). The reaction was stirred at room
temperature for 30 minutes, followed by addition of
3-(propane-1-sulfonylamino)-benzoyl chloride (77, 1.10 g, 4.20
mmol) dissolved in methylene dichloride (4.0 mL). The reaction was
stirred at room temperature for 2 hours. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give a white solid
(P-1116, 300.0 mg). MS (ESI) [M-H.sup.+].sup.-=420.1,422.1.
Example 9
Synthesis of 5-chloro-1H-pyrrolo[2,3-b]pyridine 80
[1221] Compound 80 was synthesized in two steps from
5-bromo-1-triisopropylsilyl-7-azaindole 68 as shown in Scheme
21.
##STR00678##
Step 1--Preparation
5-chloro-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (79)
[1222] To 5-bromo-1-triisopropylsilyl-7-azaindole (68, 1.60 g, 4.53
mmol, prepared as described in Example 6) in tetrahydrofuran (50.0
mL), under an atmosphere of nitrogen at -78.degree. C., was added
tert-butyllithium (1.70 M in hexane, 6.12 mL). The reaction was
stirred for 1 hour, followed by addition of hexachloroethane (1.29
g, 5.43 mmol). The reaction was stirred for 3 hours, poured into
water, and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give the crude compound (79, 1.60 g).
MS(ESI)[M+H.sup.+].sup.+=309.3.
Step 2--Preparation 5-chloro-1H-pyrrolo[2,3-b]pyridine (80)
[1223] To 5-chloro-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(79, 1.40 g, 4.53 mmol) in tetrahydrofuran (15 mL) was added
tetra-n-butylammonium fluoride (1.42 g, 5.43 mmol). The reaction
mixture was stirred at room temperature for 10 minutes. The
reaction mixture was concentrated and isolated by silica gel column
chromatography eluting with 30% ethyl acetate in hexane to give the
compound (80, 0.40 g, 58% over 2 steps).
MS(ESI)[M-H.sup.+].sup.-=153.1.
[1224] 5-Fluoro-1H-pyrrolo[2,3-b]pyridine 81
##STR00679##
was prepared using the protocol of Scheme 21, substituting
hexachloroethane with N-fluoro-N-(phenylsulfonyl)
benzenesulfonamide in Step 1. MS(ESI) [M+H.sup.+].sup.+=137.1.
Example 10
Synthesis of
2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-pheno-
l P-0078 and Related Compounds
[1225] Compound P-0078 was synthesized in two steps from
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 as shown in Scheme
24.
##STR00680##
Step 1--Preparation of
2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thyl]-phenol (P-0009)
[1226] To carbonic acid tert-butyl ester
2,4-difluoro-3-formyl-phenyl ester (39, 0.405 g, 15.7 mmol) in
methanol (36 mL), under an atmosphere of nitrogen, was added
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 288.0 mg, 14.8 mmol,
prepared as described in Example 17) and potassium hydroxide (145.0
mg, 25.9 mmol). The reaction was stirred at room temperature
overnight. Then, the reaction was poured into water, and extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous sodium sulfate and concentrated. The mixture was
purified by silica gel column chromatography eluting with 4%
methanol in methylene chloride to provide two separate compounds, a
colorless oil (P-0009, 0.23 g, 44.1%, MS(ESI)
[M+H.sup.+].sup.+=354.1), and acolorless oil (P-0042, 0.050 g,
9.2%, MS(ESI) [M+H.sup.+].sup.+=367.1).
Step 2--Preparation of
2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-pheno-
l (P-0078)
[1227] To
2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methyl]-phenol (P-0009, 34.0 mg, 0.096 mmol) in acetonitrile
(15 mL), were added trifluoroacetic acid (1.0 mL, 13.0 mmol) and
triethylsilane (2.0 mL, 12.0 mmol). The reaction was stirred at
room temperature for 48 hours. The reaction mixture was poured
sodium bicarbonate solution, and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 30% ethyl acetate in
hexane to give the compound (P-0078, 6.0 mg, 19%).
MS(ESI)[M+H.sup.+].sup.+=338.1.
[1228] Additional compounds were prepared following the protocol of
Scheme 24, replacing either or both of carbonic acid tert-butyl
ester 2,4-difluoro-3-formyl-phenyl ester 39 with an appropriate
aldehyde and/or 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with an
appropriate azaindole in Step 1. Azaindoles were purchased or
synthesized as described in Examples 6 and 17. Aldehydes were
prepared as described in Example 5 (through Step 2). Some compounds
were isolated after Step 1, as either hydroxy or methoxy
derivatives. The following compounds were made following this
procedure: [1229]
3-[(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-methoxy-methyl]-2,4-difluoro-p-
henol (P-0126), [1230]
3-[(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2,4-difluoro-p-
henol (P-1180), [1231]
3-(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenol
(P-0122), [1232]
{2,4-Difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3b]pyridin-3-yl)-me-
thyl]-phenyl}-carbamic acid benzyl ester (P-0095), [1233]
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phen-
yl]-carbamic acid benzyl ester (P-0396), [1234]
{2,4-Difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-carbamic acid methyl ester (P-0065), [1235]
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phen-
yl]-carbamic acid methyl ester (P-0257), [1236] Propane-2-sulfonic
acid
{3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-methoxy-methyl]-2,4-difluoro--
phenyl}-amide (P-0356), [1237] Propane-2-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]-ami-
de (P-0867), [1238] Propane-2-sulfonic acid
{2,4-difluoro-3-[methoxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-amide (P-0947), [1239] Propane-1-sulfonic acid
{3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2,4-difluoro--
phenyl}-amide (P-0188), [1240] Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]-ami-
de (P-0910), [1241] Dimethylamino-1-sulfonic acid
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-amide (P-0944), [1242]
3-(2-Fluoro-3-methoxy-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-0269), [1243] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenylamino-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-pheny-
l]-amide (P-0818), [1244] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-am-
ide (P-0911), [1245] Propane-1-sulfonic acid
[2,4-difluoro-3-(5-morpholin-4-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-ph-
enyl]-amide (P-0964), [1246] Propane-1-sulfonic acid
{2,4-difluoro-3-[5-(4-methyl-piperidin-1-yl)-1H-pyrrolo[2,3-b]pyridin-3-y-
lmethyl]-phenyl}-amide (P-0984), [1247]
{3-[(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2,4-difluoro--
phenyl}-carbamic acid tert-butyl ester (P-0318), [1248]
{3-[(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2,4-difluoro-
-phenyl}-carbamic acid methyl ester, [1249]
{2,4-Difluoro-3-[hydroxy-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl-
]-phenyl}-carbamic acid methyl ester, and [1250]
{2,4-Difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-phenyl}--
carbamic acid methyl ester, [1251]
5-Bromo-3-(2,3-dihydro-benzo[1,4]dioxin-6-ylmethyl)-1H-pyrrolo[2,3-b]pyri-
dine (P-1474), and [1252]
5-Bromo-3-(2-fluoro-3-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-0535). The following table indicates the aldehyde (column 2) and
azaindole (column 3) used to afford the target compound (column 4).
Column 1 provides the compound number and column 5 the observed
mass. Compounds isolated after Step 1 of Scheme 24 are so noted in
column 1.
TABLE-US-00003 [1252] MS (ESI) [M + H.sup.+].sup.+ ob- Aldehyde
Azaindole Compound served P-0126 Step 1 ##STR00681## ##STR00682##
##STR00683## 369.1 371.1 P-1180 Step 1 ##STR00684## ##STR00685##
##STR00686## 353.0 355.0 P-0122 ##STR00687## ##STR00688##
##STR00689## 339.0 341.0 P-0095 P-0065 Step 1* ##STR00690##
##STR00691## ##STR00692## P-0396 ##STR00693## ##STR00694##
##STR00695## 471.2 P-0257 ##STR00696## ##STR00697## ##STR00698##
395.1 P-0356 Step 1 ##STR00699## ##STR00700## ##STR00701## 474.1
476.1 P-0867 ##STR00702## ##STR00703## ##STR00704## 460.2 P-0947
Step 1 ##STR00705## ##STR00706## ##STR00707## 459.2 P-0188 Step 1
##STR00708## ##STR00709## ##STR00710## 460 462 P-0910 ##STR00711##
##STR00712## ##STR00713## 444.1 446.1 P-0944 Step 1 ##STR00714##
##STR00715## ##STR00716## 460.2 P-0269 ##STR00717## ##STR00718##
##STR00719## 334.2 P-0818 ##STR00720## ##STR00721## ##STR00722##
457.1 P-0911 ##STR00723## ##STR00724## ##STR00725## 442.2 P-0964
##STR00726## ##STR00727## ##STR00728## 449.3 [M - H.sup.+].sup.-
P-0984 ##STR00729## ##STR00730## ##STR00731## 463.3 P-0318 Step 1
##STR00732## ##STR00733## ##STR00734## 454.1 456.1 Isolated Step 1
##STR00735## ##STR00736## ##STR00737## Isolated Step 1 ##STR00738##
##STR00739## ##STR00740## Isolated Step 1 ##STR00741## ##STR00742##
##STR00743## Isolated Step 1 ##STR00744## ##STR00745## ##STR00746##
P-1474 ##STR00747## ##STR00748## ##STR00749## 345.1 347.1 P-0535
##STR00750## ##STR00751## ##STR00752## 335.1 337.1 *P-0095 and
P-0065 were both produced in Step 1 and isolated from mixture.
These were carried through to step 2 to provide P-0396 and P-0257,
respectively.
Example 11
Synthesis of
N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)
phenyl]-3-methoxy-benzenesulfonamide P-0971
[1253] Compound P-0971 was synthesized in four steps from
2,4-difluorophenylamine 42 as shown in Scheme 26.
##STR00753##
Step 1--Preparation of
N-(2,4-difluoro-phenyl)-3-methoxy-benzenesulfonamide (91)
[1254] To 2,4-difluoro-phenylamine(42, 0.44 mL, 4.4 mmol) in
methylene chloride (10.0 mL), under an atmosphere of nitrogen, were
added pyridine (1.00 mL, 12.4 mmol) and 3-methoxy-benzenesulfonyl
chloride (1.00 g, 4.84 mmol). After 12 hours, the reaction was
poured into cold 1 M HCl and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 20% ethyl acetate in
hexane to give light yellow solid (91, 0.90 g, 69%).
MS(ESI)[M+H.sup.+].sup.+=300.
Step 2--Preparation of
N-{2,4-difluoro-3-[hydroxy-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methyl]-phenyl}-3-methoxy-benzenesulfonamide (92)
[1255] To N-(2,4-difluoro-phenyl)-3-methoxy-benzenesulfonamide (91,
0.148 g, 0.494 mmol) in tetrahydrofuran (10.0 mL) cooled in a
-78.degree. C. acetone/dry ice bath, under an atmosphere of
nitrogen, was added lithium diisopropylamide (0.85 M in
tetrahydrofuran, 1.45 mL, 1.23 mmol) dropwise. After 30 minutes,
1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde (96,
0.15 g, 0.500 mmol, prepared as described in Example 12) was added
in tetrahydrofuran (2.0 mL) into the reaction dropwise. Then the
reaction was stirred for 1 hour at -78.degree. C. and allowed to
reach room temperature. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 40% ethyl acetate in hexane to give
light yellow solid (92, 0.080 g, 26.8%).
MS(ESI)[M+H.sup.+].sup.+=602.
Step 3--Preparation of
N-{2,4-difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)methyl]-phenyl}-
-3-methoxy-benzenesulfonamide (93)
[1256] To
N-{2,4-difluoro-3-[hydroxy-(1-triisopropylsilanyl-1H-pyrrolo[2,3-
-b]pyridin-3-yl)-methyl]-phenyl}-3-methoxy-benzenesulfonamide (92,
0.075 g, 0.12 mmol) in tetrahydrofuran (3.0 mL), was added
tetra-n-butylammonium fluoride (0.039 g, 0.15 mmol). The reaction
was stirred at room temperature for 20 minutes. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 3% methanol in
dichloromethane to give light yellow solid (93, 0.030 g, 55%).
MS(ESI)[M+H.sup.+].sup.+=446.
Step 4--Preparation of
N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)phenyl]-3-methoxy-
-benzenesulfonamide (P-0971)
[1257] To
N-{2,4-difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)methyl-
]-phenyl}-3-methoxy-benzenesulfonamide (93, 0.02 g, 0.05 mmol) in
tetrahydrofuran (3.0 mL) was added Dess-Martin periodane (0.02 g,
0.015 mmol) under an atmosphere of nitrogen. The reaction was
stirred for 10 minutes at room temperature. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 3% methanol in dichloromethane
to give light yellow solid (P-0971, 0.010 g, 50%).
MS(ESI)[M+H.sup.+].sup.+=444.
[1258] Additional compounds were prepared following the protocol of
Scheme 26, replacing 3-methoxy-benzene sulfonyl chloride with the
appropriate sulfonyl chloride in Step 1. The following compounds
were made following this procedure: [1259]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-2,5-dime-
thoxy-benzenesulfonamide (P-1131), [1260]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-methox-
y-benzenesulfonamide (P-0958), [1261] Piperidine-1-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-0952), [1262]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-triflu-
oromethyl-benzenesulfonamide (P-0931), [1263]
4-Butoxy-N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-benzenesulfonamide (P-1006), [1264]
4-Chloro-N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-
-benzenesulfonamide (P-0937), [1265]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3,4-dime-
thoxy-benzenesulfonamide, (P-1090), and [1266]
3,4-Dichloro-N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phe-
nyl]-benzenesulfonamide (P-1015). The following table indicates the
sulfonyl chloride (column 2) used to afford the target compound
(column 3). Column 1 provides the compound number and column 4
gives the observed mass.
TABLE-US-00004 [1266] MS (ESI) [M + H.sup.+].sup.+ Sulfonyl
chloride Compound observed P-1131 ##STR00754## ##STR00755## 474.2
P-0958 ##STR00756## ##STR00757## 444.2 P-0952 ##STR00758##
##STR00759## 421.2 P-0931 ##STR00760## ##STR00761## 482.2 P-1006
##STR00762## ##STR00763## 486.2 P-0937 ##STR00764## ##STR00765##
446.1 [M - H.sup.+].sup.- P-1090 ##STR00766## ##STR00767## 474.2
P-1015 ##STR00768## ##STR00769## 480.0 482.1 [M - H.sup.+].sup.- *
Piperidine-1-sulfonyl chloride prepared from sulfuryl chloride and
piperidine in acetonitrile, refluxed for 8 hours, concentrated, and
used without further purification.
Example 12
Synthesis of
1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde
96
Compound 96 was synthesized in two steps from 7-azaindole 94 as
described in Scheme 27.
##STR00770##
[1267] Step 1--Preparation of
1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde (95)
[1268] To 1H-Pyrrolo[2,3-b]pyridine (94, 16.0 g, 135 mmol) in water
(110 mL), were added hexamethylenetetramine (26.0 g, 185 mmol), and
acetic acid (55.0 mL, 967 mmol). The reaction was refluxed for 12
hours. Water (329 mL) was added and the reaction was cooled to room
temperature. The reaction was filtrated and washed with water to
give the compound (95, 15.0 g, 76%).
MS(ESI)[M+H.sup.+].sup.+=147.
Step 2--Preparation of
1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde
(96)
[1269] To 1H-Pyrrolo[2,3-b]pyridine-3-carbaldehyde (95, 4.05 g,
27.71 mmol) in tetrahydrofuran (30.0 mL) were added sodium hydride
(60% in mineral oil, 1.5 g, 38 mmol) and triisopropylsilyl chloride
(8.0 mL, 38 mmol) under an atmosphere of nitrogen. The reaction was
stirred for 2 hours at room temperature. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 10.3% ethyl acetate in hexane to
give the compound (96, 3.0 g, 36%).
MS(ESI)[M+H.sup.+].sup.+=303.
Example 13
Synthesis of 5-isopropyl-1H-pyrrolo[2,3-b]pyridine 99
[1270] Compound 98 was synthesized in three steps from
5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine 68
described in Scheme 28.
##STR00771##
Step 1--Preparation of
2-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-propan-2-ol
(97)
[1271] To 5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(68, 2.0 g, 5.66 mmol, prepared as described in Example 6) in
tetrahydrofuran (20.0 mL), cooled in a -78.degree. C. acetone/dry
ice bath, under an atmosphere of nitrogen, was added
tert-butyllithium (1.7 M in tetrahydrofuran, 7.3 mL, 12 mmol)
dropwise. After 20 minutes, acetone (0.830 mL, 11 mmol) was added
dropwise to the reaction. The reaction was stirred for 30 minutes
at -78.degree. C. and then allowed to reach room temperature. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was washed with brine, dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated and
purified by silica gel column chromatography eluting with 10% ethyl
acetate in hexane to give the compound (97, 1.30 g, 69%).
MS(ESI)[M+H.sup.+].sup.+=333.
Step 2--Preparation of 5-isopropenyl-1H-pyrrolo[2,3-b]pyridine
(98)
[1272] To
2-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-propan-2-
-ol (97, 0.500 g, 1.5 mmol) in acetonitrile (10.0 mL) were added
triethylsilane (1.00 mL, 6.3 mmol) and trifluoroacetic acid (0.50
mL, 6.5 mmol) under an atmosphere of nitrogen. The reaction was
refluxed for 3 hours, then cooled down to room temperature. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was washed with brine, dried over anhydrous
sodium sulfate and filtered. The filtrate was concentrated and
purified by silica gel column chromatography eluting with 50% ethyl
acetate in hexane to give the compound (98, 0.200 g, 84%).
MS(ESI)[M+H.sup.+].sup.+=159.
Step 3--Preparation of 5-isopropyl-1H-pyrrolo[2,3-b]pyridine
(99)
[1273] To 5-isopropenyl-1H-pyrrolo[2,3-b]pyridine (98, 0.080 g,
0.501 mmol) in tetrahydrofuran (5.0 mL) was added 20% palladium
hydroxide on carbon (5.0 mg). The reaction was stirred under
hydrogen at 40 psi for 30 minutes. The reaction mixture was
filtered and concentrated to give the compound (99, 0.078 g, 96%).
MS(ESI)[M+H.sup.+].sup.+=161.
Example 14
Synthesis of 5-Methyl-1H-pyrrolo[2,3-b]pyridine 101.
[1274] Compound 101 was synthesized in two steps from
5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine 68
described in Scheme 29.
##STR00772##
Step 1--Preparation of
5-Methyl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (100)
[1275] To PdCl.sub.2(dppf) (0.04 g, 0.05 mmol) in toluene (10.0 mL)
under an atmosphere of nitrogen were added
5-bromo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (68, 0.3 g,
0.8 mmol, prepared as described in Example 6, 1.0 mL in toluene)
and methylmagnesium bromide (1.0 M in tetrahydrofuran, 3.0 mL, 3.0
mmol). The reaction was stirred 90.degree. C. for 2 hours and then
allowed to reach to room temperature. The reaction was poured into
citric acid (0.1 M in water) and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 50% ethyl acetate in
hexane to give the compound (100, 0.16 g, 60.0%).
MS(ESI)[M+H.sup.+].sup.+=289.4.
Step 2--Preparation of 5-Methyl-1H-pyrrolo[2,3-b]pyridine (101)
[1276] To 5-Methyl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(100, 0.160 g, 0.55 mmol) in tetrahydrofuran (3.0 mL) was added
tetra-n-butyl ammonium fluoride (0.145 g, 0.55 mmol). The reaction
was stirred for 1 hour at room temperature. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 3% methanol in dichloromethane
to provide light yellow solid (101, 0.07 g, 95%).
MS(ESI)[M+H.sup.+].sup.+=133.2.
5-Methyl-1H-pyrrolo[2,3-b]pyridine
##STR00773##
[1277] was prepared following the protocol of Scheme 29,
substituting methylmagnesium bromide with ethylmagnesium bromide in
Step 1.
Example 15
Synthesis of
1-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-3-propyl-urea P-0774 and Related Compounds
[1278] Compound P-0774 was synthesized in two steps from
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethyl]-phenyl}-carbamic acid methyl ester P-0065 described in
Scheme 30.
##STR00774##
Step 1--Preparation of
1-{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3b]pyridin-3-yl)--
methyl]-phenyl}-3-propyl-urea (103)
[1279] To
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methyl]-phenyl}-carbamic acid methyl ester (P-0065, 30.0
mg, 0.07 mmol, prepared as described in Example 10) was added
1-propanamine (2.0 mL, 20 mmol) and the reaction was heated to
120.degree. C. for 20 minutes in a CEM Discover microwave
instrument. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 5% methanol in methylene chloride to give the compound
(103, 20.0 mg, 60%). MS(ESI)[M+H.sup.+].sup.+=438.51.
Step 2--Preparation of
1-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-p-
henyl]-3-propyl-urea (P-0774)
[1280] To
1-{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methyl]-phenyl}-3-propyl-urea (103, 20.0 mg, 0.04 mmol)
in tetrahydrofuran (3 mL) was added Dess-Martin periodane (23.0 mg,
0.055 mmol) under an atmosphere of nitrogen. The reaction was
stirred for 10 minutes. The reaction was concentrated and purified
by silica gel column chromatography eluting with 1% methanol in
dichloromethane to give the compound (P-0774, 6.0 mg, 30%).
MS(ESI)[M+H.sup.+].sup.+=436.5.
[1281] Additional compounds were prepared following the protocol of
Scheme 30, replacing 1-propanamine with an appropriate amine and
optionally replacing
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methyl]-phenyl}-carbamic acid methyl ester P-0065 with an
appropriate carbamic acid methyl ester (see Example 10) in Step 1.
The following compounds were made following this procedure: [1282]
1-sec-Butyl-3-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl)-phenyl]-urea (P-1289), [1283]
1-Cyclopentyl-3-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-
-3-carbonyl)-phenyl]-urea (P-1317), [1284]
1-Butyl-3-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-car-
bonyl)-phenyl]-urea (P-1318), [1285]
1-Butyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
)-phenyl]-urea (P-1567), [1286]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-(2-morpholin-4-yl-ethyl)-urea (P-1580), [1287]
1-Butyl-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-
-phenyl]-urea (P-1586), [1288] Morpholine-4-carboxylic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1606), [1289]
1-Butyl-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-
-phenyl]-1-methyl-urea (P-1612), [1290]
1-Butyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
)-phenyl]-1-methyl-urea (P-1884), [1291] Morpholine-4-carboxylic
acid
[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
amide (P-1894), [1292]
1-Butyl-3-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
1-ethyl-urea (P-1983), [1293]
1-Butyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
)-phenyl]-1-ethyl-urea (P-1994), and [1294]
3-Diethylamino-pyrrolidine-1-carboxylic acid
[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
amide (P-2015). The following table indicates the amine (column 2)
and the carbamic acid methyl ester (column 3) used to afford the
target compounds (column 4). Column 1 provides the compound number
and column 5 the observed mass.
TABLE-US-00005 [1294] MS (ESI) Carbamic acid methyl [M +
H.sup.+].sup.+ Amine ester Compound observed P-1289 ##STR00775##
##STR00776## ##STR00777## 450.4 P-1317 ##STR00778## ##STR00779##
##STR00780## 462.4 P-1318 ##STR00781## ##STR00782## ##STR00783##
450.5 P-1567 ##STR00784## ##STR00785## ##STR00786## 403.2 P-1580
##STR00787## ##STR00788## ##STR00789## 464.3 P-1586 ##STR00790##
##STR00791## ##STR00792## 407.3 P-1606 ##STR00793## ##STR00794##
##STR00795## 421.1 P-1612 ##STR00796## ##STR00797## ##STR00798##
421.3 P-1884 ##STR00799## ##STR00800## ##STR00801## 417.4 P-1894
##STR00802## ##STR00803## ##STR00804## 417.4 P-1983 ##STR00805##
##STR00806## ##STR00807## 401.4 P-1994 ##STR00808## ##STR00809##
##STR00810## 431.4 P-2015 ##STR00811## ##STR00812## ##STR00813##
472.4
[1295] The intermediate product of Step 1 of Scheme 30 can
alternatively be reacted under reducing conditions to provide
analogous compounds in which the carbonyl linker of the 3-position
of azaindole to the phenyl ring is methylene. The product of Step 1
is a mixture of the hydroxyl and methoxy methyl linker of the
3-position, which may be carried through the reduction Step as the
mixture or may be isolated as either the hydroxyl or methoxy for
use in this reaction. This reduction is exemplified as follows
using the product of Step 1 in the preparation of P-1567 to prepare
1-Butyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-y-
lmethyl)-phenyl]-urea P-1571.
##STR00814##
Step 1--Preparation of
1-Butyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-
-phenyl]-urea (P-1571)
[1296] A mixture of
1-Butyl-3-2,4-difluoro-3-[hydroxy-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-y-
l)-methyl]-phenyl-urea (41 mg, 0.000081 mol, isolated from Step 1
of Scheme 30), triethylsilane (2 mL, 0.01 mol), and trifluoroacetic
acid (1 mL, 0.01 mol) in 20 ml of acetonitrile was refluxed for 3
hrs. The mixture was concentrated and the residue was redissolved
in ethyl acetate and sodium bicarbonate solution. The organic layer
was collected and dried over MgSO.sub.4. An off-white solid
compound was obtained after chromatography (P-1571, 18 mg, 57%).
MS(ESI) [M-H.sup.+].sup.-=389.2.
[1297] Additional compounds were prepared following the protocol of
Step 1 of Scheme 30 followed by the reduction step above as Step 2,
where the product of Scheme 30 Step 1 may be isolated as either the
hydroxyl or methoxy derivative, or used as the mixture, replacing
1-propanamine with an appropriate amine and
{2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3b]pyridin-3-yl)-me-
thyl]-phenyl}-carbamic acid methyl ester P-0065 with an appropriate
carbamic acid methyl ester (see Example 10) in Step 1. The
following compounds were made following this procedure: [1298]
1-Cyclopentyl-3-[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylm-
ethyl)-phenyl]-urea (P-1572), [1299]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-cyclopentyl-urea (P-1575), [1300]
1-Butyl-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro--
phenyl]-urea (P-1587), [1301]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-(2-morpholin-4-yl-ethyl)-urea (P-1594), [1302]
Morpholine-4-carboxylic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-pheny-
l]-amide (P-1595), [1303]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-(2,2,2-trifluoro-ethyl)-urea (P-1601), [1304]
1-Cyclopentyl-3-{2,4-difluoro-3-[5-(3-methanesulfonyl-phenyl)-1H-pyrrolo[-
2,3-b]pyridin-3-ylmethyl]-phenyl}-urea (P-1615), [1305]
1-Butyl-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro--
phenyl]-1-methyl-urea (P-1625), [1306]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-cyclopropylmethyl-urea (P-1652), [1307]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-(4-fluoro-phenyl)-urea (P-1657), [1308]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenyl]--
3-(4-fluoro-benzyl)-urea (P-1654), and [1309]
3-Diethylamino-pyrrolidine-1-carboxylic acid
[2,4-difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-a-
mide (P-2014). The following table indicates the amine (column 2)
and the carbamic acid methyl ester (column 3) used to afford the
target compound (column 4). For the carbamic acid methyl ester, R
can be H or CH.sub.3, where the compound was either a mixture of
the two, or either of the isolated compounds wherein R is H or R is
CH.sub.3. Column 1 provides the compound number and column 5 the
observed mass.
TABLE-US-00006 [1309] MS (ESI) Carbamic acid methyl [M +
H.sup.+].sup.+ Amine ester Compound observed P-1572 ##STR00815##
##STR00816## ##STR00817## 401.2 P-1575 ##STR00818## ##STR00819##
##STR00820## 405.2 P-1587 ##STR00821## ##STR00822## ##STR00823##
393.3 P-1594 ##STR00824## ##STR00825## ##STR00826## 450.3 P-1595
##STR00827## ##STR00828## ##STR00829## 407.2 P-1601 ##STR00830##
##STR00831## ##STR00832## 419.2 P-1615 ##STR00833## ##STR00834##
##STR00835## 525.2 P-1625 ##STR00836## ##STR00837## ##STR00838##
407.3 P-1652 ##STR00839## ##STR00840## ##STR00841## 391.2 P-1657
##STR00842## ##STR00843## ##STR00844## 461.2 P-1654 ##STR00845##
##STR00846## ##STR00847## 445.2 P-2014 ##STR00848## ##STR00849##
##STR00850## 458.4
Example 16
Synthesis of 5-Methoxy-1H-pyrrolo[2,3-b]pyridine 104 and Related
Compounds
[1310] Compound 104 was synthesized in one step from
5-bromo-1H-pyrrolo[2,3-b]pyridine 67 as described in Scheme 31.
##STR00851##
Step 1--Preparation of 5-Methoxy-1H-pyrrolo[2,3-b]pyridine
(104)
[1311] To 5-bromo-7-azaindole (67, 500.0 mg, 2.53 mmol) in
N,N-dimethylformamide (8 mL) were added copper(I) iodide (966 mg,
5.08 mmol) and sodium methoxide in methanol (3 M, 5 mL). The
reaction was stirred overnight at 120.degree. C. under an
atmosphere of Argon. The reaction was poured into water, and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate, filtered. The filtrate was concentrated
and purified with silica gel column chromatograph eluting with 20%
ethyl acetate in hexane to give white solid (104, 140 mg, 28%).
MS(ESI)[M+H.sup.+].sup.+=149.1. In an alternative method, 2.3 g
(11.7 mmol) 5-bromo-7-azaindole (67, 2.3 g, 11.7 mmol) was
dissolved in 75 mL N,N-dimethylformamide and 50 mL methanol (50
mL), adding sodium methoxide (32 g, 0.6 mol) and copper-(I) bromide
(3.2 g, 22.4 mmol) at room temperature. The reaction was stirred
for three hours at 100.degree. C. under an atmosphere of argon. The
mixture was diluted with ethyl acetate and poured into a solution
of ammonium chloride:ammonium hydroxide (4:1). The organic layer
was extracted with ammonium chloride:ammonium hydroxide (4:1),
washed with brine, dried over anhydrous sodium sulfate, filtered
and concentrated. The desired compound was isolated by silica gel
column chromatography eluting with 30% to 70% ethyl acetate in
hexanes to give a yellow solid (104, 0.27 g, 15.6%). MS(ESI)
[M+H.sup.+].sup.+=149.2.
5-Ethoxy-1H-pyrrolo[2,3-b]pyridine 506
##STR00852##
[1312] was prepared using the protocol of Scheme 31, substituting
methanol with ethanol and sodium methoxide with sodium
ethoxide.
5-(2-Methoxy-ethoxy)-1H-pyrrolo[2,3-b]pyridine 507
##STR00853##
[1313] was prepared using the protocol of Scheme 31, substituting
methanol with 2-Methoxy-ethanol and sodium methoxide with sodium
2-Methoxy-ethoxide (prepared from 2-Methoxy-ethanol and sodium
hydride). MS(ESI) [M+H.sup.+].sup.+=193.3.
[1314] Diethyl-[2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)-ethyl]-amine
508
##STR00854##
was prepared using the protocol of Scheme 31, substituting methanol
with 2-diethylamino-ethanol and sodium methoxide with sodium
2-diethylamino-ethoxide (prepared from 2 2-diethylamino-ethanol and
sodium hydride). MS(ESI) [M+H.sup.+].sup.+=234.5.
Example 17
Synthesis of 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89
[1315] 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 was synthesized
in one step from 5-bromo-1H-pyrrolo[2,3-b]pyridine 67 as described
in Scheme 32.
##STR00855##
Step 1--Preparation of 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(89)
[1316] To 5-bromo-7-azaindole (67, 1.00 g, 5.08 mmol) in water
(13.0 mL) and acetonitrile (36 mL) were added pyridine-3-boronic
acid (609, 1.0 g, 8.1 mmol), potassium carbonate (1.79 g, 0.0130
mol) and Tetrakis(triphenylphosphine)palladium(0) (50.0 mg, 0.043
mmol) under an atmosphere of nitrogen. The reaction mixture was
heated to 170.degree. C. overnight. The reaction mixture was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over sodium sulfate, and concentrated. The
residue was purified with silica gel column chromatography eluting
with 25% ethyl acetate in hexane to provide a light yellow solid
(89, 820 mg, 82%). MS(ESI)[M+H.sup.+].sup.+=196.1.
[1317] Additional compounds were prepared following the protocol of
Scheme 32, either by substituting pyridine-3-boronic acid with an
appropriate boronic acid or by substituting the 5-bromo-7-azaindole
with
5-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridin-
e and reacting with a suitable aryl or heteroaryl halide (i.e.
coupling with the boronic acid ester on the azaindole, and the
halide on the group to be coupled to the 5-position of the
azaindole). The following compounds were prepared by this
procedure: [1318] 5-(4-Chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine
(514), [1319] 5-(4-Fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine (605),
[1320] 5-Phenyl-1H-pyrrolo[2,3-b]pyridine, [1321]
5-(6-Methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine, [1322]
5-(2-Methoxy-pyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridine, [1323]
5-Pyridin-4-yl-1H-pyrrolo[2,3-b]pyridine, [1324]
4-(1H-Pyrrolo[2,3-b]pyridin-5-yl)-benzenesulfonamide, [1325]
3-(1H-Pyrrolo[2,3-b]pyridin-5-yl)-benzenesulfonamide, [1326]
5-Pyrimidin-5-yl-1H-pyrrolo[2,3-b]pyridine, [1327]
5-(3-Methanesulfonyl-phenyl)-1H-pyrrolo[2,3-b]pyridine (P-0173),
and [1328] 3-(1H-Pyrrolo[2,3-b]pyridin-5-yl)-benzamide (P-1622).
The following table indicates either 5-bromo-7-azaindole or
5-(4,4,5,5-Tetramethyl-[1,3,2]
dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine starting material
(column 1) and the appropriate reagent to be coupled to the 5
position of the azaindole (column 2) to afford the resulting
compound (column 3), with the observed mass given in column 4.
TABLE-US-00007 [1328] MS (ESI) Reagent coupled to [M +
H.sup.+].sup.+ Starting azaindole 5 position Compound observed
##STR00856## ##STR00857## ##STR00858## 229.1 ##STR00859##
##STR00860## ##STR00861## 213.1 ##STR00862## ##STR00863##
##STR00864## 195.2 ##STR00865## ##STR00866## ##STR00867## 226.2
##STR00868## ##STR00869## ##STR00870## 227.2 ##STR00871##
##STR00872## ##STR00873## 196.2 ##STR00874## ##STR00875##
##STR00876## 274.1 ##STR00877## ##STR00878## ##STR00879## 274.1
##STR00880## ##STR00881## ##STR00882## 197.2 ##STR00883##
##STR00884## ##STR00885## 273.1 ##STR00886## ##STR00887##
##STR00888## 238.2
Example 18
Synthesis of
3-(4-(4-chlorobenzyloxy)-3-methoxybenzyl)-1H-pyrrolo[2,3-b]pyridine
P-1247.
[1329] Compound P-1247 was synthesized in three steps from
4-hydroxy-3-methoxybenzaldehyde 105 as shown in Scheme 33.
##STR00889##
Step 1--Preparation of 4-(4-chlorobenzyloxy)-3-methoxybenzaldehyde
(106)
[1330] To 4-hydroxy-3-methoxybenzaldehyde (105, 600.0 mg, 3.94
mmol) and 4-chlorobenzyl bromide (557, 1.20 g, 5.84 mmol) in
acetonitrile (6 mL) was added potassium carbonate (0.390 g, 2.82
mmol). The reaction was microwaved on 300 watts, 120.degree. C. for
10 minutes. The reaction was extracted with ethyl acetate and
water. The organic layer was washed with brine, dried over
magnesium sulfate, filtered and the volatiles removed by
evaporation. The desired compound was purified by recrystallization
from hexanes to provide 106 (1.01 g, 93%). MS(ESI)
[M-H.sup.+].sup.-=275.1.
Step 2--Preparation of
3-((4-(4-chlorobenzyloxy)-3-methoxyphenyl)(methoxy)methyl)-1H-pyrrolo[2,3-
-b]pyridine (107)
[1331] To 1H-Pyrrolo[2,3-b]pyridine (94, 0.235 g, 1.99 mmol) and
4-(4-chlorobenzyloxy)-3-methoxybenzaldehyde (106, 0.500 g, 1.81
mmol) was added 5 mL of methanol followed by the addition of solid
potassium hydroxide (0.203 g, 3.61 mmol). The reaction was allowed
to stir at ambient temperature for 18 days. The reaction mixture
was poured into water and extracted with ethyl acetate. The organic
layer was separated and volatiles removed to give a solid which was
suspended in hot ethyl acetate. The suspension was allowed to cool
and the solid collected by vacuum filtration to provide 107 (548
mg, 74%). MS(ESI) [M+H.sup.+].sup.+=409.4.
Step 3--Preparation of
3-(4-(4-chlorobenzyloxy)-3-methoxybenzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1247)
[1332] To
3-((4-(4-chlorobenzyloxy)-3-methoxyphenyl)(methoxy)methyl)-1H-py-
rrolo[2,3-b]pyridine (107, 0.548 g, 1.34 mmol) in acetonitrile (20
mL) was added trifluoroacetic acid (1.7 mL, 2.21 mmol) and
triethylsilane (3.47 mL, 2.17 mmol). The reaction was stirred at
60.degree. C. for 15 hours. The volatiles were removed and the
desired compound was purified by silica gel chromatography, eluting
with a gradient from 0% to 60% ethyl acetate in hexahcs to provide
a white solid (P-1247, 505 mg, 99%).
MS(ESI)[M+H.sup.+].sup.+=379.4.
[1333] Additional compounds were prepared using the protocol of
Scheme 33, Steps 2 and 3, replacing
4-(4-chlorobenzyloxy)-3-methoxybenzaldehyde 106 with a suitable
aldehyde (prepared as described in Example 34), and optionally
replacing 1H-Pyrrolo[2,3-b]pyridine 94 with an appropriate
substituted 7-azaindole (see Example 9 or Example 16) in Step 2.
The following compounds were made following this procedure: [1334]
3-[3-Methoxy-4-(4-trifluoromethyl-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-1721), [1335]
3-[3-Trifluoromethyl-4-(4-trifluoromethyl-benzyloxy)-benzyl]-1H-pyrrolo[2-
,3-b]pyridine (P-1797), [1336]
3-{3-Methoxy-4-[4-(4-methyl-piperazin-1-ylmethyl)-benzyloxy]-benzyl}-1H-p-
yrrolo[2,3-b]pyridine (P-1821), [1337]
3-[4-(4-Chloro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-1844), [1338]
3-[4-(3-Fluoro-4-trifluoromethyl-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[-
2,3-b]pyridine (P-1849), [1339]
3-[4-(4-Chloro-3-trifluoromethyl-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[-
2,3-b]pyridine (P-1851), [1340]
2-[2-Methoxy-4-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenoxymethyl]-1H-be-
nzoimidazole (P-1870), [1341]
3-[4-(4-Chloro-2-fluoro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-5-methoxy-1-
H-pyrrolo[2,3-b]pyridine (P-1885), [1342]
3-[4-(3,4-Dichloro-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1886), [1343]
3-[4-(4-Chloro-benzyloxy)-3-fluoro-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1896), [1344]
2-[2-Fluoro-4-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenoxymethyl]-1H-ben-
zoimidazole (P-1899), [1345]
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1901), [1346]
5-Chloro-3-[4-(4-chloro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-p-
yrrolo[2,3-b]pyridine (P-1970), [1347]
5-Chloro-3-[4-(4-chloro-2-fluoro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-
-pyrrolo[2,3-b]pyridine (P-1972), [1348]
3-[4-(4-Chloro-2-fluoro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[-
2,3-b]pyridine (P-1973), [1349]
2-[4-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-5-fluoro-2-methoxy-ph-
enoxymethyl]-1H-benzoimidazole (P-1976), [1350]
2-[5-Fluoro-2-methoxy-4-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-p-
henoxymethyl]-1H-benzoimidazole (P-1977), [1351]
2-[5-Fluoro-2-methoxy-4-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenoxymeth-
yl]-1H-benzoimidazole (P-1978), [1352]
3-{4-[2-(2-Bromo-ethoxy)-ethoxy]-2-fluoro-5-methoxy-benzyl}-5-chloro-1H-p-
yrrolo[2,3-b]pyridine (P-1984), [1353]
5-Chloro-3-[2,5-difluoro-4-(2-methoxy-ethoxy)-benzyl]-1H-pyrrolo[2,3-b]py-
ridine (P-1986), [1354]
5-Chloro-3-[2-fluoro-5-methoxy-4-(2-methoxy-ethoxy)-benzyl]-1H-pyrrolo[2,-
3-b]pyridine (P-1990), [1355]
{3-[4-(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-5-fluoro-2-methoxy-p-
henoxy]-propyl}-diethyl-amine (P-2004), [1356]
5-Chloro-3-{2-fluoro-5-methoxy-4-[2-(2-methoxy-ethoxy)-ethoxy]-benzyl}-1H-
-pyrrolo[2,3-b]pyridine (P-2002), [1357]
3-(4-Benzyloxy-2,6-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-2022), [1358]
3-{2-Fluoro-5-methoxy-4-[2-(2-methoxy-ethoxy)-ethoxy]-benzyl}-5-me-
thoxy-1H-pyrrolo[2,3-b]pyridine (P-2025), and [1359]
3-{2-Fluoro-5-methoxy-4-[2-(2-methoxy-ethoxy)-ethoxy]-benzyl}-1H-pyrrolo[-
2,3-b]pyridine (P-2026). The following table indicates the aldehyde
(column 2) and the azaindole (column 3) used to afford the target
compound (column 4). Column 1 indicates the compound number and
column 5 the observed mass.
TABLE-US-00008 [1359] MS (ESI) [M + H.sup.+].sup.+ ob- Aldehyde
Azaindole Compound served P- 1721 ##STR00890## ##STR00891##
##STR00892## 413.2 P- 1797 ##STR00893## ##STR00894## ##STR00895##
451.3 P- 1821 ##STR00896## ##STR00897## ##STR00898## 457.4 P- 1844
##STR00899## ##STR00900## ##STR00901## 397.2 P- 1849 ##STR00902##
##STR00903## ##STR00904## 432.4 P- 1851 ##STR00905## ##STR00906##
##STR00907## P- 1870 ##STR00908## ##STR00909## ##STR00910## 385.4
P- 1885 ##STR00911## ##STR00912## ##STR00913## 445.3 P- 1886
##STR00914## ##STR00915## ##STR00916## 413.3 P- 1896 ##STR00917##
##STR00918## ##STR00919## 367.3 P- 1899 ##STR00920## ##STR00921##
##STR00922## 373.4 P- 1901 ##STR00923## ##STR00924## ##STR00925##
351.4 P- 1970 ##STR00926## ##STR00927## ##STR00928## 431.2 P- 1972
##STR00929## ##STR00930## ##STR00931## 449.2 P- 1973 ##STR00932##
##STR00933## ##STR00934## 415.3 P- 1976 ##STR00935## ##STR00936##
##STR00937## 437.3 P- 1977 ##STR00938## ##STR00939## ##STR00940##
433.4 P- 1978 ##STR00941## ##STR00942## ##STR00943## 403.4 P- 1984
##STR00944## ##STR00945## ##STR00946## 457.4 459.4 P- 1986
##STR00947## ##STR00948## ##STR00949## 353.4 P- 1990 ##STR00950##
##STR00951## ##STR00952## 365.3 P- 2004 ##STR00953## ##STR00954##
##STR00955## 420.4 P- 2002 ##STR00956## ##STR00957## ##STR00958##
409.3 P- 2022 ##STR00959## ##STR00960## ##STR00961## 438.1 P- 2025
##STR00962## ##STR00963## ##STR00964## 405.2 P- 2026 ##STR00965##
##STR00966## ##STR00967## 373.2
Example 19
Synthesis of Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]-amide
P-0955 and Related Compounds
[1360] As an alternative method to that of Example 2, compound
P-0955 was synthesized in nine steps from
4-chloro-2-fluoro-phenylamine 47 as shown in Scheme 37.
##STR00968## ##STR00969##
Step 1--Preparation of 3-Amino-6-chloro-2-fluoro-benzoic acid
benzyl ester (48)
[1361] To 4-chloro-2-fluoro-phenylamine (47, 6.30 mL, 57.0 mmol) in
tetrahydrofuran (300 mL), cooled with dry ice/acetone bath under an
atmosphere of nitrogen, n-butyllithium (2.50 M in hexane, 24.4 mL)
was added slowly. After 20 minutes,
1,2-Bis-(chloro-dimethyl-silanyl)-ethane (12.9 g, 60.0 mmol)
dissolved in tetrahydrofuran (40.0 mL) was added slowly to the
reaction. After 1 hour, n-butyllithium (2.50 M in hexane, 25.0 mL)
was added slowly to the reaction. The reaction was stirred at
-78.degree. C. for 20 minutes and then allowed to warm to room
temperature over 60 minutes. The reaction was cooled to -78.degree.
C., followed by addition of n-butyllithium (2.50 M in hexane, 26.0
mL) slowly. After 80 minutes, benzyl chloroformate (10.0 mL, 70.0
mmol) was added to the reaction. The reaction mixture was stirred
at -78.degree. C. overnight followed by addition of water (80 mL)
and concentrated hydrochloric acid (25 mL). The reaction was
allowed to warm to room temperature for 2 hours. The organic layer
was separated. The aqueous layer was basified with potassium
carbonate and extracted with ethyl acetate. The organic layers were
combined and washed with brine, dried over anhydrous sodium
sulfate, filtered and concentrated. The desired compound was
isolated by silica gel column chromatography (ethyl acetate/hexane
20%) to give a colorless oil (48, 12.5 g, 78.3%).
MS(ESI)[M+H.sup.+].sup.+=280.0.
Step 2--Preparation of
6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid benzyl
ester (49)
[1362] To 3-amino-6-chloro-2-fluoro-benzoic acid benzyl ester (48,
1.20 g, 4.3 mmol) in methylene chloride (28 mL) was added pyridine
(0.52 mL, 6.4 mmol) and propanesulfonyl chloride (0.685 g, 4.8
mmol). The reaction was stirred at room temperature overnight, then
poured into water, and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate,
filtered and concentrated. The desired compound was isolated with
silica gel column chromatography to give a colorless oil (49, 960
mg, 58.0%). MS(ESI) [M-H.sup.+].sup.-=384.1.
Step 3--Preparation of
6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid
(115)
[1363] To 6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid benzyl ester (49, 6.00 g, 15.6 mmol) in tetrahydrofuran (100
mL) was added 1.0 M aqueous potassium hydroxide (100 mL). The
reaction was heated to reflux overnight. The reaction was poured
into water, acidified to pH 2 with 1 N hydrochloric acid and
extracted with ethyl acetate. The organic portion was dried over
anhydrous sodium sulfate, filtered and concentrated to give a white
solid 115 (3.95 g, 85.8%).
Step 4--Preparation of 2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid (50)
[1364] To 6-chloro-2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid (115, 0.69 g, 2.3 mmol) in methanol (10 mL) was added 20%
palladium hydroxide on carbon (200 mg). The reaction was stirred
under hydrogen at 50 psi for 2 hours. The reaction was filtered and
concentrated to give white solid 50 that was used in the next step.
MS(ESI) [M-H.sup.+].sup.-=260.1.
Step 5--Preparation of 2-fluoro-3-(propane-1-sulfonylamino)-benzoic
acid methyl ester (501)
[1365] To a 2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid (50,
5.05 g, 0.0193 mol) in methylene chloride (100 mL) was added
N,N-dimethylformamide (0.075 mL, 0.97 mmol) under an atmosphere of
nitrogen. The reaction was cooled with ice/water, followed by slow
addition of Oxalyl chloride (2.00 M of in methylene chloride, 10.8
mL, 21.6 mmol). The reaction mixture was stirred at room
temperature for 3.0 hours. The reaction was cooled with ice/water,
followed by addition of methanol (36.0 mL, 0.89 mol) slowly. The
reaction was stirred at room temperature overnight. The reaction
was concentrate and purified with silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give a crude white
solid 4.0 g.
Step 6--Preparation of Propane-1-sulfonic acid
(2-fluoro-3-hydroxymethyl-phenyl)-amide (502)
[1366] To 2-fluoro-3-(propane-1-sulfonylamino)-benzoic acid methyl
ester (501, 3.80 g, 13.8 mmol) in tetrahydrofuran (133 mL) was
added lithium tetrahydroaluminate (1.00 M in tetrahydrofuran, 20.0
mL, 20.0 mmol) under an atmosphere of nitrogen at room temperature.
The reaction was stirred at room temperature for 8 hours, followed
by addition of 10 g of NaSO.sub.4.10H.sub.2O. After 12 hours, the
reaction was filtered, concentrated and purified with silica gel
column chromatography eluting with 5% methanol in methylene
chloride to give a white solid (502, 3.0 g, 87.9%).
Step 7--Preparation of propane-1-sulfonic acid
(2-fluoro-3-formyl-phenyl)-amide (503)
[1367] To propane-1-sulfonic acid
(2-fluoro-3-hydroxymethyl-phenyl)-amide (502, 0.20 g, 0.81 mmol) in
tetrahydrofuran (5.0 mL) was added Dess-Martin periodinane (0.377
g, 0.89 mmol). The reaction was stirred at room temperature for 10
minutes, then poured into water and extracted with ethyl acetate.
The organic layer was dried over anhydrous sodium sulfate, and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 20% ethyl acetate in hexane to
give a white solid (503, 100 mg, 50.0%). MS(ESI)
[M-H.sup.+].sup.+=244.1.
Step 8--Preparation of Propane-1-sulfonic acid
{3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2-fluoro-phen-
yl}-amide (504)
[1368] To 5-bromo-7-azaindole 67 (312 mg, 1.58 mmol) in methanol
(28 mL) were added propane-1-sulfonic acid
(2-fluoro-3-formyl-phenyl)-amide (503, 370 mg, 1.5 mmol) and
potassium hydroxide (422.8 mg, 7.5 mmol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature overnight,
then poured into water and extracted with ethyl acetate. The
organic layer was dried over anhydrous sodium sulfate, and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 20% ethyl acetate in hexane to
give the desired compound as white solid (504, 300 mg, 45.0%).
Step 9--Preparation of propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0955)
[1369] To Propane-1-sulfonic acid
{3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-2-fluoro-phen-
yl}-amide (504, 0.650 g, 1.47 mmol) in tetrahydrofuran (25.0 mL)
cooled with ice/water was added Dess-Martin periodinane (0.748 g,
1.76 mmol). The reaction was stirred at room temperature for 15
minutes. The reaction was poured into water containing sodium
thiosulfate and potassium carbonate and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate,
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
and washed with ethyl acetate to give white solid. (P-0955, 0.35 g,
54.1%). MS(ESI) [M+H.sup.+].sup.+=460.0, 462.0.
Butane-1-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]-amide
P-1250
##STR00970##
[1370] was prepared following the protocol of Scheme 37,
substituting propane-2-sulfonyl chloride with butane-1-sulfonyl
chloride in Step 1 and 5-bromo-1H-pyrrolo[2,3-b]pyridine 67 with
5-chloro-1H-pyrrolo[2,3-b]pyridine 80 (see Example 9) in step
8.
[1371] MS(ESI) [M-H.sup.+].sup.-=408.1.
Propane-1-sulfonic acid
[2-fluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amid-
e P-1256
##STR00971##
[1372] was prepared following the protocol of Scheme 37,
substituting 5-bromo-1H-pyrrolo[2,3-b]pyridine 67 with
5-methoxy-1H-pyrrolo[2,3-b]pyridine 104 (see Example 16) in step
8.
[1373] MS(ESI) [M-H.sup.+].sup.-=390.1.
N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-fluoro-phenyl]
benzenesulfonamide P-1255
##STR00972##
[1374] was prepared following the protocol of Scheme 37,
substituting propane-2-sulfonyl chloride with benzenesulfonyl
chloride in Step 1 and 5-bromo-1H-pyrrolo[2,3-b]pyridine 67 with
5-chloro-1H-pyrrolo[2,3-b]pyridine 80 (see Example 9) in step 8.
MS(ESI) [M-H.sup.+].sup.-=428.0.
Example 20
Synthesis of
3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]p-
yridin-5-yl-propionic acid P-1270
[1375] Compound P-1270 was synthesized in three steps from
propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide P-0773 as shown in Scheme 38.
##STR00973##
Step 1--Preparation of
(E)-3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-
-b]pyridin-5-yl-acrylic acid methyl ester (505)
[1376] To propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-am-
ide (P-0773, 125.0 mg, 0.27 mmol, prepared as described in Example
4) in N,N-dimethylformamide (4.0 mL) were added palladium acetate
(15 mg, 0.068 mmol), triphenylphosphine (36 mg, 0.14 mmol), methyl
acrylate (0.098 mL, 1.1 mmol) and triethylamine (0.114 mL, 0.82
mmol) under an atmosphere of nitrogen. The reaction was stirred at
140.degree. C. overnight, then poured into water, acidified with
water and extracted with ethyl acetate. To the filtrate in
methylene chloride (5.0 mL) was added
1,8-diazabicyclo[5.4.0]undec-7-ene (0.50 mL, 3.3 mmol). The
reaction was stirred at room temperature for 3 hours. The reaction
was concentrated and purified with silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give a light yellow oil
that was used directly in the next step.
Step 2--Preparation of
3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]p-
yridin-5-yl-acrylic acid (P-1269)
[1377] To
(E)-3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-py-
rrolo[2,3-b]pyridin-5-yl-acrylic acid methyl ester (100.0 mg, 0.22
mmol) in tetrahydrofuran (5.0 mL) and water (1.50 mL) was added
lithium hydroxide (21 mg, 0.86 mmol). The reaction was stirred at
room temperature overnight. The reaction was poured into water,
acidified with 1N HCl to pH around 1, and then extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate,
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
to give a white solid (P-1269, 30 mg). MS(ESI)
[M-H.sup.+].sup.-=448.0.
Step
3--3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[-
2,3-b]pyridin-5-yl-propionic acid (P-1270)
[1378] To
3-3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrol-
o[2,3-b]pyridin-5-yl-acrylic acid (P-1269, 20.0 mg, 0.045 mmol) in
methanol (5.0 mL) was added 20% Pd(OH)2/C (10 mg) under an
atmosphere of hydrogen. The reaction was stirred at room
temperature for 2 hours. The reaction mixture was filtered,
concentrated and purified with silica gel column chromatography
eluting with 10% methanol in methylene chloride to give a white
solid (P-1270, 8.8 mg). MS(ESI) [M-H.sup.+].sup.-=450.1.
Example 21
Synthesis of thiophene-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide 508
[1379] Compound 512 was synthesized in four steps from
2,4-difluorophenylamine 42 as shown in Scheme 39.
##STR00974##
Step 1--Preparation of 3-amino-4,2-difluoro-benzoic acid ethyl
ester (509)
[1380] To 4,2-difluoro-phenylamine (42, 6.30 mL, 57.0 mmol) in
tetrahydrofuran (300 mL), cooled with dry ice/acetone bath under an
atmosphere of nitrogen, n-butyllithium (2.50 M in hexane, 24.4 mL)
was added slowly. After 20 minutes,
1,2-Bis-(chloro-dimethyl-silanyl)-ethane (12.9 g, 60.0 mmol)
dissolved in tetrahydrofuran (40.0 mL) was slowly added to the
reaction. After 1 hour, n-butyllithium (2.50 M in hexane, 25.0 mL)
was slowly added to the reaction. The reaction was stirred at
-78.degree. C. for 20 minutes and then allowed to warm to room
temperature over 60 minutes. The reaction was cooled to -78.degree.
C., followed by addition of n-butyllithium (2.50 M in hexane, 26.0
mL) slowly. After 80 minutes, ethyl chloroformate (6.69 mL, 70.0
mmol) was added to the reaction. The reaction mixture was stirred
at -78.degree. C. overnight followed by addition of water (80 mL)
and concentrated hydrochloric acid (25 mL). The reaction was
allowed to warm to room temperature for 2 hours. The organic layer
was separated. The aqueous layer was basified with potassium
carbonate and extracted with ethyl acetate. The organic layers were
combined and washed with brine, dried over anhydrous sodium
sulfate, filtered and concentrated. The desired compound was
isolated by silica gel column chromatography (ethyl acetate/hexane
20%) to give a colorless oil (509, 4.6 g, 39%). MS(ESI)
[M+H.sup.+].sup.+=218.1.
Step 2--Preparation of
2,6-difluoro-3-(thiophene-2-sulfonylamino)-benzoic acid ethyl ester
(510)
[1381] To 3-amino-2,4-difluoro-benzoic acid ethyl ester (509, 1.20
g, 5.93 mmol) in methylene chloride (28 mL) was added pyridine
(0.52 mL, 6.4 mmol) and thiophene-2-sulfonyl chloride (0.97 g, 5.38
mmol). The reaction was stirred at room temperature overnight, then
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate,
filtered and concentrated. The desired compound was isolated with
silica gel column chromatography (ethyl acetate/hexane 20%) to give
a colorless oil (510, 1.2 g, 65.0%). MS(ESI)
[M+H.sup.+].sup.+=348.2.
Step 3--Preparation of thiophene-2-sulfonic acid
(2,4-difluoro-3-hydroxymethyl-phenyl)-amide (511)
[1382] To 2,6-difluoro-3-(thiophene-2-sulfonylamino)-benzoic acid
ethyl ester (510, 1.6 g, 3.5 mmol) in tetrahydrofuran (25.0 mL) was
added lithium tetrahydroaluminate (1.00 M in tetrahydrofuran, 8.08
mL, 8.08 mmol) under an atmosphere of nitrogen at room temperature.
The reaction was stirred at room temperature for 8 hours, followed
by addition of 10 g of NaSO.sub.4.10H.sub.2O. After 12 hours, the
reaction was filtered, concentrated and purified with silica gel
column chromatography eluting with 5% methanol in methylene
chloride to give a white solid (511, 300.0 mg, 21.0%).
Step 4--Preparation of thiophene-2-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (512)
[1383] To thiophene-2-sulfonic acid
(2,4-difluoro-3-hydroxymethyl-phenyl)-amide (511, 0.46 g, 1.52
mmol) in tetrahydrofuran (5.0 mL) was added Dess-Martin periodinane
(0.71 g, 1.67 mmol). The reaction was stirred at room temperature
for 10 minutes, then poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate,
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
to give a white solid (512, 100 mg, 21%). MS(ESI)
[M+H.sup.+].sup.+=304.2.
[1384] Thiophene-3-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide 513
##STR00975##
was prepared following the protocol of Scheme 39, substituting
thiophene-2-sulfonyl chloride with thiophene-3-sulfonyl chloride in
Step 2. MS(ESI) [M+H.sup.+].sup.+=304.2.
N-(2,4-Difluoro-3-formyl-phenyl)-methanesulfonamide 577
##STR00976##
[1385] was prepared following the protocol of Scheme 39,
substituting thiophene-2-sulfonyl chloride with methanesulfonyl
chloride in Step 2.
N-(2,4-Difluoro-3-formyl-phenyl)-3-fluoro-benzenesulfonamide
578
##STR00977##
[1386] was prepared following the protocol of Scheme 39,
substituting thiophene-2-sulfonyl chloride with
3-fluoro-benzenesulfonyl chloride in Step 2.
Example 22
Synthesis of
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxyl-
ic acid butylamide P-1486 and Related Compounds
[1387] Compound P-1486 was synthesized in three steps from
1H-indole-4-carbaldehyde 518 as shown in Scheme 41.
##STR00978##
Step 1--Preparation of 4-Formyl-indole-1-carboxylic acid butylamide
(519)
[1388] To 1H-Indole-4-carbaldehyde (518, 1.57 g, 10.8 mmol) in
acetonitrile (20 mL) was added 1-isocyanatobutane (1.81 mL, 16.2
mmol), followed by 4-dimethylaminopyridine (130 mg, 1.1 mmol). The
reaction was refluxed for 48 hours. The reaction solution was
quenched with 1 M HCl (aq.) and extracted with ethyl acetate. The
organic layer was washed with sodium bicarbonate and brine, dried
over anhydrous magnesium sulfate, filtrated and concentrated to
give a light yellow solid (519, 2.62 g, 45%). MS(ESI)
[M+H.sup.+].sup.+=245.2.
Step 2--Preparation of
4-[Hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-indole--
1-carboxylic acid butylamide (520)
[1389] To 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 51 mg, 0.26
mmol, prepared as in Example 17) in methanol (2 mL), was added
4-Formyl-indole-1-carboxylic acid butylamide (519, 84 mg, 0.34
mmol) and potassium hydroxide (44 mg, 0.78 mmol). The reaction was
stirred at room temperature overnight. The reaction was poured into
water and extracted with ethyl acetate. The organic layer was
washed with water and brine, dried over anhydrous magnesium
sulfate, filtrated and concentrated. The desired compound was
isolated with silica gel column chromatography to give an off-white
solid (520, 7 mg, 6%). MS(ESI) [M+H.sup.+].sup.+=440.3.
Step 3--Preparation of
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxyl-
ic acid butylamide (P-1486)
[1390] To
4-[Hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl-
]-indole-1-carboxylic acid butylamide (520, 7 mg, 0.016 mmol) in
tetrahydrofuran (1 mL) was added Dess-Martin periodinane (7.4 mg,
0.017 mmol). The reaction was stirred at room temperature for 30
minutes, then poured into water and extracted with ethyl acetate.
The organic layer was washed with saturated sodium bicarbonate and
brine, dried over anhydrous magnesium sulfate, filtrated and
concentrated. The desired compound was purified by Prep HPLC using
a gradient of buffer A (5% acetonitrile, 95% water, 0.1% formic
acid) and buffer B (95% acetonitrile, 5% water, 0.1% formic acid).
P-1486 was isolated as a fluffy white solid (2.8 mg, 40%). MS(ESI)
[M+H.sup.+].sup.+=438.3.
[1391] Additional compounds were prepared following the protocol of
Scheme 41, optionally substituting 1-isocyanatobutane with an
appropriate isocyanate in Step 1 under suitable base/solvent
conditions (dimethylaminopyridine and acetonitrile per Scheme 41
Step 1 or appropriate conditions) and optionally substituting
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with a suitable
7-azaindole in Step 2. The azaindole was purchased or synthesized
as described in Examples 16 and 17. The following compounds were
made following this procedure: [1392]
(1-Benzyloxymethyl-1H-indol-4-yl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methanone (P-1523), [1393]
(1-Benzenesulfonyl-1H-indol-4-yl)-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone (P-1524), [1394]
4-(5-Phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxylic
acid butylamide (P-1576), [1395]
4-[5-(4-Chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indole-1-car-
boxylic acid butylamide (P-1602), [1396]
4-(5-Phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxylic
acid benzylamide (P-1611), [1397]
4-[5-(4-Chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indole-1-car-
boxylic acid benzylamide (P-1618), [1398]
4-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxylic
acid butylamide (P-1687), [1399]
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-sulfonic
acid dimethylamide (P-1744), [1400]
(1-But-2-ynyl-1H-indol-4-yl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-y-
l)-methanone (P-1820), [1401]
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-sulfonic
acid butyl-methyl-amide (P-1830), [1402]
4-[5-(4-Sulfamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indole-1--
carboxylic acid butylamide (P-1854), [1403]
4-[5-(3-Sulfamoyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indole-1--
carboxylic acid butylamide (P-1858), [1404]
4-[5-(2-Methoxy-pyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-ind-
ole-1-carboxylic acid butylamide (P-1860), [1405]
4-(5-Pyrimidin-5-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carbox-
ylic acid butylamide (P-1862), [1406]
4-(5-Methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxylic
acid butylamide (P-1875), and [1407]
4-[5-(6-Methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indol-
e-1-carboxylic acid (pyridin-3-ylmethyl)-amide (P-1887). The
following table indicates the base/solvent (column 2) and
isocyanate used in Step 1 (column 3), and the 7-azaindole used in
Step 2 (column 4) to afford the target compound (column 5). Column
1 provides the compound number and column 6 the observed mass.
TABLE-US-00009 [1407] Step 1 MS (ESI) Base/ Step 1 [M +
H.sup.+].sup.+ Solvent reagent Azaindole Compound observed P- 1523
NaH/ THF ##STR00979## ##STR00980## ##STR00981## 459.4 P- 1524 KOH/
CH.sub.2Cl.sub.2 water ##STR00982## ##STR00983## ##STR00984## 480.1
482.1 P- 1576 * ##STR00985## ##STR00986## ##STR00987## 437.3 P-
1602 * ##STR00988## ##STR00989## ##STR00990## 471.2 P- 1611 *
##STR00991## ##STR00992## ##STR00993## 471.3 P- 1618 * ##STR00994##
##STR00995## ##STR00996## 505.2 P- 1687 * ##STR00997## ##STR00998##
##STR00999## 439.2 441.2 P- 1744 NaH/ THF ##STR01000## ##STR01001##
##STR01002## 446.2 P- 1820 NsH/ THF ##STR01003## ##STR01004##
##STR01005## 391.2 P- 1830 KOH/ CH.sub.2Cl.sub.2 water ##STR01006##
##STR01007## ##STR01008## P- 1854 * ##STR01009## ##STR01010##
##STR01011## 514.2 [M - H.sup.+].sup.- P- 1858 * ##STR01012##
##STR01013## ##STR01014## 516.2 P- 1860 * ##STR01015## ##STR01016##
##STR01017## 469.3 P- 1862 * ##STR01018## ##STR01019## ##STR01020##
439.3 P- 1875 * ##STR01021## ##STR01022## ##STR01023## 391.4 P-
1887 * ##STR01024## ##STR01025## ##STR01026## 503.4 * per Scheme 41
(i.e. dimethylaminopyridine and acetonitrile)
[1408] The product of Step 2 of Scheme 41 can alternatively be
reacted to form the corresponding compounds with methylene linker
at the 3 position of the azaindole. For example,
4-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-indole-1-carboxyli-
c acid butylamide P-1656 was prepared from
4-[Hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-indole--
1-carboxylic acid butylamide 520 as shown in Scheme 41a.
##STR01027##
Step 1--Preparation of
4-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-indole-1-carboxyli-
c add butylamide (P-1656)
[1409] A mixture of
4-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-indole--
1-carboxylic acid butylamide (520, 18 mg, 0.041 mmol),
trifluoroacetic acid (0.5 mL), triethylsilane (1 mL), and
acetonitrile (8 mL) was refluxed for 4 hours. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was washed with saturated sodium bicarbonate and brine, dried
over anhydrous magnesium sulfate, filtrated and concentrated. The
desired compound was purified by Prep HPLC using a gradient of
buffer A (5% acetonitrile, 95% water, 0.1% formic acid) and buffer
B (95% acetonitrile, 5% water, 0.1% formic acid). P-1656 was
isolated as an off-white solid (4.8 mg, 28%). MS(ESI)
[M+H.sup.+].sup.+=424.2.
[1410] The corresponding hydroxy-methyl derivative of Scheme 41
Step 2 was reacted following the protocol of Scheme 41a to prepare
4-[5-(2-methoxy-pyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl]-indo-
le-1-carboxylic acid butylamide P-1861,
4-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-indole-1-carboxylic
acid butylamide P-1876, and
4-(5-Pyridin-4-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-indole-1-carboxyli-
c acid butylamide P-1877, with structures shown below.
##STR01028##
Example 23
Synthesis of
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methanone P-1467 and Related Compounds
[1411] Compound P-1467 was synthesized in four steps from
2,4-difluorophenol 35 as shown in Scheme 43.
##STR01029##
Step 1--Preparation of 1-Benzyloxy-2,4-difluoro-benzene (525)
[1412] To 2,4-difluoro-phenol (35, 7.60 g, 0.0584 mol) in
N,N-dimethylformamide (50.0 mL) were added benzyl bromide (8.0 mL,
0.067 mol) and potassium carbonate (9.00 g, 0.0651 mol) under an
atmosphere of nitrogen. The reaction was stirred at room
temperature overnight. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give the compound as
white solid (525, 3.20 g, 25%).
Step 2--Preparation of 3-Benzyloxy-2,6-difluoro-benzaldehyde
(526)
[1413] To 1 -Benzyloxy-2,4-difluoro-benzene (525, 3.00 g, 13.6
mmol) in Tetrahydrofuran (48 mL) under an atmosphere of nitrogen
and cooled with dry ice/acetone was added n-Butyllithium (1.60 M in
hexane, 8.94 mL). After 20 minutes, N,N-dimethylformamide (1.46 mL,
0.0189 mol) was added to the reaction. After another 20 minutes,
the flask was stirred at room temperature for 30 minutes. The
reaction mixture was poured into water, acidified to pH=1, and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and purified with
silica gel column chromatography eluting with 30% ethyl acetate in
hexane to give the compound as a yellow solid (526, 2.5 g,
74%).
Step 3--Preparation of
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methanol (527)
[1414] To 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 750.0 mg,
0.003842 mol, prepared as in Example 17) in methanol (20.0 mL) were
added 3-Benzyloxy-2,6-difluoro-benzaldehyde (526, 1.12 g, 4.5 mmol)
and potassium hydroxide (1.50 g, 0.0267 mol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature overnight
and then poured into water, acidified with 1N HCl to pH around 2
and extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give the compound (527,
700 mg, 35%).
Step 4--Preparation of
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridi-
n-3-yl)-methanone (P-1467)
[1415] To
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridin-3-yl)-methanol (527, 300.0 mg, 0.68 mmol) in
tetrahydrofuran (10.0 mL) was added Dess-Martin periodinane (344
mg, 0.81 mmol). The reaction was stirred at room temperature for 10
minutes. The reaction mixture was concentrated with silica and
purified with silica gel column chromatography eluting with 10%
methanol in dichloromethane to give the compound (P-1467, 240 mg,
80%). MS(ESI) [M+H.sup.+].sup.+=442.2.
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-[2,6-difluoro-3-(2-methoxy-ethoxy)-
-phenyl]-methanone P-1453
##STR01030##
[1416] was prepared following the protocol of Scheme 43,
substituting benzyl bromide with 1-Bromo-2-methoxy-ethane in Step 1
and 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
5-Bromo-1H-pyrrolo[2,3-b]pyridine (67) in Step 3. MS (ESI)
[M+H.sup.+].sup.+=410.1, 412.1.
[2,6-Difluoro-3-(2-methoxy-ethoxy)-phenyl]-(5-methoxy-1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-methanone P-1584
##STR01031##
[1417] was prepared following the protocol of Scheme 43,
substituting benzyl bromide with 1-Bromo-2-methoxy-ethane in Step 1
and 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
5-methoxy-1H-pyrrolo[2,3-b]pyridine (104, prepared as in Example
22) in Step 3. MS (ESI) [M+H.sup.+].sup.+=363.2.
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl-
)-methanone P-1597
##STR01032##
[1418] was prepared following the protocol of Scheme 43,
substituting 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
5-methoxy-1H-pyrrolo[2,3-b]pyridine (104, prepared as in Example
16) in Step 3. MS (ESI) [M+H.sup.+].sup.+=395.2.
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2,6-difluoro-3-methoxy-phenyl)-me-
thanone P-1386
##STR01033##
[1419] was prepared following the protocol of Steps 2, 3 and 4 of
Scheme 43, substituting 1-Benzyloxy-2,4-difluoro-benzene 525 with
2,4-Difluoro-1-methoxy-benzene in Step 2 and
5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
5-bromo-1H-pyrrolo[2,3-b]pyridine (67) in Step 3. MS (ESI)
[M+H.sup.+].sup.+=367.0, 369.0.
(3-Benzyloxy-2,6-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanon-
e P-1802
##STR01034##
[1420] was prepared following the protocol of Scheme 43,
substituting 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
7-azaindole. To a solution of
(3-benzyloxy-2,6-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone (P-1802, 0.5 g, 1.37 mol) in methanol (70 mL) and
tetrahydrofuran (30 mL) was added palladium on carbon (120 mg, 10%
wt., 0.58 mol). The mixture was stirred under hydrogenation (60
psi) for six hours. After removal of solvent, the residue was dried
under vacuum, which provided
(2,6-Difluoro-3-hydroxy-phenyl)-(1H-pyrrolo[2,3-b]pyridine-3-yl)-methanon-
e 651
##STR01035##
as a white solid (363 mg, 96%). MS (ESI)
[M+H.sup.+].sup.+=275.36.
[1421] Additional compounds were prepared following steps 3 and 4
of Scheme 43, replacing 3-benzyloxy-2,6-difluoro-benzaldehyde 526
with an appropriate aldehyde and/or
pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with an appropriate
azaindole in Step 3. The azaindoles used were synthesized as
described in Examples 9 or 16. The aldehydes used were synthesized
as described in Example 5 or 21. The following compounds were made
following this procedure: [1422]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3,5-dimethoxy-phenyl)-methanone
(P-1463), [1423]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-fluoro-5-trifluoromethyl-pheny-
l)-methanone, [1424]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethoxy-phenyl)-metha-
none, [1425]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethyl-phenyl)-methan-
one, [1426]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3,5-dichloro-phenyl)-methanone,
[1427]
(2,6-Dichloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone,
[1428]
(2,6-Difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone,
[1429]
(2,6-Dimethyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone,
[1430]
(2,6-Dichloro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-y-
l)-methanone, [1431]
(2,6-Difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone, [1432]
(2,6-Dimethyl-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone, [1433]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2,2-difluoro-benzo[1,3]dioxol-4--
yl)-methanone (P-1513), [1434]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-difluoromethoxy-phenyl)-methan-
one (P-1514), [1435]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2,2-difluoro-benzo[1,3]dioxol-5--
yl)-methanone (P-1551), [1436]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-trifluoromethyl-benzenesulfonamide (P-1541), [1437]
N-[2,4-Difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-4-trifluoromethyl-benzenesulfonamide (P-1542), [1438]
N-[2,4-Difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-3-fluoro-benzenesulfonamide (P-1581), [1439]
N-[2,4-Difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-4-fluoro-benzenesulfonamide (P-1582), [1440]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-fluoro-benzenesulfonamide (P-1583), [1441]
N-[2,4-Difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-3-trifluoromethyl-benzenesulfonamide (P-1598), [1442]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-trifluoromethyl-benzenesulfonamide (P-1599), [1443]
(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-{2-fluoro-5-methoxy-4-[2-(2-meth-
oxy-ethoxy)-ethoxy]-phenyl}-methanone (P-2003), [1444]
(4-Benzyloxy-2,6-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methano-
ne (P-2020), and [1445]
[4-(4-Chloro-benzyloxy)-3-methoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone (P-1698). The following table indicates the aldehyde
(column 2) and the azaindole (column 3) used to afford the target
compound (column 4). Column 1 provides the compound number and
column 5 the observed mass.
TABLE-US-00010 [1445] Aldehyde Azaindole P-1463 ##STR01036##
##STR01037## 567 ##STR01038## ##STR01039## 568 ##STR01040##
##STR01041## 569 ##STR01042## ##STR01043## 570 ##STR01044##
##STR01045## 571 ##STR01046## ##STR01047## 572 ##STR01048##
##STR01049## 573 ##STR01050## ##STR01051## 574 ##STR01052##
##STR01053## 575 ##STR01054## ##STR01055## 576 ##STR01056##
##STR01057## P-1513 ##STR01058## ##STR01059## P-1514 ##STR01060##
##STR01061## P-1551 ##STR01062## ##STR01063## P-1541 ##STR01064##
##STR01065## P-1542 ##STR01066## ##STR01067## P-1581 ##STR01068##
##STR01069## P-1582 ##STR01070## ##STR01071## P-1583 ##STR01072##
##STR01073## P-1598 ##STR01074## ##STR01075## P-1599 ##STR01076##
##STR01077## P-2003 ##STR01078## ##STR01079## P-2020 ##STR01080##
##STR01081## P-1698 ##STR01082## ##STR01083## MS(ESI) [M +
H.sup.+].sup.+ Compound observed P-1463 ##STR01084## 361.1 363.0
567 ##STR01085## 568 ##STR01086## 569 ##STR01087## 570 ##STR01088##
571 ##STR01089## 572 ##STR01090## 573 ##STR01091## 574 ##STR01092##
575 ##STR01093## 576 ##STR01094## P-1513 ##STR01095## 381.0 383.0
P-1514 ##STR01096## 367.1 369.1 P-1551 ##STR01097## 381.1 383.1
P-1541 ##STR01098## 516.2 P-1542 ##STR01099## 512.2 P-1581
##STR01100## 462.2 P-1582 ##STR01101## 462.2 P-1583 ##STR01102##
466.1 P-1598 ##STR01103## 510.1 P-1599 ##STR01104## 514.0 P-2003
##STR01105## 423.3 P-2020 ##STR01106## 363.1 P-1698 ##STR01107##
393.2
Example 24
Synthesis of
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyrid-
ine P-1455
[1446] Compound P-1455 was synthesized in four steps from
2,4-difluorophenol 35 as shown in Scheme 43a.
##STR01108##
[1447] Steps 1-3 are identical to Steps 1-3 of Scheme 43.
Step 4--Preparation of
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyrid-
ine (P-1455)
[1448] To
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridin-3-yl)-methanol (527, 580.0 mg, 1.3 mmol) in acetonitrile
(29.0 mL) were added trifluoroacetic acid (1.9 mL, 0.025 mol) and
triethylsilane (3.9 mL, 0.024 mol). The reaction was stirred at
80.degree. C. for 1 hour. The reaction was poured into water,
basified with 1 M potassium carbonate to pH=4, and extracted with
ethyl acetate. The organic layer was dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 50% ethyl acetate in
hexane to give as a yellow solid (P-1455, 530 mg). MS(ESI)
[M+H.sup.+].sup.+=428.3.
5-Bromo-3-[2,6-difluoro-3-(2-methoxy-ethoxy)-benzyl]-1H-pyrrolo[2,3-b]pyri-
dine P-1454
##STR01109##
[1449] was prepared following the protocol of Scheme 43a by
substituting benzyl bromide with 1-Bromo-2-methoxy-ethane in Step 1
and 5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine with
5-Bromo-1H-pyrrolo[2,3-b]pyridine (67) in Step 3. MS (ESI)
[M+H.sup.+].sup.+=410.1, 412.1.
[1450] Additional compounds were prepared following steps 3 and 4
of Scheme 43a, replacing 3-benzyloxy-2,6-difluoro-benzaldehyde 526
with an appropriate aldehyde and/or
pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with an appropriate
azaindole (see Example 9 or Example 16) in Step 3. The following
compounds were made following this procedure: [1451]
5-Bromo-3-(3,5-dimethoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1464), [1452]
3-(3,5-Bis-difluoromethoxy-benzyl)-5-bromo-1H-pyrrolo[2,3-b]pyridi-
ne (P-1538), [1453]
3-(2,6-Dichloro-benzyl)-1H-pyrrolo[2,3-b]pyridine (P-1483), [1454]
3-(2,6-Difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine (P-1482), [1455]
3-(2,6-Dimethyl-benzyl)-1H-pyrrolo[2,3-b]pyridine (P-0333), [1456]
3-(2,6-Dichloro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-1478), [1457]
3-(2,6-Difluoro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-1477), [1458]
3-(2,6-Dimethyl-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-1481), [1459]
N-[2,4-Difluoro-3-(5-methoxy-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-
-3-trifluoromethyl-benzenesulfonamide (P-1590), [1460]
N-[2,4-Difluoro-3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]--
3-trifluoromethyl-benzenesulfonamide (P-1600), [1461]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-methanesu-
lfonamide (P-1603), and [1462]
N-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-ph-
enyl]-methanesulfonamide (P-1605). [1463] The following table
indicates the aldehyde (column 2) and the azaindole (column 3) used
to afford the target compound (column 4). Column 1 indicates the
compound number and column 5 the observed mass.
TABLE-US-00011 [1463] Aldehyde Azaindole P-1464 ##STR01110##
##STR01111## P-1538 ##STR01112## ##STR01113## P-1483 ##STR01114##
##STR01115## P-1482 ##STR01116## ##STR01117## P-0333 ##STR01118##
##STR01119## P-1478 ##STR01120## ##STR01121## P-1477 ##STR01122##
##STR01123## P-1481 ##STR01124## ##STR01125## P-1590 ##STR01126##
##STR01127## P-1600 ##STR01128## ##STR01129## P-1603 ##STR01130##
##STR01131## P-1605 ##STR01132## ##STR01133## MS (ESI) [M +
H.sup.+].sup.+ ob- Product served P-1464 ##STR01134## 347.1 349.1
P-1538 ##STR01135## 419.1 421.1 P-1483 ##STR01136## P-1482
##STR01137## P-0333 ##STR01138## P-1478 ##STR01139## P-1477
##STR01140## P-1481 ##STR01141## P-1590 ##STR01142## P-1600
##STR01143## 1603 ##STR01144## 1605 ##STR01145##
Example 25
Synthesis of
3-[2,6-trifluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]py-
ridine-5-carboxylic acid ethylamide P-1630
[1464] Compound P-1630 was synthesized in six steps from
5-bromo-1-triisopropylsilyl-7-azaindole 68 as shown in Scheme
45.
##STR01146## ##STR01147##
Step 1--Preparation of
1-Triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid
methyl ester (531)
[1465] To 5-bromo-1-triisopropylsilyl-7-azaindole (68, 1.50 g, 4.2
mmol, prepared as described in Example 6) in tetrahydrofuran (20.0
mL) under an atmosphere of nitrogen, cooled with dry ice/acetone,
was slowly added n-Butyllithium (10.0 M in hexane, 0.467 mL). After
60 minutes, methyl chloroformate (0.394 mL, 5.1 mmol) was added to
the reaction. After another hour, the reaction was poured into
water and extracted with ethyl acetate. The organic layer was dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give the crude compound as a light yellow solid
that was used directly in the next step.
Step 2--Preparation of 1H-Pyrrolo[2,3-b]pyridine-5-carboxylic acid
methyl ester (532)
[1466] To
1-Triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid
methyl ester (531, 0.950 g, 2.9 mmol) in tetrahydrofuran (20.0 mL)
was added tetrabutyl-ammonium fluoride, trihydrate (1.20 g, 3.8
mmol). The reaction was stirred at room temperature for 10 minutes.
The reaction was concentrated and purified with silica gel column
chromatography eluting with 4% methanol in methylene chloride to
give the compound as a white solid (532, 300 mg, 60%). MS(ESI)
[M+H.sup.+].sup.+=177.2.
Step 3--Preparation of
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-phenyl]-hydroxy-methyl-1H-pyr-
rolo[2,3-b]pyridine-5-carboxylic acid methyl ester (P-1545)
[1467] To 1H-Pyrrolo[2,3-b]pyridine-5-carboxylic acid methyl ester
(532, 155.0 mg, 0.88 mmol) in methanol (15.0 mL) were added
propane-1-sulfonic acid (2,4-difluoro-3-formyl-phenyl)-amide (73,
260.0 mg, 0.99 mmol, prepared as described in Example 7) and
potassium hydroxide (859 mg, 15.3 mmol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature overnight.
The reaction was poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
to give the compound as white solid (P-1545, 110 mg, 28%). MS(ESI)
[M+H.sup.+].sup.+=440.2,
Step 4--Preparation of
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]pyr-
idine-5-carboxylic acid methyl ester (P-1552)
[1468] To
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-phenyl]-hydroxy-meth-
yl-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid methyl ester
(P-1545, 300.0 mg, 0.23 mmol) in tetrahydrofuran (10 mL) was added
Dess-Martin periodinane (107 mg, 2.5 mmol). The reaction was
stirred at room temperature for 10 minutes. The reaction mixture
was concentrated with silica gel and then purified with silica gel
column chromatography eluting with 30% ethyl acetate in hexane to
give the compound as white solid (P-1552, 80 mg, 80%). MS(ESI)
[M+H.sup.+].sup.+=438.2.
Step 5--Preparation of
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]pyr-
idine-5-carboxylic acid (P-1559)
[1469] To
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[-
2,3-b]pyridine-5-carboxylic acid methyl ester (P-1552, 80.0 mg,
0.18 mmol) in tetrahydrofuran (10.0 mL) were added water (3.0 mL)
and lithium hydroxide (82 mg, 3.4 mmol). The reaction was stirred
at room temperature overnight. The reaction was poured into water,
acidified with 1 N HCl to pH around 1, and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate,
filtered, concentrated, and washed with ethyl acetate to give an
off-white solid (P-1559, 60 mg, 77%) MS(ESI)
[M+H.sup.+].sup.+=424.2.
Step 6: Preparation of
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[2,3-b]pyr-
idine-5-carboxylic acid ethylamide (P-1630)
[1470] To
3-[2,6-difluoro-3-(propane-1-sulfonylamino)-benzoyl]-1H-pyrrolo[-
2,3-b]pyridine-5-carboxylic acid (P-1559, 38.0 mg, 0.090 mmol) in
tetrahydrofuran (2.3 mL) was added a solution of ethylamine (2.0 M
in tetrahydrofuran, 0.20 mL), bromo-tris-pyrrolidino-phosphonium
hexafluorophosphate (80.0 mg, 0.17 mmol) and triethylamine (0.30
mL, 2.2 mmol) under an atmosphere of nitrogen. The reaction mixture
was stirred overnight at room temperature. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 40% ethyl acetate in hexane to give the compound as a
white solid (P-1630, 13.2 mg, 33%). MS(ESI)
[M-H.sup.+].sup.-=449.0.
Example 26
Synthesis of
1-butyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-
-urea P-1445
[1471] Compound P-1445 was synthesized in six steps from
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 as shown in Scheme
49.
##STR01148## ##STR01149##
Step 1--Preparation of
(3-nitro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanol
(P-1399)
[1472] To 3-nitrobenzaldehyde (534, 1.08 g, 7.17 mmol) in methanol
(34 mL) was added 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89,
1.08 g, 5.52 mmol, prepared as described in Example 17) and
potassium hydroxide (1.55 g, 27.6 mmol). The reaction was stirred
at room temperature for four hours. The reaction was poured into
water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
concentrated. The mixture was purified by silica gel column
chromatography eluting with 4% methanol in dichloromethane to
provide two different compounds, a white solid (P-1399, R.dbd.H,
1.20 g, 63%) MS(ESI) [M+H.sup.+].sup.+=347.2, and a light yellow
solid (535, R=Me, 0.434 g, 22%).
Step 2--Preparation of
(3-nitro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
(536)
[1473] To
(3-nitro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanol (P-1399, R.dbd.H, 500 mg, 1.44 mmol) in dimethylformamide
(26 mL) was added Dess-Martin periodane (674 mg, 1.59 mmol). The
reaction was stirred for one hour and the reaction was poured into
water. All solids were filtered and purified by silica gel column
chromatography eluting with 3% methanol in dichloromethane to give
the compound (536, 295 mg, 59%).
MS(ESI)[M+H.sup.+].sup.+=345.2.
Step 3--Preparation of
(3-nitro-phenyl)-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl)-methanone (537)
[1474] To
(3-nitro-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone (536, 291 mg, 0.85 mmol) in tetrahydrofuran (7 mL) was
added 1.5 M of lithium diisopropylamide in cyclohexane (676 .mu.l,
1.59 mmol) at -78.degree. C. under an atmosphere of nitrogen. After
30 minutes, p-toluenesulfonyl chloride (209 mg, 1.10 mmol) was
added in tetrahydrofuran and the reaction was stirred for three
hours. The reaction was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. All solids were filtered
and purified by silica gel column chromatography eluting with 60%
ethyl acetate in hexane to give the compound (537, 182 mg, 43%).
MS(ESI)[M+H.sup.+].sup.+=499.2.
Step 4--Preparation of
(3-Amino-phenyl)-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]-
pyridin-3-yl]-methanone (538)
[1475] To
(3-nitro-phenyl)-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrro-
lo[2,3-b]pyridin-3yl]-methanone (537, 180 mg, 0.361 mmol) in
methanol (4 mL) was added 10% palladium on carbon (20 mg) and few
drops of concentrated aqueous hydrochloric acid. The resulting
mixture was stirred under an atmosphere of hydrogen overnight and
the catalyst was filtered out through a bed of celite. The filtrate
was concentrated and purified by silica gel column chromatography
eluting with 90% ethyl acetate in hexane to give the compound (538,
58 mg, 34%). MS(ESI)[M+H.sup.+].sup.+=469.3.
Step 5--Preparation of
1-Butyl-3-3-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyrid-
ine-3-carbonyl]-phenyl-urea (539)
[1476] To
(3-Amino-phenyl)-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrro-
lo[2,3-b]pyridin-3-yl]-methanone (538, 53 mg, 0.11 mmol) in
tetrahydrofuran (1.6 mL) was added 1-isocyanatobutane (12 mg, 0.12
mmol). The reaction was heated at 90.degree. C. overnight and it
was concentrated and purified by silica gel column chromatography
eluting with 2% methanol in dichloromethane to give the compound
(539, 39 mg, 61%). MS(ESI)[M+H.sup.+].sup.+=568.4.
Step 6--Preparation of
1-Butyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl-
]-urea (P-1445)
[1477] To
1-Butyl-3-3-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,-
3-b]pyridine-3-carbonyl]-phenyl-urea (539, 33 mg, 0.058 mmol) in
tetrahydrofuran (2 mL) was added 1.0 M tetra-n-butylammonium
fluoride in tetrahydrofuran (192 .mu.l) under an atmosphere of
nitrogen and the reaction was stirred for three hours. The reaction
was poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and concentrated. All solids were filtered and purified by silica
gel column chromatography eluting with 4% methanol in
dichloromethane to give the compound (P-1445, 8 mg, 30%).
MS(ESI)[M+H.sup.+].sup.+=414.3.
Example 27
Synthesis of
1-butyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-
-urea P-1447
[1478] Compound P-1447 was synthesized in five steps from
3-[(3-nitro-phenyl)-methoxy-methyl]-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyrid-
ine 535 as shown in Scheme 50.
##STR01150## ##STR01151##
Step 1--Preparation of
3-(3-nitro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-1402)
[1479] To
3-[(3-nitro-phenyl)-methoxy-methyl]-5-pyridin-3-yl-1H-pyrrolo[2,-
3-b]pyridine (535, 431 mg, 1.20 mmol, per Example 26, Scheme 49
Step 1) in acetonitrile (130 mL), were added trifluoroacetic acid
(18 mL, 230 mmol) and triethylsilane (36 mL, 230 mmol). The
reaction was refluxed for three hours. The reaction mixture was
poured into sodium bicarbonate solution and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 80% ethyl acetate in hexane to give the compound
(P-1402, 323 mg, 82%). MS(ESI)[M+H.sup.+].sup.+=331.2.
Step 2--Preparation of
3-(3-nitro-benzyl)-5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b-
]pyridine (552)
[1480] To
3-(3-nitro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-1402, 141 mg, 0.43 mmol) in N,N-dimethylformamide (3 mL) was
added sodium hydride (60% dispersion in mineral oil, 21 mg, 0.512
mmol) under an atmosphere of nitrogen. After thirty minutes,
p-toluenesulfonyl chloride (114 mg, 0.60 mmol) in
N,N-dimethyl-formamide was added and the reaction was stirred for
three hours. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. All solids were filtered
and purified by silica gel column chromatography eluting with 40%
ethyl acetate in hexane to give the compound (552, 120 mg, 58%).
MS(ESI)[M+H.sup.+].sup.+=485.25.
Step 3--Preparation of
3-[5-Pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridin-3-ylmet-
hyl]-phenylamine (553)
[1481] To
3-(3-nitro-benzyl)-5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrr-
olo[2,3-b]pyridine (552, 230 mg, 0.14 mmol) in methanol (5 mL) was
added 10% palladium on carbon (10 mg) and few drops of concentrated
aqueous hydrochloric acid. The resulting mixture was stirred under
an atmosphere of hydrogen overnight, and the catalyst was filtered
out through a bed of celite. The filtrate was concentrated and
purified by silica gel column chromatography eluting with 90% ethyl
acetate in hexane to give the compound (553, 88 mg, 41%).
MS(ESI)[M+H.sup.+].sup.+=455.3.
Step 4--Preparation of
1-butyl-3-3-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyrid-
in-3-ylmethyl]-phenyl-urea (554)
[1482] To
3-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,3-b]pyridi-
n-3-ylmethyl]-phenylamine (553, 14 mg, 0.031 mmol) in
tetrahydrofuran (0.5 mL) was added 1-isocyanatobutane (3.4 mg, 0.03
mmol). The reaction was heated at 90.degree. C. overnight and was
concentrated and purified by silica gel column chromatography
eluting with 2% methanol in dichloromethane to give the compound
(554, 7.2 mg, 42%). MS(ESI)[M+H.sup.+].sup.+=554.4.
Step 5--Preparation of
1-Butyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-
-urea (P-1447)
[1483] To
1-butyl-3-3-[5-pyridin-3-yl-1-(toluene-4-sulfonyl)-1H-pyrrolo[2,-
3-b]pyridin-3-ylmethyl]-phenyl-urea (554, 11 mg, 0.02 mmol) in
tetrahydrofuran (0.7 mL) was added 1.0 M tetra-n-butylammonium
fluoride in tetrahydrofuran (66 .mu.l) under an atmosphere of
nitrogen and the reaction was stirred for three hours. The reaction
was poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and concentrated. All solids were filtered and purified by silica
gel column chromatography eluting with 4% methanol in
dichloromethane to give the compound (P-1447, 2.5 mg, 31%).
MS(ESI)[M+H.sup.+].sup.+=400.3.
[1484]
1-Cyclopentyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmet-
hyl)-phenyl]-urea P-1446
##STR01152##
was prepared following the protocol of Scheme 50, substituting
1-isocyanatobutane with isocyanato-cyclopentane in Step 4. MS(ESI)
[M+H.sup.+].sup.+=412.4.
Example 28
Synthesis of
3-[3-chloro-4-(4-chloro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
P-1449.
[1485] Compound P-1449 was synthesized in three steps from
3-chloro-4-hydroxy-benzaldehyde 556 as shown in Scheme 51.
##STR01153##
Step 1--Preparation of 3-chloro-4-(4-chloro-benzyloxy)-benzaldehyde
(558)
[1486] To acetonitrile (15.0 mL) were added
3-chloro-4-hydroxy-benzaldehyde (556, 0.6 g, 4 mmol),
4-chlorobenzyl bromide (557, 1.2 g, 6 mmol), and potassium
carbonate (0.9 g, 7 mmol). The reaction was heated to 150.degree.
C. for 10 minutes in a CEM Discover microwave instrument. The
reaction was poured into water, extracted with ethyl acetate, and
washed with brine. The organic layer was dried over anhydrous
sodium sulfate, filtered and concentrated. The desired compound was
isolated by silica gel column chromatography (ethyl
acetate:hexanes) (558, 0.85 g, 76%).
Step 2--Preparation of
[3-chloro-4-(4-chlorobenzyloxy)-phenyl]-methoxy-methyl-1H-pyrrolo[2,3-b]p-
yridine (559)
[1487] 1H-Pyrrolo[2,3-b]pyridine (94, 0.3 g, 2.8 mmol) was mixed
with 3-chloro-4-(4-chloro-benzyloxy)-benzaldehyde (558, 0.8 g, 3
mmol), potassium hydroxide (0.9 g, 17 mmol) and methanol (90.0 mL).
The reaction was heated to 50.degree. C. under an atmosphere of
nitrogen for six days. After neutralization with 6N hydrochloric
acid the reaction was poured into water, extracted with ethyl
acetate, and washed with brine. The organic layer was dried over
anhydrous sodium sulfate, filtered and concentrated. The desired
compound was isolated by silica gel column chromatography (ethyl
acetate: hexanes) to give a yellow solid (559, 0.6 g, 41%). MS(EST)
[M+H.sup.+].sup.+=413.2, 415.2 [M-H.sup.+].sup.+=411.1, 413.1.
Step 3--Preparation of
3-[3-chloro-4-(4-chloro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1449)
[1488]
3-[3-Chloro-4-(4-chloro-benzyloxy)-phenyl]-methoxy-methyl-1H-pyrrol-
o[2,3-b]pyridine (559, 0.2 g, 0.6 mmol) was mixed with
trifluoroacetic acid (0.226 mL, 3 mmol), triethylsilane (0.4 mL, 3
mmol) and acetonitrile (5 mL). The reaction was heated at
50.degree. C. and stirred for two days. The reaction was
concentrated. The residue was diluted with ethyl acetate and
neutralized with 2M aqueous sodium hydroxide. The reaction was
poured into water, extracted with ethyl acetate, and washed with
brine. The organic layer was dried over anhydrous sodium sulfate,
filtered and concentrated. The desired compound was isolated by
silica gel column chromatography (ethyl acetate:hexanes) to give a
yellow solid (P-1449, 0.0744 g, 33%). MS(ESI)
[M+H.sup.+].sup.+=383.2, 385.2.
[1489] Additional compounds were prepared following the protocol of
Scheme 51, replacing 3-chloro-4-hydroxy-benzaldehyde 556 with an
appropriate aldehyde and optionally replacing 4-chlorobenzyl
bromide 557 with an appropriate benzyl halide in Step 1, and
optionally replacing 1H-pyrrolo[2,3-b]pyridine 94 with an
appropriate azaindole in Step 2. The following compounds were made
following this procedure: [1490]
3-[4-(4-chloro-benzyloxy)-2-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1450), [1491]
3-[4-(4-Chloro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1462), [1492]
3-[4-(4-Chloro-benzyloxy)-3-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,-
3-b]pyridine (P-1466), [1493]
3-[4-(4-Chloro-benzyloxy)-3-ethoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1470), [1494]
3-[2-Chloro-4-(4-chloro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1471), [1495]
3-[4-(4-Chloro-benzyloxy)-3-trifluoromethoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-1487), [1496]
3-[4-(4-Chloro-benzyloxy)-3-methoxy-benzyl]-5-methoxy-1H-pyrrolo[2,3-b]py-
ridine (P-1531), [1497]
5-Chloro-3-[4-(4-chloro-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine(P-1532), [1498]
3-[4-(4-Chloro-2-fluoro-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-1544), [1499]
3-[4-(2,4-Dichloro-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1568), [1500]
3-[3-Methoxy-4-(4-methoxy-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1569), [1501]
3-[3-Methoxy-4-(2,4,6-trifluoro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyrid-
ine (P-1578), [1502]
3-[4-(2,6-Dichloro-benzyloxy)-3-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1579), and
3-[3-Chloro-4-(4-chloro-benzyloxy)-5-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-1616). The following table indicates the aldehyde (Column
2), the benzyl halide (Column 3), and the azaindole (Column 4) used
to afford the target compound (Column 5). Column 1 indicates the
compound number and column 6 the observed mass.
TABLE-US-00012 [1502] Benzyl Aldehyde halide Azaindole P-1450
##STR01154## ##STR01155## ##STR01156## P-1462 ##STR01157##
##STR01158## ##STR01159## P-1466 ##STR01160## ##STR01161##
##STR01162## P-1470 ##STR01163## ##STR01164## ##STR01165## P-1471
##STR01166## ##STR01167## ##STR01168## P-1487 ##STR01169##
##STR01170## ##STR01171## P-1531 ##STR01172## ##STR01173##
##STR01174## P-1532 ##STR01175## ##STR01176## ##STR01177## P-1544
##STR01178## ##STR01179## ##STR01180## P-1568 ##STR01181##
##STR01182## ##STR01183## P-1569 ##STR01184## ##STR01185##
##STR01186## P-1578 ##STR01187## ##STR01188## ##STR01189## P-1579
##STR01190## ##STR01191## ##STR01192## P-1616 ##STR01193##
##STR01194## ##STR01195## MS(ESI) [M+ H.sup.+].sup.+ Compound
observed P-1450 ##STR01196## 379.2 381.2 P-1462 ##STR01197## 349.1
351.2 P-1466 ##STR01198## 397.2 399.2 P-1470 ##STR01199## 393.2
395.2 P-1471 ##STR01200## 383.1 385.1 P-1487 ##STR01201## 433.2
435.2 P-1531 ##STR01202## 409.2 P-1532 ##STR01203## 413.1 P-1544
##STR01204## 397.2 P-1568 ##STR01205## 413.1 415.1 416.2 P-1569
##STR01206## 375.2 P-1578 ##STR01207## 399.2 397.1 ([M -
H.sup.+].sup.-) 1579 ##STR01208## 413.2 415.2 416.2 ([M -
H.sup.+].sup.-) P-1616 ##STR01209## 413.1
Example 29
Synthesis of
3-(4-benzyloxy-3-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
P-1613
[1503] Compound P-1613 was synthesized in two steps from
4-benzyloxy-3-methoxy-benzaldehyde 564 as shown in Scheme 53.
##STR01210##
Step 1--Preparation of
3-[(4-benzyloxy-3-methoxy-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3-b]pyridi-
ne (565)
[1504] Methanol (125 mL) and potassium hydroxide (4.4 g, 79 mmol)
were mixed with 1H-pyrrolo[2,3-b]pyridine (94, 3.1 g, 26.6 mmol)
and 4-benzyloxy-3-methoxy-benzaldehyde (564, 12.9 g, 53.2 mmol).
The reaction was stirred at room temperature for 2 days. The
resulting white solid was filtered and washed with water. Crude
material was carried forward without further purification.
Step 2--Preparation of
3-(4-benzyloxy-3-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1613)
[1505]
3-[(4-Benzyloxy-3-methoxy-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3-b]-
pyridine (565, 0.9 g, 2.4 mmol) and acetonitrile (50 mL) were mixed
with trifluoroacetic acid (0.360 mL, 4.7 mmol) and triethylsilane
(0.746 mL, 4.7 mmol). The reaction was heated at 80.degree. C. and
stirred overnight. The reaction was concentrated. The mixture was
extracted with ethyl acetate and saturated sodium bicarbonate. The
organic layer was dried over anhydrous sodium sulfate, filtered and
concentrated. The desired compound was isolated by silica gel
column chromatography to give the compound (P-1613, 0.454 g 54.8%).
MS(ESI) [M+H.sup.+].sup.+=345.3.
Example 30
Synthesis of
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-butyl-urea
P-1596
[1506] Compound P-1596 was synthesized in one step from
5-bromo-1H-pyrrolo[2,3-b]pyridine 67 as shown in Scheme 55.
##STR01211##
Step 1--Preparation of
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-butyl-urea
(P-1596)
[1507] To aluminum trichloride (3.67 g, 0.0275 mol) in
dichloromethane (100 mL, 2 mol) under an atmosphere of nitrogen was
added 5-bromo-7-azaindole (67, 1.08 g, 0.00548 mol) at room
temperature. After one hour, 3-isocyanato-benzoyl chloride (584,
5.00 g, 0.0275 mol) was added under an atmosphere of nitrogen at
room temperature. The resulting mixture was stirred over night at
room temperature. 1-Butanamine (585, 54 mL, 0.54 mol) was added
carefully. All solvents were removed. The residue was purified by
silica gel column chromatography to give the compound (P-1596, 172
mg, 8%). MS(ESI) [M-H+]-=413.1, 415.0.
[1508] Additional compounds were prepared following the protocol of
Scheme 55, replacing 1-butanamine 585 with an appropriate amine and
optionally replacing 5-bromo-7-azaindole 67 with
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 (prepared as described
in Example 17). The following compounds were made following this
procedure: [1509]
1-Benzyl-3-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-urea (P-1553), [1510]
1-Benzyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-urea (P-1554), [1511]
1-(2-Methoxy-ethyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1566), and [1512]
1-Phenyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-urea (P-1591). The following table indicates the amine (column
2) and the azaindole (column 3) used to afford the target compound
(column 4). The compound number is provided in column 1 and the
observed mass in column 5.
TABLE-US-00013 [1512] MS (ESI) [M + H.sup.+].sup.+ ob- Amine
Azaindole Compound served P- 1553 ##STR01212## ##STR01213##
##STR01214## 447.0 449.1 P- 1554 ##STR01215## ##STR01216##
##STR01217## 448.3 P- 1566 ##STR01218## ##STR01219## ##STR01220##
416.3 P- 1591 ##STR01221## ##STR01222## ##STR01223## 434.3
Example 31
Synthesis of
1-Butyl-3-3-[5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-3-car-
bonyl]-phenyl-urea P-1880
[1513] Compound P-1880 was synthesized in one step from
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-butyl-urea
P-1596 as shown in Scheme 56.
##STR01224##
Step 1--Preparation of
1-Butyl-3-3-[5-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridine-3-car-
bonyl]-phenyl-urea (P-1880)
[1514] In a microwave tube,
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-butyl-urea
(P-1596, 0.077 g, 0.00018 mol, prepared as described in Example 30,
Scheme 55),
1-Methyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole
(0.0964 g, 0.000464 mol), and
Tetrakis(triphenylphosphine)palladium(0) (0.011 g, 0.0000093 mol)
were mixed in 1.00 M of potassium carbonate in water (1.2 mL),
acetonitrile (2.0 mL, 0.037 mol), and tetrahydrofuran (1.0 mL,
0.012 mol). The resulting mixture was heated at 100.degree. C. in
the microwave for 20 minutes, then at 120.degree. C. for 10
minutes. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography to
give the compound (P-1880, 52 mg, 67%).
MS(ESI)[M+H.sup.+].sup.+=417.4.
Example 32
Synthesis of
1-Butyl-3-[2-chloro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-urea P-1828
[1515] Compound P-1828 was synthesized in two steps from
3-amino-2-chlorobenzoic acid 586 as shown in Scheme 57.
##STR01225##
Step 1--Preparation of -(3-Butyl-ureido)-2-chloro-benzoic acid
(587)
[1516] To N,N-diisopropylamine (1.72 mL, 0.0122 mol) in
tetrahydrofuran (12 mL, 0.14 mol), was added 1.6 M n-butyllithium
in hexane (7.6 mL) at -78.degree. C. under an atmosphere of
nitrogen. After 30 minutes, 3-amino-2-chlorobenzoic acid (586, 1.00
g, 0.00583 mol) was added. After another 30 minutes,
1-isocyanatobutane (2.60 mL, 0.0233 mol) was added at -78.degree.
C. under an atmosphere of nitrogen and allowed to stir for two
hours. The reaction mixture was warmed up to room temperature and
stirred at room temperature for 30 minutes. The reaction was
quenched with 1M HCl (aqueous) solution and extracted with ethyl
acetate twice. The combined organic layers were washed with brine,
dried over anhydrous magnesium sulfate and filtered. The filtrate
was concentrated and purified by silica gel column chromatography
eluting with dichloromethane:methanol:acetic acid 40:2:1 to give
the compound as an off-white solid (587, 147 mg, 9%).
Step 2--Preparation of
1-Butyl-3-[2-chloro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-urea (P-1828)
[1517] To 3-(3-Butyl-ureido)-2-chloro-benzoic acid (587, 103 mg,
0.000380 mol) was added dichloromethane (10 mL, 0.2 mol) followed
by thionyl chloride (110 .mu.L, 0.0015 mol) and 1 drop of
dimethylformamide to give a suspension. The reaction was stirred at
room temperature for 2 hours. Solid material was still present in
the reaction mixture, so tetrahydrofuran (0.5 mL, 0.006 mol) was
added and continued stirring at room temperature. The reaction
became a clear solution after 2 hours, then stirred for another
hour. All volatiles were removed under vacuum and the residue
stripped from toluene, twice. The solid was then dried under high
vacuum for 60 minutes and dissolved in dichloromethane (5 mL). This
was added to 5-(pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine (89, 0.074
g, 0.00038 mol, prepared as described in Example 17) which had been
treated with aluminum trichloride (0.25 g, 0.0019 mol) in
dichloromethane (10 mL) for 1 hour. The reaction was stirred at
room temperature overnight, then quenched with methanol (5 mL). The
resulting solution was extracted with ethyl acetate and water with
added saturated sodium bicarbonate to adjust pH.about.8. The
organic layer was washed with sodium bicarbonate and brine and
dried over magnesium sulfate and filtered. The organic layer was
concentrated and purified by silica gel column chromatography
eluting with 2% methanol in dichloromethane followed by 5% methanol
in dichloromethane to give the compound as a white solid (P-1828,
45 mg, 26%).
[1518] MS(ESI) [M+H.sup.+].sup.+=448.3.
[1519] Additional compounds were prepared following the protocol of
Scheme 57, replacing 3-amino-2-chlorobenzoic acid 586 with an
appropriate carboxylic acid and optionally replacing
1-isocyanatobutane with an appropriate isocyanate in Step 1 and
optionally replacing 5-(pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine 89
with an appropriate substituted 7-azaindole (see Example 17) in
Step 2. The following compounds were made following this procedure:
[1520]
1-Butyl-3-[2-methyl-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-urea (P-1742), [1521]
3-Butyl-1-methyl-1-[2-methyl-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine--
3-carbonyl)-phenyl]-urea (P-1855), [1522]
[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-urea
(P-1570), [1523]
[4-Fluoro-3-(5-phenyl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-urea
(P-1589), [1524]
3-{3-[5-(3-Butyl-ureido)-2-fluoro-benzoyl]-1H-pyrrolo[2,3-b]pyridin-5-yl}-
-benzamide (P-1621), [1525] 1-Butyl-3-{4-fluoro-3-[5-(3-methane
sulfonyl-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-phenyl}-urea
(P-1627), and [1526]
1-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-3-bu-
tyl-urea (P-1637). The following table indicates the carboxylic
acid (column 2), the isocyanate (column 3), and the azaindole
(column 4) used to afford the target compound (column 5). Column 1
indicates the compound number and column 6 the observed mass.
TABLE-US-00014 [1526] Iso- Acid cyanate Azaindole P-1742
##STR01226## ##STR01227## ##STR01228## P-1855 ##STR01229##
##STR01230## ##STR01231## P-1570 ##STR01232## ##STR01233##
##STR01234## P-1589 ##STR01235## ##STR01236## ##STR01237## P-1621
##STR01238## ##STR01239## ##STR01240## P-1627 ##STR01241##
##STR01242## ##STR01243## P-1637 ##STR01244## ##STR01245##
##STR01246## MS (ESI) [M + H].sup.+ ob- Compound served P- 1742
##STR01247## 428.3 P- 1855 ##STR01248## 442.3 P- 1570 ##STR01249##
377.1 379.1 P- 1589 ##STR01250## 375.2 P- 1621 ##STR01251## 474.3
P- 1627 ##STR01252## 509.2 P- 1637 ##STR01253## 433.1 435.1
Example 33
Synthesis of
1-Butyl-3-[4-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-urea (P-1534)
[1527] Compound P-1534 was synthesized in two steps from
5-(3-butylureido)-2-fluorobenzoic acid 588 (prepared from
3-fluoro-5-aminobenzoic acid and 1-isocyanatobutane following the
protocol described in Step 1 of Scheme 57, Example 32) and
5-bromo-7-azaindole 67 as shown in Scheme 58.
##STR01254##
Step 1--Preparation of
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-3-bu-
tyl-urea (P-1637)
[1528] To aluminum trichloride (0.524 g, 0.00393 mol) and
dichloromethane (20 mL, 0.3 mol) under an atmosphere of nitrogen
was added 5-bromo-7-azaindole (67, 0.155 g, 0.000787 mol) in
dichloromethane. To 5-(3-butylureido)-2-fluorobenzoic acid (588,
0.200 g, 0.000787 mol) was added 4 mL of dichloromethane (4 mL)
followed by thionyl chloride (69 .mu.L, 0.00094 mol) and a drop of
N,N-dimethylformamide. After 1 hour, the reaction remained a
suspension so additional thionyl chloride was added along with
tetrahydrofuran. The reaction remained a suspension, so was placed
in a 50.degree. C. oil bath. After another hour, the reaction was
still a suspension, and so was left to react at 50.degree. C.
overnight. The reaction had become a clear solution. All volatiles
were removed under vacuum, then the residue dissolved in
dichloromethane and added to the 5-bromo-7-azaindole and aluminum
trichloride suspension. The reaction was allowed to stir at room
temperature for 4.5 hours, followed by the addition of water and
extraction with ethyl acetate. The organic layer was dried over
magnesium sulfate and filtered. The filtrate was concentrated and
purified by silica gel column chromatography eluting with a
gradient of methanol (0 to 10%) in dichloromethane to give the
compound (P-1637, 14 mg, 4%).
Step 2--Preparation of
1-Butyl-3-[4-fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbony-
l)-phenyl]-urea (P-1534)
[1529] To
1-[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phe-
nyl]-3-butyl-urea (P-1637, 14.0 mg, 0.0000323 mol),
3-pyridylboronic acid (5.96 mg, 0.0000485 mol), and
Tetrakis(triphenylphosphine)palladium(0) (0.820 mg, 7.09E-7 mol)
were mixed in 1.00 M potassium carbonate in water (1.00 mL) and
acetonitrile (2.00 mL, 0.0383 mol). The resulting mixture was
heated at 120.degree. C. in the microwave for 40 minutes. The
reaction was extracted with ethyl acetate and water twice, and the
combined organic layers were washed with 1M sodium bicarbonate
followed by brine and the organic layer was dried over magnesium
sulfate and filtered. The organic layer was concentrated and
purified by reverse phase HPLC (acetonitrile and water with 0.1%
formic acid) to give the compound as a white solid (P-1637, 8.5 mg,
61%). MS(ESI) [M+H.sup.+].sup.+=432.3.
[1530]
1-Butyl-3-{4-fluoro-3-[5-(3-trifluoromethoxy-phenyl)-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl]-phenyl}-urea P-1660
##STR01255##
was prepared following the protocol of Scheme 58, replacing
3-Pyridylboronic acid with 3-trifluoromethoxy-phenylboronic acid in
Step 2. MS(ESI) [M+H.sup.+].sup.+=515.2.
Example 34
Synthesis of Aldehyde Reagents for Coupling to 7-azaindoles
[1531] Aldehyde compounds for coupling to the 3-position of a
7-azaindole are shown in the following Schemes.
3-Methoxy-4-[4-(4-methyl-piperazin-1-ylmethyl)-benzyloxy]-benzaldehyde
591 was prepared in one Step as shown in Scheme 59.
##STR01256##
Step 1--Synthesis of
3-Methoxy-4-[4-(4-methyl-piperazin-1-ylmethyl)-benzyloxy]-benzaldehyde
(591)
[1532] To 4-Hydroxy-3-methoxybenzaldehyde (105, 2.1 g, 0.014 mol)
in N,N-dimethylformamide (40.0 mL) were added
1,4-bis(bromomethyl)-benzene (589, 4.00 g, 0.0152 mol) and
potassium carbonate (5.0 g, 0.036 mol) under an atmosphere of
nitrogen. After 12 hours 1-methyl-piperazine (590, 3.8 mL, 0.034
mol) was added to the reaction. After 2 hours, the reaction was
poured into water and extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 20% methanol in dichloromethane to give
the compound (589, 1.2 g, 25.0%). MS(ESI)
[M+H.sup.+].sup.+=355.3.
[1533] 2-Fluoro-4-hydroxy-5-methoxy-benzaldehyde 593 was
synthesized in one step from 2-fluoro-4,5-dimethoxy-benzaldehyde
592 as shown in Scheme 60.
##STR01257##
Step 1--Synthesis of 2-fluoro-4-hydroxy-5-methoxybenzaldehyde
(593)
[1534] To 2-fluoro-4,5-dimethoxy-benzaldehyde (592, 1.00 g, 5.43
mol) in dichloromethane (50.0 mL) was added aluminum trichloride
(4.34 g, 32.6 mmol) under an atmosphere of nitrogen. The reaction
was stirred at room temperature overnight. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and washed with ethyl acetate and hexane to give a
white solid (593, 0.70 g, 76.0%).
[1535] 2,5-Difluoro-4-hydroxy-benzaldehyde 597 was synthesized in
three steps from 2,5-difluorophenol 594 as shown in Scheme 61.
##STR01258##
Step 1--Synthesis of 4-bromo-2,5-difluoro-phenol (595)
[1536] To 2,5-difluorophenol (594, 5.50 g, 0.0423 mol) in
chloroform (110.0 mL), bromine (2.18 mL, 0.0423 mol) was added
slowly. After 3 hours, the reaction was poured into a solution of
sodium thiosulfate and extracted with ethyl acetate. The organic
layer was dried over sodium sulfate, concentrated and purified with
silica gel column chromatography eluting with 20% ethyl acetate in
hexane to give a colorless oil (595, 6.20 g, 70.2%).
Step 2--(4-Bromo-2,5-difluoro-phenoxy)-tert-butyl-dimethyl-silane
(596)
[1537] To 4-bromo-2,5-difluoro-phenol (595, 3.50 g, 0.0167 mol) in
N,N-dimethylformamide (50.0 mL) were added tert-butyldimethylsilyl
chloride (3.83 g, 0.0254 mol) and 1H-imidazole (6.00 g, 0.0529
mol). The reaction was stirred at room temperature overnight, then
poured into water and extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 20% ethyl acetate in hexane to give the
compound (596, 3.0 g, 55.4%).
Step 3--2,5-Difluoro-4-hydroxy-benzaldehyde (597)
[1538] To (4-bromo-2,5-difluoro-phenoxy)-tert-butyl-dimethyl-silane
(596, 3.00 g, 9.28 mmol) in tetrahydrofuran (37.5 mL), under an
atmosphere of nitrogen at -78.degree. C., n-butyllithium (3.90 mL,
2.50 M in hexane) was added slowly. After 30 minutes,
N,N-dimethylformamide (0.825 mL, 0.0106 mol) was added to the
reaction. One hour later, the reaction was allowed to come to room
temperature. The reaction was poured into water and 1 N HCl, then
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give the compound as an
off-white solid (597, 0.86 g, 59.0%).
[1539] 4-(4-Chloro-benzyloxy)-3-fluoro-benzaldehyde 599 was
synthesized in one step from 3-fluoro-4-hydroxy-benzaldehyde 598 as
shown in Scheme 62.
##STR01259##
Step 1--Synthesis of 4-(4-chlorobenzyloxy)-3-fluoro-benzaldehyde
(599)
[1540] To 3-fluoro-4-hydroxy-benzaldehyde (598, 0.800 g, 5.71 mmol)
in N,N-dimethylformamide (50.0 mL) was added sodium hydride (260.0
mg, 60% in mineral oil, 6.50 mmol). After 15 minutes,
4-chlorobenzyl bromide (557, 1.29 g, 6.28 mmol) was added to the
reaction mixture. The reaction was stirred at 80.degree. C. for 5
hours. The reaction was poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 30% ethyl acetate in hexane
to give the compound (599, 1.3 g, 86.0%).
[1541] Additional aldehydes were prepared using the protocol of
Scheme 62, replacing either 4-chlorobenzyl bromide 557 with a
suitable alkylating agent, and/or 3-fluoro-4-hydroxy-benzaldehyde
598 with a suitable aldehyde. The following table indicates the
alkylating agent (column 1) and the starting aldehyde (column 2)
used to afford the aldehyde (column 3) synthesized following this
protocol.
TABLE-US-00015 Alkylating agent Aldehyde Compound ##STR01260##
##STR01261## ##STR01262## ##STR01263## ##STR01264## ##STR01265##
##STR01266## ##STR01267## ##STR01268## ##STR01269## ##STR01270##
##STR01271## ##STR01272## ##STR01273## ##STR01274## ##STR01275##
##STR01276## ##STR01277## ##STR01278## ##STR01279## ##STR01280##
##STR01281## ##STR01282## ##STR01283## ##STR01284## ##STR01285##
##STR01286## ##STR01287## ##STR01288## ##STR01289## ##STR01290##
##STR01291## ##STR01292## ##STR01293## ##STR01294## ##STR01295##
##STR01296## ##STR01297## ##STR01298## ##STR01299## ##STR01300##
##STR01301## ##STR01302## ##STR01303## ##STR01304## ##STR01305##
##STR01306## ##STR01307## ##STR01308## ##STR01309##
##STR01310##
Example 35
Synthesis of
[4-(4-chloro-benzyloxy)-3-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone P-1897 and Related Compounds
[1542] Compound P-1897 was synthesized in two steps from
4-(4-chloro-benzyloxy)-3-fluoro-benzaldehyde 599 as shown in Scheme
63.
##STR01311##
Step 1--Synthesis of
[4-(4-chloro-benzyloxy)-3-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanol (P-1895)
[1543] To 1H-Pyrrolo[2,3-b]pyridine (94, 100.0 mg, 0.85 mmol) in
methanol (50.0 mL) were added
4-(4-chloro-benzyloxy)-3-fluoro-benzaldehyde (599, 250.0 mg, 0.94
mmol, prepared as described in Example 34) and potassium hydroxide
(1.00 g, 17.82 mmol) under an atmosphere of nitrogen. The reaction
was stirred at room temperature overnight. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give the compound
(P-1895, 55 mg, 17.0%). MS(ESI) [M+H.sup.+].sup.+=383.3.
Step 2--Synthesis of
[4-(4-chloro-benzyloxy)-3-fluoro-phenyl](1H-pyrrolo[2,3-b]pyridin-3-yl)-m-
ethanone (P-1897)
[1544] To
[4-(4-chloro-benzyloxy)-3-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanol (P-1895, 17.7 mg, 0.046 mmol) in tetrahydrofuran
(10.0 mL) was added Dess-Martin periodinane (23.5 mg, 0.056 mmol).
The reaction was stirred at room temperature for 15 minutes. The
reaction was concentrated, then purified with silica gel column
chromatography eluting with 50% ethyl acetate in hexane to give a
white solid (P-1897, 6.4 mg, 36.3%). MS(ESI)
[M+H.sup.+].sup.+=381.3.
[1545] Additional compounds were prepared using the protocol of
Scheme 63, replacing 4-4-(4-chloro-benzyloxy)-3-fluoro-benzaldehyde
599 with a suitable aldehyde (prepared as described in Example 34),
and optionally replacing 1H-Pyrrolo[2,3-b]pyridine 94 with an
appropriate substituted 7-azaindole (see Example 9 or Example 16)
in Step 1. The following compounds were made following this
procedure: [1546]
[4-(4-Chloro-benzyloxy)-2-fluoro-5-methoxy-phenyl]-(1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methanone (P-1845), [1547]
[4-(4-Chloro-3-trifluoromethyl-benzyloxy)-3-methoxy-phenyl]-(1H-pyrrolo[2-
,3-b]pyridin-3-yl)-methanone (P-1850), [1548]
[4-(1H-Benzoimidazol-2-ylmethoxy)-3-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methanone (P-1900), [1549]
(4-Benzyloxy-2,5-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methano-
ne (P-1903), [1550]
[4-(1H-Benzoimidazol-2-ylmethoxy)-2-fluoro-5-methoxy-phenyl]-(5-methoxy-1-
H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (P-1979), [1551]
[4-(1H-Benzoimidazol-2-ylmethoxy)-2-fluoro-5-methoxy-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanone (P-1982), [1552]
[4-(1H-Benzoimidazol-2-ylmethoxy)-2,5-difluoro-phenyl]-(1H-pyrrolo[2,3-b]-
pyridin-3-yl)-methanone (P-1987), [1553]
{4-[2-(2-Bromo-ethoxy)-ethoxy]-2-fluoro-5-methoxy-phenyl}-(5-chloro-1H-py-
rrolo[2,3-b]pyridin-3-yl)-methanone (P-1988), [1554]
(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-[2,5-difluoro-4-(2-methoxy-ethox-
y)-phenyl]-methanone (P-1989), and [1555]
(5-Chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-[2-fluoro-5-methoxy-4-(2-methoxy-
-ethoxy)-phenyl]-methanone (P-1991). The following table indicates
the aldehyde (column 2) and the azaindole (column 3) used to afford
the target compound (column 4). Column 1 indicates the compound
number and column 5 the observed mass.
TABLE-US-00016 [1555] Aldehyde Azaindole P-1845 ##STR01312##
##STR01313## P-1850 ##STR01314## ##STR01315## P-1900 ##STR01316##
##STR01317## P-1903 ##STR01318## ##STR01319## P-1979 ##STR01320##
##STR01321## P-1982 ##STR01322## ##STR01323## P-1987 ##STR01324##
##STR01325## P-1988 ##STR01326## ##STR01327## P-1989 ##STR01328##
##STR01329## P-1991 ##STR01330## ##STR01331## MS(ESI) [M +
H.sup.+].sup.+ ob Compound served P- 1845 ##STR01332## 409.6 [M -
H.sup.+].sup.- P- 1850 ##STR01333## P- 1900 ##STR01334## 387.4 P-
1903 ##STR01335## 363.3 [M - H.sup.+].sup.- P- 1979 ##STR01336##
447.4 P- 1982 ##STR01337## 417.3 P- 1987 ##STR01338## 405.3 P- 1988
##STR01339## 471.2 473.2 P- 1989 ##STR01340## 365.2 [M -
H.sup.+].sup.- P- 1991 ##STR01341## 377.2 [M - H.sup.+].sup.-
Example 36
Synthesis of
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
P-1901
[1556] Compound P-1901 was synthesized in four steps from
4-bromo-2,5-difluoro-phenol 595 as shown in Scheme 64.
##STR01342##
Step 1--Synthesis of 1-Benzyloxy-4-bromo-2,5-difluoro-benzene
(600)
[1557] To 4-bromo-2,5-difluoro-phenol (595, 0.90 g, 0.0043 mol,
prepared as described in Example 34, Scheme 61) in
N,N-dimethylformamide (30.0 mL) were added sodium hydride (0.21 g,
60% in mineral oil, 0.0052 mol) and benzyl bromide (0.563 mL,
0.00474 mol). The reaction was stirred at room temperature
overnight. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 5% ethyl acetate in
hexane to give a white solid (600, 0.84 g, 65.0%).
Step
2--(4-Benzyloxy-2,5-difluoro-phenyl)-(1-triisopropylsilanyl-1H-pyrrol-
o[2,3-b]pyridin-3-yl)-methanol (601)
[1558] To 1-Benzyloxy-4-bromo-2,5-difluoro-benzene (600, 0.84 g,
2.80 mmol) in tetrahydrofuran (15.0 mL) and ether (15.0 mL), under
an atmosphere of nitrogen at -78.degree. C., n-butyllithium (1.20
mL, 2.50 M in hexane) was added slowly. After 20 minutes,
1-Triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde (96,
0.82 g, 0.0027 mol, prepared as described in Example 18) was added
to the reaction. After 20 minutes, the reaction was allowed to warm
to room temperature for 10 minutes, then poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified with silica gel column chromatography
eluting with 20% ethyl acetate in hexane to a white solid (601, 1.0
g, 70.0%). MS(ESI) [M+H.sup.+].sup.+=523.4.
Step 3--Synthesis of
(4-Benzyloxy-2,5-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methano-
l (P-1902)
[1559] To
(4-Benzyloxy-2,5-difluoro-phenyl)-(1-triisopropylsilanyl-1H-pyrr-
olo[2,3-b]pyridin-3-yl)-methanol (601, 1.00 g, 1.91 mmol) in
tetrahydrofuran (15.0 mL) was added tetrabutylammonium fluoride,
trihydrate (0.63 g, 2.04 mmol). The reaction was stirred at room
temperature for 10 minutes. The reaction was roto-evaporated and
purified with silica gel column chromatography eluting with 50%
ethyl acetate in hexane to give the compound as a white solid
(P-1902, 0.59 g, 84.0%). MS(ESI) [M+H.sup.+].sup.+=367.4.
Step 4--Synthesis of
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1901)
[1560] To
(4-Benzyloxy-2,5-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl-
)-methanol (P-1902, 500.0 mg, 1.37 mmol) in acetonitrile (25.0 mL)
were added triethylsilane (2.00 mL, 0.0125 mol) and trifluoroacetic
acid (1.00 mL, 0.0130 mol). The reaction was heated to reflux for 2
hours. The reaction was concentrated and purified with silica gel
column chromatography eluting with 50% ethyl acetate in hexane to
give a white solid (P-1901, 60.0 mg, 94.1%). MS(ESI)
[M+H.sup.+].sup.+=351.4.
3-[3-Trifluoromethyl-4-(4-trifluoromethyl-benzyloxy)-benzyl]-1H-pyrrolo[2,-
3-b]pyridine P-1797
##STR01343##
[1561] was prepared using the protocol of Scheme 64, substituting
4-bromo-2,5-difluoro-phenol 595 with
4-bromo-2-trifluoromethyl-phenol (prepared as described in Example
34, Scheme 61, Step 1, substituting 2,5-difluoro-phenol 594 with
2-trifluoromethyl-phenol) and benzyl bromide with
1-bromomethyl-4-trifluoromethyl-benzene in Step 1. MS(ESI)
[M+H.sup.+].sup.+=451.
Example 37
Synthesis of
3-[4-(4-chloro-benzyloxy)-2,5-difluoro-benzyl]-1H-pyrrolo[2,3-b]pyridine
P-1974
[1562] Compound P-1974 was synthesized in four steps from
3-(4-benzyloxy-2,5-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
P-1901 as shown in Scheme 65.
##STR01344##
Step 1--Synthesis of
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1-triisopropylsilanyl-1H-pyrrolo[2,3--
b]pyridine (602)
[1563] To
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1901, 560.0 mg, 1.60 mmol, prepared as described in Example 18,
Scheme 33) in tetrahydrofuran (28.0 mL) was added sodium hydride
(100.0 mg, 60% in mineral oil, 2.50 mmol). After 10 minutes,
triisopropylsilyl chloride (0.500 mL, 2.36 mmol) was added to the
reaction. After 4 hours, the reaction was poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give the compound (602,
0.70 g, 86.1%).
Step 2--Synthesis of
2,5-difluoro-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl-
)-phenol (603)
[1564] To
3-(4-Benzyloxy-2,5-difluoro-benzyl)-1-triisopropylsilanyl-1H-pyr-
rolo[2,3-b]pyridine (602, 0.70 g, 0.0014 mol) in methanol (30.0 mL)
was added 50% palladium hydroxide on carbon (0.1 g) under an
atmosphere of hydrogen. The reaction was stirred at room
temperature overnight. The reaction was filtered and concentrated
to give a colorless oil (603, 0.47 g, 82.0%).
Step
3--3-[4-(4-Chloro-benzyloxy)-2,5-difluoro-benzyl]-1-triisopropylsilan-
yl-1H-pyrrolo[2,3-b]pyridine (604)
[1565] To
2,5-difluoro-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-
-ylmethyl)-phenol (603, 120.0 mg, 0.29 mmol) in
N,N-dimethylformamide (15.0 mL) was added sodium hydride (18.0 mg,
60% in mineral oil, 0.45 mol) under an atmosphere of nitrogen.
After 10 minutes, 4-chlorobenzyl bromide (65.1 mg, 0.32 mol) was
added to the reaction. The reaction was stirred at 40.degree. C.
overnight. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was dried over anhydrous sodium
sulfate, and filtered. The filtrate was concentrated to give the
crude compound (604, 0.15 g) that was used directly in the next
step.
Step 4--Synthesis of
3-[4(4-chloro-benzyloxy)-2,5-difluoro-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1974)
[1566] To
3-[4-(4-chloro-benzyloxy)-2,5-difluoro-benzyl]-1-triisopropylsil-
anyl-1H-pyrrolo[2,3-b]pyridine (604, 0.150 g, 0.28 mmol) in
tetrahydrofuran (10.0 mL) was added tetra-n-butylammonium fluoride
(80.0 mg, 0.31 mmol). After 10 minutes, the reaction was
concentrated and purified by silica gel column chromatography
eluting with 50% ethyl acetate in hexane to give the compound as a
white solid (P-1974, 30.8 mg, 28.9%). MS(ESI)
[M+H.sup.+].sup.+=385.3.
2-[2,5-Difluoro-4-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenoxymethyl]-1H--
benzoimidazole P-1975
##STR01345##
[1567] was prepared using the protocol of Scheme 65, substituting
4-chlorobenzyl bromide with 2-chloromethyl-1H-benzoimidazole in
step 3. MS(ESI) [M+H.sup.+].sup.+=391.3.
Example 38
Synthesis of
1-(4-Butoxy-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-urea P-1754
[1568] Compound P-1754 was synthesized in three steps from
5-(4-fluorophenyl)-1H-pyrrolo[2,3-b]pyridine 605 as shown in Scheme
66.
##STR01346##
Step 7--Preparation of
(3-chloro-phenyl)-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-met-
hanone (606)
[1569] To 5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine (605, 530
mg, 2.5 mmol, prepared as described in Example 23) dissolved in 20
mL of dioxane was added 3-nitrobenzaldehyde (534, 758 mg, 5 mmol)
and potassium hydroxide (4 mL of 2.5M aqueous). The vial was shaken
on an orbital shaker for 16 hrs and the dioxane was removed under
reduced pressure. The residue was partitioned between ethyl acetate
and water. The aqueous layer was neutralized with the addition of
1M HCl. The organic layer was washed with brine, dried over
magnesium sulfate and concentrated under reduced pressure to give
yellow-orange oil (1.5 g). The crude material was dissolved in
dichloromethane (150 mL) and chilled to 0.degree. C. With vigorous
stirring, pyridinium chlorochromate (3.0 g, 14 mmol) was added
slowly, maintaining the solution temperature at 0.degree. C. After
complete addition the solution was allowed to stir at ambient
temperature for 1 hour. The resulting dark brown/black solution was
diluted with chloroform and passed through a plug of silica.
Elution with methanol gave 1.5 g of crude 606 that was carried on
to the next step without further purification.
Step 2--Preparation of
(3-Amino-phenyl)-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-meth-
anone (607)
[1570] Crude
(3-chloro-phenyl)-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-met-
hanone (606, 1.5 g) was dissolved in a minimal amount of methanol
(5 mL) and Pd/C (5%, 10 mg) was added. The reaction mixture was
shaken on a Parr shaker under 70 psi H.sub.2overnight. The reaction
mixture was filtered through Celite.RTM. and concentrated under
reduced pressure to give 1.4 g of crude 607 that was earned on to
the next step without further purification.
Step 3--Preparation of
1-(4-Butoxy-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-urea (P-1754)
[1571] To a solution of
-(3-Amino-phenyl)-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-met-
hanone (607, 7.5 mg) in anhydrous pyridine (200 .mu.L) was added
neat 1-butoxy-4-isocyanato-benzene (608, 1.6 mg) and the reaction
mixture was stirred at ambient temperature for 2 hours. The
reaction mixture was concentrated under reduced pressure and the
residue was dissolved in DMSO (200 .mu.L) and purified using
reverse phase HPLC with an acetonitrile/water gradient. MS(ESI)
[M+H.sup.+].sup.+=523.5.
[1572] Additional compounds were prepared following the protocol of
Scheme 66, optionally substituting
5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine 605 with a suitable
azaindole in Step 1 and/or optionally substituting
1-butoxy-4-isocyanato-benzene 608 with a suitable isocyanate in
Step 3. Azaindoles were purchased or prepared as described in
Examples 9 or 17. The following compounds were prepared by this
procedure: [1573]
1-(2-Methoxy-ethyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1566), [1574]
1-Phenyl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-urea (P-1591), [1575]
1-Phenyl-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-urea
(P-1703), [1576]
1-(4-Fluoro-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1704), [1577]
1-(4-Methoxy-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
urea (P-1705), [1578]
1-(3,4-Difluoro-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-pheny-
l]-urea (P-1706), [1579]
1-(3-Methoxy-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
urea (P-1707), [1580]
1-(3,4-Dimethoxy-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phen-
yl]-urea (P-1708), [1581]
1-(4-Chloro-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1709), [1582]
1-(3-Chloro-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1710), [1583]
1-(4-Chloro-3-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl)-phenyl]-urea (P-1711), [1584]
1-(2-Chloro-5-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl)-phenyl]-urea (P-1712), [1585]
1-(2-Chloro-4-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl)-phenyl]-urea (P-1713), [1586]
1-(2-Fluoro-3-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl)-phenyl]-urea (P-1714), [1587]
1-(4-Butoxy-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1715), [1588]
1-(3-Fluoro-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1716), [1589]
1-[3-(1H-Pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(4-trifluoromethyl--
phenyl)-urea (P-1717), [1590]
1-[3-(1H-Pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-p-tolyl-urea
(P-1718), [1591]
1-[3-(1H-Pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-m-tolyl-urea
(P-1719), [1592]
1-[3-(1H-Pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-o-tolyl-urea
(P-1720), [1593]
1-(4-Methoxy-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-car-
bonyl)-phenyl]-urea (P-1723), [1594]
1-(3,4-Difluoro-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3--
carbonyl)-phenyl]-urea (P-1724), [1595]
1-(3,4-Dimethoxy-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl)-phenyl]-urea (P-1725), [1596]
1-(3-Chloro-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1726), [1597]
1-(2-Chloro-4-trifluoromethyl-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl)-phenyl]-urea (P-1727), [1598]
1-(2-Fluoro-3-trifluoromethyl-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl)-phenyl]-urea (P-1728), [1599]
1-(4-Butoxy-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1729), [1600]
1-(3-Fluoro-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1730), [1601]
1-[3-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(4-t-
rifluoromethyl-phenyl)-urea (P-1731), [1602]
1-(2-Chloro-5-trifluoromethyl-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl)-phenyl]-urea (P-1732), [1603]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-phenyl-ure-
a (P-1733), [1604]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(4-methoxy-
-phenyl)-urea (P-1734), [1605]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3,4-diflu-
oro-phenyl)-urea (P-1735), [1606]
1-(3-Chloro-phenyl)-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)--
phenyl]-urea (P-1736), [1607]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(4-chloro--
3-trifluoromethyl-phenyl)-urea (P-1737), [1608]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(2-chloro--
5-trifluoromethyl-phenyl)-urea (P-1738), [1609]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(2-chloro--
4-trifluoromethyl-phenyl)-urea (P-1739), [1610]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(2-fluoro--
3-trifluoromethyl-phenyl)-urea (P-1740), [1611]
1-(4-Butoxy-phenyl)-3-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)--
phenyl]-urea (P-1741), [1612]
1-[3-(1H-Pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3-trifluoromethyl--
phenyl)-urea (P-1746), [1613]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3-trifluo-
romethyl-phenyl)-urea (P-1747), [1614]
1-(4-Fluoro-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1748), [1615]
1-(3-Methoxy-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-car-
bonyl)-phenyl]-urea (P-1749), [1616]
1-(4-Chloro-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carb-
onyl)-phenyl]-urea (P-1750), [1617]
1-(4-Chloro-3-trifluoromethyl-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-
-b]pyridine-3-carbonyl)-phenyl]-urea (P-1751), [1618]
1-(2-Chloro-5-trifluoromethyl-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrol-
o[2,3-b]pyridine-3-carbonyl]-phenyl}-urea (P-1752), [1619]
1-(2-Chloro-4-trifluoromethyl-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrol-
o[2,3-b]pyridine-3-carbonyl]-phenyl}-urea (P-1753), [1620]
1-(4-Butoxy-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-urea (P-1754), [1621]
1-[3-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3-t-
rifluoromethyl-phenyl)-urea (P-1755), [1622]
1-[3-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-p-to-
lyl-urea (P-1756), [1623]
1-[3-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-m-to-
lyl-urea (P-1757), [1624]
1-{3-[5-(4-Fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-phenyl}-3-
-m-tolyl-urea (P-1758), [1625]
1-[3-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-o-to-
lyl-urea (P-1759), [1626]
1-Pyridin-4-yl-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-urea
(P-1760), [1627]
1-(2-Methoxy-ethyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-u-
rea (P-1761), [1628]
1-(3-Methoxy-5-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3--
carbonyl)-phenyl]-urea (P-1762), [1629]
1-(6-Methoxy-pyridin-3-yl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-ph-
enyl]-urea (P-1763), [1630]
1-Isoxazol-3-yl-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-urea
(P-1764), [1631]
1-(3-Methyl-isoxazol-5-yl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-ph-
enyl]-urea (P-1765), [1632]
1-(3-Chloro-4-trifluoromethyl-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-c-
arbonyl)-phenyl]-urea (P-1766), [1633]
1-(3,4-Dimethyl-isoxazol-5-yl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl-
)-phenyl]-urea (P-1767), [1634]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-pyridin-4--
yl-urea (P-1770), [1635]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-pyridin-3--
yl-urea (P-1771), [1636]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(2-methoxy-
-ethyl)-urea (P-1772), [1637]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3-methoxy-
-5-trifluoromethyl-phenyl)-urea (P-1773), [1638]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(6-methoxy-
-pyridin-3-yl)-urea (P-1774), [1639]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-isoxazol-3-
-yl-urea (P-1775), [1640]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(3,4-dimet-
hyl-isoxazol-5-yl)-urea (P-1776), [1641]
1-Pyridin-4-yl-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-
-phenyl]-urea (P-1777), [1642]
1-(3-Methoxy-5-trifluoromethyl-phenyl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,-
3-b]pyridine-3-carbonyl)-phenyl]-urea (P-1778), [1643]
1-(6-Methoxy-pyridin-3-yl)-3-[3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-
-3-carbonyl)-phenyl]-urea (P-1779), [1644]
1-(4-Dimethylamino-phenyl)-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-ph-
enyl]-urea (P-1780), [1645]
1-Pyridin-3-yl-3-[3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-urea
(P-1781), [1646]
1-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-(4-dimethy-
lamino-phenyl)-urea (P-1782), [1647]
1-(4-Fluoro-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-urea (P-1816), [1648]
1-(3,4-Difluoro-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridi-
ne-3-carbonyl]-phenyl}-urea (P-1817), [1649]
1-(3,4-Dimethoxy-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyrid-
ine-3-carbonyl]-phenyl}-urea (P-1818), and [1650]
1-(3-Fluoro-phenyl)-3-{3-[5-(4-fluoro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-
-carbonyl]-phenyl}-urea (P-1819). The following table indicates the
azaindole (column 2) and isocyanate (column 3) used to afford the
target compound (column 4). Column 1 provides the compound number
and the observed mass is given in column 5.
TABLE-US-00017 [1650] Azaindole Isocyanate P-1556 ##STR01347##
##STR01348## P-1591 ##STR01349## ##STR01350## P-1703 ##STR01351##
##STR01352## P-1704 ##STR01353## ##STR01354## P-1705 ##STR01355##
##STR01356## P-1706 ##STR01357## ##STR01358## P-1707 ##STR01359##
##STR01360## P-1708 ##STR01361## ##STR01362## P-1709 ##STR01363##
##STR01364## P-1710 ##STR01365## ##STR01366## P-1711 ##STR01367##
##STR01368## P-1712 ##STR01369## ##STR01370## P-1713 ##STR01371##
##STR01372## P-1714 ##STR01373## ##STR01374## P-1715 ##STR01375##
##STR01376## P-1716 ##STR01377## ##STR01378## P-1717 ##STR01379##
##STR01380## P-1718 ##STR01381## ##STR01382## P-1719 ##STR01383##
##STR01384## P-1720 ##STR01385## ##STR01386## P-1723 ##STR01387##
##STR01388## P-1724 ##STR01389## ##STR01390## P-1725 ##STR01391##
##STR01392## P-1726 ##STR01393## ##STR01394## P-1727 ##STR01395##
##STR01396## P-1728 ##STR01397## ##STR01398## P-1729 ##STR01399##
##STR01400## P-1730 ##STR01401## ##STR01402## P-1731 ##STR01403##
##STR01404## P-1732 ##STR01405## ##STR01406## P-1733 ##STR01407##
##STR01408## P-1734 ##STR01409## ##STR01410## P-1735 ##STR01411##
##STR01412## P-1736 ##STR01413## ##STR01414## P-1737 ##STR01415##
##STR01416## P-1738 ##STR01417## ##STR01418## P-1739 ##STR01419##
##STR01420## P-1740 ##STR01421## ##STR01422## P-1741 ##STR01423##
##STR01424## P-1746 ##STR01425## ##STR01426## P-1747 ##STR01427##
##STR01428## P-1748 ##STR01429## ##STR01430## P-1749 ##STR01431##
##STR01432## P-1750 ##STR01433## ##STR01434## P-1751 ##STR01435##
##STR01436## P-1752 ##STR01437## ##STR01438## P-1753 ##STR01439##
##STR01440## P-1754 ##STR01441## ##STR01442## P-1755 ##STR01443##
##STR01444## P-1756 ##STR01445## ##STR01446## P-1757 ##STR01447##
##STR01448## P-1758 ##STR01449## ##STR01450## P-1759 ##STR01451##
##STR01452## P-1760 ##STR01453## ##STR01454## P-1761 ##STR01455##
##STR01456## P-1762 ##STR01457## ##STR01458## P-1763 ##STR01459##
##STR01460## P-1764 ##STR01461## ##STR01462## P-1765 ##STR01463##
##STR01464## P-1766 ##STR01465## ##STR01466## P-1767 ##STR01467##
##STR01468## P-1770 ##STR01469## ##STR01470## P-1771 ##STR01471##
##STR01472## P-1772 ##STR01473## ##STR01474## P-1773 ##STR01475##
##STR01476## P-1774 ##STR01477## ##STR01478## P-1775 ##STR01479##
##STR01480## P-1776 ##STR01481## ##STR01482## P-1777 ##STR01483##
##STR01484## P-1778 ##STR01485## ##STR01486## P-1779 ##STR01487##
##STR01488## P-1780 ##STR01489## ##STR01490## P-1781 ##STR01491##
##STR01492## P-1782 ##STR01493## ##STR01494## P-1816 ##STR01495##
##STR01496## P-1817 ##STR01497## ##STR01498## P-1818 ##STR01499##
##STR01500## P-1819 ##STR01501## ##STR01502## MS(ESI) [M +
H.sup.+].sup.+ Compound observed P-1566 ##STR01503## 416.3 P-1591
##STR01504## 434.3 P-1703 ##STR01505## 357.1 P-1704 ##STR01506##
375.1 P-1705 ##STR01507## 387.1 P-1706 ##STR01508## 393.1 P-1707
##STR01509## 387.1 P-1708 ##STR01510## 417.5 P-1709 ##STR01511##
391.1 P-1710 ##STR01512## 391.1 P-1711 ##STR01513## 459.1 P-1712
##STR01514## 459.1 P-1713 ##STR01515## 459.1 P-1714 ##STR01516##
443.1 P-1715 ##STR01517## 429.1 P-1716 ##STR01518## 375.1 P-1717
##STR01519## 425.1 P-1718 ##STR01520## 371.1 P-1719 ##STR01521##
371.1 P-1720 ##STR01522## 371.1 P-1723 ##STR01523## 463.9 P-1724
##STR01524## 470.3 P-1725 ##STR01525## 494.3 P-1726 ##STR01526##
468.3 P-1727 ##STR01527## 535.9 P-1728 ##STR01528## 520.3 P-1729
##STR01529## 505.9 P-1730 ##STR01530## 451.9 P-1731 ##STR01531##
502.3 P-1732 ##STR01532## 535.9 P-1733 ##STR01533## 391.1 P-1734
##STR01534## 421.1 P-1735 ##STR01535## 427.1 P-1736 ##STR01536##
425.1 P-1737 ##STR01537## 493.1 P-1738 ##STR01538## 493.1 P-1739
##STR01539## 493.1 P-1740 ##STR01540## 477.1 P-1741 ##STR01541##
463.1 P-1746 ##STR01542## 425.1 P-1747 ##STR01543## 459.1 P-1748
##STR01544## 451.9 P-1749 ##STR01545## 463.9 P-1750 ##STR01546##
467.9
P-1751 ##STR01547## 535.9 P-1752 ##STR01548## 553.2 P-1753
##STR01549## 553.2 P-1754 ##STR01550## 523.5 P-1755 ##STR01551##
502.3 P-1756 ##STR01552## 447.9 P-1757 ##STR01553## 447.9 P-1758
##STR01554## 465.1 P-1759 ##STR01555## 447.9 P-1760 ##STR01556##
358.3 P-1761 ##STR01557## 339.1 P-1762 ##STR01558## 455.1 P-1763
##STR01559## 388.3 P-1764 ##STR01560## 348.3 P-1765 ##STR01561##
362.3 P-1766 ##STR01562## 459.1 P-1767 ##STR01563## 376.3 P-1770
##STR01564## 392.3 P-1771 ##STR01565## 392.3 P-1772 ##STR01566##
373.1 P-1773 ##STR01567## 489.1 P-1774 ##STR01568## 421.9 P-1775
##STR01569## 382.3 P-1776 ##STR01570## 410.3 P-1777 ##STR01571##
435.1 P-1778 ##STR01572## 531.9 P-1779 ##STR01573## 465.1 P-1780
##STR01574## 400.3 P-1781 ##STR01575## 358.3 P-1782 ##STR01576##
434.3 P-1816 ##STR01577## 468.1 P-1817 ##STR01578## 486.1 P-1818
##STR01579## 510.2 P-1819 ##STR01580## 468.1
Example 39
Synthesis of
N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3,5-difluorobenzenesulfonamide P-1841
[1651] Compound P-1841 was synthesized in six steps from
2,4-difluoroaniline 42 as shown in Scheme 67.
##STR01581##
Step 1--Preparation of (2,4-difluoro-phenyl)-carbamic acid benzyl
ester (613)
[1652] To 2,4-difluoroaniline (42, 7.0 mL, 0.070 mol) in 100 mL of
dichloromethane was added pyridine (11 mL, 0.14 mol) and benzyl
chloroformate (11.9 mL, 0.0834 mol). The reaction mixture was
stirred at ambient temperature for 1.5 hours. The reaction mixture
was concentrated under reduced pressure and the residue was
partitioned between ethyl acetate and KHSO.sub.4 solution. The
organic layer was dried (MgSO.sub.4), concentrated and crystallized
from hexanes to give compound 613 (15.6 g, 85%).
Step 2--Preparation of (2,4-difluoro-3-formyl-phenyl)-carbamic acid
benzyl ester (614)
[1653] Into a round bottom flask was added
(2,4-difluoro-phenyl)-carbamic acid benzyl ester (613, 3.83 g, 14.5
mmol) in tetrahydrofuran (148 mL, 1.82 mol). The solution was
chilled to -78.degree. C. and n-butyllithium (1.60 M in hexane,
19.1 mL, 30.0 mmol) was added over 30 minutes followed by the
addition of, N,N-dimethylformamide (1.12 mL, 14.5 mol). The
reaction mixture was allowed to warm to ambient temperature and was
stirred overnight. The reaction mixture was poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and crystallized
from ether to give compound 614 (3.0 g, 71%).
Step 3--Preparation of
{2,4-difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-phenyl}--
carbamic acid benzyl ester (615)
[1654] Into a round bottom flask was added
5-chloro-1H-pyrrolo[2,3-b]pyridine (80, 0.524 g, 3.43 mmol,
prepared as described in Example 9) in methanol (5.00 mL, 0.123
mol). Potassium hydroxide (0.800 g, 14.2 mmol) and
(2,4-difluoro-3-formyl-phenyl)-carbamic acid benzyl ester (614,
1.02 g, 3.5 mmol) were added and the reaction mixture was stirred
overnight. The reaction mixture was poured into 1N HCl and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and crystallized
from ethyl acetate to give compound 615 (710 mg, 46%).
MS(ESI)[M+H.sup.+].sup.+=444.
Step 4--Preparation of
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-carbamic
acid benzyl ester (616)
[1655] Into a round bottom flask was added
{2,4-difluoro-3-[hydroxy-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methyl]-phenyl}--
carbamic acid benzyl ester (615, 1.01 g, 2.28 mmol) in
tetrahydrofuran (5.00 mL, 0.0616 mol). Dess-Martin periodinane
(1.20 g, 2.89 mmol) was added in portions. The reaction mixture was
stirred at ambient temperature for 10 minutes, then poured into
water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over sodium sulfate, concentrated and
purified by silica gel chromatography to give compound 616 (914 mg,
91%). MS(ESI)[M+H.sup.+].sup.+=442.
Step 5--Preparation of
(3-Amino-2,6-difluoro-phenyl)-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanone (P-1801)
[1656]
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-carb-
amic acid benzyl ester (616, 800 mg, 1.81 mmol) was added to 10 M
NaOH (15.00 mL) and warmed to reflux overnight. The reaction
mixture was diluted with 30 mL of water and was extracted with
ethyl acetate to give compound P-1801 (450 mg, 81%).
Step 6--Preparation of
N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3,5-difluorobenzenesulfonamide (P-1841)
[1657] Into a microwave reaction vessel were combined
(3-amino-2,6-difluoro-phenyl)-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanone (P-1801, 50 mg, 0.16 mmol), 3,5-difluorobenzenesulfonyl
chloride (610, 103 mg, 0.49 mmol), pyridine (0.5 mL, 6.1820 mol)
and tetrahydrofuran (3.0 mL.). The reaction was warmed in the CEM
microwave at 300 watts, 130.degree. C. for 10 minutes. The reaction
mixture was partitioned between ethyl acetate and brine. The
organic layer was collected, dried over Na.sub.2SO.sub.4, filtered
and concentrated. The compound (P-1841) was isolated using column
chromatography (silica, hexane:ethyl acetate 70:30) to obtain 36 mg
(46%) compound. MS=482.0.
[1658] Additional compounds were prepared following the protocol of
Scheme 67 Step 6, optionally substituting
(3-Amino-2,6-difluoro-phenyl)-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanone P-1801 with
(3-Amino-2,6-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
P-2021 (prepared per Scheme 67 Steps 1-5, substituting
5-chloro-1H-pyrrolo[2,3-b]pyridine 80 with
1H-pyrrolo[2,3-b]pyridine 94 in Step 3) and/or
3,5-difluorobenzenesulfonyl chloride 610 with an appropriate
sulfonyl chloride. The following compounds were prepared by this
procedure: [1659]
N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-isopropyl-benzenesulfonamide (P-1839), [1660]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-benzenes-
ulfonamide (P-0913), [1661]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-nitro--
benzenesulfonamide (P-1937), [1662]
N-{4-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenylsulfamo-
yl]-phenyl}-acetamide (P-1938), [1663]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-methox-
y-benzenesulfonamide (P-0958), [1664]
5-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenylsulfamoyl]-
-furan-2-carboxylic acid methyl ester (P-1941), [1665]
5-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenylsulfamoyl]-
-2-methyl-furan-3-carboxylic acid methyl ester (P-1942), [1666]
5-Oxazol-5-yl-thiophene-2-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1943), [1667] 5-Isoxazol-5-yl-thiophene-2-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1948), [1668]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-2,4-dime-
thoxy-benzenesulfonamide (P-1951), [1669]
2,5-Dimethyl-thiophene-3-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1952), [1670] 2,5-Dimethyl-furan-3-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1953), [1671]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-2-methyl-
-benzenesulfonamide (P-1954), [1672]
2,3-Dihydro-benzo[1,4]dioxine-6-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1955), [1673] 2,4-Dimethyl-thiazole-5-sulfonic acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1956), [1674]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-triflu-
oromethyl-benzenesulfonamide (P-0931), [1675]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-5-fluoro-
-2-methyl-benzenesulfonamide (P-1961), [1676]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-3-methyl-
-benzenesulfonamide (P-1962), [1677]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-oxazol-
-5-yl-benzenesulfonamide (P-1963), [1678]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-2,5-dime-
thoxy-benzenesulfonamide (P-1131), [1679]
2-Cyano-N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
benzenesulfonamide (P-1965), [1680]
3-Cyano-N-[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]--
benzenesulfonamide (P-1966), [1681]
N-[2,4-Difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-4-isopro-
pyl-benzenesulfonamide (P-1968), [1682] Benzothiazole-6-sulfonic
acid
[2,4-difluoro-3-(1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-phenyl]-amide
(P-1969), [1683]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-methoxy-benzenesulfonamide (P-2011), [1684]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-benzenesulfonamide (P-0885), [1685] Thiophene-2-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1267), [1686]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-methyl-benzenesulfonamide (P-1842), [1687]
N-{4-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phen-
ylsulfamoyl]- phenyl}-acetamide (P-1905), [1688]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-methoxy-benzenesulfonamide (P-0983), [1689]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-trifluoromethyl-benzenesulfonamide (P-1599), [1690]
5-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyls-
ulfamoyl]-furan-2-carboxylic acid methyl ester (P-1907), [1691]
5-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyls-
ulfamoyl]-2-methyl-furan-3-carboxylic acid methyl ester (P-1908),
[1692] 1,2-Dimethyl-1H-imidazole-4-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1911), [1693]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2-fluoro-benzenesulfonamide (P-1912), [1694]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-4-difluoromethoxy-benzenesulfonamide (P-1916), [1695]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2,4-dimethoxy-benzenesulfonamide (P-1918), [1696]
2,5-Dimethyl-thiophene-3-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1919), [1697] 2,5-Dimethyl-furan-3-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1920), [1698]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2-methyl-benzenesulfonamide (P-1921), [1699]
2,3-Dihydro-benzo[1,4]dioxine-6-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1922), [1700] 2,4-Dimethyl-thiazole-5-sulfonic acid
[3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-a-
mide (P-1923), [1701]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2,4-difluoro-benzenesulfonamide (P-1926), [1702]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-5-fluoro-2-methyl-benzenesulfonamide (P-1927), [1703]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-methyl-benzenesulfonamide (P-1928), [1704]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2,5-dimethoxy-benzenesulfonamide (P-1929), [1705]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-2-cyano-benzenesulfonamide (P-1931), and [1706]
N-[3-(5-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl]-
-3-cyano-benzenesulfonamide (P-1932). The following table indicates
the azaindole (column 2) and the sulfonyl chloride (column 3) used
to afford the target compound (column 4). The compound number is
provided in column 1, with the observed mass given in column 5.
TABLE-US-00018 [1706] Sulfonyl Azaindole chloride P-1839
##STR01582## ##STR01583## P-0913 ##STR01584## ##STR01585## P-1937
##STR01586## ##STR01587## P-1938 ##STR01588## ##STR01589## P-0958
##STR01590## ##STR01591## P-1941 ##STR01592## ##STR01593## P-1942
##STR01594## ##STR01595## P-1943 ##STR01596## ##STR01597## P-1948
##STR01598## ##STR01599## P-1951 ##STR01600## ##STR01601## P-1952
##STR01602## ##STR01603## P-1953 ##STR01604## ##STR01605## P-1954
##STR01606## ##STR01607## P-1955 ##STR01608## ##STR01609## P-1956
##STR01610## ##STR01611## P-0931 ##STR01612## ##STR01613## P-1961
##STR01614## ##STR01615## P-1962 ##STR01616## ##STR01617## P-1963
##STR01618## ##STR01619## P-1131 ##STR01620## ##STR01621## P-1965
##STR01622## ##STR01623## P-1966 ##STR01624## ##STR01625## P-1968
##STR01626## ##STR01627## P-1969 ##STR01628## ##STR01629## P-2011
##STR01630## ##STR01631## P-0885 ##STR01632## ##STR01633## P-1267
##STR01634## ##STR01635## P-1842 ##STR01636## ##STR01637## P-1905
##STR01638## ##STR01639## P-0983 ##STR01640## ##STR01641## P-1599
##STR01642## ##STR01643## P-1907 ##STR01644## ##STR01645## P-1908
##STR01646## ##STR01647## P-1911 ##STR01648## ##STR01649## P-1912
##STR01650## ##STR01651## P-1916 ##STR01652## ##STR01653## P-1918
##STR01654## ##STR01655## P-1919 ##STR01656## ##STR01657## P-1920
##STR01658## ##STR01659## P-1921 ##STR01660## ##STR01661## P-1922
##STR01662## ##STR01663## P-1923 ##STR01664## ##STR01665## P-1926
##STR01666## ##STR01667## P-1927 ##STR01668## ##STR01669## P-1928
##STR01670## ##STR01671## P-1929 ##STR01672## ##STR01673## P-1931
##STR01674## ##STR01675## P-1932 ##STR01676## ##STR01677## MS(ESI)
[M + H.sup.+].sup.+ Compound observed P-1839 ##STR01678## 489.9
P-0913 ##STR01679## 413.9 P-1937 ##STR01680## 459.1 P-1938
##STR01681## 471.1 P-0958 ##STR01682## 444.3 P-1941 ##STR01683##
462.3 P-1942 ##STR01684## 475.9 P-1943 ##STR01685## 487.1 P-1948
##STR01686## 487.1 P-1951 ##STR01687## 473.9 P-1952 ##STR01688##
447.9 P-1953 ##STR01689## 432.3 P-1954 ##STR01690## 427.9 P-1955
##STR01691## 472.3 P-1956 ##STR01692## 448.7 P-0931 ##STR01693##
481.9 P-1961 ##STR01694## 445.9 P-1962 ##STR01695## 427.9 P-1963
##STR01696## 481.1 P-1131 ##STR01697## 473.9 P-1965 ##STR01698##
439.1 P-1966 ##STR01699## 439.1 P-1968 ##STR01700## 456.3 P-1969
##STR01701## 471.1 P-2011 ##STR01702## 477.9 P-0885 ##STR01703##
447.9 P-1267 ##STR01704## 453.9 P-1842 ##STR01705## 462.3 P-1905
##STR01706## 505.1 P-0983 ##STR01707## 477.9 P-1599 ##STR01708##
515.9 P-1907 ##STR01709## 496.3 P-1908 ##STR01710## 509.9 P-1911
##STR01711## 466.3 P-1912 ##STR01712## 465.9 P-1916 ##STR01713##
513.9 P-1918 ##STR01714## P-1919 ##STR01715## 481.9 P-1920
##STR01716## 465.9 P-1921 ##STR01717## 461.9 P-1922 ##STR01718##
505.9 P-1923 ##STR01719## 483.1 P-1926 ##STR01720## 483.9 P-1927
##STR01721## 479.9 P-1928 ##STR01722## 461.9 P-1929 ##STR01723##
507.9 P-1931 ##STR01724## 473.1 P-1932 ##STR01725## 473.1
Example 40
Synthesis of
4-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxyl-
ic acid dibutylamide P-1636
[1707] Compound P-1636 was synthesized in two steps from
1H-indole-4-carboxylic acid 611 as shown in Scheme 68.
##STR01726##
Step 1--Preparation of 1-dibutylcarbamoyl-1H-indole-4-carboxylic
acid (612)
[1708] To 1H-Indole-4-carboxylic acid (611, 251 mg, 1.56 mmol) in
tetrahydrofuran (3 mL), was added 2.5M n-butyllithium in hexane
(1.28 mL, 3.19 mmol) at -78.degree. C. After 30 minutes, dibutyl
carbamyl chloride (657 mg, 3.43 mmol) was added and stirred for two
hours. The reaction solution was quenched with 1M HCl (aq.) and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous magnesium sulfate, filtrated and
concentrated. The desired compound was isolated with silica gel
column chromatography using 10% ethyl acetate in hexane to give a
white solid (612, 88 mg, 18%). MS(ESI) [M-H.sup.+].sup.-=315.1.
Step 2--Preparation of
4-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxyl-
ic acid dibutylamide (P-1636)
[1709] To 1-dibutylcarbamoyl-1H-indole-4-carboxylic acid (612, 78
mg, 0.25 mmol) in dichloromethane (2 mL), thionyl chloride (25
.mu.L, 0.34 mmol) was added and stirred for one hour, followed by
rotary evaporation to remove solvents to provide the dried acid
chloride, which was dissolved in dichloromethane for later use.
Meanwhile, 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 55 mg,
0.28 mmol, prepared as described in Example 17) in dichloromethane
(8 mL), was mixed with aluminum trichloride (215 mg, 1.6 mmol) and
stirred for one hour, followed by addition of the dried acid
chloride in dichloromethane (3 mL). The reaction was stirred at
room temperature overnight, then quenched with methanol and all
volatiles were removed. The desired compound was isolated with
silica gel column chromatography using 10% methanol in
dichloromethane to give a solid (P-1636, 11 mg, 9%). MS(ESI)
[M+H.sup.+].sup.+=494.3.
[1710] Additional compounds were prepared following the protocol of
Scheme 68, substituting dibutyl carbamyl chloride with a suitable
reagent in Step 1 and optionally substituting
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-(6-Methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine (see Example
17) in Step 2. The following compounds were prepared following this
procedure: [1711] [1-(Butane-1-sulfonyl)-1H-indol-4
yl]-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
(P-1661), [1712]
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-c-
arboxylic acid pentylamide (P-1702), [1713]
4-(5-Pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-indole-1-carboxyl-
ic acid dipropylamide (P-1722), and [1714]
4-[5-(6-Methoxy-pyridin-3-yl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-indol-
e-1-carboxylic acid butylamide (P-1827). The following table
indicates the reagent used in place of dibutyl carbamyl chloride
(column 2) and the azaindole (column 3) used to afford the target
compound (column 3). The compound number is provided in column 1,
and the observed mass is given in column 5.
TABLE-US-00019 [1714] MS(ESI) [M + H.sup.+].sup.+ Step 1 reagent
Azaindole Compound observed P-1661 ##STR01727## ##STR01728##
##STR01729## 459.2 P-1702 ##STR01730## ##STR01731## ##STR01732##
452.3 P-1722 ##STR01733## ##STR01734## ##STR01735## 466.3 P-1827
##STR01736## ##STR01737## ##STR01738## 468.3
Example 41
Synthesis of
3-(3-Benzyloxy-2-chloro-6-fluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
P-1852,
(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin--
3-yl)-methanone P-1853 and Related Compounds
[1715] Compounds P-1852 and P-1853 were synthesized in four steps
from 2-chloro-4-fluorophenol 617 and 1 H-pyrrolo[2,3-b]pyridine 94
as shown in Scheme 69.
##STR01739## ##STR01740##
Step 1--Preparation of 1-Benzyloxy-2-chloro-4-fluoro-benzene
(618)
[1716] To a solution of 2-chloro-4-fluorophenol (617, 7 g, 0.05
mol) in tetrahydrofuran (100 mL) was added sodium hydride (1.8 g,
95% dry powder, 0.071 mol) at room temperature over 15 minutes
under an atmosphere of nitrogen. The reaction mixture was stirred
at room temperature for 30 minutes. Benzyl bromide (10 g, 0.060
mol) was added slowly to the reaction mixture, then stirred at room
temperature overnight. The reaction mixture was poured into ice
water, extracted with ethyl acetate, washed with hydrochloric acid
(10%), water, brine, and dried over magnesium sulfate. After
removal of solvent, the residue was purified by silica gel column
chromatography eluting with ethyl acetate in hexane to provide
compound as a white solid (618, 7.6 g, 60%).
Step 2--Preparation of 3-Benzyloxy-2-chloro-6-fluoro-benzaldehyde
(619)
[1717] To a solution of 1-benzyloxy-2-chloro-4-fluoro-benzene (618,
5.8 g, 0.024 mol) in tetrahydrofuran (100 mL) was added 2.50 M of
n-butyllithium (2.7 mL, 2.50 M in hexane, 0.029 mol) slowly at
-78.degree. C. over 15 minutes under nitrogen. The reaction mixture
was stirred at -78.degree. C. for 30 minutes. To the reaction
mixture was then added N,N-dimethylformamide (4.2 mL, 0.054 mol).
The reaction was allowed to warm to room temperature and was
continued at room temperature overnight. The reaction mixture was
poured into ice water, extracted With ethyl acetate, washed with
hydrochloric acid (10%), water, brine, and dried over magnesium
sulfate. After removal of solvent, the residue was purified by
silica gel column chromatography eluting with ethyl acetate in
hexane to provide the compound as a white solid (619, 2.1 g, 32%).
MS(ESI) [M+H.sup.+].sup.+=265.08.
Step 3--Preparation of
3-[(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3--
b]pyridine (P-1867) and
(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanol (P-1868)
[1718] A mixture of 1H-pyrrolo[2,3-b]pyridine (94, 0.5 g, 4 mmol),
3-benzyloxy-2-chloro-6-fluoro-benzaldehyde (619, 1.3 g, 4.9 mmol),
and potassium hydroxide (0.99 g, 18 mmol) in methanol (30 mL) was
stirred at room temperature overnight. The reaction mixture was
concentrated and the residue was dissolved in ethyl acetate and
water. The organic layer was collected and washed with brine. After
removal of solvent, the residue was purified by silica gel column
chromatography eluting with ethyl acetate in hexane to provide
compound P-1867 as a white solid (1.3 g, 70%, MS(ESI)
[M+H.sup.+].sup.+=397.16), and compound P-1868 as an off-white
solid (0.2 g, 10, MS(ESI) [M+H.sup.+].sup.+=383.14).
Step 4a--Preparation of
3-(3-Benzyloxy-2-chloro-6-fluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1852)
[1719] A mixture of
3-[(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3b-
]-pyridine (P-1867, 0.1 g, 0.2 mmol), trifluoroacetic acid (0.6 mL,
8 mmol), and triethylsilane (0.3 mL, 2 mmol) in acetonitrile (10
mL) was refluxed for 2 hours. The mixture was concentrated and the
residue was dissolved in ethyl acetate. The solution was washed
with saturated sodium bicarbonate, brine, and dried over sodium
sulfate. After removal of solvent, the residue was purified by
silica gel column chromatography eluting with methanol in
dichloromethane to provide compound as an off-white solid (P-1852,
62 mg, 70%). MS(ESI) [M+H.sup.+].sup.+=367.16.
Step 4b--Preparation of
(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanone (P-1853)
[1720] To a solution of
(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanol (P-1868, 65 mg, 0.17 mmol) in tetrahydrofuran (10 mL) was
added Dess-Martin periodinane (79 mg, 0.19 mmol) at 0.degree. C.
The reaction mixture was stirred at room temperature for 2 hours.
The reaction was quenched with a saturated solution of sodium
thiosulfate, extracted with ethyl acetate, washed with sodium
bicarbonate, brine, and dried over magnesium sulfate. After removal
of solvent, the residue was purified by silica gel column
chromatography eluting with methanol in dichloromethane to provide
the compound as a light yellow solid (P-1853, 32 mg, 50%). MS(ESI)
[M+H.sup.+].sup.+=381.13.
[1721] Additional compounds were prepared following the protocol of
Scheme 69, optionally replacing 2-chloro-4-fluorophenol 617 with
2,6-difluorophenol or 2,6-dichlorophenol, optionally replacing
benzyl bromide with an appropriate substituted benzyl bromide, and
optionally replacing 1H-pyrrolo[2,3-b]pyridine 94 with an
appropriate substituted 1H-pyrrolo[2,3-b]pyridine. Azaindoles were
purchased or prepared as described in Examples 9 or 16. The
following compounds were made following this procedure: [1722]
3-[2,6-Dichloro-3-(4-chloro-benzyloxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1768), [1723]
[2,6-Dichloro-3-(4-chloro-benzyloxy)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3--
yl) methanone (P-1769), [1724]
(3-Benzyloxy-2,6-difluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methano-
ne (P-1802), [1725]
3-(3-Benzyloxy-2,6-difluoro-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1803), [1726]
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-methoxy-1H-pyrrolo[2,3-b]pyr-
idine (P-1804), [1727]
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-chloro-1H-pyrrolo[2,3-b]pyridine
(P-1824), [1728]
(3-Benzyloxy-2,6-difluoro-phenyl)-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl-
)-methanone (P-1825), [1729]
3-[(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3--
b]pyridine (P-1867), [1730]
(3-Benzyloxy-2-chloro-6-fluoro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thanol (P-1868), [1731]
[2-Chloro-3-(3-chloro-benzyloxy)-6-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (P-1869), [1732]
[2-Chloro-3-(4-chloro-benzyloxy)-6-fluoro-phenyl]-(1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (P-1874), [1733]
3-[2,6-Difluoro-3-(pyridin-4-ylmethoxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1993), and [1734]
3-[3-(4-Chloro-2-fluoro-benzyloxy)-2,6-difluoro-benzyl]-1H-pyrrolo[2,3-b]-
pyridine (P-1992). The phenol, benzyl bromide and azaindole used in
Steps 1, 2, and 3, respectively, are indicated in columns 2, 3, and
4 of the following table, respectively, to afford the target
compound (column 5). The compound number is provided in column 1,
and the observed mass is given in column 6.
TABLE-US-00020 [1734] MS (ESI) Benzyl [M + H.sup.+].sup.+ Phenol
bromide Azaindole Compound observed P-1768 ##STR01741##
##STR01742## ##STR01743## ##STR01744## 417.14 P-1769 ##STR01745##
##STR01746## ##STR01747## ##STR01748## 431.09 P-1802 ##STR01749##
##STR01750## ##STR01751## ##STR01752## 365.23 P-1803 ##STR01753##
##STR01754## ##STR01755## ##STR01756## 351.23 P-1804 ##STR01757##
##STR01758## ##STR01759## ##STR01760## 381.26 P-1824 ##STR01761##
##STR01762## ##STR01763## ##STR01764## 385.22 P-1825 ##STR01765##
##STR01766## ##STR01767## ##STR01768## 399.21 P-1869 ##STR01769##
##STR01770## ##STR01771## ##STR01772## 415.24 P-1874 ##STR01773##
##STR01774## ##STR01775## ##STR01776## 415.23 P-1993 ##STR01777##
##STR01778## ##STR01779## ##STR01780## 352.39 P-1992 ##STR01781##
##STR01782## ##STR01783## ##STR01784## 403.32
Example 42
Synthesis of
(3-Benzyloxy-2-methyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
P-1848 and
3-(3-Benzyloxy-2-methyl-benzyl)-1H-pyrrolo[2,3-b]pyridine
P-1857
[1735] Compounds P-1848 and P-1857 were synthesized in five steps
from compounds 620 and 1H-pyrrolo[2,3-b]pyridine 94 as shown in
Scheme 70.
##STR01785## ##STR01786##
Step 1--Preparation of 3-Benzyloxy-2-methyl-benzoic acid (621)
[1736] To a solution of 3-hydroxy-2-methyl-benzoic acid (620, 5.0
g, 0.033 mol) in tetrahydrofuran (100 mL) and N,N-dimethylformamide
(50 mL), sodium hydride (4.4 g as 60% dispersion in mineral oil,
0.11 mol) was added slowly over 30 minutes and the reaction was
stirred at 0.degree. C. under an atmosphere of nitrogen. The
reaction mixture was allowed to warm to room temperature, then
stirred at room temperature for 1 hour. Benzyl bromide (9.0 mL,
0.076 mol) was added slowly into the reaction mixture, and the
reaction mixture was stirred at room temperature overnight. The
reaction mixture was poured into water, extracted with ethyl
acetate, washed with a solution of ammonium chloride and ammonium
hydroxide (4:1), brine, and dried over magnesium sulfate. After
removal of solvent, the residue was purified by silica gel column
chromatography eluting with ethyl acetate in hexane to provide the
compound as a white solid (621, 5.8 g, 73%).
Step 2--Preparation of (3-Benzyloxy-2-methyl-phenyl)-methanol
(622)
[1737] To a solution of 3-benzyloxy-2-methyl-benzoic acid (621, 3.0
g, 0.012 mol) in tetrahydrofuran (100 mL), lithium aluminum hydride
(25 mL, 1M solution in tetrahydrofuran, 0.025 mol) was added
dropwise at 0.degree. C. for 5 minutes. The reaction mixture was
then stirred at room temperature overnight under an atmosphere of
nitrogen. After sodium sulfate decahydrate (20.0 g, 0.062 mol) was
added, the reaction mixture was stirred at room temperature for 10
minutes. A white solid was collected by filtration. The solid
compound was further washed with a mixture of hexane and
dichloromethane (9:1) and dried under high-vacuum (622, 2.8 g,
91%).
Step 3--Preparation of 3-Benzyloxy-2-methyl-benzaldehyde (623)
[1738] To a solution of (3-benzyloxy-2-methyl-phenyl)-methanol
(622, 627 mg, 2.75 mmol) in tetrahydrofuran (60 mL) was added
Dess-Martin periodinane (2.9 g, 6.87 mmol) at 0.degree. C. The
resulting mixture was stirred at 0.degree. C. for 50 minutes. The
reaction mixture was quenched with a solution of saturated sodium
thiosulfate, extracted with ethyl acetate, washed with sodium
bicarbonate, brine, and dried over magnesium sulfate. After removal
of solvent, the residue was purified by silica gel column
chromatography eluting with ethyl acetate in hexane to provide the
compound as a white solid (623, 0.55 g, 84%).
Step 4--Preparation of
(3-Benzyloxy-2-methyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanol
(624) and
3-[(3-Benzyloxy-2-methyl-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3-
-b]pyridine (625)
[1739] A mixture of 1H-pyrrolo[2,3-b]pyridine (94, 0.33 g, 2.8
mmol), 3-benzyloxy-2-methyl-benzaldehyde (623, 0.55 g, 2.4 mmol),
and potassium hydroxide (0.39 g, 6.1 mmol) in methanol (40 mL) was
stirred at room temperature for 17 hours. The reaction mixture was
poured into water and then extracted with ethyl acetate. The
organic layer was collected, washed with brine, and dried over
sodium sulfate. After removal of solvent, the residue was purified
by silica gel column chromatography eluting with ethyl acetate in
hexane to provide compound 624 as an off-white solid (330 mg, 39%,
MS(ESI) [M+H.sup.+].sup.+=345.29, and compound 625 as a white solid
(24 mg, 3%, MS(ESI) [M+H.sup.+].sup.+=359.30).
Step 5a--Preparation of
(3-Benzyloxy-2-methyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
(P-1848)
[1740] To a solution of
(3-Benzyloxy-2-methyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanol
(624, 0.12 g, 0.35 mmol) in tetrahydrofuran (15 mL) was added
Dess-Martin periodinane (0.37 g, 0.89 mmol) at 0.degree. C. The
reaction mixture was stirred at 0.degree. C. for 50 minutes, then
quenched with a saturated solution of sodium thiosulfate, extracted
with ethyl acetate, washed with sodium bicarbonate, brine, and
dried over magnesium sulfate. After removal of solvent, the residue
was washed with a mixture of ethyl ether and hexanes (1:1) to
provide the compound as a yellow solid (P-1848, 108 mg, 90%).
MS(ESI) [M+H.sup.+].sup.+=343.22.
Step 5b--Preparation of
3-(3-Benzyloxy-2-methyl-benzyl)-1H-pyrrolo[2,3-b]pyridine
(P-1857)
[1741] A mixture of
3-[(3-benzyloxy-2-methyl-phenyl)-methoxy-methyl]-1H-pyrrolo[2,3-b]pyridin-
e (625, 24 mg, 0.067 mmol), trifluoroacetic acid (1 mL, 13 mmol),
and triethylsilane (2 mL, 12.5 mmol) in acetonitrile (10 mL) was
refluxed for 4 hours. The mixture was concentrated and the residue
was dissolved in ethyl acetate. The solution was washed with
saturated sodium bicarbonate, brine, and dried over sodium sulfate.
After removal of solvent, the residue was washed with a mixture of
ethyl ether and hexanes (1:1) to provide the compound as a yellow
solid (P-1857, 17 mg, 75%). MS(ESI) [M+H.sup.+].sup.+=329.24.
Example 43
Synthesis of
[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone P-1892 and
3-[3-(4-chloro-benzyloxy)-2-ethoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
P-1893
[1742] Compounds P-1892 and P-1893 were synthesized in five steps
from compounds 626, 557 and 1 H-pyrrolo[2,3-b]pyridine 94 as shown
in Scheme 71.
##STR01787## ##STR01788##
Step 1--Preparation of 2,3-Bis-(4-chloro-benzyloxy)-benzaldehyde
(627)
[1743] To a solution of 2,3-dihydroxybenzaldehyde (626, 2.0 g, 14.5
mmol) in tetrahydrofuran (100 mL) was added sodium hydride (0.52 g,
13.0 mmol) at 0.degree. C. under an atmosphere of nitrogen. The
reaction mixture was allowed to warm to room temperature and was
stirred at room temperature for 30 minutes. To the reaction mixture
was then added 4-chlorobenzyl bromide (557, 2.7 g, 13.0 mmol). The
reaction mixture was stirred at room temperature under an
atmosphere of nitrogen overnight. N,N-dimethylformamide (50 mL) was
added into the reaction mixture and it was stirred at room
temperature for 24 hours. The reaction mixture was poured into ice
water and extracted with ethyl acetate. The organic layer was
collected, washed with brine, and dried over magnesium sulfate.
After removal of solvent, the residue was purified by silica gel
column chromatography eluting with ethyl acetate in hexane to
provide the compound as an off-white solid (627, 2.3 gm, 46%).
Step 2--Preparation of
3-(4-chloro-benzyloxy)-2-hydroxy-benzaldehyde (628)
[1744] To magnesium (0.098 g, turnings, 4.0 mmol) in a mixture of
anhydrous ether (20 mL) and benzene (20 mL) at 0.degree. C.,
bromine (0.10 mL, 2.0 mmol) was added dropwise. When the reaction
had started, stirring was commenced and the addition of bromine
continued until complete. The ice bath was removed and the reaction
mixture was heated until the solution was almost colorless. After
cooling down, the reaction mixture was slowly added to a solution
of 2,3-bis-(4-chloro-benzyloxy) benzaldehyde (627, 0.78 g, 2.0
mmol) in benzene (60 mL) at room temperature while stirring
vigorously. Upon completion of the addition, the reaction mixture
was stirred at room temperature overnight, then refluxed for 36
hours. After the reaction mixture was cooled down to room
temperature, a solid was collected by filtration and washed with
benzene, then boiled in hydrochloric acid (100 mL, 1.0 M) for 30
minutes. After cool down, the solution was extracted with
dichloromethane. The organic layer was washed with brine and dried
over magnesium sulfate. An off-white solid was obtained after
removal of the solvent (628, 0.32 mg, 60%). MS(ESI)
[M-H-]-261.25.
Step 3--Preparation of 3-(4-chloro-benzyloxy)-2-ethoxy-benzaldehyde
(629)
[1745] To a mixture of
3-(4-chloro-benzyloxy)-2-hydroxy-benzaldehyde (110 mg, 0.42 mmol),
potassium carbonate (150 mg, 1.1 mmol) in acetonitrile (8 mL) was
added iodoethane (0.2 mL, 2.5 mmol) at room temperature. The
mixture was stirred at 98.degree. C. for 18 hours. The reaction
mixture was poured into a solution of saturated ammonium chloride
and was extracted with ethyl acetate. The organic layer was
collected, washed with brine, and dried over magnesium sulfate.
After removal of solvent, a light yellow solid was obtained (629,
116 mg, 95%).
Step 4--Preparation of
[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)-
methanol (630) and
3-{[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-methoxy-methyl}-1H-pyrrolo[2,-
3-b]pyridine (631)
[1746] A mixture of 1H-Pyrrolo[2,3-b]pyridine (94, 26 mg, 0.22
mmol), 3-(4-chloro-benzyloxy)-2-ethoxy-benzaldehyde (629, 54 mg,
0.19 mmol), and potassium hydroxide (30 mg, 0.4.6 mmol) in methanol
(5 mL) was stirred at room temperature for 4 days. The reaction
mixture was poured into water and extracted with ethyl acetate. The
organic layer was collected, washed with brine, and dried over
sodium sulfate. After removal of solvent, the residue was purified
by silica gel column chromatography eluting with ethyl acetate in
hexane to provide compound 630 as an off-white solid (20 mg, 26%,
MS (ESI) [M+H.sup.+].sup.+=409.32) and compound 631 as an off-white
solid (44 mg, 56%, MS (ESI) [M+H.sup.+].sup.+=423.33.
Step 5a--Preparation of
[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone (P-1892)
[1747] To a solution of
[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanol (630, 20 mg, 0.05 mmol) in tetrahydrofuran (8 mL) was
added Dess-Martin periodinane (52 mg, 0.12 mmol) at 0.degree. C.
The reaction mixture was stirred at 0.degree. C. for 50 minutes.
The reaction was quenched with a saturated solution of sodium
thiosulfate, extracted with ethyl acetate, washed with sodium
bicarbonate, brine, and dried over magnesium sulfate. After removal
of solvent, the residue was washed with a mixture of ethyl ether
and hexanes (1:1) to provide the compound as a yellow solid
(P-1892, 15 mg, 75%). MS(ESI) [M+H.sup.+].sup.+=407.38.
Step 5b--Preparation of
3-[3-(4-chloro-benzyloxy)-2-ethoxy-benzyl]-1H-pyrrolo[2,3-b]pyridine
(P-1893)
[1748] A mixture of
3-{[3-(4-chloro-benzyloxy)-2-ethoxy-phenyl]-methoxy-methyl}-1H-pyrrolo[2,-
3-b]pyridine (631, 44 mg, 0.1 mmol), trifluoroacetic acid (1 mL, 13
mmol), and triethylsilane (2 mL, 12.5 mmol) in acetonitrile (10 mL)
was refluxed for 4 hours. The mixture was concentrated and the
residue was dissolved in ethyl acetate. The solution was washed
with saturated sodium bicarbonate, brine, and dried over sodium
sulfate. After removal of solvent, the residue was washed with a
mixture of ethyl ether and hexanes (1:1) to provide the compound as
a yellow solid (P-1893, 40 mg, 98%). MS(ESI)
[M+H.sup.+].sup.+=393.39.
[1749]
[3-(4-Chloro-benzyloxy)-2-methoxy-phenyl]-(1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methanone (P-1891),
[3-(4-Chloro-benzyloxy)-2-(2,2,2-trifluoroethoxy)-phenyl]-(1H-pyrrolo[2,3-
-b]pyridin-3-yl)-methanone (P-2076), and
[3-(4-chloro-2-fluoro-benzyloxy)-2-ethoxy-phenyl]-(5-methoxy-1H-pyrrolo[2-
,3-b]pyridin-3-yl)-methanone (P-2016)
##STR01789##
were prepared following the protocol of Scheme 71, substituting
iodoethane with iodomethane in Step 3 to provide P-1891, or
substituting iodoethane with 2-iodo-1,1,1-trifluoroethane in Step 3
to provide P-2076, or substituting 4-chlorobenzyl bromide 557 with
4-chloro-2-fluoro-benzyl bromide in Step 1 and 7-azaindole 94 with
5-methoxy-7-azaindole in Step 4 to provide P-2016. MS(ESI)
[M+H.sup.+].sup.+=393.4 (P-1891), 461.08 (P-2076), and 455.2
(P-2016).
Example 44
Synthesis of Propane-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluor-
o-phenyl}-amide P-0956
[1750] Compound P-0956 was synthesized in three steps from
5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine 514 and
propane-1-sulfonic acid (2,4-difluoro-3-formyl-phenyl)-amide 73 as
shown in Scheme 72.
##STR01790##
Step 1--Preparation of Propane-1-sulfonic acid
(3-{[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-hydroxy-methyl}-2-
,4-difluoro-phenyl)-amide (632) and Propane-1-sulfonic acid
(3-{[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-methoxy-methyl}-2-
,4-difluoro-phenyl)-amide (633)
[1751] To a suspension of
5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine (514, 64.9 g, 158 mM,
prepared as described in Example 17) and propane-1-sulfonic acid
(2,4-difluoro-3-formyl-phenyl)-amide (73, 90.4 g, 191 mM, prepared
as described in Example 7) in methanol in a water bath was added
potassium hydroxide (128.8 g, 1.28 M). The reaction was stirred 72
hours at room temperature and then adjusted to pH 7 with 4N
hydrochloric acid. The resulting mixture was evaporated in vacuo to
remove methanol and extracted 3.times. with ethyl acetate. The
combined organic layers were dried over sodium sulfate and
evaporated in vacuo to give a crude oil. The crude oil was
triturated with 3:1 MTBE/heptane to give a 1:3 solid mixture of 632
and 633 that was used directly for the next step.
Step 2-Preparation of Propane-1-sulfonic acid
(3-{[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-hydroxy-methyl}-2-
,4-difluoro-phenyl)-amide (632)
[1752] To a solution of 632 and 633 (ca. 315 mM) in acetic acid was
added 48% hydrobromic acid (final 8%). The resulting mixture was
stirred overnight at room temperature and then evaporated in vacuo.
The crude residue was taken up with equal volumes of ethyl acetate
and water, and adjusted to pH 7 with solid potassium carbonate. The
layers were split and the aqueous layer was extracted 2.times. with
ethyl acetate. The combined organic layers were dried over sodium
sulfate and evaporated in vacuo to give 632 as a viscous oil that
was used directly for the next step.
Step 3--Preparation of Propane-1-sulfonic acid
{3-[5-(4-chloro-phenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluor-
o-phenyl}-amide (P-0956)
[1753] To a solution of 632 (ca. 386 mM) in 1,4-dioxane was added
2,3-dichloro-5,6-dicyanobenzoquinone (83.8 g, 502 mM) followed by
water (final 4.8%). The resulting mixture was stirred 2 hours at
room temperature and then quenched with one volume of saturated
sodium bicarbonate. The mixture was evaporated in vacuo to remove
1,4-dioxane and extracted 3.times. with ethyl acetate. The combined
organic layers were dried over sodium sulfate and evaporated in
vacuo to give a crude solid that was purified on a silica-gel
column with 94:5:1 dichloromethane/methanol/ammonium hydroxide as
eluent to give P-0956 (approximately 50% yield for 3 steps) as a
white solid.
Example 45
Synthesis of 3-Iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
635
[1754] 3-Iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine 635
was synthesized in one step from 3-Iodo-1H-pyrrolo[2,3-b]pyridine
634 as shown in Scheme 73.
##STR01791##
Step 1--Preparation of
3-Iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (635)
[1755] 3-Iodo-1H-pyrrolo[2,3-b]pyridine 634 (2.00 g, 8.20 mmol) was
dissolved in N,N-dimethylformamide (50 mL). Sodium hydride (60%
dispersion in mineral oil, 390 mg, 9.8 mmol) was added. After 20
minutes, triisopropylsilyl chloride (1.74 mL, 8.20 mmol) was added
dropwise. After 1.5 hours, the reaction was poured into water and
extracted with ethyl acetate, washed with saturated sodium
bicarbonate and brine. The organic portions were dried over
anhydrous sodium sulfate and concentrated. Purification by silica
gel chromatography, 0-25% gradient ethyl acetate/hexane gave
compound 635 as a white solid (3.224 g, 98.2%). .sup.1H-NMR was
consistent with the desired compound.
Example 46
Synthesis of
1-(tert-Butyl-dimethyl-silanyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine
636
[1756]
1-(tert-Butyl-dimethyl-silanyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine
636 was synthesized in one step from
3-Iodo-1H-pyrrolo[2,3-b]pyridine 634 as shown in Scheme 74.
##STR01792##
Step 1--Preparation of
1-(tert-Butyl-dimethyl-silanyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine
(636)
[1757] 3-Iodo-1H-pyrrolo[2,3-b]pyridine 634 (1.11 g, 4.6 mmol) was
dissolved in tetrahydrofuran (120 mL). Sodium hydride (60%
dispersion in mineral oil, 0.13 g, 5.5 mmol) was added, followed by
tert-butyldimethylsilyl chloride (0.85 g, 5.5 mmol). The reaction
was stirred at room temperature overnight. The reaction was poured
into water and extracted with ethyl acetate. The organic portion
was washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified with silica
gel column chromatography eluting with 30% ethyl acetate in hexane
to give the compound as a white solid (636, 100 mg, 15%).
Example 47
Synthesis of
[5-(4-chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-(1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methanone P-2024
[1758]
[5-(4-Chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-(1H-pyrrolo[2,3-b]p-
yridin-3-yl)-methanone P-2024 was synthesized in six steps from
Kojic acid, 3, and
3-iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine, 2, as shown
in Scheme 75.
##STR01793## ##STR01794##
Step 1--Preparation of
5-(4-chloro-benzyloxy)-2-hydroxymethyl-pyran-4-one (638)
[1759] Kojic acid (637, 5.00 g, 35.2 mmol) and 4-chlorobenzyl
bromide (557, 7.95 g, 38.7 mmol) were suspended in methanol (40 mL)
in an 80 mL sealed tube. Sodium hydroxide in water (12 M, 2.93 mL)
was added. The reaction was heated at 80.degree. C.,overnight. The
resulting suspension was concentrated. Water was added and the
mixture was filtered and washed with water to provide a brown
solid. Washing with minimal methanol on the filter removed the
brown color. A white solid (638, 7.58 g, 80%) was isolated.
.sup.1H-NMR was consistent with the desired compound.
Step 2--Preparation of
5(4-chloro-benzyloxy)-2-hydroxymethyl-1H-pyridin-4-one (639)
[1760] 5-(4-Chloro-benzyloxy)-2-hydroxymethyl-pyran-4-one (638,
8.00 g, 3.00 mmol) was suspended in ammonium hydroxide (200 mL) in
an 80 mL scaled tube. The reaction was heated at 90.degree. C.
overnight. Upon cooling, the reaction was lowered to pH 10 with 6N
HCl to provide a beige solid that was collected by filtration (639,
7.8 g, 98%).
Step 3--Preparation of
[5-(4-chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-methanol (640)
[1761] 5-(4-Chloro-benzyloxy)-2-hydroxymethyl-1H-pyridin-4-one
(639, 1.06 g, 3.99 mmol) was dissolved in methanol (8.5 mL) and
N,N-dimethylformamide (46 mL). Trimethylsilyldiazomethane in hexane
(2.00 M, 3.99 mL) was added. The reaction was stirred at room
temperature overnight, then additional trimethylsilyldiazomethane
in hexane (2.00 M, 3.99 mL) was added. The reaction was stirred at
room temperature for 2 days. The mixture was adsorbed onto silica
and purified by silica gel chromatography, methanol:dichloromethane
to provide the compound (640, 798 mg, 72%). MS(ESI)
[M+H.sup.+].sup.+=280.4, 282.4.
Step 4--Preparation of
5-(4-chloro-benzyloxy)-4-methoxy-pyridine-2-carbaldehyde (641)
[1762] [5-(4-Chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-methanol
(640, 480 mg, 1.7 mmol) was dissolved in dimethyl sulfoxide (26 mL)
and Dess-Martin periodinane (909 mg, 2.1 mmol) was added. The
reaction was allowed to stir at room temperature for 2 hours. The
reaction was concentrated under high vacuum and then poured into a
solution of NaHCO.sub.3 and Na.sub.2S.sub.2O.sub.3. The mixture was
extracted with ethyl acetate. The organic portions were dried with
anhydrous sodium sulfate and filtered. The filtrate was adsorbed
onto silica and purified by silica gel chromatography, ethyl
acetate:hexanes, to provide the desired compound as a white powder
(641, 343 mg, 72%).
Step 5--Preparation of
[5-(4-chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-(1-triisopropylsilanyl-1H-
-pyrrolo[2,3-b]pyridin-3-yl)-methanol (642)
[1763] 3-Iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (635,
180 mg, 0.450 mmol) was dissolved in tetrahydrofuran (2.5 mL) and
the reaction was cooled to -20.degree. C. under an atmosphere of
nitrogen. Isopropylmagnesium chloride in tetrahydrofuran (2.00 M,
0.243 mL) was added. The reaction was stirred for 1 hour, during
which the temperature rose to 0.degree. C. The reaction was cooled
to -20.degree. C. and
5-(4-chloro-benzyloxy)-4-methoxy-pyridine-2-carbaldehyde (641, 80.0
mg, 0.288 mmol) in tetrahydrofuran (0.75 mL) was added. The
reaction was allowed to warm to room temperature and stirred
overnight. The reaction was quenched with methanol and adsorbed
onto silica, then purified by silica gel chromatography,
methanol:dichloromethane, to provide the desired product, (642, 94
mg, 59%). .sup.1H-NMR was consistent with the desired compound.
MS(ESI) [M+H.sup.+].sup.+=552.4, 554.4, 555.4.
Step 6--Preparation of
[5-(4-chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-(1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methanone (P-2024)
[1764]
[5-(4-Chloro-benzyloxy)-4-methoxy-pyridin-2-yl]-(1-triisopropylsila-
nyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanol (642, 60.0 mg, 0.11
mmol) was dissolved in tetrahydrofuran (2.00 mL). Dess-Martin
periodinane (55.3 mg, 0.13 mmol) was added to the reaction and it
was stirred at room temperature overnight. The mixture was
extracted with ethyl acetate and saturated sodium bicarbonate. The
organic portions were dried with anhydrous sodium sulfate, filtered
and the filtrate was adsorbed onto silica and purified by silica
gel chromatography, methanol:dichloromethane, to provide the
desired compound (P-2024, 10.7 mg, 25%). .sub.1H-NMR was consistent
with the desired compound. MS(ESI) [M+H.sup.+].sup.+=394.1,
396.1.
Example 48
Synthesis of
3-4-[1-(4-chloro-phenyl)-ethoxy]-3-methoxy-benzyl-1H-pyrrolo[2,3-b]pyridi-
ne P-2000
[1765]
3-4-[1-(4-Chloro-phenyl)-ethoxy]-3-methoxy-benzyl-1H-pyrrolo[2,3-b]-
pyridine P-2000 was synthesized in three steps from vanillin 105,
4-chlorophenylmethylcarbinol 643, and
1-(tert-butyl-dimethyl-silanyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine
636, as shown in Scheme 76.
##STR01795##
Step 1--Preparation of
4-[1-(4-chloro-phenyl)-ethoxy]-3-methoxy-benzaldehyde (644)
[1766] 4-Chlorophenylmethylcarbinol (643, 0.668 mL, 6.57 mmol) was
dissolved in tetrahydrofuran (60.0 mL) at 0.degree. C. under an
atmosphere of nitrogen. 4-Hydroxy-3-methoxybenzaldehyde (105, 1.00
g, 6.57 mmol) and triphenylphosphine (2.07 g, 7.89 mmol) were added
to the reaction, followed by diisopropyl azodicarboxylate (1.55 mL,
7.89 mmol) over 10 minutes. The reaction was stirred for 2 hours.
The mixture was adsorbed onto silica and purified by silica gel
chromatography, ethyl acetate:hexanes, to provide the desired
compound, (644, 1.14 g, 60%). .sup.1H-NMR was consistent with the
desired compound.
Step 2--Preparation of
[1-(tert-Butyl-dimethyl-silanyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-4-[1-(4-c-
hloro-phenyl)-ethoxy]-3-methoxy-phenyl-methanol (645)
[1767]
1-(tert-Butyl-dimethyl-silanyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine
(636, 647.0 mg, 1.81 mmol) was dissolved in tetrahydrofuran (10.0
mL) at -20.degree. C. under an atmosphere of nitrogen.
Isopropylmagnesium chloride in tetrahydrofuran (2.0 M, 0.98 mL) was
added to the reaction. The reaction was stirred for 1 hour, during
which the temperature rose to 0.degree. C. The reaction was cooled
to -20.degree. C. and
4-[1-(4-chloro-phenyl)-ethoxy]-3-methoxy-benzaldehyde (644, 420 mg,
1.4 mmol) in tetrahydrofuran (3.00 mL) was added. The reaction was
stirred for 2 hours during which time the temperature rose to
10.degree. C. The reaction was quenched with methanol and adsorbed
onto silica, then purified by silica gel chromatography, ethyl
acetate:hexanes, to provide the desired compound, (645, 463 mg,
61%). .sup.1H-NMR was consistent with the desired compound.
Step 2--Preparation of
3-4-[1-(4-chloro-phenyl)-ethoxy]-3-methoxy-benzyl-1H-pyrrolo[2,3-b]pyridi-
ne (P-2000)
[1768]
[1-(tert-Butyl-dimethyl-silanyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-4-[-
1-(4-chloro-phenyl)-ethoxy]-3-methoxy-phenyl-methanol (645, 0.200
g, 0.382 mmol) was dissolved in acetonitrile (5.00 mL).
Trifluoroacetic acid (0.138 mL) was added and the reaction was
stirred for five minutes. Triethylsilane (0.285 mL) was added and
the reaction was heated at 80.degree. C. for 2 hours. The reaction
was concentrated, then redissolved in ethyl acetate and adsorbed
onto silica and purified by silica gel chromatography, ethyl
acetate:hexanes, to provide the desired compound (P-2000, 57 mg,
38%). .sup.1H-NMR was consistent with the desired compound.
MS(ESI): [M+H.sup.+].sup.+=393.3, 395.3.
Example 49
Synthesis of
5-[4-(2-methoxyethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine 648
[1769] 5-[4-(2-Methoxyethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine 648
was synthesized in two steps from 4-bromophenol 646 as shown in
Scheme 77.
##STR01796##
Step 1--Preparation 1-Bromo-4-(2-methoxy-ethoxy)-benzene (647)
[1770] To a solution of 4-bromophenol (646, 5.0 g, 28.9 mmol) in
dimethylformamide (15 mL) were added potassium carbonate (4.40 g,
31.8 mmol) and 1-bromo-2-methoxyethane (5.00 g, 36.0 mmol) under an
atmosphere of nitrogen. The reaction mixture was stirred at ambient
temperature overnight and concentrated under reduced pressure. The
residue was slurried in ethyl acetate (50 mL) and filtered. The
filtrate was washed with saturated sodium bicarbonate solution,
dried over magnesium sulfate and filtered. Silica gel column
chromatography (0-10% ethyl acetate in hexanes) gave the desired
compound as a colorless oil (647, 3.2 g, 48%).
Step 2--Preparation of
5-[4-(2-Methoxy-ethoxy)-phenyl]-1H-pyrrolo[2,3-b]pyridine (648)
[1771] To a solution of
5-(4,4,5,5,-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridi-
ne (1.1 g, 4.3 mmol) in tetrahydrofuran (40 mL) was added
1-bromo-4-(2-methoxy-ethoxy)-benzene (647, 1.50 g, 6.49 mmol) and
tetrakis(triphenylphosphine)palladium (0) (0.25 g, 0.21 mmol). The
reaction mixture was stirred with potassium carbonate solution (10
mL, 1.0 M) and warmed to reflux overnight. The biphasic reaction
mixture was diluted with ethyl acetate (50 mL) and saturated sodium
carbonate solution (20 mL). The organic layer was separated, washed
with brine, dried over magnesium sulfate and purified by silica gel
column chromatography (50-100% ethyl acetate in hexanes) to give
the desired compound as a colorless solid (648, 782 mg, 67%). MS
(ESI) [M+H.sup.+].sup.+=267.4.
Example 50
Synthesis of
3-[2-fluoro-5-methoxy-4-(pyridin-4-ylmethoxy)-benzyl]-1H-pyrrolo[2,3-b]py-
ridine P-2040 and Related Compounds
[1772] Compound P-2040 was synthesized in one step from
5-fluoro-2-methoxy-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-yl-
methyl)-phenol 649 and pyridin-4-yl-methanol 650 as shown in Scheme
78.
##STR01797##
Step 1--Preparation of
3-[2-fluoro-5-methoxy-4-(pyridin-4-ylmethoxy)-benzyl]-1H-pyrrolo[2,3-b]py-
ridine (P-2040)
[1773]
5-Fluoro-2-methoxy-4-(1-triiisopropylsilanyl-1H-pyrrolo[2,3-b]pyrid-
ine-3-ylmethyl)-phenol (649, 10 mg, 0.024 mmol, prepared as
described in Example 52) was combined with pyridin-4-yl-methanol
(650, 3.2 mg, 0.029 mmol) in a 4 mL vial and dissolved in dry
tetrahydrofuran (200 .mu.l). Triphenylphosphine (7.7 mg) was added
and the solution was shaken until homogenous. The mixture was
cooled to below 0.degree. C. in a liquid nitrogen bath and
diisopropyl azodicarboxylate solution (50 .mu.l of 20 mg/50 .mu.l
in THF) was added. The reaction mixture was allowed to warm to room
temperature. After 2 hours, the solvent was removed under reduced
atmosphere. The crude material was dissolved in dimethyl sulfoxide
(300 .mu.l) and potassium fluoride (10 mg, 0.18 mmol) was added.
The mixture was heated gently and allowed to react overnight at
room temperature. The vial was centrifuged and the DMSO solution
was purified by reverse phase HPLC using a YMC-Pack ODS-A C-18
column (50 mm.times.10 mm ID), and eluting with water with 0.1% TFA
and a gradient of 15%-80% acetonitrile with 0.1% TFA over 8 minutes
and a flow rate of 6 mL/minute to provide the compound (P-2040, 4.4
mg, 50%). MS (ESI) [M+H.sup.+].sup.+=364.3.
[1774] Additional compounds were prepared following the protocol of
Scheme 78, replacing pyridin-4-yl-methanol 650 with an appropriate
alcohol. The following compounds were made following this
procedure: [1775]
3-[2-Fluoro-5-methoxy-4-(2-morpholin-4-yl-ethoxy)-benzyl]-1H-pyrrolo[2,3--
b]pyridine (P-2037), [1776]
3-[2-Fluoro-5-methoxy-4-(pyridin-3-ylmethoxy)-benzyl]-1H-pyrrolo[2,3-b]py-
ridine (P-2038), [1777]
3-[2-Fluoro-5-methoxy-4-(6-methyl-pyridin-2-ylmethoxy)-benzyl]-1H-pyrrolo-
[2,3-b]pyridine (P-2039), [1778]
3-[2-Fluoro-5-methoxy-4-(pyridin-2-ylmethoxy)-benzyl]-1H-pyrrolo[2,3-b]py-
ridine (P-2041), [1779]
3-[2-Fluoro-4-(2-fluoro-4-trifluoromethyl-benzyloxy)-5-methoxy-benzyl]-1H-
-pyrrolo[2,3-b]pyridine (P-2042), [1780]
3-[4-(4-Chloro-2-fluoro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[-
2,3-b]pyridine (P-1973), [1781]
3-[4-(2,4-Dimethyl-thiazol-5-ylmethoxy)-2-fluoro-5-methoxy-benzyl]-1H-pyr-
rolo[2,3-b]pyridine (P-2043), [1782]
3-[4-(2,5-Dimethyl-2H-pyrazol-3-ylmethoxy)-2-fluoro-5-methoxy-benzyl]-1H--
pyrrolo[2,3-b]pyridine (P-2044), [1783]
3-[2-Fluoro-5-methoxy-4-(3-morpholin-4-yl-propoxy)-benzyl]-1H-pyrrolo[2,3-
-b]pyridine (P-2045), [1784]
1-{2-[5-Fluoro-2-methoxy-4-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenoxy]-
-ethyl}-pyrrolidin-2-one (P-2046), [1785]
3-[2-Fluoro-4-(2-fluoro-benzyloxy)-5-methoxy-benzyl]-1H-pyrrolo[2,3-b]pyr-
idine (P-2047), [1786]
3-[2-Fluoro-5-methoxy-4-(3-methyl-pyridin-4-ylmethoxy)-benzyl]-1H-pyrrolo-
[2,3-b]pyridine (P-2048), [1787]
3-[2-Fluoro-5-methoxy-4-(6-trifluoromethyl-pyridin-3-ylmethoxy)-benzyl]-1-
H-pyrrolo[2,3-b]pyridine (P-2049), [1788]
3-[4-(2,4-Dichloro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,3-b-
]pyridine (P-2050), [1789]
3-[2-Fluoro-4-(4-imidazol-1-yl-benzyloxy)-5-methoxy-benzyl]-1H-pyrrolo[2,-
3-b]pyridine (P-2051), [1790]
3-[4-(2,4-Difluoro-benzyloxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,3-b-
]pyridine (P-2052), [1791]
3-{2-Fluoro-4-[1-(2-fluoro-phenyl)-ethoxy]-5-methoxy-benzyl}-1H-pyrrolo[2-
,3-b]pyridine (P-2053), [1792]
3-[4-(3-Cyclopentyl-propoxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,3-b]-
pyridine (P-2054), [1793]
3-[4-(1,5-Dimethyl-1H-pyrazol-3-ylmethoxy)-2-fluoro-5-methoxy-benzyl]-1H--
pyrrolo[2,3-b]pyridine (P-2055), and [1794]
3-[4-(2-Cyclopentyl-ethoxy)-2-fluoro-5-methoxy-benzyl]-1H-pyrrolo[2,3-b]p-
yridine (P-2056) The following table indicates the alcohol (column
2) used in Scheme 78 to provide the compounds (column 4). Column 1
provides the compound number and column 4 the observed mass.
TABLE-US-00021 [1794] MS(ESI) [M + H.sup.+].sup.+ Alcohol Compound
observed P-2037 ##STR01798## ##STR01799## 386.3 P-2038 ##STR01800##
##STR01801## 364.3 P-2039 ##STR01802## ##STR01803## 378.3 P-2041
##STR01804## ##STR01805## 364.3 P-2042 ##STR01806## ##STR01807##
448.7 P-1973 ##STR01808## ##STR01809## 415.1 P-2043 ##STR01810##
##STR01811## 397.9 P-2044 ##STR01812## ##STR01813## 381.1 P-2045
##STR01814## ##STR01815## 400.3 P-2046 ##STR01816## ##STR01817##
384.3 P-2047 ##STR01818## ##STR01819## 381.1 P-2048 ##STR01820##
##STR01821## 378.3 P-2049 ##STR01822## ##STR01823## 432.3 P-2050
##STR01824## ##STR01825## 431.1 P-2051 ##STR01826## ##STR01827##
429.1 P-2052 ##STR01828## ##STR01829## 399.1 P-2053 ##STR01830##
##STR01831## 395.1 P-2054 ##STR01832## ##STR01833## 383.1 P-2055
##STR01834## ##STR01835## 381.1 P-2056 ##STR01836## ##STR01837##
369.1
Example 51
Synthesis of
[2,6-difluoro-3-(pyridin-3-ylmethoxy)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-
-yl)-methanone P-2058 and Related Compounds
[1795] Compound P-2058 was synthesized in 1 step from
(2,6-Difluoro-3-hydroxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone
651 and Pyridin-3-yl-methanol 652 as shown in Scheme 79.
##STR01838##
Step 1--Preparation of
[2,6-Difluoro-3-(pyridin-3-ylmethoxy)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-
-yl)-methanone (P-2058)
[1796] In a 4 mL vial,
(2,6-Difluoro-3-hydroxy-phenyl)-(1H-pyrrolo[2,3-b]pyridine-3-yl)-methanon-
e (651, 10 mg, 0.037 mmol, prepared as described in Example 23) was
combined with pyridin-3-yl-methanol (652, 4.9 mg 0.044 mmol). The
solids were dissolved in dry tetrahydrofuran (200 .mu.l) and
triphenylphosphine (11.5 mg, 0.044 mmol) was added. Once the
solution was homogenous, the mixture was cooled to below 0.degree.
C. in liquid nitrogen bath and diisopropyl azodicarboxylate
solution (50 .mu.l of 20 mg/50 .mu.l THF) was added. The reaction
mixture was allowed to warm to room temperature and the reaction
was continued for 2 hours. The solvents were removed under reduced
atmosphere. The resultant residue was diluted with 200 .mu.l DMSO
and the mixture purified by reverse phase HPLC using a YMC-Pack
ODS-A C-18 column (50 mm.times.10 mm ID), and eluting with water
with 0.1% TFA and a gradient of 15%-80% acetonitrile with 0.1% TFA
over 8 minutes and a flow rate of 6 mL/minute to provide P-2058
(5.9 mg, 44%). MS (ESI) [M+H.sup.+].sup.+=365.9.
[1797] Additional compounds were prepared following the protocol of
Scheme 79, replacing pyridin-3-yl-methanol 652 with an appropriate
alcohol. The following compounds were made following this
procedure: [1798]
[2,6-Difluoro-3-(1-methyl-1H-imidazol-2-ylmethoxy)-phenyl]-(1H-pyrrolo[2,-
3-b]pyridin-3-yl)-methanone (P-2033), [1799]
[2,6-Difluoro-3-(6-morpholin-4-yl-pyridin-3-ylmethoxy)-phenyl]-(1H-pyrrol-
o[2,3-b]pyridin-3-yl)-methanone (P-2034), [1800]
{2,6-Difluoro-3-[4-(5-methyl-[1,2,4]oxadiazol-3-yl)-benzyloxy]-phenyl}-(1-
H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (P-2035), [1801]
[3-(6-Diethylamino-pyridin-3-ylmethoxy)-2,6-difluoro-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl-methanone (P-2036), [1802]
[3-(2-Chloro-4-fluoro-benzyloxy)-2,6-difluoro-phenyl]-(1H-pyrrolo[2,3-b]p-
yridin-3-yl)-methanone (P-2057), [1803]
[2,6-Difluoro-3-(6-methyl-pyridin-2-ylmethoxy)-phenyl]-(1H-pyrrolo[2,3-b]-
pyridin-3-yl)-methanone (P-2059), [1804]
[2,6-Difluoro-3-(pyridin-4-ylmethoxy)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-3-
-yl)-methanone (P-2060), [1805]
[3-(4-Chloro-2-fluoro-benzyloxy)-2,6-difluoro-phenyl]-(1H-pyrrolo[2,3-b]p-
yridin-3-yl)-methanone (P-2061), [1806]
[3-(2,4-Dimethyl-thiazol-5-ylmethoxy)-2,6-difluoro-phenyl]-(1H-pyrrolo[2,-
3-b]pyridin-3-yl)-methanone (P-2062), [1807]
[3-(2,5-Dimethyl-2H-pyrazol-3-ylmethoxy)-2,6-difluoro-phenyl]-(1H-pyrrolo-
[2,3-b]pyridin-3-yl)-methanone (P-2063), and [1808]
[2,6-Difluoro-3-(3-morpholin-4-yl-propoxy)-phenyl]-(1H-pyrrolo[2,3-b]pyri-
din-3-yl)-methanone (P-2064). The following table indicates the
alcohol (column 2) used to afford the compound (column 3). Column 1
provides the compound number and column 4 the observed mass.
TABLE-US-00022 [1808] MS(ESI) [M + H.sup.+].sup.+ Alcohol Compound
observed P-2033 ##STR01839## ##STR01840## 369.1 P-2034 ##STR01841##
##STR01842## 451.1 P-2035 ##STR01843## ##STR01844## 447.1 P-2036
##STR01845## ##STR01846## 437.1 P-2057 ##STR01847## ##STR01848##
417.1 P-2059 ##STR01849## ##STR01850## 380.3 P-2060 ##STR01851##
##STR01852## 365.9 P-2061 ##STR01853## ##STR01854## 417.1 P-2062
##STR01855## ##STR01856## 399.9 P-2063 ##STR01857## ##STR01858##
383.1 P-2064 ##STR01859## ##STR01860## 402.3
Example 52
Synthesis of
5-fluoro-2-methoxy-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-yl-
methyl)-phenol 649
[1809]
5-Fluoro-2-methoxy-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridi-
n-3-ylmethyl)-phenol 649 was synthesized in five steps from
2-fluoro-4-hydroxy-5-methoxy-benzaldehyde 653 and benzyl bromide as
shown in Scheme 80.
##STR01861## ##STR01862##
Step 1--Preparation of 4-Benzyloxy-2-fluoro-5-methoxy-benzaldehyde
(654)
[1810] 2-Fluoro-4-hydroxy-5-methoxy-benzaldehyde (653, 1.62 g, 9.52
mmol) was dissolved in N,N-dimethylformamide (50 mL) and sodium
hydride (60% dispersion in mineral oil, 530 mg, 13 mmol) was added.
After 20 minutes, benzyl bromide (1.5 mL, 12 mmol) was added to the
reaction mixture. The reaction was stirred at room temperature
under an atmosphere of nitrogen for 5.5 hours. The reaction was
poured into water and extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate, and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 0-50% ethyl acetate in hexane to
provide compound as a white solid, consistent with the desired
structure by .sup.1H-NMR (654, 2.0 g, 81%).
Step 2--Preparation of
(4-Benzyloxy-2-fluoro-5-methoxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo-
[2,3-b]pyridin-3-yl)-methanol (655)
[1811] 3-Iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (635,
620 mg, 1.5 mmol, prepared as described in Example 45) was
dissolved in tetrahydrofuran (15 mL) at -20.degree. C. under an
atmosphere of nitrogen. Isopropylmagnesium chloride (2.0 M in
tetrahydrofuran, 840 .mu.L) was added to the reaction. The reaction
was stirred for 1.5 hours, during which the temperature rose to
5.degree. C. The reaction was cooled to -20.degree. C.
4-Benzyloxy-2-fluoro-5-methoxy-benzaldehyde (654, 250 mg, 0.9606
mmol) in tetrahydrofuran (5.0 mL) was added to the reaction. The
reaction was stirred for 2.5 hours during which time the
temperature rose to 5.degree. C. The reaction was poured into
water. The mixture was extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate, and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 2-25% ethyl acetate in hexane to
provide compound as a white solid (655, 501 mg, 63%). MS (ESI)
[M+H.sup.+].sup.+=535.4.
Step 3--Preparation of
3-(4-Benzyloxy-2-fluoro-5-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
(656)
[1812]
(4-Benzyloxy-2-fluoro-5-methoxy-phenyl)-(1-triisopropylsilanyl-1H-p-
yrrolo[2,3-b]pyridin-3-yl)-methanol (655, 1.49 g, 2.79 mmol) was
dissolved in acetonitrile (50 mL) and trifluoroacetic Acid (1.1 mL)
was added. The reaction was stirred for 5 minutes. Triethylsilane
(2.2 mL) was added to the reaction. The reaction was heated at
80.degree. C. for 6 hours. The reaction was concentrated and the
crude material was dissolved into ethyl acetate and washed with 1 N
HCl, saturated sodium bicarbonate, and brine. The organic portion
was dried over anhydrous sodium sulfate and concentrated. The solid
obtained was used in the next reaction without further purification
(656, 833 mg, 83%). MS (ESI) [M+H.sup.+].sup.+=363.4.
Step 4--Preparation of
3-(4-Benzyloxy-2-fluoro-5-methoxy-benzyl)-1-triisopropylsilanyl-1H-pyrrol-
o[2,3-b]pyridine (657)
[1813]
3-(4-Benzyloxy-2-fluoro-5-methoxy-benzyl)-1H-pyrrolo[2,3-b]pyridine
(656, 0.877 g, 2.42 mmol) was dissolved in N,N-dimethylformamide
(30 mL). Sodium hydride (60% dispersion in mineral oil, 140 mg, 3.6
mmol) was added at room temperature. After 20 minutes,
triisopropylsilyl chloride (513 .mu.L, 2.42 mmol) was added
dropwise. The reaction was stirred for four hours. The reaction was
poured into water and extracted with ethyl acetate. The organic
portion was washed with saturated sodium bicarbonate and brine. The
organic portion was dried over anhydrous sodium sulfate and
filtered. The filtrate was adsorbed onto silica gel and purified by
silica gel chromatography using 20-80% ethyl acetate/hexane. The
resulting material was purified a second time with 5-30% gradient
ethyl acetate/hexane to provide the desired compound (657, 831 mg,
66%). MS (ESI) [M+H.sup.+].sup.+=519.4.
Step 5--Preparation of
5-Fluoro-2-methoxy-4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-yl-
methyl)-phenol (649)
[1814]
3-(4-Benzyloxy-2-fluoro-5-methoxy-benzyl)-1-triisopropylsilanyl-1H--
pyrrolo[2,3-b]pyridine (657, 0.831 g, 1.60 mmol) was dissolved in
methanol (40 mL) and tetrahydrofuran (40 mL). 10% Palladium on
carbon (3.41 g) was added. The reaction was shaken at 50 psi for 1
hour. The reaction was filtered through Celite and washed with
methanol. The organic portion was passed through celite several
times until a clear solution was obtained. The organic portion was
concentrated under reduced pressure to provide the desired compound
as an off-white solid (649, 587 mg, 86%). MS (ESI)
[M+H.sup.+].sup.+=429.5.
Example 53
Synthesis of
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-hydroxy-phenyl)-metha-
none P-0067 and Related Compounds
[1815] Compound P-0067 was synthesized in two steps from
5-bromo-1H-pyrrolo[2,3-b]pyridine 67 as shown in Scheme 10.
##STR01863##
Step 1--Preparation of
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phenyl)-metha-
none (P-0265)
[1816] To 5-bromo-1H-pyrrolo[2,3-b]pyridine (67, 2.00 g, 0.0102
mol) in methylene chloride (120 mL), under an atmosphere of
nitrogen, was added aluminum trichloride (8.20 g, 0.0615 mol). The
reaction was stirred at room temperature for 60 minutes, then
2-fluoro-3-methoxy-benzoyl chloride (34, 2.12 g, 0.0112 mol,
prepared from the corresponding carboxylic acid using the protocol
of Example 1, Scheme 13, Step 4), dissolved in methylene chloride
(20.0 mL), was added. After 2 hours, the reaction mixture was
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over sodium sulfate,
concentrated and purified by silica gel chromatography, eluting
with 25% ethyl acetate in hexane to provide P-0265 (approx 2.6 g,
73%). MS (ESI) [M-H.sup.+].sup.-=347, 349.
Step 2--Preparation of
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-hydroxy-phenyl)-metha-
none (P-0067)
[1817] To
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phen-
yl)-methanone (P-0265, 430.0 mg, 1.23 mmol) in methylene chloride
(60 mL) was added boron tribromide (1.0 M in methylene chloride, 10
mL). The reaction mixture was stirred at room temperature
overnight. The reaction was quenched with methanol (3.0 mL), poured
into water and extracted with ethyl acetate. The organic layer was
dried, concentrated and purified by silica gel chromatography
eluting with 25% ethyl acetate in hexane to provide the compound
(P-0067, 180 mg, 44%). MS (ESI) [M-H.sup.+].sup.-=333.0, 335.0.
2-Fluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenol
P-0778
##STR01864##
[1818] was prepared using the protocol of Scheme 10, Step 2,
replacing
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phenyl)-metha-
none P-0265 with
3-(2-Fluoro-3-methoxy-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine
(P-0269, prepared as described in Example 10, Scheme 24). MS(ESI)
[M+H.sup.+].sup.+=320.2.
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-5-hydroxy-phenyl)-methan-
one P-0068
##STR01865##
[1819] was prepared using the protocol of Scheme 10, replacing
2-Fluoro-3-methoxy-benzoyl chloride 34 with
2-Fluoro-5-methoxy-benzoyl chloride (prepared from the
corresponding carboxylic acid using the protocol of Example 1,
Scheme 13, Step 4) in Step 1. MS(ESI) [M-H.sup.+].sup.-=347,
349.
Example 54
Synthesis of
(2-fluoro-3-hydroxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone P-0036
[1820] Compound P-0036 was synthesized in three steps from
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-hydroxy-phenyl)-metha-
none P-0067 as shown in Scheme 11.
##STR01866##
Step 1--Preparation of
5-bromo-3-(3-tert-butoxycarbonyloxy-2-fluoro-benzoyl)-pyrrolo[2,3-b]pyrid-
ine-1-carboxylic acid tert-butyl ester (32)
[1821] To
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-hydroxy-phen-
yl)-methanone (P-0067, 310.0 mg, 0.93 mmol, prepared as described
in Example 53, Scheme 10) in tetrahydrofuran (40 mL) were added
sodium hydride (60% dispersion, 81.4 mg, 2.04 mmol) and
di-tert-butyldicarbonate (0.50 g, 2.3 mmol). The reaction was
stirred at room temperature for 1 hour. The reaction mixture was
poured into water and extracted with ethyl acetate. The organic,
layer was washed with brine, dried over sodium sulfate,
concentrated and purified by silica gel chromatography, eluting
with 20% ethyl acetate in hexane to provide the compound (32, 0.45
g, 91%).
Step 2--Preparation of
3-(3-tert-butoxycarbonyloxy-2-fluoro-benzoyl)-5-phenyl-pyrrolo[2,3-b]pyri-
dine-1-carboxylic acid tert-butyl ester (33)
[1822] To
5-bromo-3-(3-tert-butoxycarbonyloxy-2-fluoro-benzoyl)-pyrrolo[2,-
3-b]pyridine-1-carboxylic acid tert-butyl ester (32, 55.0 mg, 0.10
mmol) in tetrahydrofuran (10 mL) were added 1.0 M potassium
carbonate in water (3.5 mL), phenylboronic acid (18.8 mg, 0.15
mmol) and tetrakis(triphenylphosphine)palladium(0) (3.9 mg, 0.0034
mmol) under an atmosphere of nitrogen. The reaction was stirred at
65.degree. C. for 3 hours, and then poured into water and extracted
with ethyl acetate. The organic layer was washed with brine, dried
over sodium sulfate, and concentrated to give crude compound 33
that was used in the next step.
Step 3--Preparation of
(2-fluoro-3-hydroxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone (P-0036)
[1823] To
3-(3-tert-butoxycarbonyloxy-2-fluoro-benzoyl)-5-phenyl-pyrrolo[2-
,3-b]pyridine-1-carboxylic acid tert-butyl ester (33, 30.0 mg,
0.056 mmol) in methylene chloride (6.0 mL) was added 4 N HCl in
dioxane (2.0 mL). The reaction was stirred at room temperature for
36 hours. Solvents were removed by evaporation. The residue was
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over sodium sulfate,
concentrated and purified by silica gel chromatography, eluting
with 25% ethyl acetate in hexane to give a white solid (P-0036, 4.8
mg, 24%). MS(ESI) [M+H.sup.+].sup.+=333.2.
[1824] Additional compounds were prepared following the protocol of
Scheme 11, optionally replacing
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-hydroxy-phenyl)-metha-
none P-0067 with
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-hydroxy-2-methyl-phenyl)-metha-
none P-0055 (prepared as described in Example 56, Scheme 22) in
Step 1 and optionally replacing phenylboronic acid with an
appropriate boronic acid in Step 2. The following compounds were
made following this procedure: [1825]
N-{3-[3-(2-Fluoro-3-hydroxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl-
]-phenyl}-acetamide (P-0035), [1826]
N-{3-[3-(2-Fluoro-3-hydroxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-pheny-
l}-methanesulfonamide (P-0038), [1827]
3-[3-(2-Fluoro-3-hydroxy-benzoyl)-1H-pyrrolo[2,3-b]pyridin-5-yl]benzamide
(P-0034), [1828]
(3-Hydroxy-2-methyl-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3b]pyridin-3-yl)-
-methanone (P-0006), [1829]
(3-Hydroxy-2-methyl-phenyl)-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-y-
l)-methonone (P-0114), [1830]
(3-Hydroxy-2-methyl-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-meth-
anone (P-0016), [1831]
(2-Fluoro-3-hydroxy-phenyl)-[5-(3-methanesulfonylphenyl)1H-pyrrolo[2,3-b]-
pyridin-3-yl]-methanone (P-0026), [1832]
(2-fluoro-3-hydroxy-phenyl)-(5-pyrimidin-5-yl-1H-pyrrolo[2,3-b]pyridin-3--
yl)-methanone (P-0744), and [1833]
[5-(3-amino-phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]-(2-fluoro-3-hydroxy-ph-
enyl)methanone (P-0004). The following table indicates the boronic
acid (column 2) and the 5-Br azaindole (column 3) used to afford
the compound (column 4). Column 1 provides the compound number and
column 5 the observed mass.
TABLE-US-00023 [1833] MS(ESI) [M + H.sup.+].sup.+ Boronic acid 5-Br
azaindole Compound observed P-0035 ##STR01867## ##STR01868##
##STR01869## 390.2 P-0038 ##STR01870## ##STR01871## ##STR01872##
426.1 P-0034 ##STR01873## ##STR01874## ##STR01875## 376.1 P-0006
##STR01876## ##STR01877## ##STR01878## 330.2 P-0114 ##STR01879##
##STR01880## ##STR01881## 335.1 P-0016 ##STR01882## ##STR01883##
##STR01884## 329.2 P-0026 ##STR01885## ##STR01886## ##STR01887##
411.0 P-0744 ##STR01888## ##STR01889## ##STR01890## 335.1 P-0004
##STR01891## ##STR01892## ##STR01893## 348.1
Example 55
Synthesis of
(2,6-Difluoro-3-hydroxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin--
3-yl)-methanone P-0066
[1834] Compound P-0066 was synthesized in six steps from
2,4-difluoro-phenol 35 as shown in Scheme 12.
##STR01894##
Step 1--Preparation of
tert-butyl-(2,4-difluoro-phenoxy)-dimethyl-silane (36)
[1835] To 2,4-difluoro-phenol (35, 5.00 g, 38.4 mmol) in
N,N-dimethylformamide (100 mL) were added tert-butyldimethylsilyl
chloride (7.00 g, 46.4 mmol) and imidazole (6.28 g, 92.2 mmol). The
reaction was stirred at room temperature overnight. The reaction
was poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and filtrated. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 10% ethyl acetate in hexane.
The appropriate fractions were combined and concentrated to provide
the compound (36, 7.50 g, 80.0%). MS(ESI)
[M+H.sup.+].sup.+=245.1.
Step 2--Preparation of
3-(tert-butyl-dimethyl-silanyloxy)-2,6-difluoro-benzaldehyde
(37)
[1836] To tert-butyl-(2,4-difluoro-phenoxy)-dimethyl-silane (36,
5.90 g, 24.1 mmol) in tetrahydrofuran (100 mL), under an atmosphere
of nitrogen, cooled in a -78.degree. C. acetone/dry ice bath, was
added n-butyllithium (2.50 M in hexane, 10.6 mL, 26.5 mmol) slowly.
The reaction was allowed to stir for 1 hour, then
N,N-dimethylformamide (2.24 mL, 29.0 mmol) was added and the
reaction was allowed to warm to room temperature overnight. The
reaction mixture was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. The compound was
isolated by silica gel column chromatography eluting with 10% ethyl
acetate in hexane to give a colorless oil (37, 5.0 g, 76.0%).
MS(ESI) [M+H.sup.+].sup.+=272.2.
Step 3--Preparation of 2,6-difluoro-3-hydroxy-benzaldehyde (38)
[1837] To
3-(tert-butyl-dimethyl-silanyloxy)-2,6-difluoro-benzaldehyde (37,
3.50 g, 12.9 mmol) in N,N-dimethylformamide (50 mL) was added
tetrabutylammonium fluoride trihydrate (4.40 g, 14.0 mmol). The
reaction was stirred at room temperature for 1 hour. The reaction
mixture was poured into water and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and concentrated to give crude compound 38 that was used in
the next step.
Step 4--Preparation of carbonic acid tert-butyl ester
2,4-difluoro-3-formyl-phenyl ester (39)
[1838] To 2,6-difluoro-3-hydroxy-benzaldehyde (38, 1.90 g, 12.0
mmol) in tetrahydrofuran (60 mL) was added sodium hydride (60% in
mineral oil, 0.58 g, 14.5 mmol) and di-tert-butyldicarbonate (3.90
g, 17.9 mmol). The reaction was stirred at room temperature
overnight. The reaction mixture was poured into water and extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous sodium sulfate and concentrated. The desired
compound was isolated by silica gel column chromatography eluting
with 10% ethyl acetate in hexane to give a colorless oil (39, 2.50
g, 80.8%). MS(ESI) [M+H.sup.+].sup.+=258.1.
Step 5--Preparation of
2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-me-
thyl]-phenol (P-0009)
[1839] To carbonic acid tert-butyl ester
2,4-difluoro-3-formyl-phenyl ester (39, 0.405 g, 15.7 mmol) in
methanol (36 mL), under an atmosphere of nitrogen, was added
5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine (89, 288.0 mg, 14.8 mmol,
prepared as in Example 17, Scheme 32) and potassium hydroxide
(145.0 mg, 25.9 mol). The reaction was stirred at room temperature
overnight. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. The desired compound was
isolated by silica gel column chromatography eluting with 4%
methanol in methylene chloride to give a colorless oil (P-0009,
0.23 g, 44.1%). MS(ESI) [M+H.sup.+].sup.+=354.1.
Step 6--Preparation of
(2,6-difluoro-3-hydroxy-phenyl)-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin--
3-yl)-methanone (P-0066)
[1840] To
2,4-difluoro-3-[hydroxy-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-
-3-yl)-methyl]-phenol (P-0009, 45.0 mg, 1.27 mmol) in
tetrahydrofuran (15 mL) and N,N-dimethylformamide (5 mL) was added
dichlorodicyanoquinone (87 mg, 3.8 mmol). The reaction was stirred
at room temperature for 50 hours. The reaction was poured into
water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous sodium sulfate and
concentrated. The desired compound was isolated by silica gel
column chromatography eluting with 5% methanol in methylene
chloride to give a white solid (P-0066, 7 mg, 15.6%). MS(ESI)
[M+H.sup.+].sup.+=352.1.
(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2,6-difluoro-3-hydroxy-phenyl)-me-
thanone P-1271
##STR01895##
[1841] was prepared following the protocol of Scheme 12,
substituting 5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridine 89 with
5-bromo-1H-pyrrolo[2,3-b]pyridine 67 in Step 5. This was reacted
further (70.0 mg, 0.20 mmol) in methylene chloride (15 mL) by
adding pyridine (0.50 mL, 6.2 mmol), propane-1-sulfonyl chloride
(0.50 g, 3.5 mmol) and 4-dimethylaminopyridine (0.10 g, 0.82 mmol).
The reaction was stirred at room temperature overnight. The
reaction mixture was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. The compound,
propane-1-sulfonic acid
3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluoro-phenyl
ester P-0919
##STR01896##
was isolated by silica gel column chromatography eluting with 10%
ethyl acetate in hexane to give a white solid (15 mg, 16%). MS(ESI)
[M-H.sup.+].sup.-=456.9, 458.9.
Example 56
Synthesis of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-hydroxy-2-methyl-phenyl)-metha-
none P-0055
[1842] Compound P-0055 was synthesized in two steps from
5-Bromo-7-azaindole 67 as shown in Scheme 22.
##STR01897##
Step 1--Preparation of acetic acid
3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-methyl-phenyl
ester (84)
[1843] To aluminum trichloride (6.14 g, 0.046 mol) in methylene
chloride (26 mL) under an atmosphere of nitrogen was added
5-Bromo-7-azaindole (67, 1.05 g, 5.31 mmol) dissolved in methylene
dichloride (20.0 mL). The reaction was stirred at room temperature
for 65.0 minutes, then acetic acid 3-chlorocarbonyl-2-methyl-phenyl
ester (1.25 g, 5.88 mmol) in methylene dichloride (7.0 mL) was
added. The reaction was stirred for 2 hours at room temperature,
poured into water and extracted with ethyl acetate. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and concentrated to give crude compound 84 that was used directly
in the next step.
Step 2--Preparation of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-hydroxy-2-methyl-phenyl)-metha-
none (P-0055)
[1844] A solution of acetic acid
3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2-methyl-phenyl
ester (84, 25.0 mg, 0.067 mmol) in 6 N HCl (4.0 mL) was heated with
a CEM Discover microwave instrument at 110.degree. C. for 10 min.
The reaction mixture was poured into water, neutralized with 1 M
aqueous potassium carbonate, and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and concentrated. The desired compound was isolated by
silica gel column chromatography eluting with 10% ethyl acetate in
hexane to give a colorless oil (P-0055, 4.7 mg, 21%). MS(ESI)
[M-H.sup.+].sup.-=329.0, 331.0.
Example 57
Synthesis of
2-Fluoro-3-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenol
P-0052
[1845] Compound P-0052 was synthesized in five steps from
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phenyl)-metha-
none P-0265 as shown in Scheme 23.
##STR01898##
Step 1--Preparation of
5-bromo-3-(2-fluoro-3-methoxy-benzoyl)-pyrrolo[2,3-b]pyridine-1-carboxyli-
c acid tert-butyl ester (86)
[1846] To
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(2-fluoro-3-methoxy-phen-
yl)-methanone (P-0265, 600.0 mg, 1.72 mmol, prepared as described
in Example 1 or 53) in tetrahydrofuran (20 mL) was added sodium
hydride (60% dispersion, 103 mg, 2.58 mmol) under an atmosphere of
nitrogen. After 15 minutes, di-tert-butyldicarbonate (648 mg, 2.97
mmol) was added. The reaction mixture was stirred at room
temperature for 1 hour, poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. The desired compound was
isolated by silica gel column chromatography eluting with 10% ethyl
acetate in hexane to give a colorless oil (86, 0.64 g, 82.3%).
Step 2--Preparation of
3-(2-fluoro-3-methoxy-benzoyl)-5-thiophen-2-yl-pyrrolo[2,3-b]pyridine-1-c-
arboxylic acid tert-butyl ester (87)
[1847] To
5-bromo-3-(2-fluoro-3-methoxy-benzoyl)-pyrrolo[2,3-b]pyridine-1--
carboxylic acid tert-butyl ester (86, 350.0 mg, 0.78 mmol) in
tetrahydrofuran (30 mL) and water (10 mL) were added 2-thiophene
boronic acid (190 mg, 1.49 mmol),
tetrakis(triphenyl-phosphine)palladium(0) (40.0 mg, 0.0346 mmol)
and potassium carbonate (780 mg, 5.64 mmol) under an atmosphere of
nitrogen. The reaction mixture was refluxed for 2 hours. The
reaction mixture was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated to give crude compound
(87, 0.30 g, 85%).
Step 3--Preparation of
(2-fluoro-3-hydroxy-phenyl)-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-y-
l)-methanone (P-0521)
[1848] To
3-(2-fluoro-3-methoxy-benzoyl)-5-thiophen-2-yl-pyrrolo[2,3-b]pyr-
idine-1-carboxylic acid tert-butyl ester (87, 200 mg, 0.44 mmol) in
methylene chloride (10 mL), under an atmosphere of nitrogen, cooled
in a -78.degree. C. acetone/dry ice bath, was added boron
tribromide (1.0 M in hexane, 2.0 mL). The reaction mixture was
allowed to warm to room temperature and stirred overnight. The
reaction was quenched with methanol (3.0 mL), poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and concentrated. The
desired compound was isolated by silica gel column chromatography
eluting with 5% methanol in methylene chloride to give a white
solid (P-0521, 119 mg, 82.6%). MS(ESI)[M-H.sup.+].sup.-=337.
Step 4--Preparation of
2-Fluoro-3-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenol
(P-0052)
[1849] To
(2-fluoro-3-hydroxy-phenyl)-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]py-
ridin-3-yl)-methanone (P-0521, 50.0 mg, 0.15 mmol) in diethylene
glycol (5.0 mL) were added hydrazine hydrate (1.0 mL, 0.020 mol)
and potassium hydroxide (200.0 mg, 3.56 mmol). The reaction mixture
was heated in a CEM Discovery microwave instrument at 180.degree.
C. for 20 minutes, poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over
anhydrous sodium sulfate and concentrated. The desired compound was
isolated by silica gel column chromatography eluting with 5%
methanol in methylene chloride to give a white solid (P-0052, 10
mg, 20%). MS(ESI) [M+H.sup.+].sup.+=325.2.
Example 58
Synthesis of (2,4-Difluoro-3-formyl-phenyl)-carbamic acid
tert-butyl ester 124.
[1850] (2,4-Difluoro-3-formyl-phenyl)-carbamic acid tert-butyl
ester 124 was synthesized in two steps from
2,4-Difluoro-phenylamine 42 as shown in Scheme 25.
##STR01899##
Step 1--Preparation of (2,4-Difluoro-phenyl)-carbamic acid
tert-butyl ester (123)
[1851] To 2,4-difluoro-phenylamine (42, 5.0 g, 0.039 mol) in
methylene chloride (100 mL) were added di-tert-butyldicarbonate
(21.1 g, 0.0968 mol), triethylamine (16 mL, 0.12 mol) and
4-dimethylaminopyridine (0.2 g, 0.002 mol). The reaction was
stirred at room temperature overnight, poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and purified by
silica gel chromatography, eluting with 20% ethyl acetate in hexane
to provide compound 123.
Step 2--Preparation of (2,4-Difluoro-3-formyl-phenyl)-carbamic acid
tert-butyl ester (124)
[1852] To 2,4-difluoro-phenyl)-carbamic acid tert-butyl ester (123,
1.41 g, 6.17 mmol) in tetrahydrofuran (60.0 mL) under an atmosphere
of nitrogen at 78.degree. C. were added n-butyllithium (2.50 M in
hexane, 2.59 mL) for 30 min. Lithium diisopropylamide (2.0 M in
hexane, 3.4 mL) was added to the reaction. After 35 min,
N,N-dimethylformamide (1.05 mL, 0.0136 mol) was added to the
reaction mixture. The reaction was allowed to cool to room
temperature overnight. The organic layer was washed with brine,
dried over sodium sulfate, concentrated and purified by silica gel
chromatography, eluting with 20% ethyl acetate in hexane to provide
compound 124.
Example 59
Synthesis of
1-[4-(1H-Pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-3-(3-trifluoromethyl-p-
henyl)-urea P-0262
[1853] Compound P-0262 was synthesized in seven steps from
1H-Pyrrolo[2,3-b]pyridine 94 as shown in Scheme 34.
##STR01900## ##STR01901##
Step 1--Preparation of
Dimethyl-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-amine (109)
[1854] 1H-Pyrrolo[2,3-b]pyridine (94, 5.0 g, 42 mmol),
dimethylamine hydrochloride (3.8 g, 46 mmol), formaldehyde (1.4 g,
46 mmol) and isopropyl alcohol (220.0 mL) were combined in a
pressure tube. The reaction mixture was stirred at 20.degree. C.
for 12 hours and then refluxed for 2 hours. The clear solution was
evaporated to dryness in vacuo. Water (40 mL) and concentrated
hydrochloric acid (4 mL) were added to the residue. The mixture was
extracted with ether and then made strongly basic with potassium
carbonate. The basified aqueous layer was extracted with ethyl
acetate. The organic layer was washed with brine, dried over sodium
sulfate, concentrated and purified by silica gel chromatography to
give the compound (109, 5.0 g, 68%).
Step 2--Preparation of
Dimethyl-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-amin-
e (110)
[1855] Dimethyl-(1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-amine 109
(5.0 g, 28 mmol), was dissolved in N,N-dimethylformamide (80.0 mL)
and sodium hydride (60% dispersion in mineral oil, 1.25 g, 31.4
mmol) was added, followed by triisopropylsilyl chloride (6.3 mL,
30.0 mmol). The reaction was stirred at room temperature for 12
hours, then poured into water and extracted with ethyl acetate. The
organic portion was washed with brine, dried over sodium sulfate,
concentrated and purified with silica gel chromatography to give
the compound (110, 6.5 g, 70%).
Step 3--Preparation of
3-(4-Bromo-benzyl)-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(111)
[1856] 1-bromo-4-iodobenzene (2.1 g, 7.4 mmol) was dissolved in
tetrahydrofuran (15.0 mL) under an atmosphere of nitrogen at
-30.degree. C. Isopropylmagnesium chloride in tetrahydrofuran (2.0
M, 4.0 mL) was added. The reaction mixture was stirred below
-20.degree. C. for 1 hour to provide a solution of
1-bromo-4-benzene magnesium chloride.
[1857]
Dimethyl-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl-
)-amine 110 (1.25 g, 3.77 mmol) was added to toluene (20.0 mL)
under an atmosphere of nitrogen at room temperature. Isopropyl
chloroformate in toluene (1.0 M, 4.0 mL) was added. The reaction
mixture was stirred at room temperature for 3 hours to provide a
solution of
3-chloromethyl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine.
[1858] CuCN.2LiCl in tetrahydrofuran (0.7 M, 10 mL) was added to
the solution of 1-bromo-4-benzene magnesium chloride at -20.degree.
C. The reaction mixture was allowed to warm to room temperature for
10 minutes. Trimethyl phosphite (1.8 mL, 0.015 mol) was added to
the reaction. After 5 minutes, the solution of
3-chloromethyl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine was
added to the reaction at room temperature. The reaction was stirred
for 12 hours at room temperature. The reaction mixture was poured
into water and extracted with ethyl acetate. The organic layer was
washed with brine, dried over sodium sulfate, concentrated and
purified with silica gel chromatography to give the compound (111,
850 mg, 51%).
Step 4--Preparation of
Benzyl-[4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phe-
nyl]-amine (112)
[1859]
3-(4-Bromo-benzyl)-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine
(111, 330.0 mg, 0.74 mmol), benzylamine (160 mg, 1.5 mmol),
2-[di(tert-butyl)phosphino]-1,1'-biphenyl (15.0 mg, 0.05 mmol),
tris(dibenzylideneacetone)dipalladium(0) (15.0 mg, 0.016 mmol),
potassium tert-butoxide (100 mg, 0.89 mmol) and toluene (10.0 mL)
were combined under an atmosphere of nitrogen. The reaction mixture
was refluxed overnight, then poured into water and extracted with
ethyl acetate. The organic layer was washed with brine, dried over
sodium sulfate, concentrated and purified with silica gel
chromatography to give the compound (112, 260 mg, 75%).
Step 5--Preparation of
4-(1-Triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenylamine
(113)
[1860]
Benzyl-[4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethy-
l)-phenyl]-amine (112, 210.0 mg, 0.447 mmol), methanol (15.0 mL)
and palladium hydroxide (30.0 mg, 0.214 mmol) were combined. The
reaction mixture was hydrogenated under under an atmosphere of
hydrogen (1 atm) at room temperature for 2 hours. Filtration and
concentration gave the compound (113, 154 mg, 91%).
Step 6--Preparation of
1-(3-Trifluoromethyl-phenyl)-3-[4-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b-
]pyridin-3-ylmethyl)-phenyl]-urea (114)
[1861]
4-(1-Triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-pheny-
lamine (113, 70.0 mg, 0.184 mmol) was dissolved in tetrahydrofuran
(12 mL) and 3-trifluoromethyl-phenyl isocyanate (41 mg, 0.22 mmol)
and triethylamine (28 mg, 0.28 mmol) were added under an atmosphere
of nitrogen. The reaction mixture was stirred at room temperature
overnight, then poured into water and extracted with ethyl acetate.
The organic layer was washed with brine, dried over sodium sulfate,
concentrated to give crude compound 114 used in the next step.
Step 7--Preparation of
1-[4-(1H-Pyrrolo[2,3-b]pyridin-3-ylmethyl)-phenyl]-3-(3-trifluoromethyl-p-
henyl)-urea (P-0262)
[1862]
1-(3-Trifluoromethyl-phenyl)-3-[4-(1-triisopropylsilanyl-1H-pyrrolo-
[2,3-b]pyridin-3-ylmethyl)-phenyl]-urea (114, 85.0 mg, 0.150 mmol)
was dissolved in tetrahydrofuran (10.0 mL) and
tetra-n-butylammonium fluoride (52.2 mg, 0.200 mmol) was added.
After 10 minutes, the reaction mixture was poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and purified with by
silica gel chromatography to give the compound (P-0262, 23 mg, 38%
over steps 6 & 7). MS(ESI)[M+H.sup.+].sup.+=411.2
Example 60
Synthesis of
(4-Hydroxy-phenyl)-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methan-
one P-0024
[1863] Compound P-0024 was synthesized in four steps from
5-bromo-7-azaindole 67 as shown in Scheme 35.
##STR01902##
Step 1--Preparation of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(4-methoxy-phenyl)-methanone
(116)
[1864] Aluminum trichloride (2.7 g, 20.0 mmol) and methylene
chloride (30 mL) were combined under an atmosphere of nitrogen and
5-bromo-7-azaindole (67, 400.0 mg, 2.30 mmol) in methylene
dichloride (20 mL) was added. The reaction was stirred at room
temperature for 70 minutes. 4-Methoxybenzoic acid chloride (0.38 g,
2.2 mmol) (prepared from the corresponding carboxylic acid using
the protocol described in Example 1, Scheme 13, Step 4) in
methylene dichloride (7.0 mL) was added. The reaction was stirred
at room temperature for 2 hours, then poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, and concentrated to give the
compound (116, 250 mg, 34%).
Step 2--Preparation of
5-Bromo-3-(4-methoxy-benzoyl)-pyrrolo[2,3-b]pyridine-1-carboxylic
acid tert-butyl ester (117)
[1865]
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(4-methoxy-phenyl)-methanon-
e (116, 220.0 mg, 0.66 mmol) was dissolved in tetrahydrofuran (15.0
mL) and sodium hydride (60% dispersion, 29 mg, 0.73 mmol) was added
under an atmosphere of nitrogen. The reaction was stirred for 15
minutes. Di-tert-butyldicarbonate (220 mg, 1.0 mmol) was added. The
reaction was stirred for one hour, then poured into water and
extracted with ethyl acetate. The organic portion was washed with
brine, dried over sodium sulfate and concentrated to give compound
117.
Step 3--Preparation of
3-(4-Methoxy-benzoyl)-5-thiophen-2-yl-pyrrolo[2,3-b]pyridine-1-carboxylic
acid tert-butyl ester (118)
[1866]
5-Bromo-3-(4-methoxy-benzoyl)-pyrrolo[2,3-b]pyridine-1-carboxylic
acid tert-butyl ester (117, 100.0 mg, 0.23 mmol),
thiophene-2-boronic acid (59 mg, 0.46 mmol),
tetrakis(triphenylphosphine)palladium(0) (20.0 mg, 0.017 mmol),
potassium carbonate (80 mg, 0.58 mmol), tetrahydrofuran (15.0 mL)
and water (5.0 mL) were combined under an atmosphere of nitrogen.
The reaction mixture was refluxed overnight, then poured into water
and extracted with ethyl acetate. The organic layer was washed with
brine, dried over sodium sulfate, concentrated and purified with
silica gel chromatography to give compound 118 (100 mg).
Step 4--Preparation of
(4-Hydroxy-phenyl)-(5-thiophen-2-yl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methan-
one (P-0024)
[1867]
3-(4-Methoxy-benzoyl)-5-thiophen-2-yl-pyrrolo[2,3-b]pyridine-1-carb-
oxylic acid tert-butyl ester (118, 100.0 mg, 0.23 mmol) and
methylene chloride (15.0 mL) were combined under an atmosphere of
nitrogen. Boron tribromide (1.0 M in methylene chloride, 3.0 mL)
was added. The reaction was stirred at room temperature overnight.
Methanol (2.0 mL) was added into the the reaction mixture. The
reaction was concentrated to remove solvents. The residue was
extracted with water and ethyl acetate. The organic layer was
washed with brine, dried over sodium sulfate, concentrated and
purified with silica gel chromatography. The product was washed
with methanol to provide compound (P-0024, 25.0 mg, 34%). MS(ESI)
[M+H.sup.+].sup.+=321.
Example 61
Synthesis of
4-Benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine-2-carboxylic acid ethyl
ester P-1248
[1868] Compound P-1248 was synthesized in four steps from
4-chloro-7-azaindole 119 as shown in Scheme 36.
##STR01903##
Step 1--Preparation of 4-Benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine
(120)
[1869] 4-Chloro-7-azaindole (119, 100 mg, 0.66 mmol, prepared as
described in Example 70, Scheme 81), 2-Benzofuranboronic acid (212
mg, 1.31 mmol) and Potassium fluoride (127 mg, 2.18 mmol) were
stirred in 3 mL of anhydrous dioxane.
Tris(dibenzylideneacetone)dipalladium(0) chloroform adduct (10.2
mg, 9.83E-3 mmol) and tri t-butylphosphine (6.6 mg, 0.033 mol) were
added and the suspension was stirred at 100.degree. C. for 4 hours
in a pressure tube under inert atmosphere. The reaction mixture was
cooled to room temperature and was adsorbed onto silica gel and
purified by chromatography to provide the compound (120, 56 mg,
36%). MS(ESI) [M+H.sup.+].sup.+=235.20.
Step 2--Preparation of
1-Benzenesulfonyl-4-benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine
(121)
[1870] Into a Round bottom flask
4-benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine (120, 100 mg, 0.43
mmol) was suspended in methylene chloride (10 mL) and 50% KOH
solution (5 mL) was added. Tetrabutylammonium hydrogen sulfate (10
mg, 0.03 mmol) and benzenesulfonyl chloride (81.4 mg, 0.46 mmol)
were added. The resulting yellow solution was stirred at ambient
temperature for 2 hours. The reaction mixture was partitioned
between ethyl acetate and water and the organic layer was separated
and washed with water and brine then dried over MgSO.sub.4,
filtered and concentrated under reduced pressure to provide
compound 121. MS(ESI) [M+H.sup.+].sup.+=375.29.
Step
3--1-Benzenesulfonyl-4-benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine-2-ca-
rboxylic acid ethyl ester (122)
[1871] To a chilled (-78.degree. C. acetone/dry ice bath) solution
of 1-benzenesulfonyl-4-benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine
(121, 66.0 mg, 0,18 mmol) in tetrahydrofuran (5.0 mL) under an
atmosphere of nitrogen, n-butylithium (1.6 M in tetrahydrofuran,
0.14 mL, 0.23 mmol) was added dropwise. The reaction mixture was
stirred for 30 minutes and ethyl chloroformate (0.02 mL, 0.19 mmol)
was added. The reaction mixture was stirred for 1 hour while
allowing it to reach room temperature, poured into water and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated under reduced pressure and purified by
silica gel column chromatography, eluting with 20% ethyl acetate in
hexane, to give light yellow solid (122, 0.015 g, 19%).
MS(ESI)[M+H.sup.+].sup.+=447.3.
Step 4--4-Benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine-2-carboxylic
acid ethyl ester (P-1248)
[1872] To a solution of
1-benzenesulfonyl-4-benzofuran-2-yl-1H-pyrrolo[2,3-b]pyridine-2-carboxyli-
c acid ethyl ester (122, 14.0 mg, 0.03 mmol) in tetrahydrofuran
(3.0 mL), was added tetra N-butylammonium fluoride (26.0 mg, 0.10
mmol). The reaction was stirred for 48 hours at room temperature,
then poured into water and extracted with ethyl acetate. The
organic layer was washed with brine, dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced
pressure and purified by silica gel column chromatography, eluting
with 50% Ethyl acetate in hexane, to give light yellow solid
(P-1248, 3.0 mg, 36%). MS(ESI) [M+H.sup.+].sup.+=307.2.
Example 62
Synthesis of N-phenyl-1H-pyrrolo[2,3-b]pyridin-6-amine P-1350 and
Related Compounds
[1873] Compound P-1350 was synthesized in five steps from
1H-pyrrolo[2,3-b]pyridine 94 as shown in Scheme 40.
##STR01904##
Step 1--Preparation of 1H-pyrrolo[2,3-b]pyridine-N-oxide (585)
[1874] To 1H-pyrrolo[2,3-b]pyridine (94, 3.0 g, 25.3 mmol)
dissolved in 175 mL of diethyl ether was added m-CPBA (1.5 equiv)
in portions over 30 minutes with vigorous stirring. The solution
turned yellow, and precipitates formed. After two hours the solid
was collected, washed with 2.times.50 mL of ether, and
recrystallized from acetone:ether. Yield was approximately 125% due
to contaminating acid. This crude material was carried through to
the next step.
Step 2--(6-bromo-1H-pyrrolo[2,3-b]pyridin-1-yl)(phenyl)methanone
(586)
[1875] 1H-pyrrolo[2,3-b]pyridine-N-oxide (585, 500 mg, 3.72 mmol)
was dissolved in 40 mL of dry benzene. In a separate dry flask,
benzoyl bromide (2.5 equiv) and 1,1,1,3,3,3-hexamethyldisilazane
(1.0 equiv) were combined in 20 mL of dry benzene. The bromide
solution was added in 5 mL aliquots over 30 minutes to the reaction
flask. The reaction was stirred at ambient temperature for two
hours. It was then washed with 3.times.30 mL NaHCO.sub.3 (aq.,
satd.) and 1.times.30 mL brine. The organic layer was dried over
sodium sulfate and evaporated. The crude material was taken
directly to the next step without further purification.
Step 3--Preparation of 6-bromo-1H-pyrrolo[2,3-b]pyridine (587)
[1876] (6-bromo-1H-pyrrolo[2,3-b]pyridin-1-yl)(phenyl)methanone,
586 was dissolved in 20 mL of dioxane and 20 mL of 2M KOH (aq).
This was stirred at ambient temperature until analysis indicated
all of the starting material had been consumed (2 to 4 hours). The
reaction was diluted with 50 mL of ethyl acetate and washed with
2.times.25 mL of NaHCO.sub.3 (aq. satd.) and 25 mL of brine. The
organic layer was dried with sodium sulfate, evaporated and
purified by column chromatography. Combined steps 2 and 3 gave
approximately 65% overall yield.
Step 4--Preparation of
6-bromo-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine (588)
[1877] 6-bromo-1H-pyrrolo[2,3-b]pyridine (587, 275 mg, 1.39 mmol)
was dissolved in 4 mL of dry dioxane. DIEA (3 equiv) and TIPS-OTf
(2.5 equiv) were added and the reaction was stirred at 50.degree.
C. overnight. The reaction was then diluted with 20 mL of ethyl
acetate and washed 2 times with 10 mL NaHCO.sub.3 (aq., 5%) and
once with 10 mL brine. The organic fraction was dried over
MgSO.sub.4, evaporated and diluted with 5.5 mL of dry toluene 10 mg
bromide per 0.2 mL of solution) to use directly in the next
reaction step.
Step 5--Preparation of N-phenyl-1H-pyrrolo[2,3-b]pyridin-6-amine
(P-1350)
[1878] A 1 dram vial was charged with aniline (2-3 equiv), and
0.200 mL of the 588 stock solution of bromide in toluene was added.
A catalyst stock solution containing 3 mmol Pd(OAc).sub.2, 3 mmol
biphenyl-2-yl-di-tert-butyl-phosphane and 15 mL of toluene was
prepared and 0.050 mL of the catalyst solution was added to the
reaction. An excess of NaOtBu was added as a solid to the reaction.
The vial was placed in an 80.degree. C. oven for 60 minutes (shaken
several times over the hour). After cooling, the reaction was
neutralized with 0.100 mL of TFA. After 30 minutes the sample was
evaporated and resolvated in 0.300 mL of DMSO. The desired compound
was isolated by preparative HPLC/MS. MS(ESI)
[M+H.sup.+].sup.+=210.4.
[1879] The following compounds were prepared following the protocol
of scheme 40, substituting aniline with a suitable amine is Step 5:
[1880] Cyclohexyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1290),
[1881] Benzyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1291),
[1882] Cyclopropylmethyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1292), [1883]
(3-Methoxy-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1293),
[1884]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(3-trifluoromethyl-benzyl)-amine
(P-1294), [1885]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(2-trifluoromethyl-benzyl)-amine
(P-1295), [1886]
Cyclohexylmethyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1296),
[1887]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(4-trifluoromethoxy-benzyl)-amine
(P-1297), [1888]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(3-trifluoromethoxy-benzyl)-amine
(P-1298), [1889]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(2-trifluoromethoxy-benzyl)-amine
(P-1299), [1890]
Pyridin-2-ylmethyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1300),
[1891]
(4-Methanesulfonyl-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-13 01), [1892]
(4-Methoxy-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1302),
[1893] Ethyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1303), [1894]
(3-Chloro-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1304),
[1895] (4-Methyl-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1305), [1896]
(1-Methyl-piperidin-4-yl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1306), [1897]
Pyridin-3-ylmethyl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1307),
[1898]
[4-(Morpholine-4-sulfonyl)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1308), [1899]
(4-Methanesulfonyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1309), [1900]
(2-Chloro-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1310),
[1901]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(tetrahydro-pyran-4-yl)-amine
(P-1311), [1902]
(4-Chloro-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1312),
[1903] (3-Methyl-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1313), [1904]
[3-(Morpholine-4-sulfonyl)-phenyl]-(1H-pyrrolo[2,3-b]pyridin-6-yl)-
-amine (P-1314), [1905]
(3-Methanesulfonyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1315), [1906] Pyridin-3-yl-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1351), [1907]
(2-Methoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1352),
[1908]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(3-trifluoromethyl-phenyl)-amine
(P-1353), [1909]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(4-trifluoromethoxy-phenyl)-amine
(P-1354), [1910]
(4-Methoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1355),
[1911]
N,N-Dimethyl-N,-(1H-pyrrolo[2,3-b]pyridin-6-yl)-benzene-1,4-diamine
(P-1356), [1912]
(3-Methoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1357),
[1913]
(4-Morpholin-4-yl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1358), [1914]
(4-Piperidin-1-yl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1359), [1915]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(3-trifluoromethoxy-phenyl)-amine
(P-1360), [1916] (1H-Pyrrolo[2,3-b]pyridin-6-yl)-p-tolyl-amine
(P-1361), [1917]
(3-tert-Butyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1362), [1918]
(3-Dimethylamino-benzyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1363), [1919]
(3,5-Dichloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1364), [1920]
(1H-Pyrrolo[2,3-b]pyridin-6-yl)-(4-trifluoromethyl-benzyl)-amine
(P-1371), [1921]
N,N-Dimethyl-N'-(1H-pyrrolo[2,3-b]pyridin-6-yl)-benzene-1,3-diamine
(P-1372), [1922]
(3-Chloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1373),
[1923] (4-Chloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1374), [1924]
(2-Chloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine (P-1375),
[1925]
(5-Methyl-isoxazol-3-yl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1376), [1926]
(2-Morpholin-4-yl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1377), and [1927]
(2-Methanesulfonyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-6-yl)-amine
(P-1378). The following table indicates the amine (Column 2) that
is substituted in place of the aniline in Step 5 to afford the
compound (Column 3). Column 1 provides the compound number and
Column 4 the observed mass.
TABLE-US-00024 [1927] Com- MS(ESI) pound [M + H.sup.+].sup.+ number
Amine Compound structure observed P-1290 ##STR01905## ##STR01906##
216.3 P-1291 ##STR01907## ##STR01908## 224.3 P-1292 ##STR01909##
##STR01910## 188.2 P-1293 ##STR01911## ##STR01912## 254.3 P-1294
##STR01913## ##STR01914## 291.9 P-1295 ##STR01915## ##STR01916##
291.9 P-1296 ##STR01917## ##STR01918## 230.3 P-1297 ##STR01919##
##STR01920## 308.3 P-1298 ##STR01921## ##STR01922## 308.3 P-1299
##STR01923## ##STR01924## 308.3 P-1300 ##STR01925## ##STR01926##
225.1 P-1301 ##STR01927## ##STR01928## 302.3 P-1302 ##STR01929##
##STR01930## 254.3 P-1303 ##STR01931## ##STR01932## 162.2 P-1304
##STR01933## ##STR01934## 258.3 P-1305 ##STR01935## ##STR01936##
238.3 P-1306 ##STR01937## ##STR01938## 231.1 P-1307 ##STR01939##
##STR01940## 225.1 P-1308 ##STR01941## ##STR01942## 359.1 P-1309
##STR01943## ##STR01944## 287.9 P-1310 ##STR01945## ##STR01946##
258.3 P-1311 ##STR01947## ##STR01948## 218.3 P-1312 ##STR01949##
##STR01950## 258.3 P-1313 ##STR01951## ##STR01952## 238.3 P-1314
##STR01953## ##STR01954## 359.1 P-1315 ##STR01955## ##STR01956##
287.9 P-1351 ##STR01957## ##STR01958## 211.0 P-1352 ##STR01959##
##STR01960## 240.3 P-1353 ##STR01961## ##STR01962## 278.3 P-1354
##STR01963## ##STR01964## 293.9 P-1355 ##STR01965## ##STR01966##
240.3 P-1356 ##STR01967## ##STR01968## 253.1 P-1357 ##STR01969##
##STR01970## 240.3 P-1358 ##STR01971## ##STR01972## 295.1 P-1359
##STR01973## ##STR01974## 293.1 P-1360 ##STR01975## ##STR01976##
293.9 P-1361 ##STR01977## ##STR01978## 224.3 P-1362 ##STR01979##
##STR01980## 266.3 P-1363 ##STR01981## ##STR01982## 267.1 P-1364
##STR01983## ##STR01984## 278.3 P-1371 ##STR01985## ##STR01986##
291.9 P-1372 ##STR01987## ##STR01988## 253.1 P-1373 ##STR01989##
##STR01990## 244.3 P-1374 ##STR01991## ##STR01992## 244.3 P-1375
##STR01993## ##STR01994## 244.3 P-1376 ##STR01995## ##STR01996##
215.0 P-1377 ##STR01997## ##STR01998## 295.1 P-1378 ##STR01999##
##STR02000## 287.9
Example 63
Synthesis of
2-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-ph-
enoxy]-ethanol P-1395 and Related Compounds
[1928] Compound P-1395 was synthesized in four steps from
4-Chloro-2-fluoro-phenol 521 as shown in Scheme 42.
##STR02001##
Step 7--Preparation of
4-Chloro-2-fluoro-1-(2,2,2-trifluoro-ethoxy)-benzene (522)
[1929] To 4-Chloro-2-fluoro-phenol (521, 5.0 g, 0.034 mol) in
methanol (50.0 mL) was added potassium fluoride (2,2 g, 0.038 mol).
The solvent was removed. The resulting salt was added to
N,N-dimethylformaldehyde (25 mL), followed by adding
1,1,1-trifluoro-2-iodo-ethane (8.60 g, 40.9 mmol). The reaction was
stirred at 50.degree. C. overnight. The reaction was poured into
water and extracted with ethyl acetate. The organic layer was dried
over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give colorless oil
(522, 2.0 g, 26%).
Step 2--Preparation of
6-Chloro-2-fluoro-3-(2,2,2-trifluoro-ethoxy)-benzaldehyde (523)
[1930] To 4-Chloro-2-fluoro-1-(2,2,2-trifluoro-ethoxy)-benzene
(522, 0.80 g, 3.5 mmol) in THF (20 mL), cooled in dry ice/acetone
bath and under an atmosphere of nitrogen, was slowly added
n-butyllithium (1.60 M in Hexane, 2.30 mL). After an hour,
N,N-dimethylformamide (0.298 mL, 3.85 mmol) was added to the
reaction. After 30 minutes, the reaction was allowed to reach room
temperature and was stirred for 10 minutes. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give the compound (523,
450 mg, 50% ).
Step 3--Preparation of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-[6-chloro-2-fluoro-3-(2,2,2-trifl-
uoro-ethoxy)-phenyl]-methanol (524)
[1931] To 5-bromo-7-azaindole (67, 291 mg, 1.48 mmol) in methanol
(22 mL) were added
6-Chloro-2-fluoro-3-(2,2,2-trifluoro-ethoxy)-benzaldehyde (523,
400.0 mg, 1.6 mmol) and potassium hydroxide (1.49 g, 26.6 mmol)
under an atmosphere of nitrogen. The reaction was stirred at room
temperature for 48 hours. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 25% ethyl acetate in hexane to give the compound (524,
300 mg, 42%). MS (ESI) [M+H.sup.+].sup.+=453.1, 455.1.
Step 4--Preparation of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-[6-chloro-2-fluoro-3-(2,2,2-trifl-
uoro-ethoxy)-phenyl]-methanol (P-1393)
[1932] To
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-[6-chloro-2-fluoro-3-(2,-
2,2-trifluoro-ethoxy)-phenyl]-methanol (524, 140.0 mg, 0.31 mmol)
in trahydrofuran (6.0 mL) was added Dess-Martin periodinane (157
mg, 0.37 mmol). The reaction was stirred at room temperature for 10
minutes. The reaction was poured into water and extracted with
ethyl acetate. The organic layer was dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting with 20% ethyl acetate in
hexane to give the compound (P-1393, 100 mg, 72%). MS (ESI)
[M-H+]-=448.9, 450.9.
Step 5--Preparation of
[6-Chloro-2-fluoro-3-(2,2,2-trifluoro-ethoxy)-phenyl]-(5-pyridin-3-yl-1H--
pyrrolo[2,3-b]pyridin-3-yl)-methanone (P-1395)
[1933] To
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-[6-chloro-2-fluoro-3-(2,-
2,2-trifluoro-ethoxy)-phenyl]-methanol (P-1393, 53.0 mg, 0.12 mmol)
in acetonitrile (4.0 mL) was added
Tetrakis(triphenylphosphine)palladium(0) (5.0 mg, 0.0043 mmol),
3-pyridylboronic acid (15.1 mg, 0.12 mmol) and 1 M potassium
carbonate solution (1.5 mL). The reaction was microwaved (300
watts) at 160.degree. C. for 7 minutes. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 60% ethyl acetate in hexane to give the compound
(P-1395, 3.8 mg, 53%) as light yellow solid. MS (ESI)
[M+H.sup.+].sup.+=450.2.
[2,6-Difluoro-3-(2-methoxy-ethoxy)-phenyl]-(5-pyridin-3-yl-1H-pyrrolo[2,3--
b]pyridin-3-yl)-methanone P-1456
##STR02002##
[1934] was prepared following the protocol of Scheme 42,
substituting 4-Chloro-2-fluoro-phenol 521 with 2,4-difluoro-phenol
and 1,1,1-Trifluoro-2-iodo-ethane with 1-Bromo-2-methoxy-ethane in
Step 1. MS (ESI) [M+H.sup.+].sup.+=410.2.
N-(3-{3-[2,6-Difluoro-3-(2-methoxy-ethoxy)-benzoyl]-1H-pyrrolo[2,3-b]pyrid-
in-5-yl}-phenyl)-methanesulfonamide P-1472
##STR02003##
[1935] was prepared following the protocol of Scheme 42,
substituting 4-Chloro-2-fluoro-phenol 521 with 2,4-difluoro-phenol
and 1,1,1-Trifluoro-2-iodo-ethane with 1-Bromo-2-methoxy-ethane in
Step 1, and substituting pyridine-3-boronic acid with
[(3-methylsulfonyl)aminophenyl]-boronic acid in step 4. MS (ESI)
[M+H.sup.+].sup.+=502.2.
Example 64
Synthesis of
(3,5-Bis-difluoromethoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone P-1521
[1936] Compound P-1521 was synthesized in four steps from
3,5-Dihydroxy-benzaldehyde 528 as shown in Scheme 44.
##STR02004##
Step 1--Preparation of 3,5-Bis-difluoromethoxy-benzaldehyde
(529)
[1937] To 3,5-Dihydroxy-benzaldehyde (528, 1.50 g, 10.9 mmol) in
N,N-Dimethylformamide (100 mL) were added sodium
chlorodifluoroacetate (5.90 g, 0.0387 mol), potassium carbonate
(6.20 g, 0.0448 mol) and water (10 mL) under an atmosphere of
nitrogen. The reaction was stirred at 100.degree. C. overnight. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was dried over anhydrous sodium sulfate and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 20% ethyl acetate in hexane to
give the compound as colorless oil (529, 50 mg, 2%).
Step 2--Preparation of
(3,5-Bis-difluoromethoxy-phenyl)-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanol (530)
[1938] To 5-bromo-7-azaindole (67, 86.4 mg, 0.000438 mol) in
methanol (10.0 mL) were added 3,5-bis-difluoromethoxy-benzaldehyde
(529, 110.0 mg, 0.46 mmol) and potassium hydroxide (200.0 mg, 3.6
mmol) under an atmosphere of nitrogen. The reaction was stirred at
room temperature overnight. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 20% ethyl acetate in hexane to give the compound as a
white solid (530, 70 mg, 35%). MS (ESI) [M+H.sup.+].sup.+=435.1,
437.1.
Step 3--Preparation of
(3,5-Bis-difluoromethoxy-phenyl)-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)--
methanone (P-1519)
[1939] To
(3,5-Bis-difluoromethoxy-phenyl)-(5-bromo-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanol (530, 70.0 mg, 0.16 mmol) in tetrahydrofuran (6.0
mL) was added Dess-Martin periodinane (75.0 mg, 0.18 mmol). The
reaction was stirred at room temperature for 10 minutes. The
reaction mixture was concentrated with silica gel and purified with
silica gel column chromatography eluting with 30% ethyl acetate in
hexane to give the compound as a white solid (P-1519, 65.0 mg,
93%.). MS (ESI) [M-H+]-=431.0, 433.0.
Step 4--Preparation of
(3,5-Bis-difluoromethoxy-phenyl)-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-
-methanone (P-1521)
[1940] To
(3,5-Bis-difluoromethoxy-phenyl)-(5-bromo-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (P-1519, 29.0 mg, 0.067 mmol) in acetonitrile
(4.0 mL) were added phenylboronic acid (12.2 mg, 0.10 mmol),
Tetrakis(triphenylphosphine)palladium(0) (10.0 mg, 8.65E-6 mol) and
1.0 M potassium carbonate in water (1.5 mL). The reaction was
microwaved (300 watts) at 160.degree. C. for 7 minutes. The
reaction mixture was poured into water and extracted with ethyl
acetate. The organic layer was washed with brine, dried over sodium
sulfate, concentrated, and purified with silica gel column
chromatography to give the compound as a white solid (P-1521, 9.4
mg, 33%). MS (ESI) [M+H.sup.+].sup.+=431.2.
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-(3-difluoromethoxy-2,6-difluoro-ph-
enyl)-methanone P-1526
##STR02005##
[1941] was prepared using Steps 1-3 of Scheme 44 by substituting
3,5-Dihydroxy-benzaldehyde 528 with N-2,4-Difluoro-phenol in Step
1. MS (ESI) [M+H.sup.+].sup.+=401.0, 403.0.
Example 65
Synthesis of
N-Benzyl-2-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylm-
ethyl)-phenoxy]-acetamide P-1469
[1942] Compound P-1469 was synthesized in four steps from
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyrid-
ine P-1455 as shown in Scheme 46.
##STR02006##
Step 1--Preparation of
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1-triisopropylsilanyl--
1H-pyrrolo[2,3-b]pyridine (548)
[1943] To
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1H-pyrrolo[2,-
3-b]pyridine (P-1455, 700.0 mg, 1.64 mmol, prepared as described in
Example 24, Scheme 43a) in tetrahydrofuran (40.0 mL) was added
sodium hydride (60% in mineral oil, 100.0 mg, 2.5 mmol). After 10
minutes, triisopropylsilyl chloride (0.60 mL, 0.0028 mol) was added
to the reaction. The reaction was stirred at room temperature for 2
hours. The reaction was poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
to give the compound as colorless oil (548, 0.85 g, 89%).
Step 2--Preparation of
2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyr-
idin-3-ylmethyl)-phenol (549)
[1944] To
3-(3-Benzyloxy-2,6-difluoro-benzyl)-5-pyridin-3-yl-1-triisopropy-
lsilanyl-1H-pyrrolo[2,3-b]pyridine (548, 510.0 mg, 0.87 mmol) in
methanol (20.0 mL) was added 20% Pd(OH).sub.2/C (50 mg). The
reaction was stirred under an atmosphere of hydrogen for 2 hours.
Filtration and concentration gave the compound as white solid (549,
427 mg, 99%).
Step 3--Preparation of
[2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]py-
ridin-3-ylmethyl)-phenoxy]-acetic acid methyl ester (550)
[1945] To
2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo[-
2,3-b]pyridin-3-ylmethyl)-phenol (549, 427.0 mg, 0.87 mmol) in
tetrahydrofuran (20.0 mL) was added sodium hydride (60% in mineral
oil, 42.0 mg, 1.1 mmol). 30 minutes later, methyl bromoacetate (146
mg, 0.95 mmol) was added to the reaction. The reaction was stirred
at room temperature for 4 hours. The reaction was poured into water
and extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give the crude compound that was used directly in
the next step.
Step 4--Preparation of
N-Benzyl-2-[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylm-
ethyl)-phenoxy]-acetamide (P-1469)
[1946] To
[2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo-
[2,3-b]pyridin-3-ylmethyl)-phenoxy]-acetic acid methyl ester (550,
210.0 mg, 0.37 mmol) in methanol (6.0 mL) was added benzylamine
(0.45 mL, 4.1 mmol). The reaction was stirred at room temperature
for 80 hours. To the reaction was added TBAF (0.31 g, 1.0 mmol).
The reaction was concentrated and purified with silical gel column
chromatography eluting with 10% methanol in methylene chloride to
give the compound as a white solid (P-1469, 60 mg, 33%). MS (ESI)
[M+H.sup.+].sup.+=423.3.
2-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phe-
noxy]-N-ethyl-acetamide P-1475
##STR02007##
[1947] was prepared using the same protocol as Scheme 46 by
substituting benzylamine with ethylamine in Step 4. MS (ESI)
[M+H.sup.+].sup.+=423.3.
Example 66
Synthesis of
[2,4-difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phen-
oxy]-acetic acid P-1468
[1948] Compound P-1468 was synthesized in two steps from
[2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]py-
ridin-3-ylmethyl)-phenoxy]-acetic acid methyl ester 550 as shown in
Scheme 47.
##STR02008##
Step 1--Preparation of
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phen-
oxy]-acetic acid methyl ester (P-1465)
[1949] To
[2,4-Difluoro-3-(5-pyridin-3-yl-1-triisopropylsilanyl-1H-pyrrolo-
[2,3-b]pyridin-3-ylmethyl)-phenoxy]-acetic acid methyl ester (550,
40.0 mg, 0.071 mmol, prepared as described in Example 65, Scheme
46) in tetrahydrofuran (5.0 mL) was added tetra-n-butylammonium
fluoride (22.2 mg, 0.085 mmol). The reaction was stirred at room
temperature for 10 minutes. The crude compound was concentrated
with silica gel and purified by silica gel column chromatography
eluting with 3% methanol in methylene chloride to give the compound
as white solid (P-1465, 20 mg, 69%). MS (ESI)
[M+H.sup.+].sup.+=410.2.
Step 2--Preparation of
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-phen-
oxy]-acetic acid (P-1468)
[1950] To
[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmet-
hyl)-phenoxy]-acetic acid methyl ester (P-1465, 20.0 mg, 0.049
mmol) in tetrahydrofuran (8.0 mL) were added potassium hydroxide
(100.0 mg, 1.8 mmol) and water (3.0 mL). The reaction was stirred
at room temperature overnight and then poured into water, acidified
to pH=5 with 1 N HCl, and extracted with ethyl acetate. The organic
layer was dried over anhydrous sodium sulfate and filtered. The
filtrate was concentrated and purified by silica gel column
chromatography eluting with 10% methanol in methylene chloride to
give the compound as light yellow solid (P-1468, 9.1 mg, 47%). MS
(ESI) [M+H.sup.+].sup.+=396.2.
Example 67
Synthesis of
2-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-ph-
enoxy]-ethanol P-1394
[1951] Compound P-1394 was synthesized in four steps from
2,6-Difluoro-3-hydroxy-benzaldehyde 540 as shown in Scheme 48.
##STR02009##
Step 1--Preparation of
2,6-Difluoro-3-[2-(tetrahydro-pyran-2-yloxy)-ethoxy]-benzaldehyde
(541)
[1952] To 2,6-difluoro-3-hydroxy-benzaldehyde (540, 0.150 g, 0.95
mmol) in N,N-dimethylformamide (8.0 mL) were added
2-(2-bromo-ethoxy)-tetrahydro-pyran (0.218 g, 1.04 mmol) and
potassium carbonate (0.52 g, 3.8 mmol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature for 72
hours. The reaction was poured into water and extracted with ethyl
acetate. The organic layer was dried over anhydrous sodium sulfate
and filtered. The filtrate was concentrated and purified by silica
gel column chromatography eluting with 20% ethyl acetate in hexane
to give the compound as colorless oil (541, 180 mg, 66%).
Step 2--Preparation of
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-2,6-difluoro-3-[2-(tetrahydro-pyr-
an-2-yloxy)-ethoxy]-phenyl-methanol (542)
[1953] To 5-bromo-7-azaindole (67, 118 mg, 0.000597 mol) in
methanol (9.0 mL) were added
2,6-difluoro-3-[2-(tetrahydro-pyran-2-yloxy)-ethoxy]-benzaldehyde
(541, 180.0 mg, 0.63 mmol) and potassium hydroxide (601.9 mg, 10.7
mmol) under an atmosphere of nitrogen. The reaction was stirred at
room temperature overnight. The reaction was poured into water and
extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate and filtered. The filtrate was
concentrated to give crude compound that was used directly in the
next step.
Step 3--Preparation of
2-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro-phenoxy]--
ethanol (P-1392)
[1954] To
(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-2,6-difluoro-3-[2-(tetra-
hydro-pyran-2-yloxy)-ethoxy]-phenyl-methanol (542, 0.22 g, 0.46
mmol) in acetonitrile (6.0 mL) were added trifluoroacetic acid
(0.14 mL, 1.8 mmol) and triethylsilane (0.29 mL, 1.8 mmol). The
reaction was stirred at room temperature overnight. The filtrate
was concentrated and purified by silica gel column chromatography
eluting with 35% ethyl acetate in hexane to give the compound as a
white solid (P-1392, 62 mg, 35%). MS (M+H.sup.+).sup.+=383.1,
385.1.
Step 4--Preparation of
2-[2,4-Difluoro-3-(5-pyridin-3-yl-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-ph-
enoxy]-ethanol (P-1394)
[1955] To
2-[3-(5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-ylmethyl)-2,4-difluoro--
phenoxy]-ethanol (P-1392, 35.0 mg, 0.091 mmol) in acetonitrile (4.0
mL) were added 3-pyridylboronic acid (14.6 mg, 0.12 mmol),
tetrakis(triphenylphosphine)palladium(0) (3.0 mg, 0.0026 mmol) and
1 M potassium carbonate solution (1.5 mL). The reaction was
microwaved (300 watts) at 160.degree. C. for 7 minutes. The
reaction was poured into water and extracted with ethyl acetate.
The organic layer was collected and dried over anhydrous sodium
sulfate and filtered. The filtrate was concentrated and purified by
silica gel column chromatography eluting 4% methanol in methylene
chloride to give the compound as white solid (P-1394, 11.0 mg,
32%). MS (ESI) [M+H.sup.+].sup.+=382.2.
Example 68
Synthesis of
3-[4-methoxy-3-(2,2,2-trifluoro-ethoxy)-benzyl]-1H-pyrrolo[2,3-b]pyridine
P-1626
[1956] Compound P-1626 was synthesized in three steps from
3-hydroxy-4-methoxy-benzaldehyde 560 as shown in Scheme 52.
##STR02010##
Step 1--Preparation of
4-Methoxy-3-(2,2,2-trifluoro-ethoxy)-benzaldehyde (562)
[1957] 3-Hydroxy-4-methoxy-benzaldehyde (560, 2.9 g, 19.5 mmol),
2-iodo-1,1,1-trifluoroethane (561, 2.2 mL, 23 mmol), and potassium
carbonate (3.7 g, 27 mmol) were dissolved in N,N-dimethylformamide
(100 mL). The solution was stirred at 90.degree. C. overnight under
an atmosphere of nitrogen. The reaction was poured into water,
extracted with ethyl acetate, and washed with brine. The organic
layer was dried over anhydrous sodium sulfate, filtered and
concentrated. The compound was isolated by silica gel column
chromatography eluting with 25% to 50% ethyl acetate in hexanes to
give white powder (562, 1.6 g, 40%).
Step 2--Preparation of
3-Methoxy-[4-methoxy-3-(2,2,2-trifluoro-ethoxy)-phenyl]-methyl-1H-pyrrolo-
[2,3-b]pyridine (563)
[1958] 1H-Pyrrolo[2,3-b]pyridine (0.2 g, 1.8 mmol),
4-methoxy-3-(2,2,2-trifluoro-ethoxy)-benzaldehyde (562, 0.4 g, 1.9
mmol) and potassium hydroxide (0.5 g, 9.6 mmol) were combined in
methanol (20 mL). The reaction was heated to 50.degree. C. under an
atmosphere of nitrogen and stirred for two days. The reaction was
poured into water, extracted with ethyl acetate, and washed with
brine. The organic layer was dried over anhydrous sodium sulfate,
filtered and concentrated. The compound was isolated by silica gel
column chromatography eluting with 40% to 75% ethyl acetate in
hexanes to give orange oil (563, 0.2 g, 31%). MS (ESI)
[M+H.sup.+].sup.+=367.2 [M-H.sup.+].sup.-=365.1.
Step 3--Preparation of
3-[4-Metlxoxy-3-(2,2,2-trifluoro-ethoxy)-benzyl]-1H-pyrrolo[2,3-b]pyridin-
e (P-1626)
[1959]
3-methoxy-[4-methoxy-3-(2,2,2-trifluoro-ethoxy)-phenyl]-methyl-1H-p-
yrrolo[2,3-b]pyridine (563) was reacted using the same protocol
described in Example 28, Scheme 51, Step 3. The compound P-1626 was
isolated by silica gel column chromatography eluting with 50% to
75% ethyl acetate in hexanes. MS (ESI) [M+H.sup.+].sup.+=337.1.
Example 69
Synthesis of N-phenyl-1H-pyrrolo[2,3-b]pyridin-3-amine P-0221
[1960] Compound P-0221 was synthesized in two or three steps from
1H-pyrrolo[2,3-b]pyridine 94 as shown in Scheme 54.
##STR02011##
Step 1--Preparation of 3-bromo-1H-pyrrolo[2,3-b]pyridine (582)
[1961] To 1H-pyrrolo[2,3-b]pyridine (94, 600 mg) dissolved in 10 mL
of 100% acetic acid was added the brominating agent DMAP-Br.sub.3
(1.001 equiv.). The reaction was heated at 75.degree. C. until the
orange color was gone and a precipitate formed (about 60 min). It
was dissolved with 50 mL of water (ppt dissolved) and adjusted to
pH 8 with NaHCO.sub.3 (aq. sat'd). The compound precipitated from
the solution and was collected. The aqueous fraction was extracted
with 3.times.30 mL with ethyl acetate. The combined organics were
dried over Na.sub.2SO.sub.4, evaporated and the solids combined
with the previously collected solid (582, 87%).
Route A: Step
2--3-bromo-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine
(583)
[1962] To 3-bromo-1H-pyrrolo[2,3-b]pyridine (582, 0.5 mmol)
dissolved in 5 mL of dry dioxane was added BEMP (2.5 equiv). The
TIPS-OTf (2.0 equiv) was then added and the reaction was stirred
overnight (15 hours). The solution was diluted with 20 mL of ethyl
acetate and washed with 5% acetic acid (2.times.10 mL), water
(2.times.10 mL), and brine (10 mL). The organic layer was dried
over Na.sub.2SO.sub.4, filtered, evaporated and dried under vacuum.
This material was used without further purification.
Route A: Step 3--Preparation of
N-phenyl-1H-pyrrolo[2,3-b]pyridin-3-amine (P-0221)
[1963] A 1 dram vial was charged with aniline (2-3 equiv), and 10
mg of 3-bromo-1-(triisopropylsilyl)-1H-pyrrolo[2,3-b]pyridine 583
in 0.4 mL toluene (dry, degassed). A catalyst stock solution
containing 3 mmol Pd(OAc).sub.2, 3 mmol
biphenyl-2-yl-di-tert-butyl-phosphane and 15 mL of toluene was
prepared. To the reaction mixture, 0.050 mL of the catalyst
solution was added. Excess NaOtBu was added as a solid to the
reaction. The vial was then placed in a 75.degree. C. oven for 60
minutes (shaken several times over an hour). After cooling, the
reaction was neutralized with 0.100 mL of TFA. After 30 minutes the
sample was evaporated and re-solvated in 0.300 mL of DMSO. The
desired compound was isolated by preparative HPLC/MS.
Rout B: Step 2--Preparation of
N-phenyl-1H-pyrrolo[2,3-b]pyridin-3-amine (P-0221)
[1964] To 10 mg of 3-bromo-1H-pyrrolo[2,3-b]pyridine 582 was added
0.4 mL of sulfolane and aniline (2-3 equiv). The vial was sealed
and heated at 180.degree. C. for 10 minutes. After cooling, the
desired compound was isolated by preparative HPLC/MS.
[1965] The following compounds were prepared by one of the
procedures described in scheme 54, substituting aniline with a
suitable amine in Step 5 [1966]
Methyl-phenyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1434),
[1967] 4-[Methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amino]-benzoic
acid (P-1435), [1968]
(4-Chloro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1436), [1969]
Methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-m-tolyl-amine (P-1437),
[1970]
(4-Methoxy-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1438), [1971]
(3-Chloro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1439), [1972]
(4-Fluoro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1440), [1973]
Methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-(4-trifluoromethoxy-phenyl)-amine
(P-1441), [1974]
(2,4-Difluoro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1442), [1975]
(3-Fluoro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1443), [1976]
(4-Methoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1488),
[1977] (3-Chloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1489), [1978]
(3-Methoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1490),
[1979] (4-Chloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1491), [1980]
(1H-Pyrrolo[2,3-b]pyridin-3-yl)-(4-trifluoromethoxy-phenyl)-amine
(P-1492), [1981]
(1H-Pyrrolo[2,3-b]pyridin-3-yl)-(3-trifluoromethoxy-phenyl)-amine
(P-1493), [1982]
(3,4-Dichloro-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1494), [1983]
(4-Phenoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1495),
[1984] (3-Phenoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1496), [1985]
(3-Benzyl-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1497),
[1986]
N-[3-(1H-Pyrrolo[2,3-b]pyridin-3-ylamino)-phenyl]-methanesulfonamide
(P-1498), [1987]
3-Fluoro-4-(1H-pyrrolo[2,3-b]pyridin-3-ylamino)-phenol (P-1499),
[1988] 3-(1H-Pyrrolo[2,3-b]pyridin-3-ylamino)-benzenesulfonamide
(P-1500), [1989]
(2-Phenoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine (P-1501),
[1990] (3,5-Dichloro-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1502), [1991]
(3,5-Dimethoxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1503), [1992]
[3-(4-Methyl-4H-[1,2,4]triazol-3-yl)-phenyl]-(1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-amine (P-1504), [1993]
(2-Benzyloxy-phenyl)-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1505), and [1994]
(3-Methoxy-phenyl)-methyl-(1H-pyrrolo[2,3-b]pyridin-3-yl)-amine
(P-1506). The following table indicates the amine (Column 2) that
is substituted in place of the aniline in Route B, Step 2, or Route
A, Step 3, to afford the compounds (Column 3). Column 1 provides
the compound number and Column 4 the observed mass.
TABLE-US-00025 [1994] MS(ESI) Compound [M + H.sup.+].sup.+ number
Amine Compound structure observed P-1434 ##STR02012## ##STR02013##
224.3 P-1435 ##STR02014## ##STR02015## 268.3 P-1436 ##STR02016##
##STR02017## 258.3 P-1437 ##STR02018## ##STR02019## 237.9 P-1438
##STR02020## ##STR02021## 254.3 P-1439 ##STR02022## ##STR02023##
258.3 P-1440 ##STR02024## ##STR02025## 242.3 P-1441 ##STR02026##
##STR02027## 308.3 P-1442 ##STR02028## ##STR02029## 260.3 P-1443
##STR02030## ##STR02031## 242.3 P-1488 ##STR02032## ##STR02033##
239.9 P-1489 ##STR02034## ##STR02035## 244.3 P-1490 ##STR02036##
##STR02037## 240.3 P-1491 ##STR02038## ##STR02039## 244.3 P-1492
##STR02040## ##STR02041## 293.9 P-1493 ##STR02042## ##STR02043##
293.9 P-1494 ##STR02044## ##STR02045## 291.9 P-1495 ##STR02046##
##STR02047## 302.3 P-1496 ##STR02048## ##STR02049## 302.3 P-1497
##STR02050## ##STR02051## 300.3 P-1498 ##STR02052## ##STR02053##
303.1 P-1499 ##STR02054## ##STR02055## 244.3 P-1500 ##STR02056##
##STR02057## 289.1 P-1501 ##STR02058## ##STR02059## 302.3 P-1502
##STR02060## ##STR02061## 277.9 P-1503 ##STR02062## ##STR02063##
270.3 P-1504 ##STR02064## ##STR02065## 291.1 P-1505 ##STR02066##
##STR02067## 316.3 P-1506 ##STR02068## ##STR02069## 254.3
Example 70
Synthesis of 4-chloro-7-azaindole 119
[1995] 4-chloro-7-azaindole 119 was synthesized in two steps from
7-azaindole according to the protocol of Scheme 81.
##STR02070##
Step-1--Synthesis of 1H-Pyrrolo[2,3-b]pyridine 7-oxide (658)
[1996] 1 H-Pyrrolo[2,3-b]pyridine 7-oxide 658 was synthesized by
reacting 7-azaindole 94 with an oxidizing agent (e.g. m-CPBA) in a
non-reactive solvent (e.g. dimethoxyethane) as described by
Schneller, S. W.; Luo, Jiann-Kuan. J. Org. Chem. 1980,
45:4045-4048. The compound was isolated by filtration of the
resulting solid that forms upon standing at 5.degree. C. for
typically 1-3 h.
Step-2--Synthesis of 4-chloro-7-azaindole (119)
[1997] 4-chloro-7-azaindole 119 was synthesized by reacting
1H-Pyrrolo[2,3-b]pyridine 7-oxide 658 with a chlorinating agent
(e.g. POCl.sub.3) neat as described by Schneller, S. W.; Luo,
Jiann-Kuan. J. Org. Chem. 1980, 45:4045-4048. The resulting
solution after heating for 3-5 h at elevated temperatures
(100-150.degree. C.) was neutralized with a base (e.g. NH.sub.4OH)
until a solid precipitated. The solid was isolated by
filtration.
Example 71
Synthesis of
[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanone P-2086 and
3-[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-benzyl]-1H-pyrrolo-
[2,3-b]pyridine P-2085
[1998] Compounds P-2086 and P-2085 were synthesized in three steps
from compounds 659 and 1H-pyrrolo[2,3-b]pyridine 94 as shown in
Scheme 82.
##STR02071##
Step 1--Preparation of
3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-benzaldehyde
(660)
[1999] To a solution of
3-(4-Chloro-2-fluoro-benzyloxy)-2-hydroxy-benzaldehyde (659, 140
mg, 0.5 mmol, prepared by protocol of Example 43, Steps 1 and 2 of
Scheme 71, using 4-chloro-2-fluoro-benzyl bromide in place of
4-chloro-benzyl bromide in Step 1) in tetrahydrofuran (8 mL) was
added dropwise a mixture of 2-fluoro-ethanol (64 mg, 1.0 mmol),
triphenylphosphine (180 mg, 0.7 mmol), and diisopropyl
azodicarboxylate (120 mg, 0.6 mol) in tetrahydrofuran (5 ml) at
0.degree. C. The reaction mixture was stirred at 0.degree. C. for
10 minutes and then at 40.degree. C. for 3 days. The reaction
mixture was dissolved in water and ethyl acetate. The organic
layers were collected, washed with brine, and dried over magnesium
sulfate. After removal of solvent, the residue was purified by
silica gel column chromatography eluting with ethyl acetate in
hexanes to provide the compound as a white solid (660, 88 mg, 54%).
MS (ESI) [M+H.sup.+].sup.+=327.12.
Step 2--Preparation of
[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanol (661) and
3-{[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-methoxy-m-
ethyl}-1H-pyrrolo[2,3-b]pyridine (662)
[2000] A solution of
3-(4-chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-benzaldehyde
(660, 88 mg, 0.27 mmol), 1H-pyrrolo[2,3-b]pyridine (94, 38 mg, 0.32
mmol), and potassium hydroxide (45 mg, 0.81 mol) in methanol (5 mL)
was stirred at room temperature for 24 hours. The reaction mixture
was poured into water and extracted with ethyl acetate. The organic
layer was collected, washed with brine, and dried over sodium
sulfate. After removal of solvent, the residue was purified by
silica gel column chromatography eluting with ethyl acetate in
hexane to provide compound 661 as a white solid (67 mg, 56%),
MS(ESI) [M+H.sup.+].sup.+=445.13 and compound 662 as a white solid
(36 mg, 29%), MS(ESI) [M+H.sup.+].sup.+=459.15.
Step 3a--Preparation of
[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanone (P-2086)
[2001] To a solution of
[3-(4-chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-(1H-pyrrolo[-
2,3-b]pyridin-3-yl)-methanol (661, 60 mg, 0.1 mmol) in
tetrahydrofuran (10 mL) was added Dess-Martin periodinane (69 mg,
0.16 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 3 hours. The reaction was quenched with a saturated
solution of sodium thiosulfate, extracted with ethyl acetate,
washed with sodium bicarbonate, brine, and dried over magnesium
sulfate. After removal of solvent, the residue was purified by
silica gel column chromatography eluting with ethyl acetate in
hexanes to provide the compound as a white solid (P-2086, 15 mg,
20%). MS(ESI) [M+H.sup.+].sup.+=441.06.
Step 3b--Preparation of
3-[3-(4-Chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-benzyl]-1H-pyrrolo-
[2,3-b]pyridine (P-2085)
[2002] A mixture of
3-{[3-(4-chloro-2-fluoro-benzyloxy)-2-(2-fluoro-ethoxy)-phenyl]-methoxy-m-
ethyl}-1H-pyrrolo[2,3-b]pyridine (662, 36 mg, 0.078 mmol),
triethylsilane (0.5 mL, 3 mmol), and trifluoroacetic acid (0.2 mL,
2 mmol) in acetonitrile (20 mL) was stirred at 80.degree. C. for 2
hours. The mixture was concentrated and the residue was dissolved
in ethyl acetate. The solution was washed with saturated sodium
bicarbonate, brine, and dried over sodium sulfate. After removal of
solvent, the residue was purified by silica gel column
chromatography eluting with ethyl acetate in hexanes to provide the
compound as a yellow solid (P-2085, 24 mg, 71%). MS(ESI)
[M+H.sup.+].sup.+=429.15.
3-(4-Chloro-benzyloxy)-2-(2,2-difluoro-ethoxy)-phenyl]-(1H-pyrrolo[2,3-b]p-
yridin-3-yl)-methanone (P-2075)
##STR02072##
[2003] was prepared following the protocol of Scheme 82,
substituting 2-fluoro-ethanol with 2,2-difluoro-ethanol and
substituting 3-(4-Chloro-2-fluoro-benzyloxy)-2-hydroxy-benzaldehyde
with 3-(4-chloro-benzyloxy)-2-hydroxy-benzaldehyde (628 of Example
43) in Step 1 to provide P-2075. MS(ESI)
[M+H.sup.+].sup.+=443.1.
Example 72
Synthesis of
[3-(2-Chloro-4-methanesulfonyl-benzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,-
3-b]pyridin-3-yl)-methanone P-2094
[2004] Compound P-2094 was synthesized in four steps from compounds
635 and 663 as shown in Scheme 83.
##STR02073## ##STR02074##
Step 7--Preparation of
(S-Benzyloxy-2-ethoxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-methanol (664)
[2005] To a solution of
3-iodo-1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyridine (635, 1.306
g, 3.26 mmol) in tetrahydrofuran (42 mL) at -20.degree. C. under
nitrogen was added isopropylmagnesium chloride (1.70 mL, 2.0 M
solution in tetrahydrofuran, 3.40 mmol). The reaction mixture was
stirred at -20.degree. C. for 1.5 hours. It was allowed to warm to
5.degree. C. and then kept at 5.degree. C. for 1 hour. The reaction
mixture was then cooled down to -20.degree. C. To this solution was
slowly added a solution of 2-ethoxy-3-benzyloxybenzaldehyde (663,
0.698 g, 2.72 mmol, prepared by protocol of Example 43, Steps 1-3
of Scheme 71, using benzyl bromide in place of 4-chloro-benzyl
bromide in Step 1) in tetrahydrofuran (42 mL). The reaction mixture
was stirred at -20.degree. C. for 2.5 hrs, and was allowed to warm
to 5.degree. C. for 2.5 hours. The reaction mixture was poured into
iced water, extracted with ethyl acetate, washed with saturated
ammonium chloride and brine, and dried over magnesium sulfate.
After removal of solvent, the residue was purified by silica gel
column chromatography eluting with ethyl acetate in hexane to
provide the compound as light-yellow oil (664, 200 mg, 13.9%).
Step 2--Preparation of
(2-ethoxy-3-hydroxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (665)
[2006] To a solution of
(3-benzyloxy-2-ethoxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyr-
idin-3-yl)-methanol (664, 195 mg, 0.37 mmol) in a mixture of
methanol (20 mL) and tetrahydrofuran (50 mL) was added palladium on
carbon (50 mg, 10% wt., 0.2 mmol). The mixture was stirred under
hydrogenation for seventeen hours. After removal of solvent, the
residue was washed with a mixture of ethyl ether and hexanes to
provide the compound as a white solid (665, 63 mg, 95%). MS(ESI)
[M+H.sup.+].sup.+=439.37.
Step 3--Preparation of
[3-(2-Chloro-4-methanesulfonyl-benzyloxy)-2-ethoxy-phenyl]-(1-triisopropy-
lsilanyl-1H-pyrrolo[2,3-b]pyridin-3-yl)-methanone (666)
[2007] To a solution of
(2-ethoxy-3-hydroxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (665, 40 mg, 0.064 mmol) in tetrahydrofuran (15
mL) was added sodium hydride (3.32 mg, 0.083 mmol) at room
temperature under an atmosphere of nitrogen. The mixture was
stirred at room temperature for 40 minutes, then
1-bromomethyl-2-chloro-4-methanesulfonyl-benzene (21.72 mg, 0.077
mmol) was added to the reaction mixture. It was stirred at room
temperature overnight. The mixture was then poured into water and
was extracted with ethyl acetate. The organic layer was collected
and washed with brine, dried over magnesium sulfate. After removal
of the solvent, a crude compound as light yellow oil was obtained
(666, 84 mg).
Step 4--Preparation of
[3-(2-Chloro-4-methanesulfonylbenzyloxy)-2-ethoxy-phenyl]-(1H-pyrrolo[2,3-
-b]pyridin-3-yl)-methanone (P-2094)
[2008] To a solution of
(2-ethoxy-3-hydroxy-phenyl)-(1-triisopropylsilanyl-1H-pyrrolo[2,3-b]pyrid-
in-3-yl)-methanone (666, 84 mg, 0.054 mmol) in methanol (10 mL) was
added potassium hydroxide (6 N solution) until pH of the solution
turned to over 10. Potassium fluoride (30 mg, 0.5 mmol) was then
added to the reaction mixture and the mixture was stirred at room
temperature for 6 hours. The reaction mixture was then poured into
saturated sodium carbonate and was extracted with ethyl acetate.
The organic layer was collected and washed with brine, dried over
magnesium sulfate. After removal of the solvent, the residue was
purified by preparative HPLC to provide as a white solid (P-2094, 5
mg, 19%). MS(ESI) [M+H.sup.+].sup.+=485.17.
Example 73
Synthesis of Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-amide
P-1403
[2009] Compound P-1403 was synthesized in seven steps from
3-fluoro-5-nitrobenzoic acid 667 as shown in Scheme 84.
##STR02075##
Step 1--Preparation of 3-fluoro-5-aminobenzoic acid (668)
[2010] Into a Parr pressure reactor were added
3-fluoro-5-nitrobenzoic acid (667, 5.0 g, 0.027 mol), methanol
(50.0 mL), 20% Pd(OH).sub.2 on carbon (300 mg). The reaction was
shaken under an atmosphere of hydrogen at 50 psi overnight. The
reaction was filtered through celite and was concentrated to
dryness to provide a white solid (668, 4.0 g, 95.0%).
Step 2--Preparation of 2-Fluoro-5-(propane-1-sulfonylamino)-benzoic
acid (669)
[2011] To 3-fluoro-5-aminobenzoic acid (668, 3.00 g, 0.0180 mol) in
methylene chloride (204 mL) were added pyridine (41 mL, 0.50 mol)
and propane-1-sulfonyl chloride (2.23 mL, 0.0198 mol) under an
atmosphere of nitrogen. The reaction was stirred at room
temperature for 5 days. The reaction was poured into water,
adjusted pH to 1 with 1N HCl, and extracted with ethyl acetate. The
organic layer was dried over anhydrous sodium sulfate, and
filtered. The filtrate was concentrated and purified by silica gel
column chromatography eluting with 5% methanol in methylene
chloride to give a white solid (669, 2.0 g, 42%). MS (ESI)
[M-H.sup.+].sup.-=260.1.
Step 3--Preparation of 2-Fluoro-5-(propane-1-sulfonylamino)-benzoic
acid methyl ester (670)
[2012] To 2-Fluoro-5-(propane-1-sulfonylamino)-benzoic acid (669,
2.0 g, 0.0076 mol) in methanol (20.0 mL) was added sulfuric acid
(0.90 mL, 0.017 mol). The reaction was stirred at room temperature
overnight. The reaction was concentrated and purified with silica
gel column chromatograph eluting with 20% ethyl acetate in hexane
to give a white solid (670, 1.27 g, 62%). MS (ESI)
[M-H.sup.+].sup.-=274.1.
Step 4--Preparation of Propane-1-sulfonic acid
(4-fluoro-3-hydroxymethyl-phenyl)-amide (671)
[2013] To 2-Fluoro-5-(propane-1-sulfonylamino )-benzoic acid methyl
ester (670, 1.20 g, 0.00436 mol) in tetrahydrofuran (100.0 mL) was
added lithium tetrahydroaluminate (1.00 M in tetrahydrofuran, 10.0
mL) under an atmosphere of nitrogen. The reaction was stirred at
room temperature overnight. To the reaction was added
Na.sub.2SO.sub.4.10H.sub.2O (5 g), and then stirred at room
temperature for 1 hour. The reaction was filtered, concentrated and
purified with silica gel column chromatography eluting with 50%
ethyl acetate in hexane to give the compound (671, 0.90 g, 83%). MS
(ESI) [M_31 H.sup.+].sup.-=246.1.
Step 5--Preparation of Propane-1-sulfonic acid
(4-fluoro-3-formyl-phenyl)-amide (672)
[2014] To propane-1-sulfonic acid
(4-fluoro-3-hydroxymethyl-phenyl)-amide (671, 0.483 g, 0.00195 mol)
in tetrahydrofuran (10.0 mL), cooled with ice/water, was added
Dess-Martin periodinane (1.00 g, 0.00236 mol). The reaction was
stirred at room temperature for 10 minutes. The reaction was poured
into water and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 30% ethyl acetate in hexane to give a white solid
(672, 360 mg, 75%). MS (ESI)[M-H.sup.+].sup.-=244.1.
Step 6--Preparation of Propane-1-sulfonic acid
3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-4-fluoro-pheny-
l-amide (673)
[2015] To 5-bromo-7-azaindole (67, 170.0 mg, 0.86 mmol) in methanol
(7.0 mL) were added propane-1-sulfonic acid
(4-fluoro-3-formyl-phenyl)-amide (672, 220.0 mg, 0.90 mmol) and
potassium hydroxide (0.50 g, 0.0089 mol) under an atmosphere of
nitrogen. The reaction was stirred at room temperature overnight.
The reaction was poured into water, acidified with 1N HCl to pH=5,
and extracted with ethyl acetate. The organic layer was dried over
anhydrous sodium sulfate, and filtered. The filtrate was
concentrated and purified by silica gel column chromatography
eluting with 10% methanol in methylene chloride to give the
compound (673, 55.0 mg, 14.0%). MS (ESI) [M+H.sup.+].sup.+=442.1,
444.1.
Step 7--Preparation of Propane-1-sulfonic acid
[3-(5-bromo-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-4-fluoro-phenyl]-amide
(P-1403)
[2016] To propane-1-sulfonic acid
3-[(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)-hydroxy-methyl]-4-fluoro-pheny-
l-amide (673, 55.0 mg, 0.12 mmol) in tetrahydrofuran (8.0 mL) was
added Dess-Martin periodinane (70.0 mg, 0.17 mmol). The reaction
was stirred at room temperature for 5 minutes. The reaction was
concentrated with silica gel and purified with silica gel column
chromatography eluting with 25% ethyl acetate in hexane to give an
off-white solid (P-1403, 26.2 mg, 47%). MS (ESI)
[M+H.sup.+].sup.+=437.9, 439.9.
Example 74
Additional Compounds
[2017] Additional compounds of the invention were synthesized
following the methods of the Examples above or similar methods, or
by methods known to those of skill in the art, and are shown in the
following Table 1.
TABLE-US-00026 TABLE 1 Additional compounds of the invention
##STR02076## P-0001 ##STR02077## P-0002 ##STR02078## P-0003
##STR02079## P-0005 ##STR02080## P-0010 ##STR02081## P-0011
##STR02082## P-0012 ##STR02083## P-0013 ##STR02084## P-0014
##STR02085## P-0015 ##STR02086## P-0017 ##STR02087## P-0018
##STR02088## P-0019 ##STR02089## P-0020 ##STR02090## P-0021
##STR02091## P-0022 ##STR02092## P-0023 ##STR02093## P-0025
##STR02094## P-0028 ##STR02095## P-0029 ##STR02096## P-0030
##STR02097## P-0031 ##STR02098## P-0032 ##STR02099## P-0033
##STR02100## P-0039 ##STR02101## P-0040 ##STR02102## P-0041
##STR02103## P-0043 ##STR02104## P-0044 ##STR02105## P-0045
##STR02106## P-0046 ##STR02107## P-0047 ##STR02108## P-0048
##STR02109## P-0049 ##STR02110## P-0050 ##STR02111## P-0051
##STR02112## P-0053 ##STR02113## P-0054 ##STR02114## P-0056
##STR02115## P-0057 ##STR02116## P-0058 ##STR02117## P-0059
##STR02118## P-0060 ##STR02119## P-0061 ##STR02120## P-0062
##STR02121## P-0063 ##STR02122## P-0064 ##STR02123## P-0069
##STR02124## P-0070 ##STR02125## P-0071 ##STR02126## P-0072
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##STR02414## P-0393 ##STR02415## P-0394 ##STR02416## P-0395
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##STR02423## P-0403 ##STR02424## P-0404 ##STR02425## P-0405
##STR02426## P-0406 ##STR02427## P-0407 ##STR02428## P-0408
##STR02429## P-0410 ##STR02430## P-0411 ##STR02431## P-0412
##STR02432## P-0413 ##STR02433## P-0414 ##STR02434## P-0415
##STR02435## P-0416 ##STR02436## P-0417 ##STR02437## P-0418
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##STR02534## P-0517 ##STR02535## P-0518 ##STR02536## P-0519
##STR02537## P-0520 ##STR02538## P-0522 ##STR02539## P-0523
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##STR02940## P-0986 ##STR02941## P-0987 ##STR02942## P-0988
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##STR02946## P-0993 ##STR02947## P-0994 ##STR02948## P-0995
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##STR02952## P-1000 ##STR02953## P-1001 ##STR02954## P-1003
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##STR02961## P-1014 ##STR02962## P-1016 ##STR02963## P-1017
##STR02964## P-1018 ##STR02965## P-1019 ##STR02966## P-1021
##STR02967## P-1022 ##STR02968## P-1023 ##STR02969## P-1024
##STR02970## P-1025 ##STR02971## P-1026 ##STR02972## P-1027
##STR02973## P-1029 ##STR02974## P-1030 ##STR02975## P-1031
##STR02976## P-1032 ##STR02977## P-1033 ##STR02978## P-1034
##STR02979## P-1035 ##STR02980## P-1036 ##STR02981## P-1037
##STR02982## P-1038 ##STR02983## P-1039 ##STR02984## P-1040
##STR02985## P-1041 ##STR02986## P-1042 ##STR02987## P-1043
##STR02988## P-1044 ##STR02989## P-1045 ##STR02990## P-1046
##STR02991## P-1047 ##STR02992## P-1048 ##STR02993## P-1049
##STR02994## P-1050 ##STR02995## P-1051 ##STR02996## P-1052
##STR02997## P-1053 ##STR02998## P-1054 ##STR02999## P-1055
##STR03000## P-1057 ##STR03001## P-1058 ##STR03002## P-1059
##STR03003## P-1060 ##STR03004## P-1061 ##STR03005## P-1062
##STR03006## P-1063 ##STR03007## P-1064 ##STR03008## P-1065
##STR03009## P-1066 ##STR03010## P-1067 ##STR03011## P-1068
##STR03012## P-1069 ##STR03013## P-1070 ##STR03014## P-1071
##STR03015## P-1072 ##STR03016## P-1073 ##STR03017## P-1074
##STR03018## P-1075 ##STR03019## P-1076 ##STR03020## P-1077
##STR03021## P-1078 ##STR03022## P-1079 ##STR03023## P-1080
##STR03024## P-1081 ##STR03025## P-1082 ##STR03026## P-1083
##STR03027## P-1084 ##STR03028## P-1085 ##STR03029## P-1086
##STR03030## P-1087 ##STR03031## P-1088 ##STR03032## P-1089
##STR03033## P-1091 ##STR03034## P-1092 ##STR03035## P-1093
##STR03036## P-1094 ##STR03037## P-1095 ##STR03038## P-1096
##STR03039## P-1097 ##STR03040## P-1098 ##STR03041## P-1099
##STR03042## P-1100 ##STR03043## P-1101 ##STR03044## P-1102
##STR03045## P-1103 ##STR03046## P-1104 ##STR03047## P-1105
##STR03048## P-1106 ##STR03049## P-1107 ##STR03050## P-1108
##STR03051## P-1109 ##STR03052## P-1110 ##STR03053## P-1111
##STR03054## P-1112 ##STR03055## P-1113 ##STR03056## P-1114
##STR03057## P-1115 ##STR03058## P-1117 ##STR03059## P-1118
##STR03060## P-1119 ##STR03061## P-1120 ##STR03062## P-1121
##STR03063## P-1122 ##STR03064## P-1123 ##STR03065## P-1124
##STR03066## P-1125 ##STR03067## P-1126 ##STR03068## P-1127
##STR03069## P-1128 ##STR03070## P-1129 ##STR03071## P-1130
##STR03072## P-1132 ##STR03073## P-1133 ##STR03074## P-1134
##STR03075## P-1135 ##STR03076## P-1136
##STR03077## P-1137 ##STR03078## P-1138 ##STR03079## P-1139
##STR03080## P-1140 ##STR03081## P-1141 ##STR03082## P-1142
##STR03083## P-1143 ##STR03084## P-1144 ##STR03085## P-1145
##STR03086## P-1146 ##STR03087## P-1147 ##STR03088## P-1148
##STR03089## P-1149 ##STR03090## P-1150 ##STR03091## P-1151
##STR03092## P-1152 ##STR03093## P-1153 ##STR03094## P-1154
##STR03095## P-1155 ##STR03096## P-1156 ##STR03097## P-1157
##STR03098## P-1158 ##STR03099## P-1159 ##STR03100## P-1160
##STR03101## P-1161 ##STR03102## P-1162 ##STR03103## P-1163
##STR03104## P-1164 ##STR03105## P-1165 ##STR03106## P-1166
##STR03107## P-1167 ##STR03108## P-1168 ##STR03109## P-1169
##STR03110## P-1170 ##STR03111## P-1171 ##STR03112## P-1172
##STR03113## P-1173 ##STR03114## P-1174 ##STR03115## P-1175
##STR03116## P-1176 ##STR03117## P-1177 ##STR03118## P-1178
##STR03119## P-1179 ##STR03120## P-1181 ##STR03121## P-1182
##STR03122## P-1183 ##STR03123## P-1184 ##STR03124## P-1185
##STR03125## P-1186 ##STR03126## P-1187 ##STR03127## P-1188
##STR03128## P-1189 ##STR03129## P-1190 ##STR03130## P-1191
##STR03131## P-1192 ##STR03132## P-1193 ##STR03133## P-1194
##STR03134## P-1195 ##STR03135## P-1196 ##STR03136## P-1197
##STR03137## P-1198 ##STR03138## P-1199 ##STR03139## P-1200
##STR03140## P-1201 ##STR03141## P-1202 ##STR03142## P-1203
##STR03143## P-1204 ##STR03144## P-1205 ##STR03145## P-1206
##STR03146## P-1207 ##STR03147## P-1208 ##STR03148## P-1209
##STR03149## P-1210 ##STR03150## P-1211 ##STR03151## P-1212
##STR03152## P-1213 ##STR03153## P-1214 ##STR03154## P-1215
##STR03155## P-1216 ##STR03156## P-1217 ##STR03157## P-1218
##STR03158## P-1219 ##STR03159## P-1220 ##STR03160## P-1221
##STR03161## P-1222 ##STR03162## P-1223 ##STR03163## P-1224
##STR03164## P-1225 ##STR03165## P-1226 ##STR03166## P-1227
##STR03167## P-1228 ##STR03168## P-1229 ##STR03169## P-1230
##STR03170## P-1231 ##STR03171## P-1232 ##STR03172## P-1233
##STR03173## P-1234 ##STR03174## P-1235 ##STR03175## P-1236
##STR03176## P-1237 ##STR03177## P-1238 ##STR03178## P-1249
##STR03179## P-1272 ##STR03180## P-1273 ##STR03181## P-1274
##STR03182## P-1275 ##STR03183## P-1276 ##STR03184## P-1277
##STR03185## P-1278 ##STR03186## P-1279 ##STR03187## P-1280
##STR03188## P-1281 ##STR03189## P-1282 ##STR03190## P-1283
##STR03191## P-1284 ##STR03192## P-1285 ##STR03193## P-1286
##STR03194## P-1287 ##STR03195## P-1288 ##STR03196## P-1316
##STR03197## P-1319 ##STR03198## P-1320 ##STR03199## P-1321
##STR03200## P-1322 ##STR03201## P-1337 ##STR03202## P-1340
##STR03203## P-1341 ##STR03204## P-1342 ##STR03205## P-1345
##STR03206## P-1346 ##STR03207## P-1347 ##STR03208## P-1348
##STR03209## P-1349 ##STR03210## P-1365 ##STR03211## P-1366
##STR03212## P-1367 ##STR03213## P-1368 ##STR03214## P-1369
##STR03215## P-1370 ##STR03216## P-1379 ##STR03217## P-1380
##STR03218## P-1381 ##STR03219## P-1382 ##STR03220## P-1383
##STR03221## P-1384 ##STR03222## P-1385 ##STR03223## P-1396
##STR03224## P-1397 ##STR03225## P-1398 ##STR03226## P-1400
##STR03227## P-1401 ##STR03228## P-1404 ##STR03229## P-1431
##STR03230## P-1433 ##STR03231## P-1444 ##STR03232## P-1448
##STR03233## P-1451 ##STR03234## P-1452 ##STR03235## P-1457
##STR03236## P-1458 ##STR03237## P-1459 ##STR03238## P-1460
##STR03239## P-1461 ##STR03240## P-1473 ##STR03241## P-1476
##STR03242## P-1479 ##STR03243## P-1480 ##STR03244## P-1484
##STR03245## P-1485 ##STR03246## P-1507 ##STR03247## P-1508
##STR03248## P-1509 ##STR03249## P-1510 ##STR03250## P-1511
##STR03251## P-1512 ##STR03252## P-1517 ##STR03253## P-1518
##STR03254## P-1525 ##STR03255## P-1533 ##STR03256## P-1535
##STR03257## P-1536 ##STR03258## P-1537 ##STR03259## P-1539
##STR03260## P-1540 ##STR03261## P-1543 ##STR03262## P-1550
##STR03263## P-1555 ##STR03264## P-1556 ##STR03265## P-1557
##STR03266## P-1560 ##STR03267## P-1561 ##STR03268## P-1562
##STR03269## P-1563 ##STR03270## P-1565 ##STR03271## P-1573
##STR03272## P-1574 ##STR03273## P-1577 ##STR03274## P-1585
##STR03275## P-1588 ##STR03276## P-1592 ##STR03277## P-1593
##STR03278## P-1604 ##STR03279## P-1607 ##STR03280## P-1608
##STR03281## P-1609 ##STR03282## P-1610 ##STR03283## P-1614
##STR03284## P-1617 ##STR03285## P-1619 ##STR03286## P-1620
##STR03287## P-1623 ##STR03288## P-1624 ##STR03289## P-1628
##STR03290## P-1629 ##STR03291## P-1631 ##STR03292## P-1632
##STR03293## P-1633 ##STR03294## P-1634 ##STR03295## P-1635
##STR03296## P-1638 ##STR03297## P-1639 ##STR03298## P-1640
##STR03299## P-1641 ##STR03300## P-1642 ##STR03301## P-1643
##STR03302## P-1644 ##STR03303## P-1645 ##STR03304## P-1646
##STR03305## P-1647 ##STR03306## P-1648 ##STR03307## P-1649
##STR03308## P-1650 ##STR03309## P-1651 ##STR03310## P-1653
##STR03311## P-1655 ##STR03312## P-1658 ##STR03313## P-1659
##STR03314## P-1662 ##STR03315## P-1663 ##STR03316## P-1664
##STR03317## P-1665 ##STR03318## P-1666 ##STR03319## P-1667
##STR03320## P-1668 ##STR03321## P-1669 ##STR03322## P-1670
##STR03323## P-1671 ##STR03324## P-1672 ##STR03325## P-1673
##STR03326## P-1674 ##STR03327## P-1675
##STR03328## P-1676 ##STR03329## P-1677 ##STR03330## P-1678
##STR03331## P-1679 ##STR03332## P-1680 ##STR03333## P-1681
##STR03334## P-1682 ##STR03335## P-1683 ##STR03336## P-1684
##STR03337## P-1685 ##STR03338## P-1686 ##STR03339## P-1688
##STR03340## P-1689 ##STR03341## P-1690 ##STR03342## P-1691
##STR03343## P-1692 ##STR03344## P-1693 ##STR03345## P-1694
##STR03346## P-1695 ##STR03347## P-1696 ##STR03348## P-1697
##STR03349## P-1699 ##STR03350## P-1701 ##STR03351## P-1743
##STR03352## P-1745 ##STR03353## P-1784 ##STR03354## P-1785
##STR03355## P-1786 ##STR03356## P-1787 ##STR03357## P-1788
##STR03358## P-1789 ##STR03359## P-1790 ##STR03360## P-1791
##STR03361## P-1805 ##STR03362## P-1806 ##STR03363## P-1807
##STR03364## P-1808 ##STR03365## P-1809 ##STR03366## P-1810
##STR03367## P-1811 ##STR03368## P-1812 ##STR03369## P-1813
##STR03370## P-1814 ##STR03371## P-1815 ##STR03372## P-1826
##STR03373## P-1829 ##STR03374## P-1831 ##STR03375## P-1832
##STR03376## P-1833 ##STR03377## P-1834 ##STR03378## P-1835
##STR03379## P-1836 ##STR03380## P-1837 ##STR03381## P-1838
##STR03382## P-1846 ##STR03383## P-1847 ##STR03384## P-1856
##STR03385## P-1863 ##STR03386## P-1866 ##STR03387## P-1883
##STR03388## P-1888 ##STR03389## P-1889 ##STR03390## P-1890
##STR03391## P-1898 ##STR03392## P-1936 ##STR03393## P-1971
##STR03394## P-1981 ##STR03395## P-1985 ##STR03396## P-1995
##STR03397## P-1999 ##STR03398## P-2001 ##STR03399## P-2007
##STR03400## P-2008 ##STR03401## P-2009 ##STR03402## P-2010
##STR03403## P-2017 ##STR03404## P-2018 ##STR03405## P-2019
##STR03406## P-2021 ##STR03407## P-2023 ##STR03408## P-2027
##STR03409## P-2028 ##STR03410## P-2029 ##STR03411## P-2030
##STR03412## P-2031 ##STR03413## P-2032 ##STR03414## P-2065
##STR03415## P-2066 ##STR03416## P-2067 ##STR03417## P-2068
##STR03418## P-2069 ##STR03419## P-2070 ##STR03420## P-2071
##STR03421## P-2072 ##STR03422## P-2073 ##STR03423## P-2074
##STR03424## P-2077 ##STR03425## P-2078 ##STR03426## P-2079
##STR03427## P-2080 ##STR03428## P-2081 ##STR03429## P-2082
##STR03430## P-2083 ##STR03431## P-2084 ##STR03432## P-2087
##STR03433## P-2088 ##STR03434## P-2089 ##STR03435## P-2090
##STR03436## P-2091 ##STR03437## P-2092 ##STR03438## P-2093
##STR03439## P-2095 ##STR03440## P-2096 ##STR03441## P-2097
##STR03442## P-2098
Example 75
Kinase Activity Assays
[2018] The effect of potential modulators of kinase activity can be
measured in a variety of different assays known in the art. For
example, direct radiometric, indirect FRET or AlphaScreen assays
may be used to assess the level of phosphorylation of a substrate
in the presence of test compounds in order to determine the
inhibitory affect of the compound on the kinase. Invitrogen
(Carlsbad, Calif.) uses a FRET based assay for Btk, EGFR, EphB2,
Flt3, Irak4, Kdr, MAP2K1, MAPKAPK2, PDGFRB, PKC theta, Stk6 and
Yes. For these assays, compounds of the invention were screened by
Invitrogen using Z'-Lyte.TM. kinase assay,
[2019] Briefly, the Invitrogen kinase assay involves use of a
specific peptide substrate optimized for each kinase, containing a
fluorophore at each end that make up the FRET pair. The peptide
sensitivity to proteolytic cleavage depends on phosphorylation of
the peptide. Non-phosphorylated peptide is cleaved by a protease
while peptide phosphorylated by the kinase is not cleaved. Cleavage
of the peptide disrupts the FRET between the donor (coumarin) and
acceptor (fluorescein) fluorophores, resulting in an increase in
the ratio of donor emission to acceptor emission. The emission
ratio of coumarin to fluorescein is used to assess the reaction
progress. The extent of phosphorylation is determined from the
emission ratio, which is low when the kinase is active
(phosphorylated peptide is not cleaved, FRET pair connected) or
higher for inhibited kinase (non-phosphorylated peptide is cleaved,
FRET pair separated). Thus, the assay involves a kinase reaction in
the presence of varying concentrations of a given compound, a
development reaction with site specific protease, and detection of
the fluorescent emission ratio of coumarin and fluorscein. The
emission ratio as a function of compound concentration was used to
determine the IC.sub.50 value. Reaction conditions for each kinase
are determined to provide optimal reaction times, incubation
temperature, kinase and ATP concentrations. Test samples are made
up in 1.times. kinase buffer (50 mM HEPES pH 7.5, 50 mM MgCls, 5 mM
EGTA, 0.05% BRIJ-35) and compound at desired concentration prepared
in DMSO such that the final DMSO is 1%. Assay controls include 0%
phosphorylation, which contains no ATP, 100% phosphorylation
control, which consists of a synthetically phosphorylated peptide,
and 0% inhibition control, which contains active kinase conditions
without compound. Typically, assay plates are prepared to 10 .mu.l
final volume per sample, adding 2.5 .mu.l of compound at 4.times.
the desired concentration in 4% DMSO (serial dilution of compound
provides concentration curve), 5 .mu.l of the desired kinase mixed
with Z'-LYTE.TM. peptide substrate (2.times. in 2.times. kinase
buffer, final peptide at 2 .mu.M), and 2.5 .mu.l of 4.times. ATP
solution. All kinase except for MAP2K1 were at 10 .mu.M ATP in the
kinase reaction, while MAP2K1 was at 100 .mu.M. Samples are mixed
and the kinase reaction is incubated 1 hour at room temperature,
after which 5 .mu.l of development solution is added and mixed.
After incubation for another 1 hour at room temperature, 5 .mu.l of
stop reagent is added to each sample and mixed. The fluorescence
signals are measured to determine the emission ratio.
[2020] AlphaScreen assay was used to screen additional kinases. The
assay is similarly dependent on the phosphorylation of a peptide
substrate. In this assay, an antibody that recognizes
phosphorylated substrate is bound to an acceptor bead. The peptide
substrate is attached to biotin, which binds to a donor bead that
contains streptavidin. Thus phosphorylated substrate is bound by
antibody and streptavidin, bringing the donor and acceptor beads
into close proximity when kinase is not inhibited. The donor
produces singlet oxygen which results in emission from the acceptor
when they are in close proximity. Conversely, when kinase is
inhibited, the donor and acceptor beads are not associated and the
emission from the acceptor is reduced. The fluorescence signal vs.
compound concentration was used to determine the IC.sub.50
values.
[2021] Genetic Engineering
[2022] For some of the kinase screens, preparation of the kinase
was required. Plasmids encoding a selection of kinase enzymes were
engineered using common polymerase chain reaction (PCR) methods.
The relevant DNA sequences and encoded protein sequences used in
the assay are shown for each (see below). Complementary DNA cloned
from various human tissues were purchased from Invitrogen, and
these were used as substrates in the PCR reactions. Specific custom
synthetic oligonucleotide primers (Invitrogen see below) were
designed to initiate the PCR product, and also to provide the
appropriate restriction enzyme cleavage sites for ligation with the
plasmids. In selected cases additional pairs of oligonucleotides
(see below) were used to introduce mutations into the coding
sequence to alter the sequence for enzyme activation (BRAF), for
removing problematic surface Cys residues (FGFR1), or for disabling
the capacity to bind ATP (MEK1 substrate).
[2023] In the case of KIT, the entire sequence encoding the enzyme
was made through a gene synthesis procedure, using custom synthetic
oligonucleotides covering the entire coding sequence
(Invitrogen).
[2024] The plasmids used for ligation with the kinase-encoding
inserts were derivatives of either pET (Novagen) for expression
using E. coli, or pFastBac (Invitrogen) for expression using
baculovirus infection of insect cell cultures. In each of these
cases the kinase was engineered to include a Histidine tag for
purification using metal affinity chromatography.
[2025] In some cases the kinase-encoding plasmids were engineered
as bicistronic mRNA to co-express a second protein that modifies
the kinase protein during its expression in the host cell. In the
cases of Abl, FGFR1, Flt1, Kdr, Kit, Met, Ret, and Src protein
tyrosine phosphatase 1B (PTP), was co-expressed for
dephosphorylation of the phospho-Tyrosines. In the case of ERK2, an
activated form of MEK1 (MEK1DD), was co-expressed for
phosphorylation and activation of the ERK2. In the case of p38a, an
activated form of MEK6 (MBCK6) was co-expressed for phosphorylation
and activation of the p38a. In the case of BRAF, the chaperone
CDC37, was co-expressed for more efficient protein folding of the
BRAF.
[2026] Plasmids encoding phosphorylation substrate proteins were
expressed as N-terminal GST fusions and C-terminal biotinylation
fusions, using pGEX vectors (Amersham) modified to include
sequences encoding a C-terminal biotinylation tag. These substrates
include MEK1, a substrate for BRAF, and BAD, a substrate for
Pim1.
[2027] Protein Expression in E. Coli and Purification.
[2028] For protein expression, plasmids containing genes of
interest were transformed into E. coli strains BL21(DE3)RIL or pLyS
(Invitrogen) and transformants selected for growth on LB agar
plates containing appropriate antibiotics. Single colonies were
grown overnight at 37.degree. C. in 200 ml TB (Terrific broth)
media. 16.times.1 L of fresh TB media in 2.8 L flasks were
inoculated with 10 ml of overnight culture and grown with constant
shaking at 37.degree. C. Once cultures reached an absorbance of 1.0
at 600 nm, IPTG was added and cultures were allowed to grow for a
further 12 to 18 hrs at temperatures ranging from 12-30.degree. C.
Cells were harvested by centrifugation and pellets frozen at
-80.degree. C. until ready for lysis.
[2029] For protein Purification; frozen E. coli cell pellets were
resuspended in lysis buffer and lysed using standard mechanical
methods. Soluble proteins were purified via poly-Histidine tags
using immobilized metal affinity purification IMAC. In the case of
all kinases described here all have been purified using a 3 step
purification process utilizing; IMAC, size exclusion chromatography
and ion exchange chromatography. In most cases the poly-Histidine
tag was removed using Thrombin (Calbiochem).
[2030] In some cases the purification protocol required
modifications in order to stabilize soluble protein during
purification and concentration. In the case of BRaf 5 mM MgCl.sub.2
was required throughout purification. In the case of Ret a
combination of 1 mM ATP and 5 mM MgCl.sub.2 was required during
cell lysis and throughout purification. In the case of Zap70 a 5M
excess of AMP-PCP over protein was required as well as 5 mM
MgCl.sub.2.
[2031] Baculovirus Expression Vector System
[2032] Virus Production:
[2033] The transfection of a monolayer of Spodoptera frugiperda
(Sf9) cells was performed utilizing a bacmid containing the gene of
interest and Cellfectin (Invitrogen) transfection reagent in
antibiotic-free, serum-free Grace's complete media (Invitrogen).
After a five hour incubation, the transfection media was removed,
and the monolayer was fed with Grace's media containing 10% FBS and
antibiotics. After a 72 to 96 hr incubation, the cell supernatant
containing the virus was harvested. The titer of the virus stock
was then determined using a baculovirus titer kit (BD). The virus
stock was then expanded using low Multiplicity of Infection (MOI of
0.1) cultures and harvested 48 hrs post-infection. The titer of the
expanded virus stock was then determined for use in recombinant
protein production.
[2034] Protein Production:
[2035] The protein expression level was optimized by varying the
MOI (1-10) and time of harvest (48-72 hrs). Sf9 cells were adapted
to SF-900 II serum-free media and grown in suspension in spinner
flasks. The cell suspension was then used to inoculate a Wave
bioreactor for 25 L production scale. Cells were harvested 48-72
hrs post-infection and stored at -80.degree. C. until ready for
lysis. Proteins were purified similarly to those expressed in E.
coli.
[2036] Kinase Assay
[2037] AlphaScreen assays were done using compounds dissolved in
DMSO to a concentration of 20 mM. The compounds were diluted
accordingly to the desired final concentrations in each sample
well, using 1:3 serial dilution for a total of 8 concentration
points. Plates were prepared such that each kinase reaction is 20
.mu.l in 1.times. kinase buffer, 5% DMSO and 10 .mu.M ATP. Kinase
and assay conditions (defined below) used for each kinase are shown
in Table 3. After incubation of the kinase reaction for 1 hour at
room temperature, 5 .mu.l of donor beads in stop buffer (50 mM EDTA
in 1.times. kinase buffer) was added, the sample was mixed and
incubated for 20 minutes at room temperature before adding 5 .mu.l
of acceptor beads in stop buffer. The samples were incubated for 60
minutes at room temperature and the signal per well was read on
AlphaQuest reader. Phosphorylated substrate results in binding of
the antibody and association of the donor and acceptor beads such
that signal correlates with kinase activity. The signal vs.
compound concentration was used to determine the IC.sub.50.
[2038] Assay Condition A:
TABLE-US-00027 Kinase buffer HEPES 50 mM, pH 7.2, 5 mM MgCl.sub.2,
5 mM MnCl.sub.2, 0.2% BSA, 0.01% NP-40 Substrate 100 nM
biotin-(E4Y)3 (Open Source Biotech, Inc.) Donor bead 1 .mu.g/ml
Streptavidin coated bead (Perkin Elmer Life Science). Acceptor bead
1 .mu.g/ml PY20 coated bead (Perkin Elmer Life Science)
[2039] Assay Condition B:
TABLE-US-00028 Kinase buffer 50 mM HEPES pH 7.0, 50 mM NaCl, 2 mM
MgCl.sub.2, 1 mM MnCl.sub.2, 1 mM DTT, 0.01% Tween-20. Substrate
100 nM biotin-MEK1 (prepared as described above). Donor bead 10
.mu.g/ml Streptavidin coated bead (Perkin Elmer Life Science).
Acceptor bead 10 .mu.g/ml Protein A coated, bound to anti phospho
MEK1/2 antibody (CellSignal)
[2040] Assay Condition C:
TABLE-US-00029 Kinase buffer 8 mM HEPES pH 7.0, 4 mM MgCl.sub.2, 1
mM DTT, 0.01% Tween-20. Substrate 100 nM biotin-MBP (Upstate
Biotechnology, Waltham, MA). Donor bead 10 .mu.g/ml Streptavidin
coated bead (Perkin Elmer Life Science). Acceptor bead 10 .mu.g/ml
Protein A coated, bound to anti phospho MBP antibody
(CellSignal)
[2041] Assay Condition D:
TABLE-US-00030 Kinase buffer 8 mM MOPS pH 7.4, 2 mM MgCl.sub.2, 8
mM MnCl.sub.2, 2 mM DTT, 0.01% Tween-20. Substrate 30 nM
biotin-(E4Y)10 (Upstate Biotechnology). Donor bead 20 .mu.g/ml
Streptavidin coated bead (Perkin Elmer Life Science). Acceptor bead
20 .mu.g/ml PY20 antibody coated (Perkin Elmer Life Science)
[2042] Assay Condition E:
TABLE-US-00031 Kinase buffer 20 mM HEPES pH 7.0, 10 mM MgCl.sub.2,
1 mM DTT, 0.01% Tween-20. Substrate 30 nM biotin-ATF2 (Upstate
Biotechnology). Donor bead 10 .mu.g/ml Streptavidin coated bead
(Perkin Elmer Life Science). Acceptor bead 10 .mu.g/ml Protein A
coated, bound to anti phospho ATF2 antibody (CellSignal)
[2043] Assay Condition F:
TABLE-US-00032 Kinase buffer 8 mM HEPES pH 7.0, 4 mM MgCl.sub.2, 1
mM DTT, 0.01% Tween-20. Substrate 100 nM biotin-BAD (prepared as
described above). Donor bead 10 .mu.g/ml Streptavidin coated bead
(Perkin Elmer Life Science). Acceptor bead 10 .mu.g/ml Protein A
coated, bound to anti phospho BAD (Ser112) antibody
(CellSignal)
[2044] Assay Condition G:
TABLE-US-00033 Kinase buffer MOPS 25 mM, pH 7.1, 0.1 mM MgCl.sub.2,
5 mM MnCl.sub.2, 0.2% BSA, 1 mM DTT, 0.01% Tween-20. Substrate 100
nM biotin-(E4Y)3 (Open Source Biotech, Inc.) Donor bead 1 .mu.g/ml
Streptavidin coated bead (Perkin Elmer Life Science). Acceptor bead
1 .mu.g/ml PY20 coated bead (Perkin Elmer Life Science)
TABLE-US-00034 TABLE 3 Kinase reaction conditions (20 .mu.l
reaction volume) for kinase screens. Plasmid Expression Kinase
Vendor* Number Host Assay Condition Abl P1121 E. coli A (1 ng
kinase) B-Raf Upstate B (0.1 ng kinase) B-Raf V600E P4254 Sf9 B
(0.1 ng kinase) c-Raf-1 Upstate B (0.1 ng kinase) Erk2 P4227 E.
coli C (4 ng kinase) Fak P1358 Sf9 A (0.1 ng kinase) FGFR1 P1351 E.
coli A (0.1 ng kinase) Flt1 P1826 E. coli A (0.1 ng kinase) Flt4
ProQinase A (0.1 ng kinase) Fms Upstate D (0.5 ng kinase) Jnk1
Upstate E (0.1 ng kinase) Jnk2 Roche E (0.05 ng kinase) Jnk3
Upstate E (0.1 ng kinase) Kit P1332 E. coli A (0.1 ng kinase) Met
P1818 E. coli A (0.1 ng kinase) p38 P4292 E. coli E (6 ng kinase)
Pim1 P1215 E. coli F (0.01 ng kinase) Pyk2 Upstate D (1 ng kinase)
Ret P1378 E. coli A (0.01 ng kinase) Src P1144 E. coli A (0.01 ng
kinase) Zap70 P1868 Sf9 G (0.1 ng kinase) *Upstate = Upstate
Biotechnology Roche = Roche Protein Expression Group (Indianapolis,
IN) ProQinase = ProQinase GmbH (Freiburg, Germany)
[2045] Kit was alternatively purchased from Cell Signalling
Technology. Kit and Fms assays were also run at 100 .mu.M ATP,
where for Fms the 1.times. buffer was 8 mM MOPS pH 7.0, 2 mM
MgCl.sub.2, 8 mM MnCl.sub.2, 2 mM DTT, 50 mM NaCl, 0.01% BSA and
0.01% Tween-20 and stop buffer was 8 mM MOPS, pH 7.0, 100 mM EDTA,
0.01% BSA and for kit, 1 ng kinase was used and the 1.times. buffer
was 8 mM MOPS pH 7.0, 1 mM MgCl.sub.2, 2 mM MnCl.sub.2, 1 mM DTT,
0.001% BSA and 0.01% Tween-20, with substrate of 30 nM
biotin-(E4Y).sub.10 (Upstate Biotechnology) and beads were at 10
.mu.g/ml in stop buffer of 8 mM MOPS, pH 7.0, 100 mM EDTA, 0.3%
BSA.
[2046] Compounds screened by at least one of the methods described
above, or by similar methods, having IC.sub.50 of less than 10
.mu.M are shown in tables 2a (Abl), 2b (B-Raf), 2c (B-Raf V600E),
2d (Btk), 2e (c-Raf-1), 2f (EGFR), 2g (EphB2), 2h (Erk2), 2i (Fak),
2j (FGFR1), 2k (Flt1), 2l (Flt3), 2m (Flt4), 2n (Fms), 2o (Irak4),
2p (Jnk1), 2q (Jnk2), 2r (Jnk3), 2s (Kdr), 2t (Kit), 2u (MAP2K1),
2v (MAPKAPK2), 2w (Met), 2x (p38), 2y (PDGFRB), 2z (Pim1), 2aa (PKC
theta), 2bb (Pyk2), 2cc (Ret), 2dd (Src), 2ee (Stk6), and 2ff
(Yes), 2gg (Zap70), 2hh (Akt3), 2ii (ALK), 2jj (Cdk2), 2kk (Csk),
2ll (EphA2), 2mm (EphB4), 2nn (Frk), 2oo (Gsk.beta.), 2pp (Hck),
2qq (MAP4K4), 2rr (IGF1R), 2ss (IKK beta), 2tt (Itk), 2uu (Jak3),
2vv (MLK1), 2ww (TrkA), 2xx (PDGFRA), 2yy (Plk1), 2zz (Brk), 2ab
(ROCK1), 2ac (Syk), 2ad (TEC), and 2ac (Tie2).
TABLE-US-00035 TABLE 2a Compounds with activity toward kinase Abl
with IC.sub.50 .ltoreq. 10 .mu.M. Abl P-0001, P-0002, P-0003,
P-0005, P-0007, P-0010, P-0011, P-0012, P-0019, P-0025, P-0028,
P-0032, P-0033, P-0040, P-0041, P-0045, P-0047, P-0048, P-0050,
P-0054, P-0056, P-0058, P-0068, P-0072, P-0074, P-0090, P-0093,
P-0134, P-0140, P-0142, P-0167, P-0169, P-0196, P-0218, P-0224,
P-0244, P-0316, P-0320, P-0448, P-0453, P-0501, P-0521, P-0529,
P-0550, P-0559, P-0562, P-0579, P-0594, P-0599, P-0604, P-0611,
P-0623, P-0624, P-0645, P-0656, P-0671, P-0675, P-0691, P-0693,
P-0708, P-0738, P-0751, P-0762, P-0781, P-0794, P-0800, P-0806,
P-0885, P-1011, P-1012, P-1115, P-1127, P-1318, P-1336, P-1394,
P-1426
TABLE-US-00036 TABLE 2b Compounds with activity toward kinase B-Raf
with IC.sub.50 .ltoreq. 10 .mu.M. B-Raf P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010, P-0016,
P-0019, P-0021, P-0024, P-0025, P-0026, P-0027, P-0032, P-0034,
P-0035, P-0036, P-0037, P-0038, P-0041, P-0042, P-0045, P-0050,
P-0052, P-0054, P-0055, P-0056, P-0059, P-0060, P-0065, P-0066,
P-0067, P-0068, P-0078, P-0079, P-0082, P-0088, P-0090, P-0093,
P-0095, P-0102, P-0112, P-0114, P-0122, P-0126, P-0140, P-0143,
P-0162, P-0165, P-0166, P-0171, P-0178, P-0180, P-0184, P-0188,
P-0192, P-0196, P-0200, P-0210, P-0228, P-0257, P-0262, P-0265,
P-0269, P-0271, P-0284, P-0293, P-0297, P-0302, P-0307, P-0310,
P-0351, P-0356, P-0369, P-0382, P-0396, P-0414, P-0448, P-0486,
P-0521, P-0550, P-0556, P-0559, P-0579, P-0594, P-0599, P-0604,
P-0613, P-0636, P-0683, P-0685, P-0693, P-0697, P-0700, P-0716,
P-0721, P-0728, P-0734, P-0744, P-0745, P-0746, P-0753, P-0763,
P-0773, P-0774, P-0776, P-0778, P-0779, P-0794, P-0798, P-0805,
P-0806, P-0807, P-0818, P-0837, P-0841, P-0842, P-0848, P-0850,
P-0851, P-0853, P-0857, P-0860, P-0861, P-0863, P-0866, P-0867,
P-0868, P-0874, P-0876, P-0877, P-0883, P-0885, P-0889, P-0894,
P-0896, P-0897, P-0898, P-0902, P-0904, P-0907, P-0909, P-0910,
P-0911, P-0912, P-0913, P-0919, P-0924, P-0928, P-0931, P-0932,
P-0933, P-0937, P-0939, P-0941, P-0944, P-0946, P-0947, P-0952,
P-0954, P-0955, P-0956, P-0958, P-0959, P-0973, P-0975, P-0978,
P-0980, P-0983, P-0984, P-0987, P-0991, P-0997, P-0998, P-1003,
P-1004, P-1006, P-1009, P-1013, P-1014, P-1020, P-1027, P-1028,
P-1056, P-1076, P-1080, P-1110, P-1116, P-1243, P-1244, P-1246,
P-1247, P-1249, P-1250, P-1251, P-1252, P-1253, P-1254, P-1255,
P-1256, P-1257, P-1258, P-1259, P-1260, P-1261, P-1262, P-1263,
P-1264, P-1265, P-1266, P-1267, P-1268, P-1269, P-1270, P-1279,
P-1280, P-1281, P-1282, P-1288, P-1289, P-1317, P-1318, P-1336,
P-1338, P-1341, P-1343, P-1346, P-1347, P-1348, P-1349, P-1365,
P-1383, P-1384, P-1385, P-1386, P-1387, P-1388, P-1389, P-1390,
P-1391, P-1395, P-1397, P-1419, P-1420, P-1429, P-1430, P-1431,
P-1432, P-1433, P-1445, P-1446, P-1447, P-1451, P-1452, P-1453,
P-1454, P-1455, P-1456, P-1457, P-1458, P-1459, P-1467, P-1469,
P-1472, P-1473, P-1475, P-1477, P-1479, P-1480, P-1481, P-1485,
P-1486, P-1526, P-1527, P-1528, P-1529, P-1532, P-1534, P-1539,
P-1541, P-1542, P-1544, P-1546, P-1547, P-1548, P-1549, P-1552,
P-1553, P-1554, P-1555, P-1556, P-1559, P-1566, P-1567, P-1568,
P-1569, P-1570, P-1576, P-1577, P-1580, P-1581, P-1582, P-1583,
P-1584, P-1585, P-1586, P-1589, P-1590, P-1591, P-1592, P-1593,
P-1596, P-1597, P-1598, P-1599, P-1600, P-1602, P-1605, P-1608,
P-1609, P-1610, P-1612, P-1613, P-1616, P-1621, P-1627, P-1630,
P-1631, P-1636, P-1637, P-1638, P-1639, P-1656, P-1660, P-1663,
P-1664, P-1665, P-1670, P-1671, P-1687, P-1700, P-1701, P-1702,
P-1703, P-1704, P-1705, P-1706, P-1707, P-1708, P-1709, P-1710,
P-1711, P-1712, P-1713, P-1714, P-1715, P-1716, P-1717, P-1718,
P-1719, P-1720, P-1721, P-1722, P-1723, P-1724, P-1725, P-1726,
P-1727, P-1728, P-1729, P-1730, P-1731, P-1732, P-1733, P-1734,
P-1735, P-1736, P-1737, P-1738, P-1739, P-1740, P-1741, P-1742,
P-1746, P-1747, P-1748, P-1749, P-1750, P-1751, P-1752, P-1753,
P-1755, P-1756, P-1757, P-1758, P-1759, P-1760, P-1762, P-1763,
P-1764, P-1765, P-1766, P-1767, P-1768, P-1769, P-1770, P-1771,
P-1772, P-1773, P-1774, P-1775, P-1776, P-1777, P-1778, P-1779,
P-1780, P-1781, P-1782, P-1783, P-1784, P-1798, P-1799, P-1800,
P-1802, P-1804, P-1816, P-1817, P-1818, P-1819, P-1822, P-1823,
P-1825, P-1827, P-1828, P-1839, P-1840, P-1841, P-1842, P-1864,
P-1865, P-1871, P-1872, P-1873, P-1878, P-1879, P-1881, P-1882,
P-1907, P-1912, P-1916, P-1980, P-1996, P-1997, P-1998, P-2005,
P-2006, P-2007, P-2012, P-2013
TABLE-US-00037 TABLE 2c Compounds with activity toward kinase B-Raf
V600E with IC.sub.50 .ltoreq. 10 .mu.M. B-Raf P-0001, P-0002,
P-0003, P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010,
V600E P-0011, P-0012, P-0013, P-0014, P-0015, P-0016, P-0017,
P-0019, P-0021, P-0022, P-0024, P-0025, P-0026, P-0027, P-0028,
P-0032, P-0033, P-0034, P-0035, P-0036, P-0037, P-0038, P-0039,
P-0040, P-0041, P-0042, P-0043, P-0044, P-0045, P-0046, P-0047,
P-0048, P-0049, P-0050, P-0051, P-0052, P-0053, P-0054, P-0055,
P-0056, P-0059, P-0060, P-0062, P-0063, P-0066, P-0067, P-0068,
P-0070, P-0071, P-0072, P-0073, P-0074, P-0075, P-0078, P-0079,
P-0082, P-0085, P-0088, P-0089, P-0090, P-0092, P-0093, P-0095,
P-0097, P-0099, P-0100, P-0102, P-0107, P-0108, P-0109, P-0110,
P-0112, P-0114, P-0118, P-0119, P-0121, P-0122, P-0124, P-0126,
P-0129, P-0134, P-0135, P-0137, P-0139, P-0140, P-0141, P-0146,
P-0147, P-0148, P-0152, P-0153, P-0156, P-0160, P-0162, P-0165,
P-0166, P-0170, P-0171, P-0174, P-0175, P-0178, P-0180, P-0181,
P-0184, P-0185, P-0186, P-0187, P-0188, P-0191, P-0192, P-0196,
P-0199, P-0200, P-0205, P-0208, P-0210, P-0211, P-0215, P-0228,
P-0231, P-0237, P-0242, P-0243, P-0244, P-0246, P-0248, P-0252,
P-0255, P-0257, P-0261, P-0262, P-0265, P-0269, P-0271, P-0274,
P-0284, P-0287, P-0293, P-0295, P-0297, P-0302, P-0307, P-0308,
P-0311, P-0318, P-0320, P-0325, P-0333, P-0344, P-0347, P-0351,
P-0352, P-0355, P-0356, P-0358, P-0369, P-0373, P-0381, P-0382,
P-0386, P-0388, P-0396, P-0409, P-0414, P-0418, P-0420, P-0421,
P-0434, P-0441, P-0447, P-0448, P-0453, P-0472, P-0483, P-0486,
P-0493, P-0515, P-0521, P-0535, P-0550, P-0552, P-0559, P-0573,
P-0579, P-0592, P-0594, P-0599, P-0600, P-0603, P-0604, P-0613,
P-0623, P-0624, P-0636, P-0638, P-0645, P-0646, P-0647, P-0651,
P-0656, P-0668, P-0671, P-0679, P-0683, P-0685, P-0691, P-0693,
P-0696, P-0698, P-0700, P-0710, P-0713, P-0716, P-0721, P-0728,
P-0730, P-0734, P-0744, P-0745, P-0746, P-0751, P-0753, P-0762,
P-0763, P-0771, P-0773, P-0774, P-0776, P-0777, P-0778, P-0779,
P-0794, P-0798, P-0803, P-0805, P-0806, P-0807, P-0810, P-0811,
P-0816, P-0818, P-0819, P-0833, P-0837, P-0841, P-0842, P-0848,
P-0850, P-0851, P-0853, P-0857, P-0860, P-0861, P-0863, P-0866,
P-0867, P-0868, P-0874, P-0876, P-0877, P-0885, P-0886, P-0889,
P-0893, P-0894, P-0896, P-0897, P-0898, P-0902, P-0904, P-0905,
P-0907, P-0910, P-0911, P-0912, P-0913, P-0919, P-0927, P-0928,
P-0931, P-0933, P-0937, P-0939, P-0944, P-0946, P-0947, P-0950,
P-0951, P-0952, P-0954, P-0955, P-0956, P-0957, P-0958, P-0959,
P-0962, P-0964, P-0971, P-0976, P-0980, P-0981, P-0983, P-0984,
P-0991, P-0997, P-0998, P-1000, P-1002, P-1003, P-1004, P-1006,
P-1007, P-1008, P-1009, P-1010, P-1013, P-1014, P-1015, P-1016,
P-1018, P-1020, P-1021, P-1022, P-1024, P-1025, P-1026, P-1028,
P-1029, P-1030, P-1031, P-1035, P-1043, P-1049, P-1056, P-1061,
P-1063, P-1064, P-1065, P-1067, P-1069, P-1070, P-1071, P-1074,
P-1082, P-1085, P-1090, P-1091, P-1112, P-1113, P-1116, P-1117,
P-1120, P-1123, P-1128, P-1131, P-1138, P-1139, P-1140, P-1160,
P-1177, P-1179, P-1180, P-1181, P-1183, P-1187, P-1194, P-1195,
P-1199, P-1243, P-1244, P-1246, P-1247, P-1249, P-1250, P-1251,
P-1252, P-1253, P-1254, P-1255, P-1256, P-1257, P-1258, P-1259,
P-1260, P-1261, P-1262, P-1263, P-1264, P-1265, P-1266, P-1267,
P-1269, P-1270, P-1279, P-1280, P-1281, P-1282, P-1283, P-1288,
P-1289, P-1316, P-1317, P-1318, P-1323, P-1329, P-1336, P-1341,
P-1343, P-1345, P-1346, P-1347, P-1348, P-1349, P-1365, P-1366,
P-1368, P-1369, P-1370, P-1381, P-1383, P-1384, P-1385, P-1386,
P-1387, P-1388, P-1389, P-1390, P-1391, P-1392, P-1394, P-1395,
P-1396, P-1397, P-1398, P-1399, P-1402, P-1403, P-1409, P-1411,
P-1413, P-1414, P-1415, P-1416, P-1417, P-1418, P-1419, P-1420,
P-1423, P-1425, P-1426, P-1429, P-1430, P-1431, P-1432, P-1433,
P-1444, P-1445, P-1446, P-1447, P-1448, P-1449, P-1450, P-1451,
P-1452, P-1453, P-1454, P-1455, P-1456, P-1457, P-1458, P-1459,
P-1462, P-1465, P-1466, P-1467, P-1469, P-1470, P-1471, P-1472,
P-1473, P-1474, P-1475, P-1477, P-1478, P-1479, P-1480, P-1481,
P-1485, P-1486, P-1495, P-1505, P-1506, P-1516, P-1526, P-1527,
P-1528, P-1529, P-1530, P-1531, P-1532, P-1534, P-1539, P-1540,
P-1541, P-1542, P-1544, P-1545, P-1546, P-1547, P-1548, P-1549,
P-1550, P-1552, P-1553, P-1554, P-1556, P-1558, P-1559, P-1561,
P-1564, P-1566, P-1567, P-1568, P-1569, P-1570, P-1572, P-1575,
P-1576, P-1577, P-1578, P-1579, P-1581, P-1582, P-1583, P-1584,
P-1585, P-1586, P-1589, P-1590, P-1591, P-1592, P-1594, P-1596,
P-1597, P-1598, P-1599, P-1600, P-1601, P-1602, P-1605, P-1606,
P-1607, P-1608, P-1609, P-1610, P-1611, P-1612, P-1613, P-1614,
P-1621, P-1627, P-1630, P-1631, P-1636, P-1637, P-1638, P-1639,
P-1656, P-1660, P-1663, P-1664, P-1665, P-1666, P-1670, P-1671,
P-1687, P-1698, P-1700, P-1701, P-1702, P-1703, P-1704, P-1705,
P-1706, P-1707, P-1708, P-1709, P-1710, P-1711, P-1712, P-1713,
P-1714, P-1715, P-1716, P-1717, P-1718, P-1719, P-1720, P-1721,
P-1722, P-1723, P-1724, P-1725, P-1726, P-1727, P-1728, P-1729,
P-1730, P-1731, P-1732, P-1733, P-1734, P-1735, P-1736, P-1737,
P-1738, P-1739, P-1740, P-1741, P-1742, P-1746, P-1747, P-1748,
P-1749, P-1750, P-1751, P-1752, P-1753, P-1755, P-1756, P-1757,
P-1758, P-1759, P-1760, P-1762, P-1763, P-1764, P-1765, P-1766,
P-1767, P-1768, P-1769, P-1770, P-1771, P-1772, P-1773, P-1774,
P-1775, P-1776, P-1777, P-1778, P-1779, P-1780, P-1781, P-1782,
P-1783, P-1784, P-1797, P-1798, P-1799, P-1800, P-1802, P-1804,
P-1816, P-1817, P-1818, P-1819, P-1822, P-1823, P-1828, P-1839,
P-1840, P-1841, P-1842, P-1843, P-1864, P-1865, P-1871, P-1872,
P-1873, P-1878, P-1879, P-1881, P-1882, P-1907, P-1912, P-1916,
P-1980, P-1996, P-1997, P-1998, P-2005, P-2006, P-2007, P-2012,
P-2013
TABLE-US-00038 TABLE 2d Compounds with activity toward kinase Btk
with IC.sub.50 .ltoreq. 10 .mu.M. Btk: P-0005, P-0006, P-0007,
P-0009, P-0010, P-0011, P-0012, P-0013, P-0014, P-0015, P-0016,
P-0017, P-0019, P-0020, P-0021, P-0022, P-0024, P-0025, P-0026,
P-0027, P-0028, P-0029, P-0031, P-0033, P-0040, P-0041, P-0042,
P-0043, P-0044, P-0045, P-0046, P-0047, P-0048, P-0049, P-0050,
P-0051, P-0052, P-0053, P-0054, P-0055, P-0059, P-0060, P-0061,
P-0062, P-0063, P-0067, P-0068, P-0070, P-0072, P-0073, P-0074,
P-0075, P-0079, P-0081, P-0082, P-0083, P-0085, P-0088, P-0089,
P-0090, P-0093, P-0094, P-0097, P-0102, P-0107, P-0108, P-0109,
P-0112, P-0113, P-0125, P-0134, P-0135, P-0138, P-0139, P-0145,
P-0148, P-0152, P-0156, P-0166, P-0171, P-0217, P-0228, P-0257,
P-0280, P-0297, P-0302, P-0304, P-0314, P-0321, P-0325, P-0351,
P-0418, P-0429, P-0763, P-0806, P-0807, P-0885, P-0897, P-0991,
P-0997, P-1020, P-1262, P-1266, P-1267, P-1269, P-1317, P-1336,
P-1343, P-1346, P-1388, P-1389, P-1390, P-1420, P-1426, P-1459,
P-1473, P-1475, P-1479, P-1480, P-1481, P-1485, P-1486
TABLE-US-00039 TABLE 2e Compounds with activity toward kinase
c-Raf-1 with IC.sub.50 .ltoreq. 10 .mu.M c-Raf-1: P-0001, P-0002,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010, P-0015,
P-0016, P-0021, P-0024, P-0025, P-0026, P-0027, P-0032, P-0034,
P-0035, P-0036, P-0037, P-0038, P-0042, P-0045, P-0052, P-0055,
P-0066, P-0078, P-0079, P-0082, P-0088, P-0090, P-0102, P-0112,
P-0114, P-0121, P-0122, P-0137, P-0156, P-0162, P-0165, P-0166,
P-0170, P-0178, P-0180, P-0184, P-0188, P-0210, P-0228, P-0257,
P-0262, P-0265, P-0269, P-0297, P-0302, P-0307, P-0356, P-0369,
P-0382, P-0396, P-0418, P-0486, P-0521, P-0535, P-0542, P-0559,
P-0604, P-0613, P-0636, P-0656, P-0685, P-0700, P-0716, P-0721,
P-0728, P-0734, P-0744, P-0745, P-0746, P-0753, P-0763, P-0773,
P-0774, P-0776, P-0778, P-0779, P-0794, P-0798, P-0805, P-0806,
P-0807, P-0811, P-0818, P-0837, P-0841, P-0842, P-0848, P-0850,
P-0851, P-0853, P-0857, P-0860, P-0861, P-0863, P-0866, P-0867,
P-0868, P-0874, P-0876, P-0877, P-0883, P-0885, P-0889, P-0890,
P-0894, P-0896, P-0897, P-0898, P-0902, P-0904, P-0907, P-0909,
P-0910, P-0911, P-0912, P-0913, P-0919, P-0924, P-0928, P-0931,
P-0933, P-0937, P-0939, P-0941, P-0944, P-0946, P-0947, P-0950,
P-0952, P-0954, P-0955, P-0956, P-0957, P-0958, P-0959, P-0964,
P-0971, P-0973, P-0974, P-0975, P-0978, P-0983, P-0987, P-0991,
P-0997, P-0998, P-1002, P-1003, P-1004, P-1006, P-1009, P-1013,
P-1014, P-1015, P-1017, P-1020, P-1027, P-1028, P-1047, P-1056,
P-1061, P-1063, P-1064, P-1065, P-1070, P-1071, P-1076, P-1077,
P-1078, P-1079, P-1118, P-1122, P-1145, P-1243, P-1244, P-1246,
P-1247, P-1249, P-1250, P-1251, P-1253, P-1254, P-1255, P-1256,
P-1257, P-1258, P-1260, P-1261, P-1262, P-1265, P-1279, P-1283,
P-1288, P-1289, P-1316, P-1317, P-1318, P-1336, P-1338, P-1365,
P-1386, P-1387, P-1388, P-1389, P-1390, P-1391, P-1395, P-1396,
P-1397, P-1398, P-1403, P-1413, P-1419, P-1431, P-1432, P-1433,
P-1448, P-1451, P-1452, P-1453, P-1454, P-1455, P-1456, P-1458,
P-1541, P-1542, P-1546, P-1547, P-1581, P-1583, P-1630, P-1671,
P-1712, P-1713, P-1714, P-1733, P-1737, P-1738, P-1739, P-1740,
P-1783, P-1839, P-1864, P-1871, P-1873, P-1878, P-1879, P-1881,
P-1882
TABLE-US-00040 TABLE 2f Compounds with activity toward kinase EGFR
with IC.sub.50 .ltoreq. 10 .mu.M EGFR: P-0001, P-0002, P-0003,
P-0004, P-0025, P-0095, P-0153, P-0877
TABLE-US-00041 TABLE 2g Compounds with activity toward kinase EphB2
with IC.sub.50 .ltoreq. 10 .mu.M EphB2: P-0001, P-0003, P-0005,
P-0006, P-0007, P-0009, P-0010, P-0011, P-0012, P-0013, P-0014,
P-0015, P-0016, P-0017, P-0019, P-0020, P-0021, P-0022, P-0025,
P-0027, P-0028, P-0029, P-0032, P-0033, P-0034, P-0035, P-0036,
P-0038, P-0040, P-0041, P-0042, P-0043, P-0044, P-0045, P-0048,
P-0050, P-0052, P-0053, P-0055, P-0056, P-0059, P-0062, P-0067,
P-0068, P-0070, P-0072, P-0074, P-0075, P-0078, P-0083, P-0088,
P-0090, P-0093, P-0102, P-0107, P-0109, P-0114, P-0124, P-0125,
P-0126, P-0139, P-0145, P-0186
TABLE-US-00042 TABLE 2h Compounds with activity toward kinase Erk2
with IC.sub.50 .ltoreq. 10 .mu.M Erk2: P-0031, P-0041, P-0058,
P-0154, P-0550, P-0611, P-1336
TABLE-US-00043 TABLE 2i Compounds with activity toward kinase Fak
with IC.sub.50 .ltoreq. 10 .mu.M Fak: P-0001, P-0002, P-0003,
P-0004, P-0006, P-0007, P-0008, P-0009, P-0016, P-0018, P-0024,
P-0025, P-0026, P-0027, P-0032, P-0034, P-0035, P-0036, P-0037,
P-0045, P-0054, P-0055, P-0067, P-0078, P-0088, P-0102, P-0112,
P-0114, P-0166, P-0196, P-0209, P-0210, P-0211, P-0224, P-0257,
P-0269, P-0276, P-0293, P-0298, P-0302, P-0310, P-0333, P-0391,
P-0396, P-0437, P-0486, P-0494, P-0501, P-0611, P-0668, P-0675,
P-0685, P-0691, P-0700, P-0721, P-0774, P-0795, P-0797, P-0806,
P-0811, P-0818, P-0837, P-0850, P-0851, P-0867, P-0885, P-0889,
P-0910, P-0911, P-0933, P-0951, P-0955, P-0956, P-0986, P-0992,
P-1002, P-1009, P-1013, P-1020, P-1054, P-1083, P-1114, P-1146,
P-1190, P-1247, P-1249, P-1250, P-1253, P-1255, P-1256, P-1263,
P-1264, P-1266, P-1267, P-1269, P-1279, P-1280, P-1281, P-1282,
P-1288, P-1289, P-1316, P-1318, P-1321, P-1323, P-1329, P-1336,
P-1341, P-1346, P-1347, P-1348, P-1349, P-1359, P-1365, P-1383,
P-1384, P-1385, P-1387, P-1388, P-1389, P-1390, P-1391, P-1392,
P-1394, P-1396, P-1397, P-1400, P-1401, P-1402, P-1403, P-1411,
P-1431, P-1432, P-1433, P-1445, P-1446, P-1447, P-1449, P-1450,
P-1451, P-1452, P-1453, P-1455, P-1456, P-1457, P-1458, P-1459,
P-1473, P-1474, P-1475, P-1477, P-1478, P-1479, P-1480, P-1481,
P-1482, P-1485, P-1486, P-1492, P-1495, P-1500, P-1502, P-1685
TABLE-US-00044 TABLE 2j Compounds with activity toward kinase FGFR
with IC.sub.50 .ltoreq. 10 .mu.M FGFR: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010, P-0011,
P-0012, P-0013, P-0015, P-0016, P-0017, P-0019, P-0020, P-0021,
P-0024, P-0025, P-0026, P-0027, P-0028, P-0029, P-0032, P-0033,
P-0034, P-0035, P-0036, P-0037, P-0038, P-0039, P-0040, P-0041,
P-0042, P-0043, P-0045, P-0050, P-0052, P-0054, P-0055, P-0056,
P-0058, P-0059, P-0066, P-0067, P-0068, P-0072, P-0073, P-0078,
P-0079, P-0082, P-0088, P-0089, P-0090, P-0093, P-0096, P-0097,
P-0099, P-0101, P-0102, P-0103, P-0112, P-0114, P-0119, P-0121,
P-0129, P-0132, P-0134, P-0137, P-0140, P-0142, P-0149, P-0150,
P-0152, P-0156, P-0158, P-0165, P-0166, P-0167, P-0168, P-0170,
P-0171, P-0175, P-0178, P-0180, P-0184, P-0189, P-0192, P-0196,
P-0198, P-0204, P-0205, P-0206, P-0210, P-0211, P-0215, P-0217,
P-0218, P-0219, P-0220, P-0227, P-0228, P-0232, P-0233, P-0239,
P-0242, P-0244, P-0246, P-0248, P-0257, P-0262, P-0265, P-0267,
P-0269, P-0271, P-0274, P-0284, P-0287, P-0289, P-0293, P-0294,
P-0297, P-0298, P-0302, P-0304, P-0308, P-0309, P-0316, P-0320,
P-0321, P-0325, P-0326, P-0329, P-0335, P-0339, P-0341, P-0344,
P-0346, P-0351, P-0363, P-0368, P-0369, P-0371, P-0373, P-0374,
P-0379, P-0383, P-0385, P-0391, P-0392, P-0396, P-0400, P-0404,
P-0409, P-0411, P-0418, P-0420, P-0421, P-0424, P-0426, P-0442,
P-0447, P-0448, P-0452, P-0453, P-0459, P-0473, P-0474, P-0479,
P-0483, P-0486, P-0489, P-0493, P-0495, P-0501, P-0507, P-0510,
P-0515, P-0520, P-0521, P-0535, P-0550, P-0556, P-0559, P-0561,
P-0562, P-0563, P-0575, P-0579, P-0594, P-0599, P-0611, P-0613,
P-0615, P-0623, P-0624, P-0632, P-0636, P-0640, P-0645, P-0647,
P-0656, P-0658, P-0668, P-0671, P-0679, P-0683, P-0685, P-0691,
P-0693, P-0697, P-0698, P-0699, P-0700, P-0708, P-0710, P-0721,
P-0728, P-0730, P-0734, P-0737, P-0744, P-0745, P-0746, P-0749,
P-0751, P-0753, P-0763, P-0771, P-0773, P-0774, P-0776, P-0798,
P-0805, P-0806, P-0807, P-0811, P-0818, P-0819, P-0826, P-0828,
P-0835, P-0837, P-0841, P-0848, P-0850, P-0851, P-0853, P-0854,
P-0857, P-0860, P-0865, P-0867, P-0868, P-0874, P-0876, P-0877,
P-0885, P-0889, P-0894, P-0896, P-0897, P-0898, P-0902, P-0904,
P-0907, P-0910, P-0911, P-0912, P-0913, P-0919, P-0927, P-0933,
P-0935, P-0937, P-0944, P-0947, P-0950, P-0951, P-0952, P-0954,
P-0955, P-0956, P-0958, P-0964, P-0974, P-0976, P-0977, P-0979,
P-0983, P-0991, P-0997, P-1002, P-1004, P-1008, P-1009, P-1015,
P-1017, P-1018, P-1020, P-1021, P-1027, P-1066, P-1074, P-1078,
P-1110, P-1111, P-1116, P-1120, P-1123, P-1125, P-1142, P-1181,
P-1182, P-1188, P-1194, P-1246, P-1249, P-1250, P-1251, P-1252,
P-1253, P-1254, P-1255, P-1256, P-1257, P-1258, P-1259, P-1260,
P-1261, P-1262, P-1263, P-1264, P-1265, P-1266, P-1267, P-1269,
P-1270, P-1272, P-1273, P-1274, P-1279, P-1280, P-1281, P-1282,
P-1283, P-1287, P-1288, P-1289, P-1316, P-1317, P-1318, P-1321,
P-1322, P-1323, P-1325, P-1326, P-1327, P-1328, P-1329, P-1330,
P-1331, P-1332, P-1333, P-1334, P-1335, P-1336, P-1337, P-1338,
P-1339, P-1340, P-1341, P-1342, P-1343, P-1344, P-1345, P-1346,
P-1347, P-1348, P-1349, P-1365, P-1366, P-1367, P-1369, P-1377,
P-1380, P-1381, P-1382, P-1383, P-1384, P-1385, P-1386, P-1387,
P-1388, P-1389, P-1390, P-1391, P-1392, P-1393, P-1394, P-1395,
P-1396, P-1397, P-1398, P-1399, P-1402, P-1403, P-1404, P-1406,
P-1407, P-1409, P-1411, P-1415, P-1416, P-1418, P-1419, P-1420,
P-1423, P-1424, P-1426, P-1428, P-1429, P-1430, P-1431, P-1433,
P-1445, P-1446, P-1447, P-1448, P-1451, P-1452, P-1453, P-1454,
P-1455, P-1456, P-1458, P-1459, P-1460, P-1461, P-1463, P-1464,
P-1465, P-1467, P-1468, P-1469, P-1472, P-1473, P-1474, P-1475,
P-1476, P-1477, P-1478, P-1479, P-1480, P-1481, P-1482, P-1485,
P-1486, P-1512, P-1516, P-1522, P-1524, P-1525, P-1526, P-1527,
P-1528, P-1529, P-1530, P-1534, P-1538, P-1539, P-1542, P-1545,
P-1546, P-1547, P-1548, P-1549, P-1550, P-1554, P-1555, P-1556,
P-1561, P-1564, P-1577, P-1581, P-1582, P-1583, P-1584, P-1585,
P-1589, P-1591, P-1592, P-1593, P-1595, P-1597, P-1603, P-1605,
P-1608, P-1609, P-1610, P-1614, P-1621, P-1622, P-1624, P-1625,
P-1685
TABLE-US-00045 TABLE 2k Compounds with activity toward kinase Flt1
with IC.sub.50 .ltoreq. 10 .mu.M Flt1: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0008, P-0009, P-0011, P-0012, P-0013, P-0016,
P-0017, P-0018, P-0019, P-0020, P-0021, P-0024, P-0026, P-0027,
P-0032, P-0033, P-0034, P-0036, P-0037, P-0038, P-0039, P-0041,
P-0054, P-0055, P-0056, P-0067, P-0068, P-0072, P-0078, P-0082,
P-0088, P-0090, P-0091, P-0101, P-0103, P-0112, P-0114, P-0127,
P-0134, P-0150, P-0154, P-0166, P-0177, P-0180, P-0184, P-0194,
P-0196, P-0206, P-0211, P-0214, P-0224, P-0244, P-0269, P-0274,
P-0278, P-0287, P-0298, P-0302, P-0315, P-0320, P-0325, P-0326,
P-0337, P-0371, P-0373, P-0383, P-0404, P-0409, P-0421, P-0448,
P-0455, P-0461, P-0470, P-0477, P-0483, P-0486, P-0491, P-0514,
P-0515, P-0521, P-0550, P-0559, P-0579, P-0603, P-0624, P-0629,
P-0632, P-0636, P-0640, P-0656, P-0668, P-0679, P-0683, P-0685,
P-0691, P-0700, P-0708, P-0721, P-0733, P-0737, P-0749, P-0751,
P-0757, P-0768, P-0771, P-0773, P-0774, P-0777, P-0805, P-0866,
P-0868, P-0951, P-0958, P-0962, P-1002, P-1008, P-1010, P-1018,
P-1021, P-1027, P-1082, P-1110, P-1112, P-1147, P-1160, P-1181,
P-1194, P-1246, P-1247, P-1250, P-1251, P-1255, P-1256, P-1259,
P-1260, P-1261, P-1262, P-1266, P-1269, P-1279, P-1289, P-1317,
P-1318, P-1365, P-1366, P-1370, P-1372, P-1373, P-1383, P-1393,
P-1395, P-1403, P-1404, P-1406, P-1411, P-1415, P-1416, P-1417,
P-1418, P-1420, P-1422, P-1423, P-1424, P-1425, P-1426, P-1427,
P-1428, P-1429, P-1430, P-1431, P-1432, P-1433, P-1445, P-1446,
P-1447, P-1457, P-1460, P-1461, P-1462, P-1463, P-1464, P-1465,
P-1467, P-1468, P-1469, P-1472, P-1475, P-1486, P-1491, P-1492,
P-1495, P-1497, P-1499, P-1502, P-1505, P-1523, P-1526, P-1527,
P-1528, P-1529, P-1530, P-1531, P-1532, P-1533, P-1534, P-1541,
P-1542, P-1544, P-1546, P-1547, P-1548, P-1549, P-1552, P-1553,
P-1554, P-1556, P-1557, P-1559, P-1564, P-1566, P-1567, P-1569,
P-1570, P-1571, P-1572, P-1575, P-1576, P-1577, P-1580, P-1581,
P-1582, P-1583, P-1584, P-1585, P-1586, P-1587, P-1589, P-1590,
P-1591, P-1592, P-1593, P-1594, P-1595, P-1596, P-1597, P-1598,
P-1599, P-1600, P-1601, P-1602, P-1603, P-1605, P-1606, P-1608,
P-1609, P-1610, P-1612, P-1613, P-1614, P-1615, P-1618, P-1619,
P-1621, P-1622, P-1624, P-1625, P-1686
TABLE-US-00046 TABLE 2l Compounds with activity toward kinase Flt3
with IC.sub.50 .ltoreq. 10 .mu.M Flt3: P-0088, P-0262, P-0409,
P-0636, P-0806, P-1244, P-1280, P-1318, P-1336, P-1394, P-1426
TABLE-US-00047 TABLE 2m Compounds with activity toward kinase Flt4
with IC.sub.50 .ltoreq. 10 .mu.M Flt4: P-0001, P-0003, P-0005,
P-0011, P-0012, P-0019, P-0024, P-0029, P-0032, P-0039, P-0040,
P-0041, P-0090, P-0093, P-0097, P-0117, P-0142, P-0167, P-0190,
P-0196, P-0204, P-0205, P-0206, P-0211, P-0215, P-0218, P-0224,
P-0265, P-0316, P-0325, P-0335, P-0352, P-0418, P-0448, P-0452,
P-0453, P-0495, P-0521, P-0544, P-0550, P-0559, P-0579, P-0594,
P-0599, P-0617, P-0624, P-0632, P-0645, P-0656, P-0668, P-0679,
P-0691, P-0703, P-0708, P-0737, P-0738, P-0751, P-0762, P-0771,
P-0813
TABLE-US-00048 TABLE 2n Compounds with activity toward kinase Fms
with IC.sub.50 .ltoreq. 10 .mu.M Fms: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010, P-0012,
P-0016, P-0019, P-0020, P-0021, P-0025, P-0026, P-0027, P-0032,
P-0033, P-0034, P-0035, P-0036, P-0037, P-0038, P-0042, P-0045,
P-0048, P-0054, P-0067, P-0068, P-0072, P-0078, P-0079, P-0082,
P-0088, P-0090, P-0093, P-0101, P-0102, P-0103, P-0112, P-0114,
P-0121, P-0129, P-0134, P-0145, P-0152, P-0166, P-0170, P-0175,
P-0180, P-0184, P-0189, P-0196, P-0210, P-0211, P-0228, P-0233,
P-0237, P-0239, P-0244, P-0257, P-0262, P-0269, P-0274, P-0284,
P-0287, P-0291, P-0293, P-0297, P-0298, P-0302, P-0317, P-0325,
P-0327, P-0333, P-0351, P-0369, P-0373, P-0383, P-0391, P-0396,
P-0409, P-0418, P-0420, P-0421, P-0461, P-0483, P-0486, P-0491,
P-0501, P-0515, P-0535, P-0559, P-0563, P-0613, P-0636, P-0656,
P-0685, P-0721, P-0733, P-0753, P-0763, P-0771, P-0773, P-0774,
P-0778, P-0798, P-0805, P-0806, P-0807, P-0811, P-0818, P-0835,
P-0837, P-0848, P-0850, P-0851, P-0853, P-0854, P-0857, P-0867,
P-0874, P-0876, P-0883, P-0885, P-0898, P-0911, P-0913, P-0919,
P-0927, P-0931, P-0933, P-0952, P-0954, P-0955, P-0956, P-0958,
P-1002, P-1008, P-1009, P-1013, P-1110, P-1112, P-1124, P-1194,
P-1246, P-1247, P-1249, P-1250, P-1251, P-1252, P-1253, P-1255,
P-1259, P-1260, P-1262, P-1263, P-1264, P-1265, P-1266, P-1267,
P-1269, P-1279, P-1280, P-1281, P-1282, P-1289, P-1316, P-1317,
P-1318, P-1321, P-1323, P-1324, P-1325, P-1326, P-1327, P-1328,
P-1329, P-1330, P-1331, P-1332, P-1333, P-1334, P-1335, P-1336,
P-1337, P-1338, P-1339, P-1340, P-1341, P-1342, P-1343, P-1344,
P-1346, P-1365, P-1366, P-1368, P-1369, P-1370, P-1372, P-1376,
P-1380, P-1381, P-1384, P-1385, P-1386, P-1387, P-1388, P-1389,
P-1390, P-1391, P-1392, P-1393, P-1394, P-1395, P-1397, P-1399,
P-1400, P-1402, P-1403, P-1404, P-1405, P-1406, P-1409, P-1410,
P-1411, P-1415, P-1416, P-1419, P-1420, P-1421, P-1423, P-1425,
P-1426, P-1427, P-1428, P-1429, P-1430, P-1431, P-1432, P-1435,
P-1443, P-1447, P-1448, P-1449, P-1450, P-1451, P-1452, P-1453,
P-1454, P-1455, P-1456, P-1457, P-1458, P-1459, P-1460, P-1461,
P-1462, P-1463, P-1464, P-1465, P-1466, P-1467, P-1468, P-1469,
P-1470, P-1471, P-1472, P-1473, P-1474, P-1475, P-1476, P-1477,
P-1478, P-1479, P-1480, P-1481, P-1482, P-1483, P-1485, P-1486,
P-1488, P-1489, P-1490, P-1491, P-1492, P-1493, P-1494, P-1495,
P-1496, P-1497, P-1498, P-1499, P-1500, P-1501, P-1502, P-1503,
P-1504, P-1505, P-1506, P-1511, P-1522, P-1525, P-1526, P-1527,
P-1528, P-1529, P-1530, P-1532, P-1534, P-1541, P-1542, P-1544,
P-1545, P-1546, P-1547, P-1548, P-1549, P-1552, P-1553, P-1554,
P-1555, P-1556, P-1558, P-1559, P-1564, P-1566, P-1567, P-1568,
P-1569, P-1570, P-1571, P-1572, P-1575, P-1576, P-1580, P-1581,
P-1583, P-1586, P-1587, P-1589, P-1591, P-1594, P-1595, P-1596,
P-1597, P-1598, P-1599, P-1602, P-1606, P-1608, P-1609, P-1610,
P-1611, P-1612, P-1613, P-1615, P-1616, P-1618, P-1621, P-1625,
P-1627, P-1630, P-1631, P-1636, P-1637, P-1638, P-1639, P-1652,
P-1653, P-1654, P-1656, P-1657, P-1660, P-1663, P-1664, P-1665,
P-1670, P-1671, P-1685, P-1687, P-1700, P-1701, P-1702, P-1703,
P-1704, P-1705, P-1706, P-1707, P-1708, P-1709, P-1710, P-1711,
P-1712, P-1713, P-1714, P-1715, P-1716, P-1717, P-1718, P-1719,
P-1720, P-1721, P-1722, P-1723, P-1724, P-1725, P-1726, P-1727,
P-1728, P-1729, P-1730, P-1731, P-1732, P-1733, P-1734, P-1735,
P-1736, P-1737, P-1738, P-1739, P-1740, P-1741, P-1742, P-1746,
P-1747, P-1748, P-1749, P-1750, P-1751, P-1753, P-1754, P-1755,
P-1756, P-1757, P-1758, P-1759, P-1760, P-1761, P-1762, P-1763,
P-1764, P-1765, P-1766, P-1767, P-1768, P-1769, P-1771, P-1772,
P-1773, P-1774, P-1775, P-1776, P-1778, P-1779, P-1780, P-1781,
P-1782, P-1783, P-1784, P-1796, P-1798, P-1799, P-1800, P-1802,
P-1803, P-1804, P-1816, P-1817, P-1818, P-1819, P-1821, P-1822,
P-1827, P-1828, P-1839, P-1840, P-1864, P-1871, P-1872, P-1873,
P-1878, P-1879, P-1881, P-1882, P-1907, P-1912, P-1916, P-1980,
P-1996, P-1997, P-1998, P-2005, P-2006, P-2007, P-2012, P-2013
TABLE-US-00049 TABLE 2o Compounds with activity toward kinase Irak4
with IC.sub.50 .ltoreq. 10 .mu.M Irak4: P-0002, P-0020, P-0076,
P-0087, P-0091, P-0130
TABLE-US-00050 TABLE 2p Compounds with activity toward kinase Jnk1
with IC.sub.50 .ltoreq. 10 .mu.M Jnk1: P-0001, P-0002, P-0003,
P-0004, P-0006, P-0008, P-0009, P-0010, P-0015, P-0016, P-0021,
P-0025, P-0026, P-0027, P-0032, P-0033, P-0034, P-0035, P-0036,
P-0037, P-0038, P-0040, P-0042, P-0045, P-0052, P-0054, P-0056,
P-0066, P-0078, P-0079, P-0082, P-0088, P-0090, P-0102, P-0112,
P-0114, P-0121, P-0134, P-0140, P-0156, P-0184, P-0196, P-0204,
P-0228, P-0244, P-0257, P-0269, P-0285, P-0297, P-0302, P-0308,
P-0431, P-0448, P-0486, P-0521, P-0559, P-0579, P-0599, P-0624,
P-0636, P-0668, P-0685, P-0691, P-0700, P-0721, P-0728, P-0734,
P-0745, P-0753, P-0763, P-0774, P-0807, P-0848, P-0850, P-0851,
P-0853, P-0860, P-0876, P-0897, P-0956, P-0958, P-0991, P-0997,
P-1002, P-1008, P-1009, P-1021, P-1251, P-1253, P-1256, P-1260,
P-1262, P-1266, P-1279, P-1280, P-1281, P-1288, P-1289, P-1317,
P-1318, P-1336, P-1338, P-1343, P-1346, P-1347, P-1348, P-1349,
P-1356, P-1359, P-1365, P-1366, P-1370, P-1384, P-1385, P-1390,
P-1394, P-1400, P-1402, P-1432, P-1433, P-1445, P-1446, P-1447,
P-1456, P-1458, P-1459, P-1465, P-1468, P-1473, P-1475, P-1486,
P-1523, P-1534, P-1546, P-1547, P-1548, P-1549, P-1553, P-1554,
P-1556, P-1566, P-1567, P-1570, P-1576, P-1577, P-1585, P-1589,
P-1591, P-1592, P-1596, P-1602, P-1610, P-1611, P-1618, P-1621,
P-1627, P-1631, P-1636, P-1637, P-1638, P-1639, P-1656, P-1660,
P-1687, P-1702, P-1706, P-1707, P-1708, P-1720, P-1722, P-1723,
P-1724, P-1725, P-1727, P-1730, P-1731, P-1742, P-1748, P-1749,
P-1750, P-1751, P-1755, P-1756, P-1757, P-1759, P-1760, P-1764,
P-1765, P-1767, P-1770, P-1775, P-1776, P-1777, P-1778, P-1779,
P-1827, P-1828, P-1839, P-1842, P-1864, P-1873, P-1878, P-1879,
P-1896, P-1897, P-1898, P-2007
TABLE-US-00051 TABLE 2q Compounds with activity toward kinase Jnk2
with IC.sub.50 .ltoreq. 10 .mu.M Jnk2: P-0001, P-0005, P-0006,
P-0009, P-0013, P-0015, P-0016, P-0025, P-0027, P-0033, P-0034,
P-0035, P-0040, P-0042, P-0052, P-0054, P-0056, P-0066, P-0069,
P-0079, P-0082, P-0088, P-0090, P-0121, P-0140, P-0142, P-0156,
P-0184, P-0196, P-0204, P-0228, P-0238, P-0269, P-0285, P-0297,
P-0308, P-0448, P-0486, P-0521, P-0579, P-0594, P-0599, P-0623,
P-0685, P-0700, P-0721, P-0734, P-0744, P-0746, P-0774, P-1253,
P-1318, P-1445, P-1447, P-1486, P-1547, P-1548, P-1554, P-1566,
P-1567, P-1570, P-1575, P-1576, P-1589, P-1591, P-1602, P-1611,
P-1621, P-1627, P-1656, P-1671, P-1687, P-1700, P-1702, P-1711,
P-1720, P-1722, P-1723, P-1724, P-1727, P-1728, P-1729, P-1730,
P-1731, P-1732, P-1737, P-1742, P-1748, P-1749, P-1750, P-1751,
P-1753, P-1755, P-1756, P-1757, P-1759, P-1760, P-1764, P-1765,
P-1767, P-1770, P-1776, P-1777, P-1778, P-1779, P-1827, P-1828,
P-1864, P-2007
TABLE-US-00052 TABLE 2r Compounds with activity toward kinase Jnk3
with IC.sub.50 .ltoreq. 10 .mu.M Jnk3: P-0001, P-0002, P-0003,
P-0004, P-0006, P-0008, P-0009, P-0010, P-0015, P-0016, P-0024,
P-0025, P-0026, P-0027, P-0031, P-0032, P-0033, P-0034, P-0035,
P-0036, P-0037, P-0038, P-0040, P-0041, P-0045, P-0047, P-0052,
P-0054, P-0056, P-0058, P-0059, P-0066, P-0078, P-0079, P-0080,
P-0088, P-0089, P-0090, P-0093, P-0102, P-0112, P-0114, P-0115,
P-0117, P-0122, P-0132, P-0133, P-0134, P-0165, P-0166, P-0167,
P-0176, P-0179, P-0184, P-0189, P-0190, P-0196, P-0204, P-0211,
P-0213, P-0218, P-0228, P-0238, P-0244, P-0257, P-0263, P-0269,
P-0279, P-0285, P-0300, P-0308, P-0313, P-0320, P-0371, P-0378,
P-0448, P-0483, P-0521, P-0550, P-0559, P-0562, P-0579, P-0594,
P-0599, P-0604, P-0624, P-0625, P-0632, P-0636, P-0640, P-0645,
P-0656, P-0659, P-0668, P-0671, P-0675, P-0682, P-0683, P-0691,
P-0697, P-0698, P-0703, P-0710, P-0716, P-0734, P-0738, P-0753,
P-0755, P-0757, P-0763, P-0774, P-0778, P-0807, P-0822, P-0851,
P-0951, P-0962, P-0991, P-1002, P-1005, P-1008, P-1010, P-1011,
P-1016, P-1018, P-1021, P-1022, P-1032, P-1082, P-1087, P-1088,
P-1253, P-1279, P-1280, P-1289, P-1317, P-1318, P-1346, P-1347,
P-1348, P-1349, P-1355, P-1356, P-1359, P-1372, P-1375, P-1384,
P-1385, P-1394, P-1400, P-1445, P-1447, P-1458, P-1465, P-1468,
P-1473, P-1475, P-1477, P-1485, P-1486, P-1489, P-1490, P-1505,
P-1529, P-1534, P-1546, P-1547, P-1548, P-1554, P-1561, P-1566,
P-1567, P-1570, P-1576, P-1577, P-1585, P-1589, P-1591, P-1592,
P-1610, P-1611, P-1618, P-1621, P-1636, P-1687, P-1702, P-1703,
P-1704, P-1706, P-1707, P-1713, P-1716, P-1720, P-1722, P-1724,
P-1742, P-1748, P-1749, P-1750, P-1753, P-1756, P-1757, P-1759,
P-1764, P-1765, P-1767, P-1770, P-1775, P-1776, P-1777, P-1827,
P-1828, P-1864, P-2007
TABLE-US-00053 TABLE 2s Compounds with activity toward kinase Kdr
with IC.sub.50 .ltoreq. 10 .mu.M Kdr: P-0001, P-0003, P-0005,
P-0006, P-0007, P-0009, P-0011, P-0012, P-0013, P-0014, P-0015,
P-0016, P-0017, P-0018, P-0019, P-0020, P-0021, P-0022, P-0023,
P-0024, P-0025, P-0027, P-0028, P-0029, P-0030, P-0031, P-0032,
P-0033, P-0039, P-0040, P-0041, P-0042, P-0043, P-0044, P-0045,
P-0046, P-0048, P-0049, P-0050, P-0051, P-0052, P-0053, P-0054,
P-0055, P-0056, P-0057, P-0059, P-0060, P-0061, P-0063, P-0064,
P-0065, P-0067, P-0068, P-0069, P-0070, P-0071, P-0073, P-0074,
P-0075, P-0076, P-0077, P-0078, P-0080, P-0081, P-0082, P-0084,
P-0085, P-0086, P-0087, P-0088, P-0089, P-0090, P-0091, P-0092,
P-0093, P-0096, P-0097, P-0098, P-0100, P-0101, P-0102, P-0103,
P-0105, P-0106, P-0107, P-0108, P-0110, P-0111, P-0112, P-0113,
P-0114, P-0115, P-0117, P-0120, P-0122, P-0125, P-0126, P-0127,
P-0128, P-0129, P-0130, P-0131, P-0133, P-0134, P-0135, P-0136,
P-0137, P-0139, P-0140, P-0141, P-0142, P-0143, P-0144, P-0145,
P-0147, P-0149, P-0152, P-0157, P-0158, P-0161, P-0162, P-0163,
P-0164, P-0165, P-0167, P-0168, P-0169, P-0170, P-0172, P-0173,
P-0174, P-0175, P-0176, P-0177, P-0179, P-0180, P-0182, P-0184,
P-0189, P-0190, P-0192, P-0194, P-0195, P-0196, P-0197, P-0198,
P-0199, P-0202, P-0203, P-0204, P-0206, P-0212, P-0213, P-0216,
P-0218, P-0224, P-0225, P-0226, P-0230, P-0234, P-0242, P-0243,
P-0260, P-0261, P-0262, P-0267, P-0268, P-0269, P-0270, P-0274,
P-0279, P-0287, P-0288, P-0289, P-0293, P-0295, P-0296, P-0299,
P-0308, P-0316, P-0319, P-0320, P-0321, P-0322, P-0326, P-0337,
P-0339, P-0369, P-0376, P-0379, P-0391, P-0409, P-0418, P-0427,
P-0448, P-0455, P-0458, P-0473, P-0482, P-0495, P-0521, P-0550,
P-0559, P-0562, P-0579, P-0611, P-0623, P-0624, P-0632, P-0640,
P-0645, P-0668, P-0679, P-0683, P-0691, P-0703, P-0708, P-0730,
P-0737, P-0738, P-0751, P-0762, P-0771, P-0777, P-0796, P-0806,
P-0813, P-0933, P-0951, P-0956, P-0962, P-0981, P-1023, P-1244,
P-1280, P-1318, P-1394, P-1426
TABLE-US-00054 TABLE 2t Compounds with activity toward kinase Kit
with IC.sub.50 .ltoreq. 10 .mu.M Kit: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0011, P-0012,
P-0013, P-0016, P-0017, P-0018, P-0019, P-0020, P-0021, P-0024,
P-0026, P-0027, P-0028, P-0029, P-0030, P-0031, P-0032, P-0033,
P-0034, P-0035, P-0036, P-0037, P-0038, P-0039, P-0040, P-0041,
P-0042, P-0050, P-0054, P-0056, P-0059, P-0064, P-0067, P-0069,
P-0072, P-0078, P-0079, P-0080, P-0082, P-0086, P-0088, P-0090,
P-0093, P-0097, P-0101, P-0102, P-0103, P-0112, P-0114, P-0115,
P-0117, P-0120, P-0121, P-0122, P-0123, P-0127, P-0129, P-0132,
P-0133, P-0134, P-0136, P-0140, P-0142, P-0152, P-0154, P-0157,
P-0166, P-0167, P-0168, P-0171, P-0175, P-0176, P-0179, P-0180,
P-0184, P-0189, P-0190, P-0195, P-0196, P-0204, P-0205, P-0206,
P-0210, P-0211, P-0213, P-0216, P-0217, P-0218, P-0224, P-0233,
P-0235, P-0237, P-0244, P-0248, P-0253, P-0257, P-0262, P-0263,
P-0265, P-0269, P-0274, P-0279, P-0284, P-0287, P-0289, P-0293,
P-0298, P-0300, P-0302, P-0304, P-0315, P-0316, P-0321, P-0323,
P-0325, P-0326, P-0333, P-0341, P-0346, P-0352, P-0367, P-0369,
P-0371, P-0373, P-0378, P-0384, P-0385, P-0391, P-0392, P-0396,
P-0402, P-0404, P-0409, P-0411, P-0418, P-0420, P-0421, P-0427,
P-0447, P-0448, P-0450, P-0453, P-0459, P-0461, P-0473, P-0483,
P-0491, P-0495, P-0499, P-0501, P-0511, P-0519, P-0521, P-0535,
P-0550, P-0554, P-0556, P-0559, P-0561, P-0562, P-0563, P-0579,
P-0594, P-0599, P-0604, P-0611, P-0613, P-0617, P-0623, P-0624,
P-0625, P-0626, P-0632, P-0636, P-0640, P-0644, P-0645, P-0647,
P-0656, P-0658, P-0659, P-0668, P-0671, P-0679, P-0682, P-0683,
P-0685, P-0690, P-0691, P-0693, P-0697, P-0699, P-0700, P-0703,
P-0708, P-0721, P-0733, P-0736, P-0737, P-0738, P-0749, P-0751,
P-0753, P-0755, P-0757, P-0762, P-0763, P-0771, P-0773, P-0774,
P-0776, P-0778, P-0794, P-0796, P-0798, P-0800, P-0806, P-0810,
P-0813, P-0815, P-0818, P-0825, P-0835, P-0837, P-0848, P-0850,
P-0851, P-0853, P-0854, P-0857, P-0860, P-0861, P-0865, P-0866,
P-0867, P-0874, P-0876, P-0877, P-0885, P-0889, P-0898, P-0905,
P-0907, P-0910, P-0911, P-0913, P-0919, P-0924, P-0927, P-0931,
P-0933, P-0935, P-0937, P-0951, P-0952, P-0954, P-0955, P-0956,
P-0958, P-0962, P-0964, P-0978, P-0983, P-1002, P-1008, P-1009,
P-1010, P-1013, P-1016, P-1018, P-1021, P-1033, P-1082, P-1084,
P-1096, P-1110, P-1112, P-1160, P-1181, P-1194, P-1246, P-1247,
P-1249, P-1250, P-1251, P-1252, P-1253, P-1254, P-1255, P-1256,
P-1257, P-1259, P-1260, P-1261, P-1262, P-1263, P-1264, P-1266,
P-1267, P-1268, P-1269, P-1275, P-1279, P-1280, P-1281, P-1289,
P-1316, P-1317, P-1318, P-1320, P-1321, P-1323, P-1329, P-1336,
P-1338, P-1341, P-1343, P-1346, P-1347, P-1348, P-1349, P-1365,
P-1366, P-1367, P-1368, P-1369, P-1370, P-1372, P-1376, P-1380,
P-1382, P-1383, P-1384, P-1385, P-1386, P-1387, P-1388, P-1389,
P-1390, P-1391, P-1392, P-1393, P-1394, P-1395, P-1396, P-1397,
P-1399, P-1400, P-1402, P-1403, P-1404, P-1406, P-1407, P-1408,
P-1409, P-1410, P-1411, P-1413, P-1414, P-1415, P-1416, P-1417,
P-1419, P-1420, P-1422, P-1423, P-1424, P-1425, P-1426, P-1427,
P-1428, P-1429, P-1430, P-1431, P-1432, P-1433, P-1445, P-1446,
P-1447, P-1449, P-1450, P-1451, P-1452, P-1453, P-1454, P-1455,
P-1456, P-1457, P-1458, P-1460, P-1461, P-1462, P-1463, P-1464,
P-1465, P-1466, P-1467, P-1468, P-1469, P-1470, P-1471, P-1472,
P-1474, P-1475, P-1476, P-1478, P-1479, P-1480, P-1481, P-1482,
P-1483, P-1484, P-1486, P-1488, P-1489, P-1490, P-1493, P-1495,
P-1497, P-1498, P-1499, P-1500, P-1501, P-1502, P-1503, P-1505,
P-1506, P-1514, P-1521, P-1522, P-1525, P-1526, P-1527, P-1528,
P-1529, P-1530, P-1531, P-1532, P-1534, P-1538, P-1541, P-1542,
P-1543, P-1544, P-1545, P-1546, P-1547, P-1548, P-1549, P-1550,
P-1551, P-1552, P-1553, P-1554, P-1557, P-1559, P-1562, P-1564,
P-1565, P-1566, P-1567, P-1568, P-1569, P-1574, P-1575, P-1576,
P-1578, P-1580, P-1581, P-1582, P-1583, P-1590, P-1591, P-1593,
P-1598, P-1599, P-1605, P-1630, P-1671, P-1685, P-1700, P-1703,
P-1704, P-1705, P-1706, P-1707, P-1708, P-1709, P-1711, P-1712,
P-1713, P-1714, P-1718, P-1719, P-1720, P-1733, P-1737, P-1739,
P-1740, P-1767, P-1776, P-1783, P-1798, P-1822, P-1839, P-1840,
P-1864, P-1865, P-1871, P-1872, P-1873, P-1878, P-1879, P-1881,
P-1882, P-1980, P-1996, P-1997, P-1998
TABLE-US-00055 TABLE 2u Compounds with activity toward kinase
MAP2K1 with IC.sub.50 .ltoreq. 10 .mu.M MAP2K1: P-0001, P-0002,
P-0003, P-0005, P-0006, P-0008, P-0010, P-0011, P-0012, P-0014,
P-0015, P-0017, P-0018, P-0022, P-0023, P-0029, P-0031, P-0041,
P-0046, P-0051, P-0057, P-0058, P-0061, P-0076, P-0079, P-0081,
P-0087, P-0098, P-0099, P-0105, P-0108, P-0111, P-0120, P-0149,
P-0152, P-0158, P-0167, P-0170, P-0177, P-0194, P-0198, P-0331,
P-0337, P-0568, P-0806
TABLE-US-00056 TABLE 2v Compounds with activity towards kinase
MAPKAPK2 with IC.sub.50 .ltoreq. 10 .mu.M MAPKAPK2: P-0007, P-0041,
P-0057, P-0058, P-0077, P-0086, P-0104, P-0106, P-0151, P-0226
TABLE-US-00057 TABLE 2w Compounds with activity toward kinase Met
with IC.sub.50 .ltoreq. 10 .mu.M Met: P-0001, P-0002, P-0004,
P-0006, P-0008, P-0009, P-0015, P-0016, P-0020, P-0026, P-0027,
P-0028, P-0034, P-0035, P-0037, P-0038, P-0041, P-0052, P-0054,
P-0058, P-0066, P-0076, P-0078, P-0082, P-0101, P-0114, P-0117,
P-0140, P-0146, P-0149, P-0156, P-0165, P-0184, P-0228, P-0262,
P-0269, P-0320, P-0325, P-0369, P-0419, P-0542, P-0550, P-0675,
P-0685, P-0700, P-0716, P-0721, P-0746, P-0761, P-0763, P-0773,
P-0778, P-0781, P-0811, P-0822, P-0842, P-0951, P-0962, P-1008,
P-1012, P-1023, P-1037, P-1068, P-1092, P-1114, P-1132, P-1280,
P-1394, P-1465, P-1527
TABLE-US-00058 TABLE 2x Compounds with activity toward kinase p38
with IC.sub.50 .ltoreq. 10 .mu.M p38: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0008, P-0010, P-0016, P-0019, P-0025,
P-0026, P-0027, P-0028, P-0032, P-0034, P-0035, P-0036, P-0037,
P-0038, P-0041, P-0045, P-0050, P-0056, P-0058, P-0059, P-0066,
P-0068, P-0079, P-0082, P-0088, P-0090, P-0093, P-0112, P-0114,
P-0122, P-0134, P-0155, P-0156, P-0158, P-0163, P-0167, P-0170,
P-0184, P-0190, P-0196, P-0204, P-0205, P-0210, P-0228, P-0244,
P-0257, P-0262, P-0267, P-0297, P-0302, P-0308, P-0369, P-0442,
P-0448, P-0511, P-0519, P-0521, P-0550, P-0559, P-0579, P-0594,
P-0599, P-0604, P-0611, P-0624, P-0636, P-0645, P-0668, P-0671,
P-0679, P-0685, P-0691, P-0699, P-0700, P-0703, P-0708, P-0716,
P-0721, P-0728, P-0734, P-0738, P-0744, P-0745, P-0746, P-0751,
P-0753, P-0757, P-0763, P-0773, P-0774, P-0776, P-0798, P-0806,
P-0807, P-0841, P-0842, P-0868, P-0884, P-0887, P-0933, P-1042,
P-1046
TABLE-US-00059 TABLE 2y Compounds with activity toward kinase
PDGFRB with IC.sub.50 .ltoreq. 10 .mu.M PDGFRB: P-0088, P-0262
TABLE-US-00060 TABLE 2z Compounds with activity toward kinase Pim1
with IC.sub.50 .ltoreq. 10 .mu.M Pim1: P-0024, P-0090
TABLE-US-00061 TABLE 2aa Compounds with activity toward kinase PKC
theta with IC.sub.50 .ltoreq. 10 .mu.M PKC theta: P-0001, P-0002,
P-0003, P-0004, P-0006, P-0008, P-0011, P-0012, P-0013, P-0014,
P-0017, P-0019, P-0020, P-0021, P-0022, P-0024, P-0025, P-0026,
P-0030, P-0031, P-0039, P-0044, P-0046, P-0049, P-0051, P-0055,
P-0057, P-0060, P-0063, P-0069, P-0070, P-0109, P-0112, P-0238,
P-0270
TABLE-US-00062 TABLE 2bb Compounds with activity toward kinase Pyk2
with IC.sub.50 .ltoreq. 10 .mu.M Pyk2: P-0001, P-0010, P-0015,
P-0018, P-0021, P-0024, P-0032, P-0041, P-0052, P-0056, P-0058,
P-0069, P-0078, P-0086, P-0088, P-0095, P-0112, P-0116, P-0122,
P-0137, P-0142, P-0166, P-0178, P-0188, P-0257, P-0262, P-0269,
P-0287, P-0318, P-0320, P-0356, P-0369, P-0396, P-0486, P-0501,
P-0521, P-0529, P-0550, P-0559, P-0562, P-0611, P-0636, P-0685,
P-0697, P-0700, P-0728, P-0753, P-0806, P-0818, P-0837, P-0848,
P-0850, P-0851, P-0857, P-0861, P-0866, P-0867, P-0874, P-0883,
P-0897, P-0898, P-0910, P-0911, P-0912, P-0919, P-0924, P-0928,
P-0944, P-0946, P-0947, P-0957, P-0964, P-0978, P-0991, P-0997,
P-1002, P-1010, P-1018, P-1069, P-1280, P-1281, P-1282, P-1288,
P-1316, P-1349, P-1365, P-1367, P-1368, P-1370, P-1372, P-1373,
P-1375, P-1376, P-1377, P-1379, P-1380, P-1381
TABLE-US-00063 TABLE 2cc Compounds with activity toward kinase Ret
with IC.sub.50 .ltoreq. 10 .mu.M Ret: P-0001, P-0001, P-0002,
P-0003, P-0003, P-0004, P-0005, P-0005, P-0006, P-0008, P-0009,
P-0010, P-0011, P-0012, P-0013, P-0014, P-0015, P-0016, P-0017,
P-0019, P-0020, P-0022, P-0024, P-0025, P-0026, P-0027, P-0028,
P-0028, P-0029, P-0032, P-0032, P-0033, P-0034, P-0035, P-0036,
P-0037, P-0038, P-0039, P-0040, P-0040, P-0041, P-0041, P-0042,
P-0043, P-0044, P-0045, P-0046, P-0047, P-0048, P-0049, P-0050,
P-0051, P-0052, P-0053, P-0054, P-0055, P-0056, P-0058, P-0059,
P-0060, P-0061, P-0062, P-0063, P-0064, P-0065, P-0066, P-0069,
P-0070, P-0071, P-0072, P-0073, P-0075, P-0078, P-0079, P-0082,
P-0083, P-0085, P-0088, P-0089, P-0090, P-0094, P-0095, P-0096,
P-0097, P-0099, P-0100, P-0101, P-0102, P-0103, P-0107, P-0108,
P-0109, P-0110, P-0113, P-0114, P-0115, P-0116, P-0117, P-0119,
P-0121, P-0122, P-0123, P-0124, P-0134, P-0135, P-0137, P-0138,
P-0139, P-0140, P-0141, P-0142, P-0148, P-0152, P-0156, P-0158,
P-0159, P-0165, P-0167, P-0171, P-0175, P-0179, P-0181, P-0184,
P-0186, P-0190, P-0196, P-0196, P-0204, P-0205, P-0206, P-0210,
P-0211, P-0215, P-0218, P-0224, P-0228, P-0231, P-0232, P-0236,
P-0244, P-0244, P-0245, P-0246, P-0248, P-0250, P-0257, P-0262,
P-0265, P-0269, P-0280, P-0286, P-0289, P-0293, P-0297, P-0302,
P-0304, P-0307, P-0308, P-0314, P-0316, P-0316, P-0320, P-0320,
P-0321, P-0325, P-0329, P-0339, P-0341, P-0344, P-0347, P-0351,
P-0352, P-0363, P-0367, P-0369, P-0371, P-0378, P-0385, P-0392,
P-0396, P-0412, P-0418, P-0434, P-0448, P-0448, P-0452, P-0453,
P-0453, P-0469, P-0472, P-0486, P-0495, P-0501, P-0501, P-0517,
P-0520, P-0521, P-0521, P-0533, P-0536, P-0542, P-0550, P-0550,
P-0559, P-0559, P-0561, P-0579, P-0594, P-0596, P-0599, P-0599,
P-0604, P-0608, P-0611, P-0623, P-0623, P-0624, P-0624, P-0632,
P-0636, P-0638, P-0640, P-0644, P-0645, P-0645, P-0647, P-0656,
P-0659, P-0668, P-0668, P-0671, P-0675, P-0678, P-0679, P-0682,
P-0683, P-0691, P-0693, P-0697, P-0698, P-0699, P-0700, P-0703,
P-0708, P-0710, P-0716, P-0721, P-0726, P-0728, P-0730, P-0734,
P-0735, P-0736, P-0737, P-0738, P-0744, P-0745, P-0746, P-0749,
P-0751, P-0753, F-0757, P-0761, P-0762, P-0763, P-0771, P-0778,
P-0794, P-0795, P-0796, P-0807, P-0810, P-0811, P-0813, P-0822,
P-0825, P-0826, P-0835, P-0841, P-0863, P-0865, P-0881, P-0939,
P-0976, P-0977, P-0985, P-0998, P-1000, P-1005, P-1007, P-1011,
P-1019, P-1024, P-1025, P-1026, P-1029, P-1031, P-1033, P-1036,
P-1066, P-1072, P-1073, P-1075, P-1081, P-1085, P-1089, P-1097,
P-1111, P-1113, P-1115, P-1117, P-1119, P-1121, P-1125, P-1126,
P-1129, P-1130, P-1133, P-1134, P-1135, P-1137, P-1178, P-1181,
P-1185, P-1188, P-1189, P-1191, P-1192, P-1193, P-1196, P-1198,
P-1200, P-1201
TABLE-US-00064 TABLE 2dd Compounds with activity toward kinase Src
with IC.sub.50 .ltoreq. 10 .mu.M Src: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0007, P-0008, P-0009, P-0010, P-0016,
P-0017, P-0021, P-0025, P-0026, P-0027, P-0028, P-0032, P-0034,
P-0035, P-0036, P-0037, P-0038, P-0040, P-0041, P-0045, P-0055,
P-0067, P-0068, P-0072, P-0078, P-0079, P-0082, P-0088, P-0090,
P-0102, P-0112, P-0114, P-0134, P-0152, P-0166, P-0171, P-0196,
P-0209, P-0210, P-0237, P-0244, P-0269, P-0302, P-0316, P-0320,
P-0373, P-0396, P-0448, P-0453, P-0483, P-0501, P-0515, P-0521,
P-0550, P-0559, P-0562, P-0599, P-0623, P-0624, P-0645, P-0647,
P-0668, P-0685, P-0700, P-0721, P-0753, P-0763, P-0771, P-0774,
P-0805, P-0806, P-0807, P-0818, P-0837, P-0848, P-0850, P-0851,
P-0853, P-0857, P-0866, P-0874, P-0876, P-0877, P-0885, P-0889,
P-0898, P-0907, P-0910, P-0933, P-0950, P-0952, P-0955, P-0956,
P-0958, P-0997, P-1009, P-1010, P-1013, P-1020, P-1021, P-1181,
P-1247, P-1249, P-1250, P-1251, P-1252, P-1253, P-1254, P-1255,
P-1256, P-1257, P-1258, P-1261, P-1262, P-1263, P-1264, P-1265,
P-1266, P-1267, P-1269, P-1280, P-1281, P-1288, P-1289, P-1316,
P-1317, P-1318, P-1336, P-1338, P-1343, P-1345, P-1346, P-1347,
P-1348, P-1349, P-1366, P-1383, P-1384, P-1385, P-1387, P-1388,
P-1390, P-1391, P-1394, P-1396, P-1397, P-1398, P-1399, P-1403,
P-1416, P-1417, P-1431, P-1432, P-1433, P-1445, P-1446, P-1447,
P-1448, P-1451, P-1457, P-1459, P-1469, P-1472, P-1473, P-1475,
P-1476, P-1477, P-1478, P-1479, P-1480, P-1481, P-1484, P-1485,
P-1486, P-1495, P-1496, P-1506, P-1527, P-1530
TABLE-US-00065 TABLE 2ee Compounds with activity toward kinase Stk6
with IC.sub.50 .ltoreq. 10 .mu.M Stk6: P-0001, P-0002, P-0003,
P-0007, P-0008, P-0009, P-0010, P-0011, P-0013, P-0014, P-0015,
P-0018, P-0020, P-0022, P-0023, P-0029, P-0030, P-0031, P-0032,
P-0033, P-0035, P-0040, P-0041, P-0042, P-0043, P-0044, P-0045,
P-0046, P-0049, P-0051, P-0052, P-0054, P-0056, P-0057, P-0058,
P-0060, P-0061, P-0063, P-0064, P-0065, P-0069, P-0070, P-0071,
P-0076, P-0077, P-0080, P-0081, P-0082, P-0086, P-0087, P-0088,
P-0091, P-0092, P-0093, P-0094, P-0098, P-0099, P-0100, P-0101,
P-0104, P-0105, P-0106, P-0110, P-0111, P-0115, P-0117, P-0118,
P-0119, P-0120, P-0123, P-0127, P-0128, P-0129, P-0131, P-0132,
P-0133, P-0136, P-0140, P-0143, P-0146, P-0147, P-0148, P-0153,
P-0154, P-0155, P-0157, P-0160, P-0162, P-0163, P-0164, P-0169,
P-0172, P-0173, P-0174, P-0176, P-0177, P-0179, P-0181, P-0185,
P-0187, P-0188, P-0189, P-0191, P-0193, P-0199, P-0201, P-0202,
P-0203, P-0206, P-0207, P-0208, P-0212, P-0213, P-0214, P-0221,
P-0225, P-0235, P-0237, P-0249, P-0250, P-0251, P-0253, P-0260,
P-0261, P-0269, P-0272, P-0276, P-0279, P-0281, P-0283, P-0287,
P-0290, P-0295, P-0300, P-0313, P-0317, P-0319, P-0322, P-0345,
P-0348, P-0355, P-0370, P-0372, P-0406, P-0407, P-0417, P-0419,
P-0426, P-0436, P-0441, P-0445, P-0469, P-0471, P-0489, P-0546,
P-0806, P-0885, P-0933, P-0955, P-1013, P-1280, P-1336, P-1394,
P-1426
TABLE-US-00066 TABLE 2ff Compounds with activity toward kinase Yes
with IC.sub.50 .ltoreq. 10 .mu.M Yes: P-0005, P-0007, P-0010,
P-0011, P-0012, P-0013, P-0014, P-0015, P-0016, P-0017, P-0019,
P-0020, P-0021, P-0022, P-0024, P-0027, P-0028, P-0029, P-0031,
P-0033, P-0036, P-0040, P-0042, P-0043, P-0044, P-0045, P-0046,
P-0047, P-0048, P-0049, P-0050, P-0051, P-0052, P-0053, P-0055,
P-0056, P-0059, P-0060, P-0061, P-0062, P-0063, P-0065, P-0067,
P-0068, P-0070, P-0072, P-0074, P-0075, P-0081, P-0082, P-0083,
P-0088, P-0090, P-0093, P-0095, P-0097, P-0101, P-0102, P-0107,
P-0109, P-0112, P-0122, P-0124, P-0125, P-0126, P-0129, P-0134,
P-0138, P-0139, P-0145, P-0152, P-0153, P-0161, P-0162, P-0166,
P-0171, P-0175, P-0188, P-0202, P-0209, P-0210, P-0230, P-0237,
P-0271, P-0283, P-0310, P-0327, P-0483, P-0636
TABLE-US-00067 TABLE 2gg Compounds with activity toward kinase
Zap70 with IC.sub.50 .ltoreq. 10 .mu.M Zap70: P-0001, P-0004,
P-0015, P-0030, P-0032, P-0033, P-0034, P-0035, P-0037, P-0038,
P-0040, P-0041, P-0047, P-0058, P-0123, P-0193, P-0195, P-0205,
P-0218, P-0228, P-0249, P-0275, P-0296, P-0310, P-0320, P-0342,
P-0348, P-0359, P-0360, P-0378, P-0379, P-0387, P-0394, P-0434,
P-0442, P-0456, P-0476, P-0484, P-0495, P-0500, P-0507, P-0523,
P-0550, P-0586, P-0602, P-0607, P-0611, P-0624, P-0642, P-0649,
P-0675, P-0676, P-0694, P-0698, P-0703, P-0716, P-0724, P-0727,
P-0755, P-0795, P-0808, P-0836, P-0842, P-0856, P-0859, P-0865,
P-0875, P-0878, P-0880, P-0888, P-0929, P-0930, P-0953, P-0982,
P-0996, P-1000, P-1005, P-1019, P-1051, P-1073, P-1081, P-1089,
P-1119, P-1184, P-1197
TABLE-US-00068 TABLE 2hh Compounds with activity toward kinase Akt3
with IC.sub.50 .ltoreq. 10 .mu.M Akt3: P-0001, P-0002, P-0003,
P-0004, P-0005, P-0006, P-0008, P-0019, P-0021, P-0024, P-0025,
P-0026, P-0027, P-0034, P-0035, P-0036, P-0037, P-0038, P-0041,
P-0048, P-0055, P-0057, P-0058, P-0060, P-0067, P-0099, P-0112,
P-0114, P-0122, P-0127, P-0154, P-0196, P-0270, P-0278, P-0307,
P-0329, P-0404, P-0436, P-0573, P-0861, P-0866, P-0883, P-0890,
P-0924, P-0936, P-0940, P-0951, P-0952, P-0958, P-0962, P-0978,
P-0990, P-0995, P-1010, P-1016, P-1018, P-1021, P-1022, P-1027,
P-1034, P-1039, P-1040, P-1043, P-1047, P-1060, P-1062, P-1082,
P-1095, P-1139, P-1145, P-1147, P-1150, P-1155, P-1158, P-1172,
P-1234, P-1288, P-1459
TABLE-US-00069 TABLE 2ii Compounds with activity toward kinase ALK
with IC.sub.50 .ltoreq. 10 .mu.M ALK: P-0806, P-1280, P-1244,
P-1336, P-1394, P-1426
TABLE-US-00070 TABLE 2jj Compounds with activity toward kinase Cdk2
with IC.sub.50 .ltoreq. 10 .mu.M Cdk2: P-0806, P-1280, P-1244,
P-1394
TABLE-US-00071 TABLE 2kk Compounds with activity toward kinase Csk
with IC.sub.50 .ltoreq. 10 .mu.M Csk: P-0007, P-0805, P-0806,
P-0885, P-0933, P-0955, P-0956, P-1013, P-1020, P-1336, P-1394,
P-1426
TABLE-US-00072 TABLE 20ll Compounds with activity toward kinase
EphA2 with IC.sub.50 .ltoreq. 10 .mu.M EphA2: P-1336, P-1394
TABLE-US-00073 TABLE 2mm Compounds with activity toward kinase
EphB4 with IC.sub.50 .ltoreq. 10 .mu.M EphB4: P-0806, P-1336
TABLE-US-00074 TABLE 2nn Compounds with activity toward kinase Frk
with IC.sub.50 .ltoreq. 10 .mu.M Frk: P-0007, P-0805, P-0885,
P-0933, P-0955, P-0956, P-1013, P-1020, P-1244, P-1318, P-1336,
P-1394
TABLE-US-00075 TABLE 2oo Compounds with activity toward kinase
Gsk3.beta. with IC.sub.50 .ltoreq. 10 .mu.M Gsk3.beta.: P-0007,
P-0015, P-0017, P-0053, P-0057, P-0079, P-0081, P-0085, P-0086,
P-0094, P-0104, P-0106, P-0109, P-0123, P-0135, P-0148, P-0154,
P-0159, P-0169, P-0180, P-0207, P-0226, P-0236, P-0252, P-0273,
P-0462, P-0700, P-0728, P-0763, P-0850, P-0902, P-0913, P-0969,
P-1002, P-1142, P-1181, P-1252, P-1317, P-1336, P-1372, P-1426
TABLE-US-00076 TABLE 2pp Compounds with activity toward kinase Hck
with IC.sub.50 .ltoreq. 10 .mu.M Hck: P-0007, P-0806, P-0885,
P-0933, P-1318, P-1336, P-1394, P-1426
TABLE-US-00077 TABLE 2qq Compounds with activity toward kinase
MAP4K4 with IC.sub.50 .ltoreq. 10 .mu.M MAP4K4: P-0007, P-0057,
P-0069, P-0079, P-0082, P-0088, P-0130, P-0131, P-0152, P-0174,
P-0176, P-0198, P-0202, P-0214, P-0220, P-0256, P-0269, P-0287,
P-0300, P-0317, P-0357, P-0367, P-0369, P-0391, P-0402, P-0442,
P-0449, P-0477, P-0488, P-0495, P-0518, P-0527, P-0537, P-0573,
P-0601, P-0685, P-0695, P-0700, P-0728, P-0734, P-0753, P-0800,
P-0806, P-0811, P-0850, P-0851, P-0853, P-0862, P-0885, P-0896,
P-0902, P-0904, P-0909, P-0913, P-0931, P-0933, P-0937, P-0954,
P-0958, P-0971, P-0986, P-1017, P-1042, P-1056, P-1252, P-1253,
P-1279, P-1280, P-1289, P-1317, P-1318, P-1336, P-1372, P-1383,
P-1394, P-1406, P-1411, P-1414, P-1415, P-1417, P-1418, P-1426,
P-1429, P-1685
TABLE-US-00078 TABLE 2rr Compounds with activity toward kinase
IGF1R with IC.sub.50 .ltoreq. 10 .mu.M IGF1R: P-0002, P-0003,
P-0004, P-0009, P-0031, P-0079, P-0080, P-0084, P-0115, P-0136,
P-0154, P-0157, P-0212, P-0213, P-0700, P-0716, P-0746, P-0850,
P-1336, P-1337, P-1390, P-1394
TABLE-US-00079 TABLE 2ss Compounds with activity toward kinase IKK
beta with IC.sub.50 .ltoreq. 10 .mu.M IKK beta: P-0007, P-0013,
P-0014, P-0029, P-0057, P-0073, P-0084, P-0085, P-0086, P-0087,
P-0096, P-0098, P-0106, P-0111, P-0115, P-0120, P-0127, P-0128,
P-0133, P-0135, P-0163, P-0164, P-0172, P-0177, P-0179, P-0216,
P-0270, P-0272, P-0315, P-0376, P-0404, P-0410, P-0436, P-0629,
P-0682, P-0690, P-0790, P-0896, P-0920, P-0962, P-1223
TABLE-US-00080 TABLE 2tt Compounds with activity toward kinase Itk
with IC.sub.50 .ltoreq. 10 .mu.M Itk: P-0002, P-0003, P-0004,
P-0006, P-0008, P-0009, P-0013, P-0016, P-0019, P-0020, P-0024,
P-0025, P-0027, P-0031, P-0034, P-0035, P-0036, P-0038, P-0067,
P-0173, P-0196, P-0521, P-0579, P-0716, P-0778, P-0883, P-0951,
P-1016, P-1067, P-1337, P-1385
TABLE-US-00081 TABLE 2uu Compounds with activity toward kinase Jak3
with IC.sub.50 .ltoreq. 10 .mu.M Jak3: P-0003, P-0004, P-0009,
P-0013, P-0014, P-0019, P-0020, P-0022, P-0024, P-0034, P-0039,
P-0044, P-0046, P-0049, P-0051, P-0060, P-0061, P-0063, P-0070,
P-0084, P-0101, P-0106, P-0108, P-0109, P-0119, P-0122, P-0124,
P-0138, P-0141, P-0146, P-0171, P-0178, P-0187, P-0215, P-0318,
P-0521, P-0730, P-0863, P-01367, P-01385
TABLE-US-00082 TABLE 2vv Compounds with activity toward kinase MLK1
with IC.sub.50 .ltoreq. 10 .mu.M MLK1: P-1336, P-1426
TABLE-US-00083 TABLE 2ww Compounds with activity toward kinase TrkA
with IC.sub.50 .ltoreq. 10 .mu.M TrkA: P-0409, P-0806, P-1244,
P-1426
TABLE-US-00084 TABLE 2xx Compounds with activity toward kinase
PDGFRA with IC.sub.50 .ltoreq. 10 .mu.M PDGFRA: P-0007, P-0409,
P-0806, P-0885, P-0933, P-1280, P-1336, P-1394, P-1426
TABLE-US-00085 TABLE 2yy Compounds with activity toward kinase PIk1
with IC.sub.50 .ltoreq. 10 .mu.M Plk1: P-0018, P-0022, P-0031,
P-0044, P-0046, P-0067, P-0075, P-0083, P-0085, P-0099, P-0113,
P-0123, P-0128, P-0135, P-0146, P-0148, P-0154, P-0178, P-0286,
P-0332, P-0345, P-0366, P-0480, P-0490, P-0581, P-0863, P-0954,
P-1138
TABLE-US-00086 TABLE 2zz Compounds with activity toward kinase Brk
with IC.sub.50 .ltoreq. 10 .mu.M Brk: P-0007, P-0805, P-0806,
P-0885, P-0933, P-0955, P-0956, P-1013, P-1020, P-1244, P-1318,
P-1336, P-1394
TABLE-US-00087 TABLE 2ab Compounds with activity toward kinase
ROCK1 with IC.sub.50 .ltoreq. 10 .mu.M ROCK1: P-0057
TABLE-US-00088 TABLE 2ac Compounds with activity toward kinase Syk
with IC.sub.50 .ltoreq. 10 .mu.M Syk: P-0002, P-0010, P-0033,
P-0054, P-0056, P-0057, P-0089, P-0196, P-0448, P-0521, P-0599,
P-1336
TABLE-US-00089 TABLE 2ad Compounds with activity toward kinase TEC
with IC.sub.50 .ltoreq. 10 .mu.M TEC: P-0017, P-0033, P-0044,
P-0088, P-0156, P-0166, P-0228, P-0257, P-0297, P-0429, P-0897,
P-0954, P-0983, P-0991, P-0997, P-1020, P-1317
TABLE-US-00090 TABLE 2ae Compounds with activity toward kinase Tie2
with IC.sub.50 .ltoreq. 10 .mu.M Tie2: P-0806, P-1280, P-1336,
P-1394, P-1426
[2047] Plasmid Sequence and PCR Primer Information
TABLE-US-00091 Abl PCR primers Abl ABL-227
CACCACGGTGTGTCCCCCAACTACGA 1424 (SEQ ID NO: _) CABL-A
GTCACGTCGACTCAGACGCCTTGTTTCCCCAGCT 736 (SEQ ID NO: _) P1121.
pET-SPEC BI-PTP Abl G227-V515-X
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccaccacggtgtgtcc M G H
H H H H H G V S
cccaactacgacaagtgggagatggaacgcacggacatcaccatgaagcacaagctgggc P N Y
D K W E M E R T D I T M K H K L G
gggggccagtacggggaggtgtacgagggcgtgtggaagaaatacagcctgacggtggcc G G Q
Y G E V Y E G V W K K Y S L T V A
gtgaagaccttgaaggaggacaccatggaggtggaagagttcttgaaagaagctgcagtc V K T
L K E D T M E V E E F L K E A A V
atgaaagagatcaaacaccctaacctggtgcagctccttggggtctgcacccgggagccc M K E
I K H P N L V Q L L G V C T R E P
ccgttctatatcatcactgagttcatgacctacgggaacctcctggactacctgagggag P F Y
I I T E F M T Y G N L L D Y L R E
tgcaaccggcaggaggtgaacgccgtggtgctgctgtacatggccactcagatctcgtca C N R
Q E V N A V V L L Y M A T Q I S S
gccatggagtacctggagaagaaaaacttcatccacagagatcttgctgcccgaaactgc A M E
Y L E K K N F I H R D L A A R N C
ctggtaggggagaaccacttggtgaaggtagctgattttggcctgagcaggttgatgaca L V G
E N H L V K V A D F G L S R L M T
ggggacacctacacagcccatgctggagccaagttccccatcaaatggactgcacccgag G D T
Y T A H A G A K F P I K W T A P E
agcctggcctacaacaagttctccatcaagtccgacgtctgggcatttggagtattgctt S L A
Y N K F S I K S D V W A F G V L L
tgggaaattgctacctatggcatgtccccttacccgggaattgacctgtcccaggtgtat W E I
A T Y G M S P Y P G I D L S Q V Y
gagctgctagagaaggactaccgcatggagcgcccagaaggctgcccagagaaggtctat E L L
E K D Y R M E R P E G C P E K V Y
gaactcatgcgagcatgttggcagtggaatccctctgaccggccctcctttgctgaaatc E L M
R A C W Q W N P S D R P S F A E I
caccaagcctttgaaacaatgttccaggaatccagtatctcagacgaagtggaaaaggag H Q A
F E T M F Q E S S I S D E V E K E ctggggaaacaaggcgtctgagtcgac (SEQ
ID NO: _) L G K Q G V - (SEQ ID NO: _)
TABLE-US-00092 B-Raf V600E PCR primers BRAF BRAF437D-S
ACGGGACCATATGGATGATTGGGAGATTCCTGA 4783 (SEQ ID NO: _) BRAF722K-A
CACTGGTCGACTATTTTGGCAATGAGCGGGCCA 4784 (SEQ ID NO: _) BRAFV599E-S
GGTCTAGCTACAGAAAAATCTCGATGGAG 893 (SEQ ID NO: _) BRAFV599E-A
CTCCATCGAGATTTTTCTGTAGCTAGACC 894 (SEQ ID NO: _) P4254.
pFastBacBD-CDC37 BRAF D437-K722-X, V600E
tattccggattattcataccgtcccaccatcgggcgcggatctcggtccgaaacc
atgtcgtactaccatcaccatcaccatcacgattacgatatcccaacgaccgaaaacctg M S Y
Y H H H H H H D Y D I P T T E N L
tattttcagggccatatggatgattgggagattcctgatgggcagattacagtgggacaa Y F Q
G H M D D W E I P D G Q I T V G Q
agaattggatctggatcatttggaacagtctacaagggaaagtggcatggtgatgtggca R I G
S G S F G T V Y K G K W H G D V A
gtgaaaatgttgaatgtgacagcacctacacctcagcagttacaagccttcaaaaatgaa V K M
L N V T A P T P Q Q L Q A F K N E
gtaggagtactcaggaaaacacgacatgtgaatatcctactcttcatgggctattccaca V G V
L R K T R H V N I L L F M G Y S T
aagccacaactggctattgttacccagtggtgtgagggctccagcttgtatcaccatctc K P Q
L A I V T Q W C E G S S L Y H H L
catatcattgagaccaaatttgagatgatcaaacttatagatattgcacgacagactgca H I I
E T K F E M I K L I D I A R Q T A
cagggcatggattacttacacgccaagtcaatcatccacagagacctcaagagtaataat Q G M
D Y L H A K S I I H R D L K S N N
atatttcttcatgaagacctcacagtaaaaataggtgattttggtctagctacagaaaaa I F L
H E D L T V K I G D F G L A T E K
tctcgatggagtgggtcccatcagtttgaacagttgtctggatccattttgtggatggca S R W
S G S H Q F E Q L S G S I L W M A
ccagaagtcatcagaatgcaagataaaaatccatacagctttcagtcagatgtatatgca P E V
I R M Q D K N P Y S F Q S D V Y A
tttggaattgttctgtatgaattgatgactggacagttaccttattcaaacatcaacaac F G I
V L Y E L M T G Q L P Y S N I N N
agggaccagataatttttatggtgggacgaggatacctgtctccagatctcagtaaggta R D Q
I I F M V G R G Y L S P D L S K V
cggagtaactgtccaaaagccatgaagagattaatggcagagtgcctcaaaaagaaaaga R S N
C P K A M K R L M A E C L K K K R
gatgagagaccactctttccccaaattctcgcctctattgagctgctggcccgctcattg D E R
P L F P Q I L A S I E L L A R S L
ccaaaatagtcgactagagcctgcagtctcgaggcatgcggtaccaagctt (SEQ ID NO: _)
P K - (SEQ ID NO: _)
TABLE-US-00093 Erk2 PCR primers ERK2 ERK2-S
GGCAGCCCATATGGCGGCGGCGGCGGCGGC 748 (SEQ ID NO: _) ERK2-A
TGTCCGTCGACATTTAAGATCTGTATCCTGG 749 P4227.pET15S ERK2/MEK1DD
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggcagcagccatcatcatcatcatcacagc M G S
S H H H H H H S
agcggcctggtgccgcgcggcagccatatggcggcggcggcgggcgcgggcccggagatg S G L
V P R G S H M A A A A G A G P E M
gtccgcgggcaggtgttcgacgtggggccgcgctacaccaacctctcgtacatcggcgag V R G
Q V F D V G P R Y T N L S Y I G E
ggcgcctacggcatggtgtgctctgcttatgataatgtcaacaaagttcgagtagctatc G A Y
G M V C S A Y D N V N K V R V A I
aagaaaatcagcccctttgagcaccagacctactgccagagaaccctgagggagataaaa K K I
S P F E H Q T Y C Q R T L R E I K
atcttactgcgcttcagacatgagaacatcattggaatcaatgacattattcgagcacca I L L
R F R H E N I I G I N D I I R A P
accatcgagcaaatgaaagatgtatatatagtacaggacctcatggaaacagatctttac T I E
Q M K D V Y I V Q D L M E T D L Y
aagctcttgaagacacaacacctcagcaatgaccatatctgctattttctctaccagatc K L L
K T Q H L S N D H I C Y F L Y Q I
ctcagagggttaaaatatatccattcagctaacgttctgcaccgtgacctcaagccttcc L R G
L K Y I H S A N V L H R D L K P S
aacctgctgctcaacaccacctgtgatctcaagatctgtgactttggcctggcccgtgtt N L L
L N T T C D L K I C D F G L A R V
gcagatccagaccatgatcacacagggttcctgacagaatatgtggccacacgttggtac A D P
D H D H T G F L T E Y V A T R W Y
agggctccagaaattatgttgaattccaagggctacaccaagtccattgatatttggtct R A P
E I M L N S K G Y T K S I D I W S
gtaggctgcattctggcagaaatgctttctaacaggcccatctttccagggaagcattat V G C
I L A E M L S N R P I F P G K H Y
cttgaccagctgaaccacattttgggtattcttggatccccatcacaagaagacctgaat L D Q
L N H I L G I L G S P S Q E D L N
tgtataataaatttaaaagctaggaactatttgctttctcttccacacaaaaataaggtg C I I
N L K A R N Y L L S L P H K N K V
ccatggaacaggctgttcccaaatgctgactccaaagctctggacttattggacaaaatg P W N
R L F P N A D S K A L D L L D K M
ttgacattcaacccacacaagaggattgaagtagaacaggctctggcccacccatatctg L T F
N P H K R I E V E Q A L A H P Y L
gagcagtattacgacccgagtgacgagcccatcgccgaagcaccattcaagttcgacatg E Q Y
Y D P S D E P I A E A P F K F D M
gaattggatgacttgcctaaggaaaagctcaaagaactaatttttgaagagactgctaga E L D
D L P K E K L K E L I F E E T A R ttccagccaggatacagatcttaaatgtcgac
(SEQ ID NO: _) F Q P G Y R S - (SEQ ID NO: _)
TABLE-US-00094 Fak PCR primers FAK FAK411
GCTGGATCCACCAGGGATTATGAGATTCAAAG 2156 (SEQ ID NO: _) FAK686
GTTCTTGTCGACTACTGAGCCTTCTCTTCCTCCA 2157 (SEQ ID NO: _)
P1358.pFastBacHtb FAK S411-Q686-X
tattccggattattcataccgtcccaccatcgggcgcggatctcggtccgaaacc
atgtcgtactaccatcaccatcaccatcacgattacgatatcccaacgaccgaaaacctg M S Y
Y H H H H H H D Y D I P T T E N L
tattttcagggcgccatgggatccaccagggattatgagattcaaagagaaagaatagaa Y F Q
G A M G S T R D Y E I Q R E R I E
cttggacgatgtattggagaaggccaatttggagatgtacatcaaggcatttatatgagt L G R
C I G E G Q F G D V H Q G I Y M S
ccagagaatccagctttggcggttgcaattaaaacatgtaaaaactgtacttcggacagc P E N
P A L A V A I K T C K N C T S D S
gtgagagagaaatttcttcaagaagccttaacaatgcgtcagtttgaccatcctcatatt V R E
K F L Q E A L T M R Q F D H P H I
gtgaagctgattggagtcatcacagagaatcctgtctggataatcatggagctgtgcaca V K L
I G V I T E N P V W I I M E L C T
cttggagagctgaggtcatttttgcaagtaaggaaatacagtttggatctagcatctttg L G E
L R S F L Q V R K Y S L D L A S L
atcctgtatgcctatcagcttagtacagctcttgcatatctagagagcaaaagatttgta I L Y
A Y Q L S T A L A Y L E S K R F V
cacagggacattgctgctcggaatgttctggtgtcctcaaatgattgtgtaaaattagga H R D
I A A R N V L V S S N D C V K L G
gactttggattatcccgatatatggaagatagtacttactacaaagcttccaaaggaaaa D F G
L S R Y M E D S T Y Y K A S K G K
ttgcctattaaatggatggctccagagtcaatcaattttcgacgttttacctcagctagt L P I
K W M A P E S I N F R R F T S A S
gacgtatggatgtttggtgtgtgtatgtgggagatactgatgcatggtgtgaagcctttt D V W
M F G V C M W E I L M H G V K P F
caaggagtgaagaacaatgatgtaatcggtcgaattgaaaatggggaaagattaccaatg Q G V
K N N D V I G R I E N G E R L P M
cctccaaattgtcctcctaccctctacagccttatgacgaaatgctgggcctatgacccc P P N
C P P T L Y S L M T K C W A Y D P
agcaggcggcccaggtttactgaacttaaagctcagctcagcacaatcctggaggaagag S R R
P R F T E L K A Q L S T I L E E E
aaggctcagtagtcgacgagctcactagtcgcggccgctttcgaatctagagcctgcagt K A Q
- S T S S L V A A A F E S R A C S
ctcgaggcatgcggtaccaagcttgtcgagaagtactagaggatcataatc (SEQ ID NO: _)
L E A C G T K L V E K Y - (SEQ ID NO: _)
TABLE-US-00095 FGFR1 PCR primers FGFR1 FGFR1-S
GACTCCTCATATGGCAGGGGTCTCTGAGTATGA 1237 (SEQ ID NO: _) FGFR-SAL
CAGGTCGTCGACTACTCCTGGTTGGAGGTCAAGG 1611 (SEQ ID NO: _) C488A-1
CTGGGAGAGGGCGCGTTTGGGCAGGTGG 2038 (SEQ ID NO: _) C488A-2
CCACCTGCCCAAACGCGCCCTCTCCCAG 2039 (SEQ ID NO: _) C584S-1
CAGGGCTGGAATACAGCTACAACCCCAGC 2041 (SEQ ID NO: _) C584S-2
GCTGGGGTTGTAGCTGTATTCCAGCCCTG 2042 (SEQ ID NO: _) P1351.pET N6
BI-PTP FGFR A458-E765-X C488A, C584S
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccatcatatggcaggg M G H
H H H H H M A G
gtctctgagtatgagcttcccgaagaccctcgctgggagctgcctcgggacagactggtc V S E
Y E L P E D P R W E L P R D R L V
ttaggcaaacccctgggagagggcgcgtttgggcaggtggtgttggcagaggctatcggg L G K
P L G E G A F G Q V V L A E A I G
ctggacaaggacaaacccaaccgtgtgaccaaagtggctgtgaagatgttgaagtcggac L D K
D K P N R V T K V A V K M L K S D
gcaacagagaaagacttgtcagacctgatctcagaaatggagatgatgaagatgatcggg A T E
K D L S D L I S E M E M M K M I G
aagcataagaatatcatcaacctgctgggggcctgcacgcaggatggtcccttgtatgtc K H K
N I I N L L G A C T Q D G P L Y V
atcgtggagtatgcctccaagggcaacctgcgggagtacctgcaggcccggaggccccca I V E
Y A S K G N L R E Y L Q A R R P P
gggctggaatacagctacaaccccagccacaacccagaggagcagctctcctccaaggac G L E
Y S Y N P S H N P E E Q L S S K D
ctggtgtcctgcgcctaccaggtggcccgaggcatggagtatctggcctccaagaagtgc L V S
C A Y Q V A R G M E Y L A S K K C
atacaccgagacctggcagccaggaatgtcctggtgacagaggacaatgtgatgaagata I H R
D L A A R N V L V T E D N V M K I
gcagactttggcctcgcacgggacattcaccacatcgactactataaaaagacaaccaac A D F
G L A R D I H H I D Y Y K K T T N
ggccgactgcctgtgaagtggatggcacccgaggcattatttgaccggatctacacccac G R L
P V K W M A P E A L F D R I Y T H
cagagtgatgtgtggtctttcggggtgctcctgtgggagatcttcactctgggcggctcc Q S D
V W S F G V L L W E I F T L G G S
ccataccccggtgtgcctgtggaggaacttttcaagctgctgaaggagggtcaccgcatg P Y P
G V P V E E L F K L L K E G H R M
gacaagcccagtaactgcaccaacgagctgtacatgatgatgcgggactgctggcatgca D K P
S N C T N E L Y M M M R D C W H A
gtgccctcacagagacccaccttcaagcagctggtggaagacctggaccgcatcgtggcc V P S
Q R P T F K Q L V E D L D R I V A
ttgacctccaaccaggagtagtcgacgaaggagatatatcc (SEQ ID NO: _) L T S N Q
E - (SEQ ID NO: _)
TABLE-US-00096 Flt1 PCR primers FLT1 FLT1-S
ATCAATTCATATGGACCCAGATGAAGTTCC 737 (SEQ ID NO: _) FLT1-A
ATGTAGTCGACCTAATCCTGTTGTACATTTGCTT 738 (SEQ ID NO: _) P1826.pETN6
BI-PTP FLT1 M799-D1165-X WT
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccatcatatggaccca M G H
H H H H H M D P
gatgaagttcctttggatgagcagtgtgagcggctcccttatgatgccagcaagtgggag D E V
P L D E Q C E R L P Y D A S K W E
tttgcccgggagagacttaaactgggcaaatcacttggaagaggggcttttggaaaagtg F A R
E R L K L G K S L G R G A F G K V
gttcaagcatcagcatttggcattaagaaatcacctacgtgccggactgtggctgtgaaa V Q A
S A F G I K K S P T C R T V A V K
atgctgaaagagggggccacggccagcgagtacaaagctctgatgactgagctaaaaatc M L K
E G A T A S E Y K A L M T E L K I
ttgacccacattggccaccatctgaacgtggttaacctgctgggagcctgcaccaagcaa L T H
I G H H L N V V N L L G A C T K Q
ggagggcctctgatggtgattgttgaatactgcaaatatggaaatctctccaactacctc G G P
L M V I V E Y C K Y G N L S N Y L
aagagcaaacgtgacttattttttctcaacaaggatgcagcactacacatggagcctaag K S K
R D L F F L N K D A A L H M E P K
aaagaaaaaatggagccaggcctggaacaaggcaagaaaccaagactagatagcgtcacc K E K
M E P G L E Q G K K P R L D S V T
agcagcgaaagctttgcgagctccggctttcaggaagataaaagtctgagtgatgttgag S S E
S F A S S G F Q E D K S L S D V E
gaagaggaggattctgacggtttctacaaggagcccatcactatggaagatctgatttct E E E
D S D G F Y K E P I T M E D L I S
tacagttttcaagtggccagaggcatggagttcctgtcttccagaaagtgcattcatcgg Y S F
Q V A R G M E F L S S R K C I H R
gacctggcagcgagaaacattcttttatctgagaacaacgtggtgaagatttgtgatttt D L A
A R N I L L S E N N V V K I C D F
ggccttgcccgggatatttataagaaccccgattatgtgagaaaaggagatactcgactt G L A
R D I Y K N P D Y V R K G D T R L
cctctgaaatggatggctcccgaatctatctttgacaaaatctacagcaccaagagcgac P L K
W M A P E S I F D K I Y S T K S D
gtgtggtcttacggagtattgctgtgggaaatcttctccttaggtgggtctccataccca V W S
Y G V L L W E I F S L G G S P Y P
ggagtacaaatggatgaggacttttgcagtcgcctgagggaaggcatgaggatgagagct G V Q
M D E D F C S R L R E G M R M R A
cctgagtactctactcctgaaatctatcagatcatgctggactgctggcacagagaccca P E Y
S T P E I Y Q I M L D C W H R D P
aaagaaaggccaagatttgcagaacttgtggaaaaactaggtgatttgcttcaagcaaat K E R
P R F A E L V E K L G D L L Q A N
gtacaacaggattaggtcgaccaccaccaccaccaccactgagatccggctggccctact V Q Q
D - (SEQ ID NO: _)
ggccgaaaggaattcgaggccagcagggccaccgctgagcaataactagcataacccctt
ggggcctctaaacgggtcttgaggggttttttg (SEQ ID NO: _)
TABLE-US-00097 Kit PCR primers KIT 8K1A
ATGTACGAAGTTCAGTGGAAAGTTGTTGAAGAAATCAACGG 1776 (SEQ ID NO: _) 8K1B
GGTCGATGTAAACGTAGTTGTTACCGTTGATTTCTTCAACAACTTT 1777 (SEQ ID NO: _)
8K2A AACAACTACGTTTACATCGACCCGACCCAGCTGCCGTACGAC 1779 (SEQ ID NO: _)
8K2B GTTACGCGGGAACTCCCATTTGTGGTCGTACGGCAGCTGGGTC 1781 (SEQ ID NO:
_) 8K3A AAATGGGAGTTCCCGCGTAACCGTCTGTCTTTCGGTAAAACCC 1782 (SEQ ID
NO: _) 8K3B ACCGAACGCACCCGCACCCAGGGTTTTACCGAAAGACAGAC 1783 (SEQ ID
NO: _) 8K4A GGTGCGGGTGCGTTCGGTAAAGTTGTTGAAGCGACCGCGTACG 1784 (SEQ
ID NO: _) 8K4B GCCGCGTCAGATTTGATCAGACCGTACGCGGTCGCTTCAAC 1785 (SEQ
ID NO: _) 8K5A CTGATCAAATCTGACGCGGCGATGACCGTTGCGGTTAAAATGC 1786
(SEQ ID NO: _) 8K5B GTCAGGTGCGCAGACGGTTTCAGCATTTTAACCGCAACGGTCA
1787 (SEQ ID NO: _) 8K6A
AAACCGTCTGCGCACCTGACCGAACGTGAAGCGCTGATGTCTG 1788 (SEQ ID NO: _)
8K6B CCAGGTAAGACAGAACTTTCAGTTCAGACATCAGCGCTTCACGT 1789 (SEQ ID NO:
_) 8K7A CTGAAAGTTCTGTCTTACCTGGGTAACCACATGAACATCGTTAA 1791 (SEQ ID
NO: _) 8K7B GGTGCACGCACCCAGCAGGTTAACGATGTTCATGTGGTTAC 1792 (SEQ ID
NO: _) 8K8A CTGCTGGGTGCGTGCACCATCGGTGGTCCGACCCTGGTTATCA 1793 (SEQ
ID NO: _) 8K8B GTCACCGTAGCAGCAGTATTCGGTGATAACCAGGGTCGGACCA 1794
(SEQ ID NO: _) 8K9A GAATACTGCTGCTACGGTGACCTGCTGAACTTCCTGCGTCGTA
1795 (SEQ ID NO: _) 8K9B
AGAGCAGATGAAAGAGTCACGTTTACGACGCAGGAAGTTCAGC 1796 (SEQ ID NO: _)
8K10A CGTGACTCTTTCATCTGCTCTAAACAGGAAGACCACGCGGAAG 1797 (SEQ ID NO:
_) 8K10B CAGCAGGTTTTTGTACAGCGCCGCTTCCGCGTGGTCTTCCTGT 1798 (SEQ ID
NO: _) 8K11A GCGCTGTACAAAAACCTGCTGCACTCTAAAGAATCTTCTTGCTC 1799 (SEQ
ID NO: _) 8K11B CCATGTATTCGTTGGTAGAGTCAGAGCAAGAAGATTCTTTAGAGT 1811
(SEQ ID NO: _) 8K11A GACTCTACCAACGAATACATGGACATGAAACCGGGTGTTTCTTA
1812 (SEQ ID NO: _) 8K11B
TCCGCTTTGGTCGGAACAACGTAAGAAACACCCGGTTTCATGT 1813 (SEQ ID NO: _)
8K12A GTTGTTCCGACCAAAGCGGACAAACGTCGTTCTGTTCGTATCG 1814 (SEQ ID NO:
_) 8K12B TAACGTCACGTTCGATGTAAGAACCGATACGAACAGAACGACGTTT 1815 (SEQ
ID NO: _) 8K13A TCTTACATCGAACGTGACGTTACCCCGGCGATCATGGAAGACG 1816
(SEQ ID NO: _) 8K13B CCAGGTCCAGCGCCAGTTCGTCGTCTTCCATGATCGCCGG 1817
(SEQ ID NO: _) 8K14A GAACTGGCGCTGGACCTGGAAGACCTGCTGTCTTTCTCTTACC
1818 (SEQ ID NO: _) 8K14B
GAACGCCATACCTTTCGCAACCTGGTAAGAGAAAGACAGCAGGT 1819 (SEQ ID NO: _)
8K15A GTTGCGAAAGGTATGGCGTTCCTGGCGTCTAAAAACTGCATCCA 1821 (SEQ ID NO:
_) 8K15B CGCGCCGCCAGGTCACGGTGGATGCAGTTTTTAGACGCC 1822 (SEQ ID NO:
_) 8K16A CGTGACCTGGCGGCGCGTAACATCCTGCTGACCCACGGTCG 1823 (SEQ ID NO:
_) 8K16B ACCGAAGTCGCAGATTTTGGTGATACGACCGTGGGTCAGCAGG 1824 (SEQ ID
NO: _) 8K17A ACCAAAATCTGCGACTTCGGTCTGGCGCGTGACATCAAAAACG 1825 (SEQ
ID NO: _) 8K17B GTTACCTTTAACAACGTAGTTAGAGTCGTTTTTGATGTCACGCGCC 1826
(SEQ ID NO: _) 8K18A TCTAACTACGTTGTTAAAGGTAACGCGCGTCTGCCGGTTAAATG
1827 (SEQ ID NO: _) 8K18B
GAAGATAGATTCCGGCGCCATCCATTTAACCGGCAGACGCGC 1829 (SEQ ID NO: _)
8K19A ATGGCGCCGGAATCTATCTTCAACTGCGTTTACACCTTCGAATC 1831 (SEQ ID NO:
_) 8K19B GATACCGTAAGACCAAACGTCAGATTCGAAGGTGTAAACGCAG 1832 (SEQ ID
NO: _) 8K20A GACGTTTGGTCTTACGGTATCTTCCTGTGGGAACTGTTCTCTC 1833 (SEQ
ID NO: _) 8K20B CCTGTGGGAACTGTTCTCTCTGGGTTCTTCTCCGTACCCGG 1834 (SEQ
ID NO: _) 8K21A GGTTCTTCTCCGTACCCGGGTATGCCGGTTGACTCTAAATTCTAT 1835
(SEQ ID NO: _) 8K21B CGGAAACCTTCTTTGATCATTTTGTAGAATTTAGAGTCAACCGGC
1836 (SEQ ID NO: _) 8K22A
AAAATGATCAAAGAAGGTTTCCGTATGCTGTCTCCGGAACACG 1837 (SEQ ID NO: _)
8K22B ATGTCGTACATTTCCGCCGGCGCGTGTTCCGGAGACAGCATA 1838 (SEQ ID NO:
_) 8K23A CCGGCGGAAATGTACGACATCATGAAAACCTGCTGGGACGCG 1839 (SEQ ID
NO: _) 8K23B AAGGTCGGACGTTTCAGCGGGTCCGCGTCCCAGCAGGTTTTC 1841 (SEQ
ID NO: _) 8K24A CCGCTGAAACGTCCGACCTTCAAACAGATCGTTCAGCTGATCG 1842
(SEQ ID NO: _) 8K24B TTGGTAGATTCAGAGATCTGTTTTTCGATCAGCTGAACGATCTGTT
1843 (SEQ ID NO: _) 8K25A
AAACAGATCTCTGAATCTACCAACCACATCTACTCTAACCTGGC 1844 (SEQ ID NO: _)
8K25B TGACGGTTCGGAGAGCAGTTCGCCAGGTTAGAGTAGATGTGG 1845 (SEQ ID NO:
_) 8K26A AACTGCTCTCCGAACCGTCAGAAACCGGTTGTTGACCACTCTG 1846 (SEQ ID
NO: _) 8K26B GTAGAACCAACAGAGTTGATACGAACAGAGTGGTCAACAACCGGT 1847
(SEQ ID NO: _) 8K27A CGTATCAACTCTGTTGGTTCTACCGCGTCTTCTTCTCAGCCG
1848 (SEQ ID NO: _) 8K27B AACGTCGTCGTGAACCAGCAGCGGCTGAGAAGAAGACGCG
1849 (SEQ ID NO: _) 8K-F GTTGTTTCATATGTACGAAGTTCAGTGGAAAG 1851 (SEQ
ID NO: _) 8K-R GTTGTTTGTCGACTAAACGTCGTCGTGAACCAGCAG 1852 (SEQ ID
NO: _) KIT COD-K948X GTTCTTGTCGACTATTTCTGACGGTTCGGAGAGC 3411 (SEQ
ID NO: _)
TABLE-US-00098 P1332.N6 BI PTP KIT M552-K948-X COD
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccatcatatgtacgaa M G H
H H H H H M Y E
gttcagtggaaagttgttgaagaaatcaacggtaacaactacgtttacatcgacccgacc V Q W
K V V E E I N G N N Y V Y I D P T
cagctgccgtacgaccacaaatgggagttcccgcgtaaccgtctgtctttcggtaaaacc Q L P
Y D H K W E F P R N R L S F G K T
ctgggtgcgggtgcgttcggtaaagttgttgaagcgaccgcgtacggtctgatcaaatct L G A
G A F G K V V E A T A Y G L I K S
gacgcggcgatgaccgttgcggttaaaatgctgaaaccgtctgcgcacctgaccgaacgt D A A
M T V A V K M L K P S A H L T E R
gaagcgctgatgtctgaactgaaagttctgtcttacctgggtaaccacatgaacatcgtt E A L
M S E L K V L S Y L G N H M N I V
aacctgctgggtgcgtgcaccatcggtggtccgaccctggttatcaccgaatactgctgc N L L
G A C T I G G P T L V I T E Y C C
tacggtgacctgctgaacttcctgcgtcgtaaacgtgactctttcatctgctctaaacag Y G D
L L N F L R R K R D S F I C S K Q
gaagaccacgcggaagcggcgctgtacaaaaacctgctgcactctaaagaatcttcttgc E D H
A E A A L Y K N L L H S K E S S C
tctgactctaccaacgaatacatggacatgaaaccgggtgtttcttacgttgttccgacc S D S
T N E Y M D M K P G V S Y V V P T
aaagcggacaaacgtcgttctgttcgtatcggttcttacatcgaacgtgacgttaccccg K A D
K R R S V R I G S Y I E R D V T P
gcgatcatggaagacgacgaactggcgctggacctggaagacctgctgtctttctcttac A I M
E D D E L A L D L E D L L S F S Y
caggttgcgaaaggtatggcgttcctggcgtctaaaaactgcatccaccgtgacctggcg Q V A
K G M A F L A S K N C I H R D L A
gcgcgtaacatcctgctgacccacggtcgtatcaccaaaatctgcgacttcggtctggcg A R N
I L L T H G R I T K I C D F G L A
cgtgacatcaaaaacgactctaactacgttgttaaaggtaacgcgcgtctgccggttaaa R D I
K N D S N Y V V K G N A R L P V K
tggatggcgccggaatctatcttcaactgcgtttacaccttcgaatctgacgtttggtct W M A
P E S I F N C V Y T F E S D V W S
tacggtatcttcctgtgggaactgttctctctgggttcttctccgtacccgggtatgccg Y G I
F L W E L F S L G S S P Y P G M P
gttgactctaaattctacaaaatgatcaaagaaggtttccgtatgctgtctccggaacac V D S
K F Y K M I K E G F R M L S P E H
gcgccggcggaaatgtacgacatcatgaaaacctgctgggacgcggacccgctgaaacgt A P A
E M Y D I M K T C W D A D P L K R
ccgaccttcaaacagatcgttcagctgatcgaaaaacagatctctgaatctaccaaccac P T F
K Q I V Q L I E K Q I S E S T N H
atctactctaacctggcgaactgctctccgaaccgtcagaaatagtcgactgaaaaagga I Y S
N L A N C S P N R Q K - (SEQ ID NO: _) agagt (SEQ ID NO: _)
TABLE-US-00099 Met PCR primers MET G1056
CATCCTACATATGGGGGACTCTGATATATCCAGTC 1223 (SEQ ID NO: _) G-1364
CTAGCAGGTCGACTACCCAATGAAAGTAGAGAAGATCGC 1318 (SEQ ID NO: _)
P1818.pETN6 BI-PTP MET G1056-G1364-X WT
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccatcatatgggggac M G H
H H H H H M G D
tctgatatatccagtccattactgcaaaatactgtccacattgacctcagtgctctaaat S D I
S S P L L Q N T V H I D L S A L N
ccagagctggtccaggcagtgcagcatgtagtgattgggcccagtagcctgattgtgcat P E L
V Q A V Q H V V I G P S S L I V H
ttcaatgaagtcataggaagagggcattttggttgtgtatatcatgggactttgttggac F N E
V I G R G H F G C V Y H G T L L D
aatgatggcaagaaaattcactgtgctgtgaaatccttgaacagaatcactgacatagga N D G
K K I H C A V K S L N R I T D I G
gaagtttcccaatttctgaccgagggaatcatcatgaaagattttagtcatcccaatgtc E V S
Q F L T E G I I M K D F S H P N V
ctctcgctcctgggaatctgcctgcgaagtgaagggtctccgctggtggtcctaccatac L S L
L G I C L R S E G S P L V V L P Y
atgaaacatggagatcttcgaaatttcattcgaaatgagactcataatccaactgtaaaa M K H
G D L R N F I R N E T H N P T V K
gatcttattggctttggtcttcaagtagccaaaggcatgaaatatcttgcaagcaaaaag D L I
G F G L Q V A K G M K Y L A S K K
tttgtccacagagacttggctgcaagaaactgtatgctggatgaaaaattcacagtcaag F V H
R D L A A R N C M L D E K F T V K
gttgctgattttggtcttgccagagacatgtatgataaagaatactatagtgtacacaac V A D
F G L A R D M Y D K E Y Y S V H N
aaaacaggtgcaaagctgccagtgaagtggatggctttggaaagtctgcaaactcaaaag K T G
A K L P V K W M A L E S L Q T Q K
tttaccaccaagtcagatgtgtggtcctttggcgtgctcctctgggagctgatgacaaga F T T
K S D V W S F G V L L W E L M T R
ggagccccaccttatcctgatgtaaacacctttgatataactgtttacttgttgcaaggg G A P
P Y P D V N T F D I T V Y L L Q G
agaagactcctacaacccgaatactgcccagaccccttatatgaagtaatgctaaaatgc R R L
L Q P E Y C P D P L Y E V M L K C
tggcaccctaaagccgaaatgcgcccatccttttctgaactggtgtcccggatatcagcg W H P
K A E M R P S F S E L V S R I S A atcttctctactttcattgggtagtcgac
(SEQ ID NO: _) I F S T F I G - (SEQ ID NO: _)
TABLE-US-00100 p38 PCR primers p38A P38A-S
CCGGATCCATATGTCTCAGGAGAGGCCCAC 253 (SEQ ID NO: _) P38A-A
GAAACCCTCGAGTCAGGACTCCATCTCTTCTTG 254 P4292.pET15S P38A M1-S360-X
MKK6DD taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggcagcagccatcatcatcatcatcacagc M G S
S H H H H H H S
agcggcctggtgccgcgcggcagccatatgtctcaggagaggcccacgttctaccggcag S G L
V P R G S H M S Q E R P T F Y R Q
gagctgaacaagacaatctgggaggtgcccgagcgttaccagaacctgtctccagtgggc E L N
K T I W E V P E R Y Q N L S P V G
tctggcgcctatggctctgtgtgtgctgcttttgacacaaaaacggggttacgtgtggca S G A
Y G S V C A A F D T K T G L R V A
gtgaagaagctctccagaccatttcagtccatcattcatgcgaaaagaacctacagagaa V K K
L S R P F Q S I I H A K R T Y R E
ctgcggttacttaaacacatgaaacatgaaaatgtgattggtctgttggacgtttttaca L R L
L K H M K H E N V I G L L D V F T
cctgcaaggtctctggaggaattcaatgatgtgtatctggtgacccatctcatgggggca P A R
S L E E F N D V Y L V T H L M G A
gatctgaacaacattgtgaaatgtcagaagcttacagatgaccatgttcagttccttatc D L N
N I V K C Q K L T D D H V Q F L I
taccaaattctccgaggtctaaagtatatacattcagctgacataattcacagggaccta Y Q I
L R G L K Y I H S A D I I H R D L
aaacctagtaatctagctgtgaatgaagactgtgagctgaagattctggattttggactg K P S
N L A V N E D C E L K I L D F G L
gctcggcacacagatgatgaaatgacaggctacgtggccactaggtggtacagggctcct A R H
T D D E M T G Y V A T R W Y R A P
gagatcatgctgaactggatgcattacaaccagacagttgatatttggtcagtgggatgc E I M
L N W M H Y N Q T V D I W S V G C
ataatggccgagctgttgactggaagaacattgtttcctggtacagaccatattgatcag I M A
E L L T G R T L F P G T D H I D Q
ttgaagctcattttaagactcgttggaaccccaggggctgagcttttgaagaaaatctcc L K L
I L R L V G T P G A E L L K K I S
tcagagtctgcaagaaactatattcagtctttgactcagatgccgaagatgaactttgcg S E S
A R N Y I Q S L T Q M P K M N F A
aatgtatttattggtgccaatcccctggctgtcgacttgctggagaagatgcttgtattg N V F
I G A N P L A V D L L E K M L V L
gactcagataagagaattacagcggcccaagcccttgcacatgcctactttgctcagtac D S D
K R I T A A Q A L A H A Y F A Q Y
cacgatcctgatgatgaaccagtggccgatccttatgatcagtcctttgaaagcagggac H D P
D D E P V A D P Y D Q S F E S R D
ctccttatagatgagtggaaaagcctgacctatgatgaagtcatcagctttgtgccacca L L I
D E W K S L T Y D E V I S F V P P
ccccttgaccaagaagagatggagtcctgactcgac (SEQ ID NO: _) P L D Q E E M E
S - (SEQ ID NO: _)
TABLE-US-00101 Pim1 PCR primers PIM1 PIM-1S
GCTGGCGCATATGAAGGAGAAGGAGCCCCTGGAG 233 (SEQ ID NO: _) PIM-1A
GAAAGGGTCGACTTTGCTGGGCCCCGGCGACAG 234 (SEQ ID NO: _) P1215.pET29SRI
PIM1 E29-K313 HIS WT
agatcgatctcgatcccgcgaaattaatacgactcactataggggaattgtgagcggataa
caattcccctctagaaataattttgtttaactttaagaaggagatatacatatgaaggag M K E
aaggagcccctggagtcgcagtaccaggtgggcccgctactgggcagcggcggcttcggc K E P
L E S Q Y Q V G P L L G S G G F G
tcggtctactcaggcatccgcgtctccgacaacttgccggtggccatcaaacacgtggag S V Y
S G I R V S D N L P V A I K H V E
aaggaccggatttccgactggggagagctgcctaatggcactcgagtgcccatggaagtg K D R
I S D W G E L P N G T R V P M E V
gtcctgctgaagaaggtgagctcgggtttctccggcgtcattaggctcctggactggttc V L L
K K V S S G F S G V I R L L D W F
gagaggcccgacagtttcgtcctgatcctggagaggcccgagccggtgcaagatctcttc E R P
D S F V L I L E R P E P V Q D L F
gacttcatcacggaaaggggagccctgcaagaggagctggcccgcagcttcttctggcag D F I
T E R G A L Q E E L A R S F F W Q
gtgctggaggccgtgcggcactgccacaactgcggggtgctccaccgcgacatcaaggac V L E
A V R H C H N C G V L H R D I K D
gaaaacatccttatcgacctcaatcgcggcgagctcaagctcatcgacttcgggtcgggg E N I
L I D L N R G E L K L I D F G S G
gcgctgctcaaggacaccgtctacacggacttcgatgggacccgagtgtatagccctcca A L L
K D T V Y T D F D G T R V Y S P P
gagtggatccgctaccatcgctaccatggcaggtcggcggcagtctggtccctggggatc E W I
R Y H R Y H G R S A A V W S L G I
ctgctgtatgatatggtgtgtggagatattcctttcgagcatgacgaagagatcatcagg L L Y
D M V C G D I P F E H D E E I I R
ggccaggttttcttcaggcagagggtctcttcagaatgtcagcatctcattagatggtgc G Q V
F F R Q R V S S E C Q H L I R W C
ttggccctgagaccatcagataggccaaccttcgaagaaatccagaaccatccatggatg L A L
R P S D R P T F E E I Q N H P W M
caagatgttctcctgccccaggaaactgctgagatccacctccacagcctgtcgccgggg Q D V
L L P Q E T A E I H L H S L S P G
cccagcaaagtcgaccaccaccaccaccaccactgagatccggctgctaacaaagcccga P S K
V D H H H H H H - (SEQ ID NO: _)
aaggaattcgagttggctgctgccaccgctgagcaataactagcataaccccttggggcc
tctaaacgggtcttgaggggttttttg (SEQ ID NO: _)
TABLE-US-00102 Ret PCR primers RET RETH661
GTTCTTCATATGCACAAGTTTGCCCACAAGCCA 2184 (SEQ ID NO: _) RE-1012-HIS
GTTCTTGTCGACCCTCTTAACCATCATCTTCTCCAGGTCT 2431 P1378.pET-SF BI-PTP
RET H661-R1012 HIS
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataatttt
gtttaactttaagaaggagatatacatatgcacaagtttgcccacaagccacccatctcc M H K
F A H K P P I S
tcagctgagatgaccttccggaggcccgcccaggccttcccggtcagctactcctcttcc S A E
M T F R R P A Q A F P V S Y S S S
ggtgcccgccggccctcgctggactccatggagaaccaggtctccgtggatgccttcaag G A R
R P S L D S M E N Q V S V D A F K
atcctggaggatccaaagtgggaattccctcggaagaacttggttcttggaaaaactcta I L E
D P K W E F P R K N L V L G K T L
ggagaaggcgaatttggaaaagtggtcaaggcaacggccttccatctgaaaggcagagca G E G
E F G K V V K A T A F H L K G R A
gggtacaccacggtggccgtgaagatgctgaaagagaacgcctccccgagtgagcttcga G Y T
T V A V K M L K E N A S P S E L R
gacctgctgtcagagttcaacgtcctgaagcaggtcaaccacccacatgtcatcaaattg D L L
S E F N V L K Q V N H P H V I K L
tatggggcctgcagccaggatggcccgctcctcctcatcgtggagtacgccaaatacggc Y G A
C S Q D G P L L L I V E Y A K Y G
tccctgcggggcttcctccgcgagagccgcaaagtggggcctggctacctgggcagtgga S L R
G F L R E S R K V G P G Y L G S G
ggcagccgcaactccagctccctggaccacccggatgagcgggccctcaccatgggcgac G S R
N S S S L D H P D E R A L T M G D
ctcatctcatttgcctggcagatctcacaggggatgcagtatctggccgagatgaagctc L I S
F A W Q I S Q G M Q Y L A E M K L
gttcatcgggacttggcagccagaaacatcctggtagctgaggggcggaagatgaagatt V H R
D L A A R N I L V A E G R K M K I
tcggatttcggcttgtcccgagatgtttatgaagaggattcctacgtgaagaggagccag S D F
G L S R D V Y E E D S Y V K R S Q
ggtcggattccagttaaatggatggcaattgaatccctttttgatcatatctacaccacg G R I
P V K W M A I E S L F D H I Y T T
caaagtgatgtatggtcttttggtgtcctgctgtgggagatcgtgaccctagggggaaac Q S D
V W S F G V L L W E I V T L G G N
ccctatcctgggattcctcctgagcggctcttcaaccttctgaagaccggccaccggatg P Y P
G I P P E R L F N L L K T G H R M
gagaggccagacaactgcagcgaggagatgtaccgcctgatgctgcaatgctggaagcag E R P
D N C S E E M Y R L M L Q C W K Q
gagccggacaaaaggccggtgtttgcggacatcagcaaagacctggagaagatgatggtt E P D
K R P V F A D I S K D L E K M M V
aagagggtcgaccaccaccaccaccaccactgagatccggctggccctactggccgaaag K R V
D H H H H H H - (SEQ ID NO: _)
gaattcgaggccagcagggccaccgctgagcaataactagcataaccccttggggcctct
aaacgggtcttgaggggttttttg (SEQ ID NO: _)
TABLE-US-00103 Src PCR primers SRC-86
GCTGGCCCATATGGTGACCACCTTTGTGGCCCT 1452 (SEQ ID NO: _) SRC-452
(L536) GCTACTAGTCGACCTAGAGGTTCTCCCCGGGCT 1453 (SEQ ID NO: _) P1144.
pET-N6 BI-PTP SRC V86-L536-X
taatacgactcactataggggaattgtgagcggataacaattcccctctagaaataattt
tgtttaactttaagaaggagatataccatgggtcaccaccatcaccatcatatggtgacc M G H
H H H H H M V T
acctttgtggccctctatgactatgagtctaggacggagacagacctgtccttcaagaaa T F V
A L Y D Y E S R T E T D L S F K K
ggcgagcggctccagattgtcaacaacacagagggagactggtggctggcccactcgctc G E R
L Q I V N N T E G D W W L A H S L
agcacaggacagacaggctacatccccagcaactacgtggcgccctccgactccatccag S T G
Q T G Y I P S N Y V A P S D S I Q
gctgaggagtggtattttggcaagatcaccagacgggagtcagagcggttactgctcaat A E E
W Y F G K I T R R E S E R L L L N
gcagagaacccgagagggaccttcctcgtgcgagaaagtgagaccacgaaaggtgcctac A E N
P R G T F L V R E S E T T K G A Y
tgcctctcagtgtctgacttcgacaacgccaagggcctcaacgtgaagcactacaagatc C L S
V S D F D N A K G L N V K H Y K I
cgcaagctggacagcggcggcttctacatcacctcccgcacccagttcaacagcctgcag R K L
D S G G F Y I T S R T Q F N S L Q
cagctggtggcctactactccaaacacgccgatggcctgtgccaccgcctcaccaccgtg Q L V
A Y Y S K H A D G L C H R L T T V
tgccccacgtccaagccgcagactcagggcctggccaaggatgcctgggagatccctcgg C P T
S K P Q T Q G L A K D A W E I P R
gagtcgctgcggctggaggtcaagctgggccagggctgctttggcgaggtgtggatgggg E S L
R L E V K L G Q G C F G E V W M G
acctggaacggtaccaccagggtggccatcaaaaccctgaagcctggcacgatgtctcca T W N
G T T R V A I K T L K P G T M S P
gaggccttcctgcaggaggcccaggtcatgaagaagctgaggcatgagaagctggtgcag E A F
L Q E A Q V M K K L R H E K L V Q
ttgtatgctgtggtttcagaggagcccatttacatcgtcacggagtacatgagcaagggg L Y A
V V S E E P I Y I V T E Y M S K G
agtttgctggactttctcaagggggagacaggcaagtacctgcggctgcctcagctggtg S L L
D F L K G E T G K Y L R L P Q L V
gacatggctgctcagatcgcctcaggcatggcgtacgtggagcggatgaactacgtccac D M A
A Q I A S G M A Y V E R M N Y V H
cgggaccttcgtgcagccaacatcctggtgggagagaacctggtgtgcaaagtggccgac R D L
R A A N I L V G E N L V C K V A D
tttgggctggctcggctcattgaagacaatgagtacacggcgcggcaaggtgccaaattc F G L
A R L I E D N E Y T A R Q G A K F
cccatcaagtggacggctccagaagctgccctctatggccgcttcaccatcaagtcggac P I K
W T A P E A A L Y G R F T I K S D
gtgtggtccttcgggatcctgctgactgagctcaccacaaagggacgggtgccctaccct V W S
F G I L L T E L T T K G R V P Y P
gggatggtgaaccgcgaggtgctggaccaggtggagcggggctaccggatgccctgcccg G M V
N R E V L D Q V E R G Y R M P C P
ccggagtgtcccgagtccctgcacgacctcatgtgccagtgctggcggaaggagcctgag P E C
P E S L H D L M C Q C W R K E P E
gagcggcccaccttcgagtacctgcaggccttcctggaggactacttcacgtccaccgag E R P
T F E Y L Q A F L E D Y F T S T E
ccccagtaccagccccggggagaacctctaggtcgacgaaggagatatatcc (SEQ ID NO: _)
P Q Y Q P G E N L - (SEQ ID NO: _)
TABLE-US-00104 Sap70 PCR primers ZAP70 ZAP70-D327-BAMHI-N
AGAGGGATCCGCCACCATGGACAAGAAGCTCTTCCTGAA 5172 (SEQ ID NO: _)
ZAP70-G606-HIS ACGAATTCTAGTGGTGGTGGTGGTGGTGGTGCCCTTCCACCT 5171
TGCTG (SEQ ID NO: _) P1868.pFastBac1 ZAP70 D327-G606 HIS, WT
agatcatggagataattaaaatgataaccatctcgcaaataaataagtattttactgtttt
cgtaacagttttgtaataaaaaaacctataaatattccggattattcataccgtcccacc
atcgggcgcggatccgccaccatggacaagaagctcttcctgaagcgcgataacctcctc M D K
K L F L K R D N L L
atagctgacattgaacttggctgcggcaactttggctcagtgcgccagggcgtgtaccgc I A D
I E L G C G N F G S V R Q G V Y R
atgcgcaagaagcagatcgacgtggccatcaaggtgctgaagcagggcacggagaaggca M R K
K Q I D V A I K V L K Q G T E K A
gacacggaagagatgatgcgcgaggcgcagatcatgcaccagctggacaacccctacatc D T E
E M M R E A Q I M H Q L D N P Y I
gtgcggctcattggcgtctgccaggccgaggccctcatgctggtcatggagatggctggg V R L
I G V C Q A E A L M L V M E M A G
ggcgggccgctgcacaagttcctggtcggcaagagggaggagatccctgtgagcaatgtg G G P
L H K F L V G K R E E I P V S N V
gccgagctgctgcaccaggtgtccatggggatgaagtacctggaggagaagaactttgtg A E L
L H Q V S M G M K Y L E E K N F V
caccgtgacctggcggcccgcaacgtcctgctggttaaccggcactacgccaagatcagc H R D
L A A R N V L L V N R H Y A K I S
gactttggcctctccaaagcactgggtgccgacgacagctactacactgcccgctcagca D F G
L S K A L G A D D S Y Y T A R S A
gggaagtggccgctcaagtggtacgcacccgaatgcatcaacttccgcaagttctccagc G K W
P L K W Y A P E C I N F R K F S S
cgcagcgatgtctggagctatggggtcaccatgtgggaggccttgtcctacggccagaag R S D
V W S Y G V T M W E A L S Y G Q K
ccctacaagaagatgaaagggccggaggtcatggccttcatcgagcagggcaagcggatg P Y K
K M K G P E V M A F I E Q G K R M
gaatgcccaccagagtgtccacccgaactgtacgcactcatgagtgactgctggatctac E C P
P E C P P E L Y A L M S D C W I Y
aagtgggaggatcgccccgacttcctgaccgtggagcagcgcatgcgagcctgttactac K W E
D R P D F L T V E Q R M R A C Y Y
agcctggccagcaaggtggaagggcaccaccaccaccaccaccactagaattc(SEQ ID NO: _)
S L A S K V E G H H H H H H H - (SEQ ID NO: _)
TABLE-US-00105 BAD substrate PCR primers BAD BAD-N
GTTGTGACATATGTTCCAGATCCCAGAGTTTG 1613 (SEQ ID NO: _) BAD-S
GTTGTGAGTCGACTCACTGGGAGGGGGCGGA 1614 (SEQ ID NO: _) P963.pET-BH BAD
M1-Q168-X
tacgactcactataggggaattgtgagcggataacaattcccctctagaaataattttgt
ttaactttaagaaggagatataccatggctggttgcctgaacgacatcttcgaagctcag M A G
C L N D I F E A Q
aaaatcgaatggcaccatcaccatcaccatatgttccagatcccagagtttgagccgagt K I E
W H H H H H H M F Q I P E F E P S
gagcaggaagactccagctctgcagagaggggcctgggccccagccccgcaggggacggg E Q E
D S S S A E R G L G P S P A G D G
ccctcaggctccggcaagcatcatcgccaggccccaggcctcctgtgggacgccagtcac P S G
S G K H H R Q A P G L L W D A S H
cagcaggagcagccaaccagcagcagccatcatggaggcgctggggctgtggagatccgg Q Q E
Q P T S S S H H G G A G A V E I R
agtcgccacagctcctaccccgcggggacggaggacgacgaagggatgggggaggagccc S R H
S S Y P A G T E D D E G M G E E P
agcccctttcggggccgctcgcgctcggcgccccccaacctctgggcagcacagcgctat S P F
R G R S R S A P P N L W A A Q R Y
ggccgcgagctccggaggatgagtgacgagtttgtggactcctttaagaagggacttcct G R E
L R R M S D E F V D S F K K G L P
cgcccgaagagcgcgggcacagcaacgcagatgcggcaaagctccagctggacgcgagtc R P K
S A G T A T Q M R Q S S S W T R V
ttccagtcctggtgggatcggaacttgggcaggggaagctccgccccctcccagtgagtc F Q S
W W D R N L G R G S S A P S Q -
gaccaccaccaccaccaccactgagatccggctggccctactggccgaaaggaattcgag
gccagcagggccaccgctgagcaataactagcataaccccttggggcctctaaacgggtc
ttgaggggttttttg (SEQ ID NO: _) (polypeptide SEQ ID NO: _)
TABLE-US-00106 MEKI substrate PCR primers MEK1 MEK1-S
CGGGTCCCATATGCCCAAGAAGAAGCCGAC 755 (SEQ ID NO: _) MEK-HIS
GTTCGTTGTCGACGACGCCAGCAGCATGGGTTG 2127 (SEQ ID NO: _) K97A-1
(K104A) CTAATTCATCTGGAGATCGCGCCCGCAATCCGG 2023 (SEQ ID NO: _)
K97A-2 (K104A) CCGGATTGCGGGCGCGATCTCCAGATGAATTAG 2024 (SEQ ID NO:
_) P1277.pGEX-BIO MEK1 K97A
atgtcccctatactaggttattggaaaattaagggccttgtgcaacccactcgacttctt M S P
I L G Y W K I K G L V Q P T R L L
ttggaatatcttgaagaaaaatatgaagagcatttgtatgagcgcgatgaaggtgataaa L E Y
L E E K Y E E H L Y E R D E G D K
tggcgaaacaaaaagtttgaattgggtttggagtttcccaatcttccttattatattgat W R N
K K F E L G L E F P N L P Y Y I D
ggtgatgttaaattaacacagtctatggccatcatacgttatatagctgacaagcacaac G D V
K L T Q S M A I I R Y I A D K H N
atgttgggtggttgtccaaaagagcgtgcagagatttcaatgcttgaaggagcggttttg M L G
G C P K E R A E I S M L E G A V L
gatattagatacggtgtttcgagaattgcatatagtaaagactttgaaactctcaaagtt D I R
Y G V S R I A Y S K D F E T L K V
gattttcttagcaagctacctgaaatgctgaaaatgttcgaagatcgtttatgtcataaa D F L
S K L P E M L K M F E D R L C H K
acatatttaaatggtgatcatgtaacccatcctgacttcatgttgtatgacgctcttgat T Y L
N G D H V T H P D F M L Y D A L D
gttgttttatacatggacccaatgtgcctggatgcgttcccaaaattagtttgttttaaa V V L
Y M D P M C L D A F P K L V C F K
aaacgtattgaagctatcccacaaattgataagtacttgaaatccagcaagtatatagca K R I
E A I P Q I D K Y L K S S K Y I A
tggcctttgcagggctggcaagccacgtttggtggtggcgaccatcctccaaaatcggat W P L
Q G W Q A T F G G G D H P P K S D
ctggttccgcgtggatctcatatgcccaagaagaagccgacgcccatccagctgaacccg L V P
R G S H M P K K K P T P I Q L N P
gcccccgacggctctgcagttaacgggaccagctctgcggagaccaacttggaggccttg A P D
G S A V N G T S S A E T N L E A L
cagaagaagctggaggagctagagcttgatgagcagcagcgaaagcgccttgaggccttt Q K K
L E E L E L D E Q Q R K R L E A F
cttacccagaagcagaaggtgggagaactgaaggatgacgactttgagaagatcagtgag L T Q
K Q K V G E L K D D D F E K I S E
ctgggggctggcaatggcggtgtggtgttcaaggtctcccacaagccttctggcctggtc L G A
G N G G V V F K V S H K P S G L V
atggccagagcgctaattcatctggagatcaaacccgcaatccggaaccagatcataagg M A R
A L I H L E I K P A I R N Q I I R
gagctgcaggttctgcatgagtgcaactctccgtacatcgtgggcttctatggtgcgttc E L Q
V L H E C N S P Y I V G F Y G A F
tacagcgatggcgagatcagtatctgcatggagcacatggatggaggttctctggatcaa Y S D
G E I S I C M E H M D G G S L D Q
gtcctgaagaaagctggaagaattcctgaacaaattttaggaaaagttagcattgctgta V L K
K A G R I P E Q I L G K V S I A V
ataaaaggcctgacatatctgagggagaagcacaagatcatgcacagagatgtcaagccc I K G
L T Y L R E K H K I M H R D V K P
tccaacatcctagtcaactcccgtggggagatcaagctctgtgactttggggtcagcggg S N I
L V N S R G E I K L C D F G V S G
cagctcatcgactccatggccaactccttcgtgggcacaaggtcctacatgtcgccagaa Q L I
D S M A N S F V G T R S Y M S P E
agactccaggggactcattactctgtgcagtcagacatctggagcatgggactgtctctg R L Q
G T H Y S V Q S D I W S M G L S L
gtagagatggcggttgggaggtatcccatccctcctccagatgccaaggagctggagctg V E M
A V G R Y P I P P P D A K E L E L
atgtttgggtgccaggtggaaggagatgcggctgagaccccacccaggccaaggaccccc M F G
C Q V E G D A A E T P P R P R T P
gggaggccccttagctcatacggaatggacagccgacctcccatggcaatttttgagttg G R P
L S S Y G M D S R P P M A I F E L
ttggattacatagtcaacgagcctcctccaaaactgcccagtggagtgttcagtctggaa L D Y
I V N E P P P K L P S G V F S L E
tttcaagattttgtgaataaatgcttaataaaaaaccccgcagagagagcagatttgaag F Q D
F V N K C L I K N P A E R A D L K
caactcatggttcatgcttttatcaagagatctgatgctgaggaagtggattttgcaggt Q L M
V H A F I K R S D A E E V D F A G
tggctctgctccaccatcggccttaaccagcccagcacaccaacccatgctgctggcgtc W L C
S T I G L N Q P S T P T H A A G V
gtcgacctgaacgacatcttcgaagctcagaaaatcgaatggcaccgttagaattc V D L N D
I F E A Q K I E W H R - (SEQ ID NO: _) (nucleic acid SEQ ID NO:
_)
Example 76
Efficacy of Compounds in Combination with Standard-of-Care
Chemotherapeutic Agents in Four Human Cancer Cell Lines
[2048] Compounds of the invention, such as compounds of Formula
III, in combination with a standard chemotherapeutic agent, such as
5-fluorouracil, carboplatin, dacarbazine, gefitinib, oxaliplatin,
paclitaxel, SN-38, temozolomide, or vinblastine, can be assessed
for their effectiveness in killing human tumor cells. Human tumor
cell lines, such as A-375 (malignant melanoma), SK-MEL-2 (malignant
melanoma, skin metastasis), COLO 205 (colorectal adenocarcinoma,
ascites metastasis) or SW-620 (colorectal adenocarcinoma, lymph
node metastasis) can be treated with a compound of Formula III
alone, or in combination with one of the above-mentioned
chemotherapeutic agents.
[2049] Tumor cells are grown as a monolayer at 37.degree. C. in a
humidified atmosphere (5% CO.sub.2, 95% air). Cells are grown in a
suitable culture medium, e.g. RPMI 1640 (Ref BE12-702F, Cambrex,
Verviers, Belgium) containing 2 mM L-glutamine and supplemented
with 10% fetal bovine serum (RefDE14-801E, Cambrex). For
experimental use, the tumor cells are detached from the culture
flask by a 5-minute treatment with trypsin-versene (Ref 02-007E,
Cambrex), diluted in Hanks' medium without calcium or magnesium
(Ref BE10-543F, Cambrex). Trypsin treatment is neutralized by
culture medium addition. The cells are counted in a hemocytometer
and their viability assessed by 0.25% trypan blue exclusion.
[2050] The cell lines are checked for mycoplasma contamination with
the Mycotect assay kit (Ref 15672-017, Invitrogen, Cergy-Pontoise,
France) in accordance with the manufacturer's instructions. The
mycoplasma test is assayed from the culture supernatants of the
cell lines and compared to negative and positive controls.
[2051] The tumor cells (10,000 per well) are plated in 96-well
flat-bottom microtitration plates (Ref 055260, Nunc, Dutscher,
Brumath, France) and incubated at 37.degree. C. for 24 hours before
treatment in 100 .mu.l of drug-free culture medium supplemented
with 10% FBS. In order to assess the IC.sub.50 of each compound to
be used for each cell line, the tumor cells are incubated in a 200
.mu.l final volume of RPMI 1640 supplemented with 10% FBS and
containing either a compound of Formula III or one of
5-fluorouracil, carboplatin, dacarbazine, gefitinib, oxaliplatin,
paclitaxel, SN-38, temozolomide, or vinblastine. The compounds are
tested in a suitable concentration range, such as 10.sup.-8 to
10.sup.-3 M for a compound of Formula III, 5-fluorouracil,
dacarbazine or gefitinib, 10.sup.-9 to 10.sup.-4 M for carboplatin,
oxaliplatin, or temozolomide, 10.sup.-11 to 10.sup.-6 M for
paclitaxel or SN-38, and 10.sup.-15 to 10.sup.-10 M for
vinblastine. Compounds of Formula III are dissolved in DMSO and
diluted with culture medium to the desired concentrations.
5-fluorouracil (50 mg/ml, Dakota Pharm, LePlessis Robinson,
France), carboplatin (10 mg/ml, Aguettant, Lyon, France), and
paclitaxel (6 mg/ml, Bristol-Myers Squibb SpA, Rueil Malmaison,
France), are diluted with culture medium to the desired
concentrations. Dacarbazine (Sigma, Saint Quentin Fallavier,
France) and vinblastine (Lilly France S.A., Saint Cloud, France)
are dissolved in NaCl 0.9% and diluted with culture medium to the
desired concentrations. Gefitinib is dissolved in a mixed solution
of RPMI 1640 and DMSO and diluted with culture medium to the
desired concentrations (maximum final DMSO of 0.1% v/v). SN-38 (LKT
Laboratories. Inc., St. Paul, Minn.) is dissolved in DMSO and
diluted with culture medium to the desired concentrations (maximum
final DMSO of 0.1% v/v). Temozolomide (LKT Laboratories. Inc., St.
Paul, Minn.) is dissolved in water for injection and diluted with
culture medium to the desired concentrations. Cells are incubated
for 96 hours in the presence of test substances at 37.degree. C.
under 5% CO.sub.2. At the end of treatments, the cytotoxic activity
is evaluated by an MTT assay.
[2052] For the MTT assay, at the end of the cells treatment, 20
.mu.l of a 5 mg/ml solution 0.22 .mu.m filtered tetrazolium reagent
(MTT, Ref M2128, Sigma) in Phosphate Buffered Saline (PBS, Ref
BE17-517Q, Cambrex), is added in each well. Culture plates are
incubated for 2 h at 37.degree. C. The resulting supernatant is
removed and formazan crystals dissolved with 200 .mu.l of DMSO per
well. Absorbency (OD) is measured at 570 nm in each well using
VICTOR.sup.3.TM. 1420 multilabeled counter (Wallac, PerkinElmer,
Courtaboeuf, France).
[2053] The IC.sub.50 for each compound on each cell line is
determined from the OD measurements of each sample. The dose
response inhibition of cell proliferation is expressed as:
IC=(OD of drug exposed cells/OD of drug free wells).times.100.
The mean of multiple measurements for each Concentration is plotted
vs. the drug concentration. The dose-response curves are plotted
using XLFit 3 (IDBS, United Kingdom). The IC.sub.50 (drug
concentration to obtain 50% inhibition of cell proliferation)
determination values are calculated using the XLFit 3 from semi-log
curves. The IC.sub.50 value determined for each compound in each
cell line is used to determine the concentration of a compound of
Formula III and of the standard chemotherapeutic to be used in
combination.
[2054] The cells are treated with a combination of five
concentrations of a compound of Formula III and five concentrations
of one of 5-fluorouracil, carboplatin, dacarbazine, gefitinib,
oxaliplatin, paclitaxel, SN-38, temozolomide, or vinblastine, based
on the IC.sub.50 results. The compounds and cells are treated per
the IC.sub.50 determination described above and assayed by the MTT
assay.
[2055] The results are assessed to determine whether the
combination is synergistic or antagonistic. The compound
interactions are calculated by multiple drug effect analysis and
are performed by the median equation principle according to the
methodology described by Chou and Talalay (Adv. Enzyme Regul. 1984,
22: 27-55).
[2056] The combination index (CI) will be calculated by the Chou et
al. equation (Adv. Enzyme Regul. 1984, 22: 27-55; Encyclopaedia of
human biology, Academic Press, 1991, 2: 371-9; Synergism and
Antagonism in Chemotherapy, Academic Press, 1991, 61-102) which
takes into account both the potency (D.sub.m or IC.sub.50) and the
shape of the dose-effect curve (the m value). The general equation
for the CI of the two compounds is given by:
CI = ( D ) 1 ( D x ) 1 + ( D ) 2 ( D x ) 2 + ( D ) 1 ( D ) 2 ( D x
) 1 ( D x ) 2 ##EQU00002##
where: [2057] (D.sub.x).sub.1 and (D.sub.x).sub.2 in the
denominators are the doses (or concentrations) for compound 1 and
compound 2 alone which demonstrate x % of inhibition, whereas
(D).sub.1 and (D).sub.2 in the numerators are doses of both
compounds (1 and 2) in combination that also inhibit x %
(iso-effective). CI<1, =1, and >1 indicate synergism,
additive effect and antagonism, respectively.
[2058] The (D.sub.x).sub.1 and (D.sub.x).sub.2 can be calculated
from the median-effect equation of Chou et al. (J. Natl. Cancer
Inst. 1994, 86: 1517-24):
D x = D m ( f a ( 1 - f a ) ) 1 / m ##EQU00003##
where: [2059] D.sub.m is the median-effect dose that is obtained
from the anti-log of x-intercept of the median-effect plot,
x=log(D) versus y=log{f.sub.a/(1-f.sub.a)}, or
D.sub.m=10.sup.(y-intercept)/m; and m is the slope of the
median-effect plot and f.sub.a is the fraction of cells affected by
the treatment. [2060] Each CI will be calculated with CalcuSyn
software (Biosoft, UK) from the mean affected fraction at each drug
ratio concentration.
[2061] Additional examples. Unless specifically indicated
otherwise, the Formula enumeration and R group enumeration used in
the following examples is not related to such enumeration in other
sections of this application. The reagents and solvents used in
these examples can be readily substituted with appropriate
alternatives as are known in the art and isolation of products is
readily achieved by methods known in the art, including, but not
limited to, extraction, crystallization, and chromatographic
methods. For the following examples, the following formulae are
defined:
##STR03443##
wherein: [2062] R.sup.4 and R.sup.5 are as defined in paragraph
[0008]; [2063] R.sup.12, R.sup.13 and R.sup.16 are as defined in
paragraph [0103]; [2064] R.sup.28 is --SO.sub.2R.sup.25,
--C(.dbd.O)R.sup.25, --C(.dbd.O)NR.sup.25, --C(.dbd.O)OR.sup.25,
--C(.dbd.S)R.sup.25, --C(.dbd.S)NR.sup.25, --C(.dbd.S)OR.sup.25, or
--SO.sub.2NR.sup.25R.sup.26; wherein R.sup.25 and R.sup.26 are
hydrogen, optionally substituted lower alkyl, optionally
substituted lower alkenyl, optionally substituted lower alkynyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, or optionally
substituted heteroaryl, or [2065] R.sup.25 and R.sup.26 together
with the nitrogen form optionally substituted 5-7 membered
heterocycloalkyl or optionally substituted 5 or 7 membered nitrogen
containing heteroaryl. [2066] R.sup.29 is hydrogen or optionally
substituted lower alkyl; and [2067] R.sup.30 is optionally
substituted lower alkyl or optionally substituted benzyl, wherein
optionally substituted benzyl refers to an optionally substituted
aralkyl with unsubstituted structure --CH.sub.2-phenyl.
Example 77
Synthesis of Compound of Formula II, where R.sup.12 and R.sup.16
are Independently fluoro or chloro
##STR03444##
[2068] Step 1--Synthesis of Compound of Formula II
[2069] Compound of Formula II, where R.sup.12 and R.sup.16 are
fluoro or chloro, may be synthesized by reacting a compound of
Formula III with an organolithium reagent (e.g. n-butyllithium,
lithium diisopropylamine) in an inert solvent (e.g. THF), followed
by the addition of a formylating reagent (e.g. DMF). The reaction
is allowed to proceed, typically at -78.degree. C., for 1-2 hours
and the desired product is isolated by standard procedures (e.g.
extraction, silica gel chromatography).
Example 78
Synthesis of Compound of Formula III, where R.sup.13 is
NR.sup.28R.sup.29
##STR03445##
[2070] Step-1--Synthesis of Compound of Formula III where R.sup.13
is NR.sup.28R.sup.29
[2071] Compound of Formula III, where R.sup.13 is
NR.sup.28R.sup.29, may be synthesized by reacting a compound of
Formula IV with a base (e.g. pyridine, sodium hydride) in an inert
solvent (e.g. DMF, CH.sub.2Cl.sub.2), followed by an appropriate
reagent (R.sup.25SO.sub.2Cl, e.g. propane-1-sulfonyl chloride;
R.sup.25C(.dbd.O)Cl, e.g. acetyl chloride; R.sup.25NCO, e.g. propyl
isocyanate; R.sup.25OC(.dbd.O)Cl, e.g. benzyl chloroformate;
R.sup.26R.sup.25NSO.sub.2Cl, e.g. dimethylsulfamoyl chloride.) The
reaction is allowed to proceed, typically at room temperature, for
8-12 hours and the desired product is isolated by standard
procedures (e.g. extraction and silica gel chromatography).
Example 79
Synthesis of Compound of Formula III, where R.sup.13 is
CONR.sup.25R.sup.26
##STR03446##
[2072] Step-1--Synthesis of Compound of Formula III where R.sup.13
is C(.dbd.O)NR.sup.25R.sup.26
[2073] Compound of Formula III, where R.sup.13 is
C(.dbd.O)NR.sup.25R.sup.26, may be synthesized by reacting a
compound of Formula V with a base (e.g. N,N-diisopropylethylamine)
and condensing reagents (e.g.
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and
1-hydroxybenzotriazole) in an inert solvent (e.g. tetrahydrofuran),
followed by an amine (R.sup.25R.sup.26NH, e.g. 1-propanamine). The
reaction is allowed to proceed, typically at room temperature, for
8-12 hours and the desired product is isolated by standard
procedures (e.g. extraction, silica gel chromatography).
Example 80
Synthesis of Compound of Formula III, where R.sup.13 is
SO.sub.2NR.sup.25R.sup.26
##STR03447##
[2074] Step-1--Synthesis of Compound of Formula III
[2075] Compound of Formula III, where R.sup.13 is
--SO.sub.2NR.sup.25R.sup.26, is synthesized by reacting a compound
of Formula VI with a base (e.g. triethylamine) in an inert solvent
(e.g. CH.sub.2Cl.sub.2), followed by an appropriate reagent
(R.sup.25R.sup.25NH.sub.2, e.g. 1-propanamine). The reaction is
allowed to proceed, typically at room temperature, for 2-5 hours
and the desired product is isolated by standard procedures (e.g.
extraction, silica gel chromatography).
Example 81
Synthesis of Compound of Formula III, where R.sup.13 is OP where P
is a Protecting Group
##STR03448##
[2076] Step-1--Synthesis of compound of Formula III, where R.sup.13
is OP and P is a Protecting Group
[2077] Compound of Formula III, where R.sup.13 is --OP and P is a
protecting group, is synthesized by reacting a compound of Formula
VII with a base (e.g. imidazole) in a polar solvent (e.g. DMF),
followed by an appropriate reagent (e.g.
tert-butyl-chloro-dimethyl-silane) to introduce the protecting
group. The reaction is allowed to proceed, typically at room
temperature, for 8-12 hours, and the desired product is isolated by
standard procedures (e.g. extraction, silica gel
chromatography).
Example 82
Synthesis of Compound of Formula VIII
##STR03449##
[2078] Step-1--Synthesis of Compound of Formula VIII
[2079] Compound of Formula VIII is synthesized by reacting a
compound of Formula IX with a base (e.g. pyridine) in an inert
solvent (e.g. CH.sub.2Cl.sub.2), followed by sulfuryl chloride. The
reaction is allowed to proceed, typically under reflux, for 8-12
hours and the desired product is isolated by standard procedures
(e.g. evaporation).
Example 83
Synthesis of Compounds of Formula X
##STR03450##
[2080] Step 1--Preparation of Compounds of Formula XIIa and
XIIb
[2081] To a compound of Formula XVIII and a compound of Formula II
is added an appropriate solvent (e.g. methanol) followed by an
appropriate base (e.g. potassium hydroxide, sodium methoxide). The
reaction is typically allowed to stir at room temperature
overnight. Isolation by convention means (e.g. extraction, washing
and filtering) affords a mixture of compounds of Formula XIIa and
XIIb which may be separated by silica gel chromatography if
desired.
Step 2--Preparation of Compounds of Formula X
[2082] To a compound of Formula XIIa or XIIb in an appropriate
solvent (e.g. acetonitrile) is added a reducing agent (e.g.
trifluoroacetic acid and triethylsilane). Typically, the reaction
is allowed to stir at room temperature overnight. Isolation by
conventional means (e.g. extraction and silica gel column
chromatography) affords compounds of Formula X.
Example 84
Synthesis of Compounds of Formula I
##STR03451##
[2083] Step 1--Preparation of Compounds of Formula I
[2084] To a compound of Formula XIIa in an appropriate solvent
(e.g. THF) is added an oxidizing agent (e.g. Dess-Martin periodane,
TEMPO, DDQ). Typically, the reaction is allowed to stir at room
temperature for 20 minutes. Isolation by conventional means (e.g.
extraction and silica gel column chromatography) affords compounds
of Formula I.
Example 85
Synthesis of Compounds of Formula XIII where R.sup.12 and R.sup.16
are Independently chloro or fluoro
##STR03452##
[2085] Step -1--Synthesis of compound of Formula XIII, where
R.sup.12 and R.sup.16 are chloro or fluoro
[2086] Compound of Formula XIII, where R.sup.12 and R.sup.16 are
chloro or fluoro, is synthesized by reacting a compound of Formula
XIV with an organolithium reagent (e.g. n-butyllithium) and a
temporary protecting group (e.g.
1,2-Bis-(chloro-dimethyl-silanyl)-ethane) and DMF in a inert
solvent (e.g. THF) under inert atmosphere (e.g. argon) at
-78.degree. C. for 2-4 hours, followed by removal of the temporary
protecting group using an acid (e.g. 1N HCl). The product is
isolated by extraction and silica gel column chromatography.
Example 86
Synthesis of Compounds of Formula XV, where R.sup.13 is
NHR.sup.28
##STR03453##
[2087] Step 1--Synthesis of Compound of Formula XVI
[2088] Compound of Formula XVI may be synthesized by reacting a
compound of Formula XIII with a base (e.g. pyridine, sodium
hydride) in an inert solvent (e.g. DMF, CH.sub.2Cl.sub.2), followed
by an appropriate reagent (R.sup.25SO.sub.2Cl, e.g.,
propane-1-sulfonyl chloride; R.sup.25C(.dbd.O)Cl, e.g., acetyl
chloride; R.sup.25NCO, e.g., propyl isocyanate;
R.sup.25OC(.dbd.O)Cl, e.g., benzyl chloroformate;
R.sup.26R.sup.25NSO.sub.2Cl, e.g., dimethylsulfamoyl chloride). The
reaction is allowed to proceed, typically at room temperature, for
8-12 hours and the desired product is isolated by standard
procedures (e.g. extraction and silica gel chromatography).
Step 2--Synthesis of Compound of Formula XVII
[2089] Compound of Formula XVII, may be synthesized by hydrolyzing
a compound of Formula XVI with an aqueous base solution (e.g.
sodium hydroxide). The reaction is allowed to proceed, typically
under reflux, for 8-12 hours and the desired product is isolated by
standard procedures (e.g. extraction).
Step 3--Synthesis of Compound of Formula XV, where R.sup.13 is
NHR.sup.28
[2090] Compound of Formula XV, where R.sup.13 is NHR.sup.28, may be
synthesized by reacting a compound of Formula XVII with thionyl
chloride. The reaction is allowed to proceed, typically under
reflux, for 3 hours and the desired product is isolated by standard
procedures (e.g. evaporation).
Example 87
Synthesis of Compounds of Formula XV
##STR03454##
[2091] Step 1--Synthesis of Compound of Formula XV
[2092] Compound of Formula XV may be synthesized by reacting a
compound of Formula XIX with thionyl chloride. The reaction is
allowed to proceed, typically under reflux, for 3 hours and the
desired product is isolated by standard procedures (e.g.
evaporation).
Example 88
Synthesis of Compounds of Formula I
##STR03455##
[2093] Step-1--Synthesis of Compound of Formula I
[2094] Compound of Formula I is synthesized by reacting a compound
of Formula XVIII with a compound of Formula XV (Example 83, e.g.
benzoyl chloride) in the presence of a Lewis acid (e.g. aluminum
trichloride) in an inert solvent (e.g. methylene chloride) under an
inert atmosphere (e.g. argon) at room temperature or with heating
up to reflux for 1-18 hours. The product is isolated by extraction
and silica gel column chromatography.
Example 89
Synthesis of Compounds of Formula I where R.sup.5 is aryl or
heteroaryl
##STR03456##
[2095] Step 1: Synthesis of Compound of Formula I, where R.sup.5 is
aryl or heteraryl
[2096] Compound of the Formula I, where R.sup.5 is aryl or
heteroaryl, is prepared by reacting a compound of Formula I, where
R.sup.5 is bromo, under Suzuki coupling conditions, with boronic
acid (e.g. phenyl boronic acid) in the presence of a base (e.g.,
potassium carbonate) and catalyst (e.g., Pd(Ph.sub.3P.sub.4) in
aqueous/THF solvent system. After 4-12 hours with heating to
80.degree. C. or heating in a microwave instrument at 120.degree.
C. for 15 minutes, the product is isolated by standard workup
procedures (e.g. silica gel column chromatography).
Example 90
Synthesis of Compounds of Formula I
##STR03457##
[2097] Step-1--Synthesis of Compound XX
[2098] Compound of Formula XX can be synthesized by reacting a
compound of Formula XVIII with hexamethyltetramine and acetic acid
in water with heating to reflux for two hours. After cooling, the
desired product precipitates and may be collected by
filtration.
Step-2--Synthesis of Compound of Formula XXI
[2099] Compound of Formula XXI, where P is a protecting group, is
synthesized by reacting a compound XX with an appropriate reagent
to introduce a protecting group (P--X, e.g.
triisopropylsilylchloride) and a base (e.g. sodium hydride) in a
solvent (e.g. THF) typically at room temperature for 8-12 hours.
The product is isolated by conventional means (e.g.
extraction).
Step-3--Synthesis of Compound of Formula XXII
[2100] Compound of Formula XXII is synthesized by reading a
compound of Formula XXI in a solvent (e.g. THF) with an
organolithium reagent (e.g. phenyl lithium) in a solvent (e.g. THF)
under an inert atmosphere, cooled to -78.degree. C. An appropriate
organolithium reagent can also be prepared by reacting compounds of
Formula III, where R.sup.12 and R.sup.16 are independently fluoro
or chloro, with an organolithium reagent (e.g. butyllithium) in a
solvent (e.g. THF) under an inert atmosphere, cooled to -78.degree.
C. The reaction is typically allowed to warm to room temperature
and stirred for 30 minutes. The product is isolated by conventional
means (e.g. extraction).
Step-4--Synthesis of an Intermediate of Compound of Formula I
[2101] An intermediate of compound of Formula I is synthesized by
reacting a compound of Formula XXII with an appropriate reagent to
remove the protecting group, P, (e.g. tetra-n-butyl ammonium
fluoride) in an appropriate solvent (e.g. THF). The final product
is isolated by standard procedures (e.g. extraction).
Step-5--Synthesis of Compound of Formula I
[2102] Compound of Formula I is synthesized by reacting the
intermediate from Step 4 with an oxidizing agent (e.g. Dess-Martin
periodane, TEMPO) in an aprotic solvent (e.g. THF) typically at
room temperature for 20 minutes. The product is isolated by
conventional means (e.g. extraction and silica gel
chromatography).
Example 91
Synthesis of Compounds of Formula I, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26
##STR03458##
[2103] Step 1--Preparation of Compounds of Formula XIIa where
R.sup.13 is NHC(.dbd.O)OR.sup.28
[2104] To a compound of Formula XVIII and a compound of Formula II,
where R.sup.13 is NHC(.dbd.O)OR.sup.28, is added an appropriate
solvent (e.g. methanol) followed by an appropriate base (e.g.
potassium hydroxide). The reaction is typically allowed to stir at
room temperature overnight. Isolation by convention means (e.g.
extraction, washing and filtering) affords compound of Formula XIIa
where R.sup.13 is NHC(.dbd.O)OR.sup.28.
Step 2--Preparation of Compounds of Formula XIIa, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26
[2105] Compound of Formula XIIa, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26, is synthesized by reacting a compound
of Formula XIIa, where R.sup.13 is NHC(.dbd.O)OR.sup.28, with an
amine of the formula NHR.sup.25R.sup.26 (e.g. propylamine) in an
appropriate solvent (e.g. dioxane) followed by an appropriate base
(e.g. triethylamine). The reaction is typically heated at
140.degree. C. for 15 minutes in a CEM Discover microwave
instrument. Isolation by convention means (e.g. extraction, washing
and filtering) affords compounds of Formula XIIa, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26.
Step 3--Preparation of Compounds of Formula I, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26
[2106] Compound of Formula I, where R.sup.13 is
NHC(.dbd.O)NR.sup.25R.sup.26, is synthesized by reacting a compound
of Formula XIIa, where R.sup.13 is NHC(.dbd.O)NR.sup.25R.sup.26,
with an oxidizing agent (e.g. Dess-Martin periodane, TEMPO) in an
appropriate solvent (e.g. THF). The reaction is typically stirred
at room temperature for 15 minutes. Isolation by convention means
(e.g. extraction, washing and filtering) affords compounds of
Formula I, where R.sup.13 is NHC(.dbd.O)NR.sup.25R.sup.26.
Example 92
Synthesis of Compounds of Formula I, where R.sup.13 is
NHC(.dbd.O)OR.sup.25
##STR03459##
[2107] Step 1--Preparation of Compounds of Formula XIIa, where
R.sup.13 is NHC(O)OR.sup.25
[2108] Compound of Formula XIIa, where R.sup.13 is
NHC(.dbd.O)OR.sup.25, is synthesized by reacting a compound of
Formula XIIa, where R.sup.13 is NHC(.dbd.O)OR.sup.28, with an
alcohol of the formula R.sup.25OH (e.g. methanol) in an appropriate
solvent (e.g. dioxane) followed by an appropriate base (e.g.
triethylamine). The reaction is typically heated at 140.degree. C.
for 15 minutes in a CEM Discover microwave instrument. Isolation by
convention means (e.g. extraction, washing and filtering) affords
compounds of Formula XIIa, where R.sup.13 is NHC(K))OR.sup.25.
Step 2--Preparation of Compounds of Formula I, where R.sup.13 is
NHC(.dbd.O)OR.sup.25
[2109] Compound of Formula I, where R.sup.13 is
NHC(.dbd.O)OR.sup.25, is synthesized by reacting a compound of
Formula XIIa, where R.sup.13 is NHC(.dbd.O)OR.sup.25, with an
oxidizing agent (e.g. Dess-Martin periodane, TEMPO) in an
appropriate solvent (e.g. THF). The reaction is typically stirred
at room temperature for 15 minutes. Isolation by convention means
(e.g. extraction, washing and filtering) affords compounds of
Formula I, where R.sup.13 is NHC(.dbd.O)OR.sup.25.
Example 93
Synthesis of Compounds of Formula I, where R.sup.4 and R.sup.5 are
hydrogen
##STR03460##
[2110] Step 1--Preparation of Compounds of Formula I where R.sup.4
and R.sup.5 are hydrogen
[2111] Compound of Formula I, where R.sup.4 and R.sup.5 are
hydrogen, is synthesized by hydrogenating compound of Formula I,
where R.sup.4 is hydrogen and R.sup.5 is bromo, in the presence of
an appropriate solvent (e.g methanol) and catalyst (e.g. 10% Pd/C),
under an atmosphere of hydrogen gas. The reaction is typically
allowed to stir at room temperature for 8-12 hours. Isolation by
convention means (e.g. extraction, washing and filtering) affords
compounds of Formula I, where R.sup.4 and R.sup.5 are hydrogen.
Synthesis of Compound of Formula Ia:
[2112] Compounds of Formula Ia are compounds of Formula XVIII in
which R.sup.4 is hydrogen and R.sup.5 is the only substituent on
the core structure. Exemplary synthetic schemes for groups of
compounds within Formula Ia are shown in Examples 91 to 99 for
different selections of R.sup.5.
##STR03461##
Example 94
Synthesis of Compounds of Formula Ia where R.sup.5 is aryl or
heteroaryl
##STR03462##
[2114] Compound of Formula Ia, where R.sup.5 is aryl or heteroaryl,
is synthesized from compound 1 under Suzuki reaction conditions
using aryl or heteroaryl bornonic acids (e.g. phenyl bornonic acid)
in the presence of a base (e.g. potassium carbonate) and a catalyst
(e.g. Pd(PPh.sub.3).sub.4) in aqueous/THF system with thermal
heating (e.g. 80.degree. C. for 12 hours) or microwave heating
(e.g. 120.degree. C. for 15 minutes). The product is isolated by
conventional means (e.g. silica gel column chromatography).
Example 95
Synthesis of Compounds of Formula Ia where R.sup.5 is alkyl or
cycloalkyl
##STR03463##
[2115] Step-1--Synthesis of Compound of Formula XXIII
[2116] Compound of Formula XXIII, where P is a protecting group, is
synthesized by reacting compound 1 with a base (e.g. sodium
hydride) in an inert solvent (e.g. THF), followed by an appropriate
reagent (P--X, e.g., triisopropylsilylchloride) for introduction of
a protecting group. The reaction is allowed to proceed, typically
at room temperature, for 8-12 hours and the desired product is
isolated by standard procedures (e.g. extraction) (Greene, T. W.;
Wuts, P.G.M. Protective Groups in Organic Synthesis I, 3.sup.rd
ed.; John Wiley & Sons: New York, 1981).
Step-2--Synthesis of an Intermediate of Compound of Formula Ia,
where R.sup.5 is alkyl and cycloalkyl
[2117] An intermediate of compound of Formula Ia, where R.sup.5 is
alkyl or cycloalkyl, is synthesized by reacting a compound of
Formula XXIII with an alkyl or cycloalkyl Grignard reagent (e.g.
ethyl magnesium bromide) in the presence of catalyst (e.g.
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(11)) in an
inert solvent (e.g. toluene) at low temperature (e.g. -78.degree.
C.) or under reflux for 2-8 hours. The product is isolated by
standard procedures (e.g. extraction and silica gel column
chromatography) as described by literature. (T. Hayashi, M.
Konishi, Y. Kobori, M. Kumada, T. Higuchi, K. Hirotsu; J. Am. Chem.
Soc. 1984, 106, 158-163).
Step-3--Synthesis of Compound of Formula Ia, where R.sup.5 is alkyl
and cycloalkyl
[2118] Compound of Formula Ia, where R.sup.5 is alkyl or
cycloalkyl, is synthesized by reacting an intermediate of compound
Formula Ia from Step 2 with an appropriate reagent to remove the
protecting group (e.g. tetrabutylammonium fluoride) in an
appropriate solvent (e.g. tetrahydrofuran). The product is isolated
by standard procedures (e.g. extraction and silica gel column
chromatography).
Example 96
Synthesis of Compounds of Formula Ia where R.sup.5 is
NR.sup.22R.sup.23
##STR03464##
[2119] Step-1--Synthesis of an Intermediate of Compound of Formula
XXIV
[2120] An intermediate of compound of Formula XXIV, is synthesized
by reacting a compound of Formula XXIII, with an amine of the
formula NHR.sup.22R.sup.23 (e.g. aniline) in a solvent (e.g.
toluene), in the presence of a base (e.g. sodium tert-butoxide) and
a catalyst composed of a metal (e.g.,
Tris(dibenzylideneacetone)dipalladium(0)) and a ligand (e.g.,
tri-tert-butylphosphine) with heating, typically to 95.degree. C.,
for 8-12 hours as described (Thomas, et. al., J. Am. Chem. Soc.,
2001, 123, 9404) by substituting a compound of Formula XXIII for
the N-substituted-3,6-dibromocarbazole. The desired compound is
purified by silica gel column chromatography. This intermediate is
used directly in Step 3 to provide compound of Formula Ia where
R.sup.5 is NR.sup.22R.sup.23 and R.sup.22 and R.sup.23 are not
--C(X)R.sup.20, --C(X)NR.sup.17R.sup.18, --S(O).sub.2R.sup.21, or
S(O).sub.2NR.sup.17R.sup.18 or alternatively, it can be
additionally substituted as described in Step 2.
Step-2--Synthesis of Compound of Formula XXIV
[2121] The intermediate from Step 1 can be further modified when
R.sup.22 or R.sup.23 is hydrogen. In this case, the intermediate
from Step 1 can be reacted with a base (e.g. sodium hydride) in a
solvent (e.g. N,N-dimethylformamide), followed by reaction with an
alkylating reagent (e.g. benzyl bromide) or an acylating reagent
(e.g. benzoyl chloride, phenyl isocyanate, phenyl isothiocyanate,
phenylsulfonyl chloride) typically at room temperature or with
heating up to 80.degree. C. for 1-12 hours. The desired product can
be purified by conventional means (e.g. silica gel column
chromatography). Alternatively, when R.sup.22 or R.sup.23 is a
suitable protecting group (e.g. benzyl), it may be removed by
appropriate treatment (e.g. hydrogenation) to provide a compound
where R.sup.22 and/or R.sup.23 are hydrogen, which is suitable for
further modification with an alkylating reagent or acylating
reagent as described herein.
Step-3--Synthesis of Compound of Formula Ia, where R.sup.5 is
--NR.sup.22R.sup.23
[2122] Compound of Formula Ia, where R.sup.5 is
--NR.sup.22R.sup.23, is synthesized by reacting a compound of
Formula XXIV with an appropriate reagent to remove the protecting
group (e.g. tetra-n-butylammonium fluoride) in an appropriate
solvent (e.g. methanol). The final product can be isolated by
standard procedures (e.g. extraction).
Example 97
Synthesis of Compounds of Formula Ia where R.sup.5 is
C(O)NR.sup.25R.sup.26
##STR03465##
[2123] Step-1--Synthesis of Compound of Formula XXIII
[2124] Compound of Formula XIII, where P is a protecting group, is
synthesized by reacting compound 1 with a base (e.g. sodium
hydride) in a solvent (e.g. THF), followed by an appropriate
reagent (P--X, e.g. triisopropylsilylchloride) for introduction of
a protecting group. The reaction is allowed to proceed, typically
at room temperature for 8-12 hours, and the desired product is
isolated by standard procedures (e.g. extraction and silica gel
column chromatography) (Greene, T. W.; Wuts, P. G. M. Protective
Groups in Organic Synthesis I, 3.sup.rd ed.; John Wiley & Sons:
New York, 1981).
Step -2--Synthesis of Compound of Formula XXVI
[2125] Compound of Formula XXVI may be synthesized by reacting a
compound of Formula XXIII with sodium cyanidc in a polar aprotic
solvent (e.g. DMF) in an inert atmosphere (e.g. argon), in the
presence of a catalyst (e.g. cuprous iodide or
Tris(dibenzylideneacetone)dipalladium(0)) following the procedure
described by Buchwald et. al., J. Am. Chem. Soc., 2003, 125,
2890-2891, by substituting 5-bromo-7-azaindole for
5-bromo-indole.
Step -3--Synthesis of Compound of Formula XXVII
[2126] Compound of Formula XXVII may be synthesized by heating a
compound of Formula XXVI with aqueous base (e.g. aq. KOH) in the
presence of an alcohol (e.g. ethanol) at higher temperatures (e.g.
90.degree. C.) for required time, typically 24 h, as described in
Org. Syn. Collective Volume 2, 292 (1943). Alternatively, compounds
of Formula XXVII may be synthesized directly from a compound of
Formula XXIII by reacting a compound of Formula XXIII with a strong
base (e.g. n-butyllithium) and benzyl chloroformate in an inert
solvent (e.g. THF), and further dcbenzylation by hydrogenating the
obtained benzyl ester with hydrogen, in the presence of a catalyst
(e.g. 20% Pd(OH).sub.2/C) at room temperature. The product may be
isolated by filtration and evaporation.
Step-4--Synthesis of Compound of Formula XXVIII
[2127] Compound of Formula XXVIII may be synthesized by reacting a
compound of Formula XXVII with an amine (e.g. benzylamine) in a
polar aprotic solvent (e.g. DMF) in an inert atmosphere, in
presence of an activating agent (e.g. PyBroP
(Bromotri(pyrrolidino)phosphonium hexafluorophosphate) following
the procedure described by Coste et. al., J. Org. Chem., 1994, 59,
2437.
Step-5--Synthesis of Compound of Formula Ia
[2128] Compound of Formula Ia, where R.sup.5 is
C(O)NR.sup.25R.sup.26, may be synthesized by cleaving the
protective group (e.g. TIPS) of a compound of Formula XXVIII with
appropriate reagents (e.g. TBAF) and isolating the product (e.g.
extraction and silica gel column chromatography).
Example 98
Synthesis of Compounds of Formula Ia where R.sup.5 is
CH.sub.2NHR.sup.15R.sup.26
##STR03466##
[2129] Step-2--Synthesis of Compound of Formula XXIII
[2130] Compound of Formula XXIII, where P is a protecting group, is
synthesized by reacting compound 1 with a base (e.g. sodium
hydride) in a solvent (e.g. THF), followed by an appropriate
reagent (P--X, e.g. triisopropylsilylchloride) for introduction of
a protecting group. The reaction is allowed to proceed, typically
at room temperature, for 8-12 hours and the desired product is
isolated by standard procedures (e.g. extraction and silica gel
column chromatography) (Greene, T. W.; Wuts, P.G.M. Protective
Groups in Organic Synthesis I, 3.sup.rd ed.; John Wiley & Sons:
New York, 1981).
Step-2--Synthesis of Compound of Formula XXVI
[2131] Compound of Formula XXVI may be synthesized by reacting a
compound of Formula XXIII with sodium cyanidc in a polar aprotic
solvent (e.g. DMF) in an inert atmosphere, in presence of a
catalyst (e.g. Tris(dibenzylideneacetone)dipalladium(0) or cuprous
iodide) following the procedure described by Buchwald et. al., J
Am. Chem. Soc., 2003, 125, 2890-2891, by substituting
5-bromo-7-azaindole for 5-bromo-indole.
Step-3--Synthesis of Compound of Formula XXIX
[2132] Compound of Formula XXIX can be synthesized from a compound
of Formula XXVI under hydrogenation conditions using a catalyst
(e.g. PtO.sub.2) in an atmosphere of H.sub.2as described by Secrist
III et. al., J Org. Chem., 1972, 37, 335-336.
Step-4--Synthesis of Compound of Formula XXX
[2133] Compound of Formula XXX can be synthesized from a compound
of Formula XXIX with an electrophilic reagent (e.g. benzyl bromide,
benzenesulfonyl chloride, benzoyl chloride, phenyl isocyanate,
phenyl isothiocyanate) in a polar aprotic solvent (e.g. DMF) in an
inert atmosphere, in the presence of a base (e.g. K.sub.2CO.sub.3,
Et.sub.3N). The product can be isolated by standard methods (e.g.
aqueous work up and silica gel column chromatography).
Step-5--Synthesis of Compound of Formula Ia
[2134] Compound of Formula Ia, where R.sup.5 is
CH.sub.2NHR.sup.25R.sup.26, can be synthesized from a compound of
Formula XXX with an electrophilic reagent (e.g. benzyl bromide,
benzenesulfonyl chloride, benzoyl chloride, phenyl isocyanate,
phenyl isothiocyanate) in a polar aprotic solvent (e.g. DMF) in an
inert atmosphere, in the presence of a base (e.g. K.sub.2CO.sub.3,
Et.sub.3N), followed by deprotection of the protecting group with
appropriate conditions (e.g. tetra-n-butylammonium fluoride) and
purification by conventional means (e.g. silica gel
chromatography).
Example 99
Synthesis of Compounds of Formula Ia where R.sup.5 is OR.sup.25
##STR03467##
[2135] Step-1--Synthesis of Compound of Formula Ia, where R.sup.5
is OR.sup.25
[2136] A compound of Formula Ia, where R.sup.5 is OR.sup.25, is
synthesized by reacting compound 1 with a reagent of formula
R.sup.25OH (e.g. methanol) in the presence of base (e.g. sodium
methoxide) and copper (I) bromide in a solvent (e.g.
N,N-dimethylformamide) typically with heating to reflux for 2-8
hours as described by Mazeas, et. al. in Heterocycles, 1999,
50:1065. The desired intermediate is purified by conventional means
(e.g. silica gel column chromatography).
Example 100
Synthesis of Compounds of Formula Ia where R.sup.5 is SR.sup.25
##STR03468##
[2138] Compound of Formula Ia, where R.sup.5 is SR.sup.25, can be
prepared by reacting compound 1 with a strong base (e.g. potassium
hydride or t-butyl lithium) and dialkyldisulfides (e.g.
dimethyldisulfane) or thiophenols (e.g. 4-methoxythiophenol) in a
polar aprotic solvent (e.g. N,N-dimethylformamide) in an inert
atmosphere following the procedure described by Yang et. al.,
Heterocycles, 1992, 34, 1169, by substituting 5-bromo-7-azaindole
for 5-bromo-indole.
Example 101
Synthesis of Compounds of Formula Ia where R.sup.5 is S(O)R.sup.25
or S(O).sub.2R.sup.25
##STR03469##
[2140] Compounds of Formula Ia, where R.sup.5 is S(O)R.sup.2 or
S(O).sub.2R.sup.25 can be prepared by reacting compound of Formula
Ia, where R.sup.5 is SR.sup.25, with 1 or 2 equivalents of
oxidizing agent (e.g. Oxone), respectively, in a polar solvent
(e.g. DMF), using standard procedures.
Example 102
Synthesis of Compound of Formula Ic, where Y.sup.3 is C(O) or
CH.sub.2; Y.sup.6 is NH; and R.sup.27 is as Defined in Paragraph
[0020]
##STR03470##
[2141] Step-1 Synthesis of Compound of Formula XXXI
[2142] Compound of Formula XXXI in turn can be prepared by reacting
6-bromo-7-azaindole (Minakata, S., et al, Synthesis, 1992, p
661-663) with an appropriate nucleophile (e.g. acid chlorides,
alkyl halides, and the like) in polar aprotic solvents (e.g. THF,
DMF) in the presence of a Lewis acid (e.g. AlCl.sub.3, InCl.sub.3,
etc.). The product may be isolated by following standard procedures
(work up and silica gel column chromatography).
Step-2 Synthesis of Compound of Formula Ic
[2143] Compound of Formula Ic can be prepared by reacting compound
of Formula XXXI with R.sup.27NH.sub.2under Buckwald reaction
conditions (Palladium tetrakis trialkylphosphine and a base). The
product may be isolated by following standard procedures (work up
and silica gel column chromatography).
Example 103
Synthesis of Compound of Formula Id, where Y.sup.3 is C(O) or
CH.sub.2; Y.sup.2 is CH.sub.2; and R.sup.27 is as Defined in
Paragraph [0024]
##STR03471##
[2144] Step-1 Synthesis of Compound of Formula XXXII
[2145] Compound of Formula XXXII can be prepared from
2-methyl-7-azaindole (Clemo, S; J. Chem. Soc., 1945, p 603-607) by
reacting with N-bromosuccinimide in carbon tetrachloride followed
by the reaction of the di bromo compound with a nucleophile (e.g.
amine, alkoxy, phenoxy, etc). The product may be isolated by
following standard procedures (work up and silica gel column
chromatography).
Step-2 Synthesis of Compound of Formula Id
[2146] Compound of Formula XXXII can be reacted with a lithium
reagent (e.g. butyl lithium) followed by the reaction with an
aldehyde to provide the key intermediate to compound of Formula Id.
This intermediate can be reacted with a reducing agent (e.g.
trifluoroacetic acid and triethylsilane) at room temperature
overnight to provide a compound of Formula Id where Y.sup.3 is
CH.sub.2as described in Example 79 step-2, or with an oxidizing
agent (e.g. Dess-Martin periodane, TEMPO) at room temperature for
20 minutes to provide a compound of Formula Id where Y.sup.3 is
C(O) as described in Example 80.
Example 104
Synthesis of Compound of Formula Ie, where Y.sup.4 is NH; Y.sup.2
is CH.sub.2; and R.sup.27 is as Defined in Paragraph [0027]
##STR03472##
[2147] Step-1 Synthesis of Compound of Formula XXXIII
[2148] Compound of Formula XXXIII can be prepared by reacting
4-chloro-1-phenylsulfonyl-7-azaindole (Mendiola, J., et al.; J.
Org. Chem., 2004, p 4974-4983) with lithium reagent (e.g. LDA) at
low temperature (e.g. -40.degree. C.) followed by the addition of
an alkyl halide (e.g. benzyl bromide). The product may be isolated
by following standard procedures (work up and silica gel column
chromatography).
Step-2 Synthesis of Compound of Formula Ie
[2149] Compound of Formula Ie can be prepared by reacting a
compound of Formula XXXIII with an amine of formula
R.sup.27NH.sub.2under Buckwald reaction conditions followed by
deprotection and isolating the product by following standard
procedures (work up and silica gel column chromatography).
Example 105
Synthesis of Compound of Formula If, where Y.sup.5 is NH; Y.sup.2
is CH.sub.2; R.sup.27 is as Defined in Paragraph [0030]
##STR03473##
[2150] Step-1 Synthesis of Compound of Formula XXXIV
[2151] Compound of Formula XXXIV can be prepared by reacting a
compound of Formula XXXV with a strong base (e.g. KH, potassium
tert-butoxide) in a suitable solvent (e.g. NMP) at ambient
temperature (Koradin, C., et.al.; Tetrahedron, 2003, p 1571-1588).
The product may be isolated by following standard procedures (work
up and silica gel column chromatography).
Step-2 Synthesis of Compound of Formula If
[2152] Compound of Formula If can be prepared by reacting a
compound of Formula XXXIV with an amine of formula
R.sup.27NH.sub.2under Buckwald reaction conditions followed by
deprotection and isolating the product by following standard
procedures (work up and silica gel column chromatography).
Example 106
Synthesis of Compound of Formula Ij, where Y.sup.4 is NH; Y.sup.5
is O; and R.sup.27 is as Defined in Paragraph [0042]
##STR03474##
[2153] Step-1 Synthesis of Compound of Formula XXXVI
[2154] Compound of Formula XXXVI can be synthesized by reacting
N-triisopopylsilyl-4-chloro-5-hydroxy-7-azaindole (L'Heureux, A.,
Thibault, C., and Ruel, R.; Tetrahedron Letters, 2004, p 2317-2319)
with an alcohol (e.g. benzyl alcohol) under Mitsunobu reaction
conditions and isolating the product by following standard
procedures (work up and silica gel column chromatography).
Step-2 Synthesis of Compound of Formula Ij
[2155] Compound of Formula Ij can be prepared by reacting a
compound of Formula XXXVI with an amine of formula
R.sup.27NH.sub.2under Buckwald reaction conditions followed by
deprotection and isolating the product by following standard
procedures (work up and silica gel column chromatography).
[2156] Additional examples of certain methods contemplated by the
present invention may be found in the following applications: U.S.
Prov. App. No. 60/580,898, filed Jun. 17, 2004; U.S. Prov. App. No.
60/682,076, filed May 17, 2005; U.S. Prov. App. No. 60/682,058,
filed May 17, 2005; U.S. Prov. App. No. 60/682,063, filed May 17,
2005; U.S. Prov. App. No. 60/682,051, filed May 17, 2005; U.S.
Prov. App. No. 60/682,042, filed May 17, 2005; U.S. Prov. App. No.
60/692,750, filed Jun. 22, 2005; and U.S. Prov. App. No.
60/692,960, filed Jun. 22, 2005; each of which are hereby
incorporated by reference herein in their entireties including all
specifications, figures, and tables, and for all purposes.
[2157] All patents and other references cited in the specification
are indicative of the level of skill of those skilled in the art to
which the invention pertains, and are incorporated by reference in
their entireties, including any tables and figures, to the same
extent as if each reference had been incorporated by reference in
its entirety individually.
[2158] One skilled in the art would readily appreciate that the
present invention is well adapted to obtain the ends and advantages
mentioned, as well as those inherent therein. The methods,
variances, and compositions described herein as presently
representative of preferred embodiments are exemplary and are not
intended as limitations on the scope of the invention. Changes
therein and other uses will occur to those skilled in the art,
which are encompassed within the spirit of the invention, are
defined by the scope of the claims.
[2159] It will be readily apparent to one skilled in the art that
varying substitutions and modifications may be made to the
invention disclosed herein Without departing from the scope and
spirit of the invention. For example, variations can be made to
crystallization or co-crystallization conditions for Ret and Ret
surrogate proteins and/or various kinase domain sequences can be
used. Thus, such additional embodiments are within the scope of the
present invention and the following claims.
[2160] The invention illustratively described herein suitably may
be practiced in the absence of any element or elements, limitation
or limitations which is not specifically disclosed herein. Thus,
for example, in each instance herein any of the terms "comprising",
"consisting essentially of" and "consisting of" may be replaced
with either of the other two terms. The terms and expressions which
have been employed are used as terms of description and not of
limitation, and there is no intention that in the use of such terms
and expressions of excluding any equivalents of the features shown
and described or portions thereof, but it is recognized that
various modifications are possible within the scope of the
invention claimed. Thus, it should be understood that although the
present invention has been specifically disclosed by preferred
embodiments and optional features, modification and variation of
the concepts herein disclosed may be resorted to by those skilled
in the art, and that such modifications and variations are
considered to be within the scope of this invention as defined by
the appended claims.
[2161] In addition, where features or aspects of the invention are
described in terms of Markush groups or other grouping of
alternatives, those skilled in the art will recognize that the
invention is also thereby described in terms of any individual
member or subgroup of members of the Markush group or other
group.
[2162] Also, unless indicated to the contrary, where various
numerical values are provided for embodiments, additional
embodiments are described by taking any 2 different values as the
endpoints of a range. Such ranges are also within the scope of the
described invention.
[2163] Thus, additional embodiments are within the scope of the
invention and within the following claims.
Sequence CWU 1
1
125126DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 1caccacggtg tgtcccccaa ctacga 26234DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
2gtcacgtcga ctcagacgcc ttgtttcccc agct 343987DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(978) 3taatacgact cactataggg gaattgtgag cggataacaa
ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc atg ggt cac
cac cat cac cac cac ggt 114 Met Gly His His His His His His Gly 1
5gtg tcc ccc aac tac gac aag tgg gag atg gaa cgc acg gac atc acc
162Val Ser Pro Asn Tyr Asp Lys Trp Glu Met Glu Arg Thr Asp Ile
Thr10 15 20 25atg aag cac aag ctg ggc ggg ggc cag tac ggg gag gtg
tac gag ggc 210Met Lys His Lys Leu Gly Gly Gly Gln Tyr Gly Glu Val
Tyr Glu Gly 30 35 40gtg tgg aag aaa tac agc ctg acg gtg gcc gtg aag
acc ttg aag gag 258Val Trp Lys Lys Tyr Ser Leu Thr Val Ala Val Lys
Thr Leu Lys Glu 45 50 55gac acc atg gag gtg gaa gag ttc ttg aaa gaa
gct gca gtc atg aaa 306Asp Thr Met Glu Val Glu Glu Phe Leu Lys Glu
Ala Ala Val Met Lys 60 65 70gag atc aaa cac cct aac ctg gtg cag ctc
ctt ggg gtc tgc acc cgg 354Glu Ile Lys His Pro Asn Leu Val Gln Leu
Leu Gly Val Cys Thr Arg 75 80 85gag ccc ccg ttc tat atc atc act gag
ttc atg acc tac ggg aac ctc 402Glu Pro Pro Phe Tyr Ile Ile Thr Glu
Phe Met Thr Tyr Gly Asn Leu90 95 100 105ctg gac tac ctg agg gag tgc
aac cgg cag gag gtg aac gcc gtg gtg 450Leu Asp Tyr Leu Arg Glu Cys
Asn Arg Gln Glu Val Asn Ala Val Val 110 115 120ctg ctg tac atg gcc
act cag atc tcg tca gcc atg gag tac ctg gag 498Leu Leu Tyr Met Ala
Thr Gln Ile Ser Ser Ala Met Glu Tyr Leu Glu 125 130 135aag aaa aac
ttc atc cac aga gat ctt gct gcc cga aac tgc ctg gta 546Lys Lys Asn
Phe Ile His Arg Asp Leu Ala Ala Arg Asn Cys Leu Val 140 145 150ggg
gag aac cac ttg gtg aag gta gct gat ttt ggc ctg agc agg ttg 594Gly
Glu Asn His Leu Val Lys Val Ala Asp Phe Gly Leu Ser Arg Leu 155 160
165atg aca ggg gac acc tac aca gcc cat gct gga gcc aag ttc ccc atc
642Met Thr Gly Asp Thr Tyr Thr Ala His Ala Gly Ala Lys Phe Pro
Ile170 175 180 185aaa tgg act gca ccc gag agc ctg gcc tac aac aag
ttc tcc atc aag 690Lys Trp Thr Ala Pro Glu Ser Leu Ala Tyr Asn Lys
Phe Ser Ile Lys 190 195 200tcc gac gtc tgg gca ttt gga gta ttg ctt
tgg gaa att gct acc tat 738Ser Asp Val Trp Ala Phe Gly Val Leu Leu
Trp Glu Ile Ala Thr Tyr 205 210 215ggc atg tcc cct tac ccg gga att
gac ctg tcc cag gtg tat gag ctg 786Gly Met Ser Pro Tyr Pro Gly Ile
Asp Leu Ser Gln Val Tyr Glu Leu 220 225 230cta gag aag gac tac cgc
atg gag cgc cca gaa ggc tgc cca gag aag 834Leu Glu Lys Asp Tyr Arg
Met Glu Arg Pro Glu Gly Cys Pro Glu Lys 235 240 245gtc tat gaa ctc
atg cga gca tgt tgg cag tgg aat ccc tct gac cgg 882Val Tyr Glu Leu
Met Arg Ala Cys Trp Gln Trp Asn Pro Ser Asp Arg250 255 260 265ccc
tcc ttt gct gaa atc cac caa gcc ttt gaa aca atg ttc cag gaa 930Pro
Ser Phe Ala Glu Ile His Gln Ala Phe Glu Thr Met Phe Gln Glu 270 275
280tcc agt atc tca gac gaa gtg gaa aag gag ctg ggg aaa caa ggc gtc
978Ser Ser Ile Ser Asp Glu Val Glu Lys Glu Leu Gly Lys Gln Gly Val
285 290 295tgagtcgac 9874297PRTArtificial SequenceDescription of
Artificial Sequence Synthetic Plasmid 4Met Gly His His His His His
His Gly Val Ser Pro Asn Tyr Asp Lys1 5 10 15Trp Glu Met Glu Arg Thr
Asp Ile Thr Met Lys His Lys Leu Gly Gly 20 25 30Gly Gln Tyr Gly Glu
Val Tyr Glu Gly Val Trp Lys Lys Tyr Ser Leu 35 40 45Thr Val Ala Val
Lys Thr Leu Lys Glu Asp Thr Met Glu Val Glu Glu 50 55 60Phe Leu Lys
Glu Ala Ala Val Met Lys Glu Ile Lys His Pro Asn Leu65 70 75 80Val
Gln Leu Leu Gly Val Cys Thr Arg Glu Pro Pro Phe Tyr Ile Ile 85 90
95Thr Glu Phe Met Thr Tyr Gly Asn Leu Leu Asp Tyr Leu Arg Glu Cys
100 105 110Asn Arg Gln Glu Val Asn Ala Val Val Leu Leu Tyr Met Ala
Thr Gln 115 120 125Ile Ser Ser Ala Met Glu Tyr Leu Glu Lys Lys Asn
Phe Ile His Arg 130 135 140Asp Leu Ala Ala Arg Asn Cys Leu Val Gly
Glu Asn His Leu Val Lys145 150 155 160Val Ala Asp Phe Gly Leu Ser
Arg Leu Met Thr Gly Asp Thr Tyr Thr 165 170 175Ala His Ala Gly Ala
Lys Phe Pro Ile Lys Trp Thr Ala Pro Glu Ser 180 185 190Leu Ala Tyr
Asn Lys Phe Ser Ile Lys Ser Asp Val Trp Ala Phe Gly 195 200 205Val
Leu Leu Trp Glu Ile Ala Thr Tyr Gly Met Ser Pro Tyr Pro Gly 210 215
220Ile Asp Leu Ser Gln Val Tyr Glu Leu Leu Glu Lys Asp Tyr Arg
Met225 230 235 240Glu Arg Pro Glu Gly Cys Pro Glu Lys Val Tyr Glu
Leu Met Arg Ala 245 250 255Cys Trp Gln Trp Asn Pro Ser Asp Arg Pro
Ser Phe Ala Glu Ile His 260 265 270Gln Ala Phe Glu Thr Met Phe Gln
Glu Ser Ser Ile Ser Asp Glu Val 275 280 285Glu Lys Glu Leu Gly Lys
Gln Gly Val 290 295533DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 5acgggaccat atggatgatt
gggagattcc tga 33633DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 6cactggtcga ctattttggc aatgagcggg cca
33729DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 7ggtctagcta cagaaaaatc tcgatggag
29829DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 8ctccatcgag atttttctgt agctagacc
2991006DNAArtificial SequenceDescription of Artificial Sequence
Synthetic PlasmidCDS(56)..(961) 9tattccggat tattcatacc gtcccaccat
cgggcgcgga tctcggtccg aaacc atg 58 Met 1tcg tac tac cat cac cat cac
cat cac gat tac gat atc cca acg acc 106Ser Tyr Tyr His His His His
His His Asp Tyr Asp Ile Pro Thr Thr 5 10 15gaa aac ctg tat ttt cag
ggc cat atg gat gat tgg gag att cct gat 154Glu Asn Leu Tyr Phe Gln
Gly His Met Asp Asp Trp Glu Ile Pro Asp 20 25 30ggg cag att aca gtg
gga caa aga att gga tct gga tca ttt gga aca 202Gly Gln Ile Thr Val
Gly Gln Arg Ile Gly Ser Gly Ser Phe Gly Thr 35 40 45gtc tac aag gga
aag tgg cat ggt gat gtg gca gtg aaa atg ttg aat 250Val Tyr Lys Gly
Lys Trp His Gly Asp Val Ala Val Lys Met Leu Asn50 55 60 65gtg aca
gca cct aca cct cag cag tta caa gcc ttc aaa aat gaa gta 298Val Thr
Ala Pro Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu Val 70 75 80gga
gta ctc agg aaa aca cga cat gtg aat atc cta ctc ttc atg ggc 346Gly
Val Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met Gly 85 90
95tat tcc aca aag cca caa ctg gct att gtt acc cag tgg tgt gag ggc
394Tyr Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu Gly
100 105 110tcc agc ttg tat cac cat ctc cat atc att gag acc aaa ttt
gag atg 442Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe
Glu Met 115 120 125atc aaa ctt ata gat att gca cga cag act gca cag
ggc atg gat tac 490Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln
Gly Met Asp Tyr130 135 140 145tta cac gcc aag tca atc atc cac aga
gac ctc aag agt aat aat ata 538Leu His Ala Lys Ser Ile Ile His Arg
Asp Leu Lys Ser Asn Asn Ile 150 155 160ttt ctt cat gaa gac ctc aca
gta aaa ata ggt gat ttt ggt cta gct 586Phe Leu His Glu Asp Leu Thr
Val Lys Ile Gly Asp Phe Gly Leu Ala 165 170 175aca gaa aaa tct cga
tgg agt ggg tcc cat cag ttt gaa cag ttg tct 634Thr Glu Lys Ser Arg
Trp Ser Gly Ser His Gln Phe Glu Gln Leu Ser 180 185 190gga tcc att
ttg tgg atg gca cca gaa gtc atc aga atg caa gat aaa 682Gly Ser Ile
Leu Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp Lys 195 200 205aat
cca tac agc ttt cag tca gat gta tat gca ttt gga att gtt ctg 730Asn
Pro Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val Leu210 215
220 225tat gaa ttg atg act gga cag tta cct tat tca aac atc aac aac
agg 778Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn
Arg 230 235 240gac cag ata att ttt atg gtg gga cga gga tac ctg tct
cca gat ctc 826Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser
Pro Asp Leu 245 250 255agt aag gta cgg agt aac tgt cca aaa gcc atg
aag aga tta atg gca 874Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met
Lys Arg Leu Met Ala 260 265 270gag tgc ctc aaa aag aaa aga gat gag
aga cca ctc ttt ccc caa att 922Glu Cys Leu Lys Lys Lys Arg Asp Glu
Arg Pro Leu Phe Pro Gln Ile 275 280 285ctc gcc tct att gag ctg ctg
gcc cgc tca ttg cca aaa tagtcgacta 971Leu Ala Ser Ile Glu Leu Leu
Ala Arg Ser Leu Pro Lys290 295 300gagcctgcag tctcgaggca tgcggtacca
agctt 100610302PRTArtificial SequenceDescription of Artificial
Sequence Synthetic Plasmid 10Met Ser Tyr Tyr His His His His His
His Asp Tyr Asp Ile Pro Thr1 5 10 15Thr Glu Asn Leu Tyr Phe Gln Gly
His Met Asp Asp Trp Glu Ile Pro 20 25 30Asp Gly Gln Ile Thr Val Gly
Gln Arg Ile Gly Ser Gly Ser Phe Gly 35 40 45Thr Val Tyr Lys Gly Lys
Trp His Gly Asp Val Ala Val Lys Met Leu 50 55 60Asn Val Thr Ala Pro
Thr Pro Gln Gln Leu Gln Ala Phe Lys Asn Glu65 70 75 80Val Gly Val
Leu Arg Lys Thr Arg His Val Asn Ile Leu Leu Phe Met 85 90 95Gly Tyr
Ser Thr Lys Pro Gln Leu Ala Ile Val Thr Gln Trp Cys Glu 100 105
110Gly Ser Ser Leu Tyr His His Leu His Ile Ile Glu Thr Lys Phe Glu
115 120 125Met Ile Lys Leu Ile Asp Ile Ala Arg Gln Thr Ala Gln Gly
Met Asp 130 135 140Tyr Leu His Ala Lys Ser Ile Ile His Arg Asp Leu
Lys Ser Asn Asn145 150 155 160Ile Phe Leu His Glu Asp Leu Thr Val
Lys Ile Gly Asp Phe Gly Leu 165 170 175Ala Thr Glu Lys Ser Arg Trp
Ser Gly Ser His Gln Phe Glu Gln Leu 180 185 190Ser Gly Ser Ile Leu
Trp Met Ala Pro Glu Val Ile Arg Met Gln Asp 195 200 205Lys Asn Pro
Tyr Ser Phe Gln Ser Asp Val Tyr Ala Phe Gly Ile Val 210 215 220Leu
Tyr Glu Leu Met Thr Gly Gln Leu Pro Tyr Ser Asn Ile Asn Asn225 230
235 240Arg Asp Gln Ile Ile Phe Met Val Gly Arg Gly Tyr Leu Ser Pro
Asp 245 250 255Leu Ser Lys Val Arg Ser Asn Cys Pro Lys Ala Met Lys
Arg Leu Met 260 265 270Ala Glu Cys Leu Lys Lys Lys Arg Asp Glu Arg
Pro Leu Phe Pro Gln 275 280 285Ile Leu Ala Ser Ile Glu Leu Leu Ala
Arg Ser Leu Pro Lys 290 295 3001130DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
11ggcagcccat atggcggcgg cggcggcggc 301231DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
12tgtccgtcga catttaagat ctgtatcctg g 31131232DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(1221) 13taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggc agc agc cat cat cat cat cat 114 Met Gly Ser Ser His His His
His His 1 5cac agc agc ggc ctg gtg ccg cgc ggc agc cat atg gcg gcg
gcg gcg 162His Ser Ser Gly Leu Val Pro Arg Gly Ser His Met Ala Ala
Ala Ala10 15 20 25ggc gcg ggc ccg gag atg gtc cgc ggg cag gtg ttc
gac gtg ggg ccg 210Gly Ala Gly Pro Glu Met Val Arg Gly Gln Val Phe
Asp Val Gly Pro 30 35 40cgc tac acc aac ctc tcg tac atc ggc gag ggc
gcc tac ggc atg gtg 258Arg Tyr Thr Asn Leu Ser Tyr Ile Gly Glu Gly
Ala Tyr Gly Met Val 45 50 55tgc tct gct tat gat aat gtc aac aaa gtt
cga gta gct atc aag aaa 306Cys Ser Ala Tyr Asp Asn Val Asn Lys Val
Arg Val Ala Ile Lys Lys 60 65 70atc agc ccc ttt gag cac cag acc tac
tgc cag aga acc ctg agg gag 354Ile Ser Pro Phe Glu His Gln Thr Tyr
Cys Gln Arg Thr Leu Arg Glu 75 80 85ata aaa atc tta ctg cgc ttc aga
cat gag aac atc att gga atc aat 402Ile Lys Ile Leu Leu Arg Phe Arg
His Glu Asn Ile Ile Gly Ile Asn90 95 100 105gac att att cga gca cca
acc atc gag caa atg aaa gat gta tat ata 450Asp Ile Ile Arg Ala Pro
Thr Ile Glu Gln Met Lys Asp Val Tyr Ile 110 115 120gta cag gac ctc
atg gaa aca gat ctt tac aag ctc ttg aag aca caa 498Val Gln Asp Leu
Met Glu Thr Asp Leu Tyr Lys Leu Leu Lys Thr Gln 125 130 135cac ctc
agc aat gac cat atc tgc tat ttt ctc tac cag atc ctc aga 546His Leu
Ser Asn Asp His Ile Cys Tyr Phe Leu Tyr Gln Ile Leu Arg 140 145
150ggg tta aaa tat atc cat tca gct aac gtt ctg cac cgt gac ctc aag
594Gly Leu Lys Tyr Ile His Ser Ala Asn Val Leu His Arg Asp Leu Lys
155 160 165cct tcc aac ctg ctg ctc aac acc acc tgt gat ctc aag atc
tgt gac 642Pro Ser Asn Leu Leu Leu Asn Thr Thr Cys Asp Leu Lys Ile
Cys Asp170 175 180 185ttt ggc ctg gcc cgt gtt gca gat cca gac cat
gat cac aca ggg ttc 690Phe Gly Leu Ala Arg Val Ala Asp Pro Asp His
Asp His Thr Gly Phe 190 195 200ctg aca gaa tat gtg gcc aca cgt tgg
tac agg gct cca gaa att atg 738Leu Thr Glu Tyr Val Ala Thr Arg Trp
Tyr Arg Ala Pro Glu Ile Met 205 210 215ttg aat tcc aag ggc tac acc
aag tcc att gat att tgg tct gta ggc 786Leu Asn Ser Lys Gly Tyr Thr
Lys Ser Ile Asp Ile Trp Ser Val Gly 220 225 230tgc att ctg gca gaa
atg ctt tct aac agg ccc atc ttt cca ggg aag 834Cys Ile Leu Ala Glu
Met Leu Ser Asn Arg Pro Ile Phe Pro Gly Lys 235 240 245cat tat ctt
gac cag ctg aac cac att ttg ggt att ctt gga tcc cca 882His Tyr Leu
Asp Gln Leu Asn His Ile Leu Gly Ile Leu Gly Ser Pro250 255 260
265tca caa gaa gac ctg aat tgt ata ata aat tta aaa gct agg aac tat
930Ser Gln Glu Asp Leu Asn Cys Ile Ile Asn Leu Lys Ala Arg Asn Tyr
270 275 280ttg ctt tct ctt cca cac aaa aat aag gtg cca tgg aac agg
ctg ttc 978Leu Leu Ser Leu Pro His Lys Asn Lys Val Pro Trp Asn Arg
Leu Phe 285 290 295cca aat gct gac tcc aaa gct ctg gac tta ttg gac
aaa atg ttg aca 1026Pro Asn Ala Asp Ser Lys Ala Leu Asp Leu Leu Asp
Lys Met Leu Thr 300 305 310ttc aac cca cac aag agg att gaa gta gaa
cag gct ctg gcc cac cca 1074Phe Asn Pro His Lys Arg Ile Glu Val Glu
Gln Ala Leu Ala His Pro 315 320 325tat ctg gag cag tat tac gac ccg
agt gac gag ccc atc gcc gaa gca 1122Tyr Leu Glu Gln Tyr Tyr Asp Pro
Ser Asp Glu Pro Ile Ala Glu Ala330 335 340 345cca ttc aag ttc gac
atg gaa ttg gat gac ttg cct aag gaa aag ctc 1170Pro Phe Lys Phe Asp
Met Glu Leu Asp Asp Leu Pro Lys Glu Lys Leu 350 355 360aaa gaa cta
att ttt gaa gag act gct aga ttc cag cca gga tac aga 1218Lys Glu Leu
Ile Phe
Glu Glu Thr Ala Arg Phe Gln Pro Gly Tyr Arg 365 370 375tct
taaatgtcga c 1232Ser14378PRTArtificial SequenceDescription of
Artificial Sequence Synthetic Plasmid 14Met Gly Ser Ser His His His
His His His Ser Ser Gly Leu Val Pro1 5 10 15Arg Gly Ser His Met Ala
Ala Ala Ala Gly Ala Gly Pro Glu Met Val 20 25 30Arg Gly Gln Val Phe
Asp Val Gly Pro Arg Tyr Thr Asn Leu Ser Tyr 35 40 45Ile Gly Glu Gly
Ala Tyr Gly Met Val Cys Ser Ala Tyr Asp Asn Val 50 55 60Asn Lys Val
Arg Val Ala Ile Lys Lys Ile Ser Pro Phe Glu His Gln65 70 75 80Thr
Tyr Cys Gln Arg Thr Leu Arg Glu Ile Lys Ile Leu Leu Arg Phe 85 90
95Arg His Glu Asn Ile Ile Gly Ile Asn Asp Ile Ile Arg Ala Pro Thr
100 105 110Ile Glu Gln Met Lys Asp Val Tyr Ile Val Gln Asp Leu Met
Glu Thr 115 120 125Asp Leu Tyr Lys Leu Leu Lys Thr Gln His Leu Ser
Asn Asp His Ile 130 135 140Cys Tyr Phe Leu Tyr Gln Ile Leu Arg Gly
Leu Lys Tyr Ile His Ser145 150 155 160Ala Asn Val Leu His Arg Asp
Leu Lys Pro Ser Asn Leu Leu Leu Asn 165 170 175Thr Thr Cys Asp Leu
Lys Ile Cys Asp Phe Gly Leu Ala Arg Val Ala 180 185 190Asp Pro Asp
His Asp His Thr Gly Phe Leu Thr Glu Tyr Val Ala Thr 195 200 205Arg
Trp Tyr Arg Ala Pro Glu Ile Met Leu Asn Ser Lys Gly Tyr Thr 210 215
220Lys Ser Ile Asp Ile Trp Ser Val Gly Cys Ile Leu Ala Glu Met
Leu225 230 235 240Ser Asn Arg Pro Ile Phe Pro Gly Lys His Tyr Leu
Asp Gln Leu Asn 245 250 255His Ile Leu Gly Ile Leu Gly Ser Pro Ser
Gln Glu Asp Leu Asn Cys 260 265 270Ile Ile Asn Leu Lys Ala Arg Asn
Tyr Leu Leu Ser Leu Pro His Lys 275 280 285Asn Lys Val Pro Trp Asn
Arg Leu Phe Pro Asn Ala Asp Ser Lys Ala 290 295 300Leu Asp Leu Leu
Asp Lys Met Leu Thr Phe Asn Pro His Lys Arg Ile305 310 315 320Glu
Val Glu Gln Ala Leu Ala His Pro Tyr Leu Glu Gln Tyr Tyr Asp 325 330
335Pro Ser Asp Glu Pro Ile Ala Glu Ala Pro Phe Lys Phe Asp Met Glu
340 345 350Leu Asp Asp Leu Pro Lys Glu Lys Leu Lys Glu Leu Ile Phe
Glu Glu 355 360 365Thr Ala Arg Phe Gln Pro Gly Tyr Arg Ser 370
3751532DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 15gctggatcca ccagggatta tgagattcaa ag
321634DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 16gttcttgtcg actactgagc cttctcttcc tcca
34171066DNAArtificial SequenceDescription of Artificial Sequence
Synthetic PlasmidCDS(56)..(964)CDS(968)..(1051) 17tattccggat
tattcatacc gtcccaccat cgggcgcgga tctcggtccg aaacc atg 58 Met 1tcg
tac tac cat cac cat cac cat cac gat tac gat atc cca acg acc 106Ser
Tyr Tyr His His His His His His Asp Tyr Asp Ile Pro Thr Thr 5 10
15gaa aac ctg tat ttt cag ggc gcc atg gga tcc acc agg gat tat gag
154Glu Asn Leu Tyr Phe Gln Gly Ala Met Gly Ser Thr Arg Asp Tyr Glu
20 25 30att caa aga gaa aga ata gaa ctt gga cga tgt att gga gaa ggc
caa 202Ile Gln Arg Glu Arg Ile Glu Leu Gly Arg Cys Ile Gly Glu Gly
Gln 35 40 45ttt gga gat gta cat caa ggc att tat atg agt cca gag aat
cca gct 250Phe Gly Asp Val His Gln Gly Ile Tyr Met Ser Pro Glu Asn
Pro Ala50 55 60 65ttg gcg gtt gca att aaa aca tgt aaa aac tgt act
tcg gac agc gtg 298Leu Ala Val Ala Ile Lys Thr Cys Lys Asn Cys Thr
Ser Asp Ser Val 70 75 80aga gag aaa ttt ctt caa gaa gcc tta aca atg
cgt cag ttt gac cat 346Arg Glu Lys Phe Leu Gln Glu Ala Leu Thr Met
Arg Gln Phe Asp His 85 90 95cct cat att gtg aag ctg att gga gtc atc
aca gag aat cct gtc tgg 394Pro His Ile Val Lys Leu Ile Gly Val Ile
Thr Glu Asn Pro Val Trp 100 105 110ata atc atg gag ctg tgc aca ctt
gga gag ctg agg tca ttt ttg caa 442Ile Ile Met Glu Leu Cys Thr Leu
Gly Glu Leu Arg Ser Phe Leu Gln 115 120 125gta agg aaa tac agt ttg
gat cta gca tct ttg atc ctg tat gcc tat 490Val Arg Lys Tyr Ser Leu
Asp Leu Ala Ser Leu Ile Leu Tyr Ala Tyr130 135 140 145cag ctt agt
aca gct ctt gca tat cta gag agc aaa aga ttt gta cac 538Gln Leu Ser
Thr Ala Leu Ala Tyr Leu Glu Ser Lys Arg Phe Val His 150 155 160agg
gac att gct gct cgg aat gtt ctg gtg tcc tca aat gat tgt gta 586Arg
Asp Ile Ala Ala Arg Asn Val Leu Val Ser Ser Asn Asp Cys Val 165 170
175aaa tta gga gac ttt gga tta tcc cga tat atg gaa gat agt act tac
634Lys Leu Gly Asp Phe Gly Leu Ser Arg Tyr Met Glu Asp Ser Thr Tyr
180 185 190tac aaa gct tcc aaa gga aaa ttg cct att aaa tgg atg gct
cca gag 682Tyr Lys Ala Ser Lys Gly Lys Leu Pro Ile Lys Trp Met Ala
Pro Glu 195 200 205tca atc aat ttt cga cgt ttt acc tca gct agt gac
gta tgg atg ttt 730Ser Ile Asn Phe Arg Arg Phe Thr Ser Ala Ser Asp
Val Trp Met Phe210 215 220 225ggt gtg tgt atg tgg gag ata ctg atg
cat ggt gtg aag cct ttt caa 778Gly Val Cys Met Trp Glu Ile Leu Met
His Gly Val Lys Pro Phe Gln 230 235 240gga gtg aag aac aat gat gta
atc ggt cga att gaa aat ggg gaa aga 826Gly Val Lys Asn Asn Asp Val
Ile Gly Arg Ile Glu Asn Gly Glu Arg 245 250 255tta cca atg cct cca
aat tgt cct cct acc ctc tac agc ctt atg acg 874Leu Pro Met Pro Pro
Asn Cys Pro Pro Thr Leu Tyr Ser Leu Met Thr 260 265 270aaa tgc tgg
gcc tat gac ccc agc agg cgg ccc agg ttt act gaa ctt 922Lys Cys Trp
Ala Tyr Asp Pro Ser Arg Arg Pro Arg Phe Thr Glu Leu 275 280 285aaa
gct cag ctc agc aca atc ctg gag gaa gag aag gct cag tag tcg 970Lys
Ala Gln Leu Ser Thr Ile Leu Glu Glu Glu Lys Ala Gln Ser290 295
300acg agc tca cta gtc gcg gcc gct ttc gaa tct aga gcc tgc agt ctc
1018Thr Ser Ser Leu Val Ala Ala Ala Phe Glu Ser Arg Ala Cys Ser
Leu305 310 315 320gag gca tgc ggt acc aag ctt gtc gag aag tac
tagaggatca taatc 1066Glu Ala Cys Gly Thr Lys Leu Val Glu Lys Tyr
325 33018331PRTArtificial SequenceDescription of Artificial
Sequence Synthetic Plasmid 18Met Ser Tyr Tyr His His His His His
His Asp Tyr Asp Ile Pro Thr1 5 10 15Thr Glu Asn Leu Tyr Phe Gln Gly
Ala Met Gly Ser Thr Arg Asp Tyr 20 25 30Glu Ile Gln Arg Glu Arg Ile
Glu Leu Gly Arg Cys Ile Gly Glu Gly 35 40 45Gln Phe Gly Asp Val His
Gln Gly Ile Tyr Met Ser Pro Glu Asn Pro 50 55 60Ala Leu Ala Val Ala
Ile Lys Thr Cys Lys Asn Cys Thr Ser Asp Ser65 70 75 80Val Arg Glu
Lys Phe Leu Gln Glu Ala Leu Thr Met Arg Gln Phe Asp 85 90 95His Pro
His Ile Val Lys Leu Ile Gly Val Ile Thr Glu Asn Pro Val 100 105
110Trp Ile Ile Met Glu Leu Cys Thr Leu Gly Glu Leu Arg Ser Phe Leu
115 120 125Gln Val Arg Lys Tyr Ser Leu Asp Leu Ala Ser Leu Ile Leu
Tyr Ala 130 135 140Tyr Gln Leu Ser Thr Ala Leu Ala Tyr Leu Glu Ser
Lys Arg Phe Val145 150 155 160His Arg Asp Ile Ala Ala Arg Asn Val
Leu Val Ser Ser Asn Asp Cys 165 170 175Val Lys Leu Gly Asp Phe Gly
Leu Ser Arg Tyr Met Glu Asp Ser Thr 180 185 190Tyr Tyr Lys Ala Ser
Lys Gly Lys Leu Pro Ile Lys Trp Met Ala Pro 195 200 205Glu Ser Ile
Asn Phe Arg Arg Phe Thr Ser Ala Ser Asp Val Trp Met 210 215 220Phe
Gly Val Cys Met Trp Glu Ile Leu Met His Gly Val Lys Pro Phe225 230
235 240Gln Gly Val Lys Asn Asn Asp Val Ile Gly Arg Ile Glu Asn Gly
Glu 245 250 255Arg Leu Pro Met Pro Pro Asn Cys Pro Pro Thr Leu Tyr
Ser Leu Met 260 265 270Thr Lys Cys Trp Ala Tyr Asp Pro Ser Arg Arg
Pro Arg Phe Thr Glu 275 280 285Leu Lys Ala Gln Leu Ser Thr Ile Leu
Glu Glu Glu Lys Ala Gln Ser 290 295 300Thr Ser Ser Leu Val Ala Ala
Ala Phe Glu Ser Arg Ala Cys Ser Leu305 310 315 320Glu Ala Cys Gly
Thr Lys Leu Val Glu Lys Tyr 325 3301933DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
19gactcctcat atggcagggg tctctgagta tga 332034DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
20caggtcgtcg actactcctg gttggaggtc aagg 342128DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
21ctgggagagg gcgcgtttgg gcaggtgg 282228DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
22ccacctgccc aaacgcgccc tctcccag 282329DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
23cagggctgga atacagctac aaccccagc 292429DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
24gctggggttg tagctgtatt ccagccctg 29251061DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(1038) 25taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggt cac cac cat cac cat cat atg 114 Met Gly His His His His His
His Met 1 5gca ggg gtc tct gag tat gag ctt ccc gaa gac cct cgc tgg
gag ctg 162Ala Gly Val Ser Glu Tyr Glu Leu Pro Glu Asp Pro Arg Trp
Glu Leu10 15 20 25cct cgg gac aga ctg gtc tta ggc aaa ccc ctg gga
gag ggc gcg ttt 210Pro Arg Asp Arg Leu Val Leu Gly Lys Pro Leu Gly
Glu Gly Ala Phe 30 35 40ggg cag gtg gtg ttg gca gag gct atc ggg ctg
gac aag gac aaa ccc 258Gly Gln Val Val Leu Ala Glu Ala Ile Gly Leu
Asp Lys Asp Lys Pro 45 50 55aac cgt gtg acc aaa gtg gct gtg aag atg
ttg aag tcg gac gca aca 306Asn Arg Val Thr Lys Val Ala Val Lys Met
Leu Lys Ser Asp Ala Thr 60 65 70gag aaa gac ttg tca gac ctg atc tca
gaa atg gag atg atg aag atg 354Glu Lys Asp Leu Ser Asp Leu Ile Ser
Glu Met Glu Met Met Lys Met 75 80 85atc ggg aag cat aag aat atc atc
aac ctg ctg ggg gcc tgc acg cag 402Ile Gly Lys His Lys Asn Ile Ile
Asn Leu Leu Gly Ala Cys Thr Gln90 95 100 105gat ggt ccc ttg tat gtc
atc gtg gag tat gcc tcc aag ggc aac ctg 450Asp Gly Pro Leu Tyr Val
Ile Val Glu Tyr Ala Ser Lys Gly Asn Leu 110 115 120cgg gag tac ctg
cag gcc cgg agg ccc cca ggg ctg gaa tac agc tac 498Arg Glu Tyr Leu
Gln Ala Arg Arg Pro Pro Gly Leu Glu Tyr Ser Tyr 125 130 135aac ccc
agc cac aac cca gag gag cag ctc tcc tcc aag gac ctg gtg 546Asn Pro
Ser His Asn Pro Glu Glu Gln Leu Ser Ser Lys Asp Leu Val 140 145
150tcc tgc gcc tac cag gtg gcc cga ggc atg gag tat ctg gcc tcc aag
594Ser Cys Ala Tyr Gln Val Ala Arg Gly Met Glu Tyr Leu Ala Ser Lys
155 160 165aag tgc ata cac cga gac ctg gca gcc agg aat gtc ctg gtg
aca gag 642Lys Cys Ile His Arg Asp Leu Ala Ala Arg Asn Val Leu Val
Thr Glu170 175 180 185gac aat gtg atg aag ata gca gac ttt ggc ctc
gca cgg gac att cac 690Asp Asn Val Met Lys Ile Ala Asp Phe Gly Leu
Ala Arg Asp Ile His 190 195 200cac atc gac tac tat aaa aag aca acc
aac ggc cga ctg cct gtg aag 738His Ile Asp Tyr Tyr Lys Lys Thr Thr
Asn Gly Arg Leu Pro Val Lys 205 210 215tgg atg gca ccc gag gca tta
ttt gac cgg atc tac acc cac cag agt 786Trp Met Ala Pro Glu Ala Leu
Phe Asp Arg Ile Tyr Thr His Gln Ser 220 225 230gat gtg tgg tct ttc
ggg gtg ctc ctg tgg gag atc ttc act ctg ggc 834Asp Val Trp Ser Phe
Gly Val Leu Leu Trp Glu Ile Phe Thr Leu Gly 235 240 245ggc tcc cca
tac ccc ggt gtg cct gtg gag gaa ctt ttc aag ctg ctg 882Gly Ser Pro
Tyr Pro Gly Val Pro Val Glu Glu Leu Phe Lys Leu Leu250 255 260
265aag gag ggt cac cgc atg gac aag ccc agt aac tgc acc aac gag ctg
930Lys Glu Gly His Arg Met Asp Lys Pro Ser Asn Cys Thr Asn Glu Leu
270 275 280tac atg atg atg cgg gac tgc tgg cat gca gtg ccc tca cag
aga ccc 978Tyr Met Met Met Arg Asp Cys Trp His Ala Val Pro Ser Gln
Arg Pro 285 290 295acc ttc aag cag ctg gtg gaa gac ctg gac cgc atc
gtg gcc ttg acc 1026Thr Phe Lys Gln Leu Val Glu Asp Leu Asp Arg Ile
Val Ala Leu Thr 300 305 310tcc aac cag gag tagtcgacga aggagatata
tcc 1061Ser Asn Gln Glu 31526317PRTArtificial SequenceDescription
of Artificial Sequence Synthetic Plasmid 26Met Gly His His His His
His His Met Ala Gly Val Ser Glu Tyr Glu1 5 10 15Leu Pro Glu Asp Pro
Arg Trp Glu Leu Pro Arg Asp Arg Leu Val Leu 20 25 30Gly Lys Pro Leu
Gly Glu Gly Ala Phe Gly Gln Val Val Leu Ala Glu 35 40 45Ala Ile Gly
Leu Asp Lys Asp Lys Pro Asn Arg Val Thr Lys Val Ala 50 55 60Val Lys
Met Leu Lys Ser Asp Ala Thr Glu Lys Asp Leu Ser Asp Leu65 70 75
80Ile Ser Glu Met Glu Met Met Lys Met Ile Gly Lys His Lys Asn Ile
85 90 95Ile Asn Leu Leu Gly Ala Cys Thr Gln Asp Gly Pro Leu Tyr Val
Ile 100 105 110Val Glu Tyr Ala Ser Lys Gly Asn Leu Arg Glu Tyr Leu
Gln Ala Arg 115 120 125Arg Pro Pro Gly Leu Glu Tyr Ser Tyr Asn Pro
Ser His Asn Pro Glu 130 135 140Glu Gln Leu Ser Ser Lys Asp Leu Val
Ser Cys Ala Tyr Gln Val Ala145 150 155 160Arg Gly Met Glu Tyr Leu
Ala Ser Lys Lys Cys Ile His Arg Asp Leu 165 170 175Ala Ala Arg Asn
Val Leu Val Thr Glu Asp Asn Val Met Lys Ile Ala 180 185 190Asp Phe
Gly Leu Ala Arg Asp Ile His His Ile Asp Tyr Tyr Lys Lys 195 200
205Thr Thr Asn Gly Arg Leu Pro Val Lys Trp Met Ala Pro Glu Ala Leu
210 215 220Phe Asp Arg Ile Tyr Thr His Gln Ser Asp Val Trp Ser Phe
Gly Val225 230 235 240Leu Leu Trp Glu Ile Phe Thr Leu Gly Gly Ser
Pro Tyr Pro Gly Val 245 250 255Pro Val Glu Glu Leu Phe Lys Leu Leu
Lys Glu Gly His Arg Met Asp 260 265 270Lys Pro Ser Asn Cys Thr Asn
Glu Leu Tyr Met Met Met Arg Asp Cys 275 280 285Trp His Ala Val Pro
Ser Gln Arg Pro Thr Phe Lys Gln Leu Val Glu 290 295 300Asp Leu Asp
Arg Ile Val Ala Leu Thr Ser Asn Gln Glu305 310 3152730DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
27atcaattcat atggacccag atgaagttcc 302834DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
28atgtagtcga cctaatcctg ttgtacattt gctt 34291353DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(1212) 29taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggt cac cac cat cac cat cat atg 114 Met
Gly His His His His His His Met 1 5gac cca gat gaa gtt cct ttg gat
gag cag tgt gag cgg ctc cct tat 162Asp Pro Asp Glu Val Pro Leu Asp
Glu Gln Cys Glu Arg Leu Pro Tyr10 15 20 25gat gcc agc aag tgg gag
ttt gcc cgg gag aga ctt aaa ctg ggc aaa 210Asp Ala Ser Lys Trp Glu
Phe Ala Arg Glu Arg Leu Lys Leu Gly Lys 30 35 40tca ctt gga aga ggg
gct ttt gga aaa gtg gtt caa gca tca gca ttt 258Ser Leu Gly Arg Gly
Ala Phe Gly Lys Val Val Gln Ala Ser Ala Phe 45 50 55ggc att aag aaa
tca cct acg tgc cgg act gtg gct gtg aaa atg ctg 306Gly Ile Lys Lys
Ser Pro Thr Cys Arg Thr Val Ala Val Lys Met Leu 60 65 70aaa gag ggg
gcc acg gcc agc gag tac aaa gct ctg atg act gag cta 354Lys Glu Gly
Ala Thr Ala Ser Glu Tyr Lys Ala Leu Met Thr Glu Leu 75 80 85aaa atc
ttg acc cac att ggc cac cat ctg aac gtg gtt aac ctg ctg 402Lys Ile
Leu Thr His Ile Gly His His Leu Asn Val Val Asn Leu Leu90 95 100
105gga gcc tgc acc aag caa gga ggg cct ctg atg gtg att gtt gaa tac
450Gly Ala Cys Thr Lys Gln Gly Gly Pro Leu Met Val Ile Val Glu Tyr
110 115 120tgc aaa tat gga aat ctc tcc aac tac ctc aag agc aaa cgt
gac tta 498Cys Lys Tyr Gly Asn Leu Ser Asn Tyr Leu Lys Ser Lys Arg
Asp Leu 125 130 135ttt ttt ctc aac aag gat gca gca cta cac atg gag
cct aag aaa gaa 546Phe Phe Leu Asn Lys Asp Ala Ala Leu His Met Glu
Pro Lys Lys Glu 140 145 150aaa atg gag cca ggc ctg gaa caa ggc aag
aaa cca aga cta gat agc 594Lys Met Glu Pro Gly Leu Glu Gln Gly Lys
Lys Pro Arg Leu Asp Ser 155 160 165gtc acc agc agc gaa agc ttt gcg
agc tcc ggc ttt cag gaa gat aaa 642Val Thr Ser Ser Glu Ser Phe Ala
Ser Ser Gly Phe Gln Glu Asp Lys170 175 180 185agt ctg agt gat gtt
gag gaa gag gag gat tct gac ggt ttc tac aag 690Ser Leu Ser Asp Val
Glu Glu Glu Glu Asp Ser Asp Gly Phe Tyr Lys 190 195 200gag ccc atc
act atg gaa gat ctg att tct tac agt ttt caa gtg gcc 738Glu Pro Ile
Thr Met Glu Asp Leu Ile Ser Tyr Ser Phe Gln Val Ala 205 210 215aga
ggc atg gag ttc ctg tct tcc aga aag tgc att cat cgg gac ctg 786Arg
Gly Met Glu Phe Leu Ser Ser Arg Lys Cys Ile His Arg Asp Leu 220 225
230gca gcg aga aac att ctt tta tct gag aac aac gtg gtg aag att tgt
834Ala Ala Arg Asn Ile Leu Leu Ser Glu Asn Asn Val Val Lys Ile Cys
235 240 245gat ttt ggc ctt gcc cgg gat att tat aag aac ccc gat tat
gtg aga 882Asp Phe Gly Leu Ala Arg Asp Ile Tyr Lys Asn Pro Asp Tyr
Val Arg250 255 260 265aaa gga gat act cga ctt cct ctg aaa tgg atg
gct ccc gaa tct atc 930Lys Gly Asp Thr Arg Leu Pro Leu Lys Trp Met
Ala Pro Glu Ser Ile 270 275 280ttt gac aaa atc tac agc acc aag agc
gac gtg tgg tct tac gga gta 978Phe Asp Lys Ile Tyr Ser Thr Lys Ser
Asp Val Trp Ser Tyr Gly Val 285 290 295ttg ctg tgg gaa atc ttc tcc
tta ggt ggg tct cca tac cca gga gta 1026Leu Leu Trp Glu Ile Phe Ser
Leu Gly Gly Ser Pro Tyr Pro Gly Val 300 305 310caa atg gat gag gac
ttt tgc agt cgc ctg agg gaa ggc atg agg atg 1074Gln Met Asp Glu Asp
Phe Cys Ser Arg Leu Arg Glu Gly Met Arg Met 315 320 325aga gct cct
gag tac tct act cct gaa atc tat cag atc atg ctg gac 1122Arg Ala Pro
Glu Tyr Ser Thr Pro Glu Ile Tyr Gln Ile Met Leu Asp330 335 340
345tgc tgg cac aga gac cca aaa gaa agg cca aga ttt gca gaa ctt gtg
1170Cys Trp His Arg Asp Pro Lys Glu Arg Pro Arg Phe Ala Glu Leu Val
350 355 360gaa aaa cta ggt gat ttg ctt caa gca aat gta caa cag gat
1212Glu Lys Leu Gly Asp Leu Leu Gln Ala Asn Val Gln Gln Asp 365 370
375taggtcgacc accaccacca ccaccactga gatccggctg gccctactgg
ccgaaaggaa 1272ttcgaggcca gcagggccac cgctgagcaa taactagcat
aaccccttgg ggcctctaaa 1332cgggtcttga ggggtttttt g
135330375PRTArtificial SequenceDescription of Artificial Sequence
Synthetic Plasmid 30Met Gly His His His His His His Met Asp Pro Asp
Glu Val Pro Leu1 5 10 15Asp Glu Gln Cys Glu Arg Leu Pro Tyr Asp Ala
Ser Lys Trp Glu Phe 20 25 30Ala Arg Glu Arg Leu Lys Leu Gly Lys Ser
Leu Gly Arg Gly Ala Phe 35 40 45Gly Lys Val Val Gln Ala Ser Ala Phe
Gly Ile Lys Lys Ser Pro Thr 50 55 60Cys Arg Thr Val Ala Val Lys Met
Leu Lys Glu Gly Ala Thr Ala Ser65 70 75 80Glu Tyr Lys Ala Leu Met
Thr Glu Leu Lys Ile Leu Thr His Ile Gly 85 90 95His His Leu Asn Val
Val Asn Leu Leu Gly Ala Cys Thr Lys Gln Gly 100 105 110Gly Pro Leu
Met Val Ile Val Glu Tyr Cys Lys Tyr Gly Asn Leu Ser 115 120 125Asn
Tyr Leu Lys Ser Lys Arg Asp Leu Phe Phe Leu Asn Lys Asp Ala 130 135
140Ala Leu His Met Glu Pro Lys Lys Glu Lys Met Glu Pro Gly Leu
Glu145 150 155 160Gln Gly Lys Lys Pro Arg Leu Asp Ser Val Thr Ser
Ser Glu Ser Phe 165 170 175Ala Ser Ser Gly Phe Gln Glu Asp Lys Ser
Leu Ser Asp Val Glu Glu 180 185 190Glu Glu Asp Ser Asp Gly Phe Tyr
Lys Glu Pro Ile Thr Met Glu Asp 195 200 205Leu Ile Ser Tyr Ser Phe
Gln Val Ala Arg Gly Met Glu Phe Leu Ser 210 215 220Ser Arg Lys Cys
Ile His Arg Asp Leu Ala Ala Arg Asn Ile Leu Leu225 230 235 240Ser
Glu Asn Asn Val Val Lys Ile Cys Asp Phe Gly Leu Ala Arg Asp 245 250
255Ile Tyr Lys Asn Pro Asp Tyr Val Arg Lys Gly Asp Thr Arg Leu Pro
260 265 270Leu Lys Trp Met Ala Pro Glu Ser Ile Phe Asp Lys Ile Tyr
Ser Thr 275 280 285Lys Ser Asp Val Trp Ser Tyr Gly Val Leu Leu Trp
Glu Ile Phe Ser 290 295 300Leu Gly Gly Ser Pro Tyr Pro Gly Val Gln
Met Asp Glu Asp Phe Cys305 310 315 320Ser Arg Leu Arg Glu Gly Met
Arg Met Arg Ala Pro Glu Tyr Ser Thr 325 330 335Pro Glu Ile Tyr Gln
Ile Met Leu Asp Cys Trp His Arg Asp Pro Lys 340 345 350Glu Arg Pro
Arg Phe Ala Glu Leu Val Glu Lys Leu Gly Asp Leu Leu 355 360 365Gln
Ala Asn Val Gln Gln Asp 370 3753141DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
31atgtacgaag ttcagtggaa agttgttgaa gaaatcaacg g 413246DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
32ggtcgatgta aacgtagttg ttaccgttga tttcttcaac aacttt
463342DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 33aacaactacg tttacatcga cccgacccag ctgccgtacg ac
423443DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 34gttacgcggg aactcccatt tgtggtcgta cggcagctgg gtc
433543DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 35aaatgggagt tcccgcgtaa ccgtctgtct ttcggtaaaa ccc
433641DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 36accgaacgca cccgcaccca gggttttacc gaaagacaga c
413743DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 37ggtgcgggtg cgttcggtaa agttgttgaa gcgaccgcgt acg
433841DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 38gccgcgtcag atttgatcag accgtacgcg gtcgcttcaa c
413943DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 39ctgatcaaat ctgacgcggc gatgaccgtt gcggttaaaa tgc
434043DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 40gtcaggtgcg cagacggttt cagcatttta accgcaacgg tca
434143DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 41aaaccgtctg cgcacctgac cgaacgtgaa gcgctgatgt ctg
434244DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 42ccaggtaaga cagaactttc agttcagaca tcagcgcttc acgt
444344DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 43ctgaaagttc tgtcttacct gggtaaccac atgaacatcg ttaa
444441DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 44ggtgcacgca cccagcaggt taacgatgtt catgtggtta c
414543DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 45ctgctgggtg cgtgcaccat cggtggtccg accctggtta tca
434643DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 46gtcaccgtag cagcagtatt cggtgataac cagggtcgga cca
434743DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 47gaatactgct gctacggtga cctgctgaac ttcctgcgtc gta
434843DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 48agagcagatg aaagagtcac gtttacgacg caggaagttc agc
434943DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 49cgtgactctt tcatctgctc taaacaggaa gaccacgcgg aag
435043DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 50cagcaggttt ttgtacagcg ccgcttccgc gtggtcttcc tgt
435144DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 51gcgctgtaca aaaacctgct gcactctaaa gaatcttctt gctc
445245DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 52ccatgtattc gttggtagag tcagagcaag aagattcttt
agagt 455344DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 53gactctacca acgaatacat ggacatgaaa
ccgggtgttt ctta 445443DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 54tccgctttgg tcggaacaac
gtaagaaaca cccggtttca tgt 435543DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 55gttgttccga ccaaagcgga
caaacgtcgt tctgttcgta tcg 435646DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 56taacgtcacg ttcgatgtaa
gaaccgatac gaacagaacg acgttt 465743DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
57tcttacatcg aacgtgacgt taccccggcg atcatggaag acg
435840DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 58ccaggtccag cgccagttcg tcgtcttcca tgatcgccgg
405943DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 59gaactggcgc tggacctgga agacctgctg tctttctctt acc
436044DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 60gaacgccata cctttcgcaa cctggtaaga gaaagacagc aggt
446144DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 61gttgcgaaag gtatggcgtt cctggcgtct aaaaactgca tcca
446239DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 62cgcgccgcca ggtcacggtg gatgcagttt ttagacgcc
396341DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 63cgtgacctgg cggcgcgtaa catcctgctg acccacggtc g
416443DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 64accgaagtcg cagattttgg tgatacgacc gtgggtcagc agg
436543DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 65accaaaatct gcgacttcgg tctggcgcgt gacatcaaaa acg
436646DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 66gttaccttta acaacgtagt tagagtcgtt tttgatgtca
cgcgcc 466744DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 67tctaactacg ttgttaaagg taacgcgcgt
ctgccggtta aatg 446842DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 68gaagatagat tccggcgcca
tccatttaac cggcagacgc gc 426944DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 69atggcgccgg aatctatctt
caactgcgtt tacaccttcg aatc 447043DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 70gataccgtaa gaccaaacgt
cagattcgaa ggtgtaaacg cag 437143DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 71gacgtttggt cttacggtat
cttcctgtgg gaactgttct ctc 437241DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 72cctgtgggaa ctgttctctc
tgggttcttc tccgtacccg g 417345DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 73ggttcttctc cgtacccggg
tatgccggtt gactctaaat tctat 457445DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 74cggaaacctt ctttgatcat
tttgtagaat ttagagtcaa ccggc 457543DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 75aaaatgatca aagaaggttt
ccgtatgctg tctccggaac acg 437642DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 76atgtcgtaca tttccgccgg
cgcgtgttcc ggagacagca ta 427742DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 77ccggcggaaa tgtacgacat
catgaaaacc tgctgggacg cg 427842DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 78aaggtcggac gtttcagcgg
gtccgcgtcc cagcaggttt tc 427943DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 79ccgctgaaac gtccgacctt
caaacagatc gttcagctga tcg 438046DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 80ttggtagatt cagagatctg
tttttcgatc agctgaacga tctgtt 468144DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
81aaacagatct ctgaatctac caaccacatc tactctaacc tggc
448242DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 82tgacggttcg gagagcagtt cgccaggtta gagtagatgt gg
428343DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 83aactgctctc cgaaccgtca gaaaccggtt gttgaccact ctg
438445DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 84gtagaaccaa cagagttgat acgaacagag tggtcaacaa
ccggt 458542DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 85cgtatcaact ctgttggttc taccgcgtct
tcttctcagc cg 428640DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 86aacgtcgtcg tgaaccagca gcggctgaga
agaagacgcg 408732DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 87gttgtttcat atgtacgaag ttcagtggaa ag
328836DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 88gttgtttgtc gactaaacgt cgtcgtgaac cagcag
368934DNAArtificial SequenceDescription of Artificial Sequence
Synthetic Primer 89gttcttgtcg actatttctg acggttcgga gagc
34901325DNAArtificial SequenceDescription of Artificial Sequence
Synthetic PlasmidCDS(88)..(1302) 90taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggt cac cac cat cac cat cat atg 114 Met Gly His His His His His
His Met 1 5tac gaa gtt cag tgg aaa gtt gtt gaa gaa atc aac ggt aac
aac tac 162Tyr Glu Val Gln Trp Lys Val Val Glu Glu Ile Asn Gly Asn
Asn Tyr10 15 20 25gtt tac atc gac ccg acc cag ctg ccg tac gac cac
aaa tgg gag ttc 210Val Tyr Ile Asp Pro Thr Gln Leu Pro Tyr Asp His
Lys Trp Glu Phe 30 35 40ccg cgt aac cgt ctg tct ttc ggt aaa acc ctg
ggt gcg ggt gcg ttc 258Pro Arg Asn Arg Leu Ser Phe Gly Lys Thr Leu
Gly Ala Gly Ala Phe 45 50 55ggt aaa gtt gtt gaa gcg acc gcg tac ggt
ctg atc aaa tct gac gcg 306Gly Lys Val Val Glu Ala Thr Ala Tyr Gly
Leu Ile Lys Ser Asp Ala
60 65 70gcg atg acc gtt gcg gtt aaa atg ctg aaa ccg tct gcg cac ctg
acc 354Ala Met Thr Val Ala Val Lys Met Leu Lys Pro Ser Ala His Leu
Thr 75 80 85gaa cgt gaa gcg ctg atg tct gaa ctg aaa gtt ctg tct tac
ctg ggt 402Glu Arg Glu Ala Leu Met Ser Glu Leu Lys Val Leu Ser Tyr
Leu Gly90 95 100 105aac cac atg aac atc gtt aac ctg ctg ggt gcg tgc
acc atc ggt ggt 450Asn His Met Asn Ile Val Asn Leu Leu Gly Ala Cys
Thr Ile Gly Gly 110 115 120ccg acc ctg gtt atc acc gaa tac tgc tgc
tac ggt gac ctg ctg aac 498Pro Thr Leu Val Ile Thr Glu Tyr Cys Cys
Tyr Gly Asp Leu Leu Asn 125 130 135ttc ctg cgt cgt aaa cgt gac tct
ttc atc tgc tct aaa cag gaa gac 546Phe Leu Arg Arg Lys Arg Asp Ser
Phe Ile Cys Ser Lys Gln Glu Asp 140 145 150cac gcg gaa gcg gcg ctg
tac aaa aac ctg ctg cac tct aaa gaa tct 594His Ala Glu Ala Ala Leu
Tyr Lys Asn Leu Leu His Ser Lys Glu Ser 155 160 165tct tgc tct gac
tct acc aac gaa tac atg gac atg aaa ccg ggt gtt 642Ser Cys Ser Asp
Ser Thr Asn Glu Tyr Met Asp Met Lys Pro Gly Val170 175 180 185tct
tac gtt gtt ccg acc aaa gcg gac aaa cgt cgt tct gtt cgt atc 690Ser
Tyr Val Val Pro Thr Lys Ala Asp Lys Arg Arg Ser Val Arg Ile 190 195
200ggt tct tac atc gaa cgt gac gtt acc ccg gcg atc atg gaa gac gac
738Gly Ser Tyr Ile Glu Arg Asp Val Thr Pro Ala Ile Met Glu Asp Asp
205 210 215gaa ctg gcg ctg gac ctg gaa gac ctg ctg tct ttc tct tac
cag gtt 786Glu Leu Ala Leu Asp Leu Glu Asp Leu Leu Ser Phe Ser Tyr
Gln Val 220 225 230gcg aaa ggt atg gcg ttc ctg gcg tct aaa aac tgc
atc cac cgt gac 834Ala Lys Gly Met Ala Phe Leu Ala Ser Lys Asn Cys
Ile His Arg Asp 235 240 245ctg gcg gcg cgt aac atc ctg ctg acc cac
ggt cgt atc acc aaa atc 882Leu Ala Ala Arg Asn Ile Leu Leu Thr His
Gly Arg Ile Thr Lys Ile250 255 260 265tgc gac ttc ggt ctg gcg cgt
gac atc aaa aac gac tct aac tac gtt 930Cys Asp Phe Gly Leu Ala Arg
Asp Ile Lys Asn Asp Ser Asn Tyr Val 270 275 280gtt aaa ggt aac gcg
cgt ctg ccg gtt aaa tgg atg gcg ccg gaa tct 978Val Lys Gly Asn Ala
Arg Leu Pro Val Lys Trp Met Ala Pro Glu Ser 285 290 295atc ttc aac
tgc gtt tac acc ttc gaa tct gac gtt tgg tct tac ggt 1026Ile Phe Asn
Cys Val Tyr Thr Phe Glu Ser Asp Val Trp Ser Tyr Gly 300 305 310atc
ttc ctg tgg gaa ctg ttc tct ctg ggt tct tct ccg tac ccg ggt 1074Ile
Phe Leu Trp Glu Leu Phe Ser Leu Gly Ser Ser Pro Tyr Pro Gly 315 320
325atg ccg gtt gac tct aaa ttc tac aaa atg atc aaa gaa ggt ttc cgt
1122Met Pro Val Asp Ser Lys Phe Tyr Lys Met Ile Lys Glu Gly Phe
Arg330 335 340 345atg ctg tct ccg gaa cac gcg ccg gcg gaa atg tac
gac atc atg aaa 1170Met Leu Ser Pro Glu His Ala Pro Ala Glu Met Tyr
Asp Ile Met Lys 350 355 360acc tgc tgg gac gcg gac ccg ctg aaa cgt
ccg acc ttc aaa cag atc 1218Thr Cys Trp Asp Ala Asp Pro Leu Lys Arg
Pro Thr Phe Lys Gln Ile 365 370 375gtt cag ctg atc gaa aaa cag atc
tct gaa tct acc aac cac atc tac 1266Val Gln Leu Ile Glu Lys Gln Ile
Ser Glu Ser Thr Asn His Ile Tyr 380 385 390tct aac ctg gcg aac tgc
tct ccg aac cgt cag aaa tagtcgactg 1312Ser Asn Leu Ala Asn Cys Ser
Pro Asn Arg Gln Lys 395 400 405aaaaaggaag agt
132591405PRTArtificial SequenceDescription of Artificial Sequence
Synthetic Plasmid 91Met Gly His His His His His His Met Tyr Glu Val
Gln Trp Lys Val1 5 10 15Val Glu Glu Ile Asn Gly Asn Asn Tyr Val Tyr
Ile Asp Pro Thr Gln 20 25 30Leu Pro Tyr Asp His Lys Trp Glu Phe Pro
Arg Asn Arg Leu Ser Phe 35 40 45Gly Lys Thr Leu Gly Ala Gly Ala Phe
Gly Lys Val Val Glu Ala Thr 50 55 60Ala Tyr Gly Leu Ile Lys Ser Asp
Ala Ala Met Thr Val Ala Val Lys65 70 75 80Met Leu Lys Pro Ser Ala
His Leu Thr Glu Arg Glu Ala Leu Met Ser 85 90 95Glu Leu Lys Val Leu
Ser Tyr Leu Gly Asn His Met Asn Ile Val Asn 100 105 110Leu Leu Gly
Ala Cys Thr Ile Gly Gly Pro Thr Leu Val Ile Thr Glu 115 120 125Tyr
Cys Cys Tyr Gly Asp Leu Leu Asn Phe Leu Arg Arg Lys Arg Asp 130 135
140Ser Phe Ile Cys Ser Lys Gln Glu Asp His Ala Glu Ala Ala Leu
Tyr145 150 155 160Lys Asn Leu Leu His Ser Lys Glu Ser Ser Cys Ser
Asp Ser Thr Asn 165 170 175Glu Tyr Met Asp Met Lys Pro Gly Val Ser
Tyr Val Val Pro Thr Lys 180 185 190Ala Asp Lys Arg Arg Ser Val Arg
Ile Gly Ser Tyr Ile Glu Arg Asp 195 200 205Val Thr Pro Ala Ile Met
Glu Asp Asp Glu Leu Ala Leu Asp Leu Glu 210 215 220Asp Leu Leu Ser
Phe Ser Tyr Gln Val Ala Lys Gly Met Ala Phe Leu225 230 235 240Ala
Ser Lys Asn Cys Ile His Arg Asp Leu Ala Ala Arg Asn Ile Leu 245 250
255Leu Thr His Gly Arg Ile Thr Lys Ile Cys Asp Phe Gly Leu Ala Arg
260 265 270Asp Ile Lys Asn Asp Ser Asn Tyr Val Val Lys Gly Asn Ala
Arg Leu 275 280 285Pro Val Lys Trp Met Ala Pro Glu Ser Ile Phe Asn
Cys Val Tyr Thr 290 295 300Phe Glu Ser Asp Val Trp Ser Tyr Gly Ile
Phe Leu Trp Glu Leu Phe305 310 315 320Ser Leu Gly Ser Ser Pro Tyr
Pro Gly Met Pro Val Asp Ser Lys Phe 325 330 335Tyr Lys Met Ile Lys
Glu Gly Phe Arg Met Leu Ser Pro Glu His Ala 340 345 350Pro Ala Glu
Met Tyr Asp Ile Met Lys Thr Cys Trp Asp Ala Asp Pro 355 360 365Leu
Lys Arg Pro Thr Phe Lys Gln Ile Val Gln Leu Ile Glu Lys Gln 370 375
380Ile Ser Glu Ser Thr Asn His Ile Tyr Ser Asn Leu Ala Asn Cys
Ser385 390 395 400Pro Asn Arg Gln Lys 4059235DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
92catcctacat atgggggact ctgatatatc cagtc 359339DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
93ctagcaggtc gactacccaa tgaaagtaga gaagatcgc 39941049DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(1041) 94taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggt cac cac cat cac cat cat atg 114 Met Gly His His His His His
His Met 1 5ggg gac tct gat ata tcc agt cca tta ctg caa aat act gtc
cac att 162Gly Asp Ser Asp Ile Ser Ser Pro Leu Leu Gln Asn Thr Val
His Ile10 15 20 25gac ctc agt gct cta aat cca gag ctg gtc cag gca
gtg cag cat gta 210Asp Leu Ser Ala Leu Asn Pro Glu Leu Val Gln Ala
Val Gln His Val 30 35 40gtg att ggg ccc agt agc ctg att gtg cat ttc
aat gaa gtc ata gga 258Val Ile Gly Pro Ser Ser Leu Ile Val His Phe
Asn Glu Val Ile Gly 45 50 55aga ggg cat ttt ggt tgt gta tat cat ggg
act ttg ttg gac aat gat 306Arg Gly His Phe Gly Cys Val Tyr His Gly
Thr Leu Leu Asp Asn Asp 60 65 70ggc aag aaa att cac tgt gct gtg aaa
tcc ttg aac aga atc act gac 354Gly Lys Lys Ile His Cys Ala Val Lys
Ser Leu Asn Arg Ile Thr Asp 75 80 85ata gga gaa gtt tcc caa ttt ctg
acc gag gga atc atc atg aaa gat 402Ile Gly Glu Val Ser Gln Phe Leu
Thr Glu Gly Ile Ile Met Lys Asp90 95 100 105ttt agt cat ccc aat gtc
ctc tcg ctc ctg gga atc tgc ctg cga agt 450Phe Ser His Pro Asn Val
Leu Ser Leu Leu Gly Ile Cys Leu Arg Ser 110 115 120gaa ggg tct ccg
ctg gtg gtc cta cca tac atg aaa cat gga gat ctt 498Glu Gly Ser Pro
Leu Val Val Leu Pro Tyr Met Lys His Gly Asp Leu 125 130 135cga aat
ttc att cga aat gag act cat aat cca act gta aaa gat ctt 546Arg Asn
Phe Ile Arg Asn Glu Thr His Asn Pro Thr Val Lys Asp Leu 140 145
150att ggc ttt ggt ctt caa gta gcc aaa ggc atg aaa tat ctt gca agc
594Ile Gly Phe Gly Leu Gln Val Ala Lys Gly Met Lys Tyr Leu Ala Ser
155 160 165aaa aag ttt gtc cac aga gac ttg gct gca aga aac tgt atg
ctg gat 642Lys Lys Phe Val His Arg Asp Leu Ala Ala Arg Asn Cys Met
Leu Asp170 175 180 185gaa aaa ttc aca gtc aag gtt gct gat ttt ggt
ctt gcc aga gac atg 690Glu Lys Phe Thr Val Lys Val Ala Asp Phe Gly
Leu Ala Arg Asp Met 190 195 200tat gat aaa gaa tac tat agt gta cac
aac aaa aca ggt gca aag ctg 738Tyr Asp Lys Glu Tyr Tyr Ser Val His
Asn Lys Thr Gly Ala Lys Leu 205 210 215cca gtg aag tgg atg gct ttg
gaa agt ctg caa act caa aag ttt acc 786Pro Val Lys Trp Met Ala Leu
Glu Ser Leu Gln Thr Gln Lys Phe Thr 220 225 230acc aag tca gat gtg
tgg tcc ttt ggc gtg ctc ctc tgg gag ctg atg 834Thr Lys Ser Asp Val
Trp Ser Phe Gly Val Leu Leu Trp Glu Leu Met 235 240 245aca aga gga
gcc cca cct tat cct gat gta aac acc ttt gat ata act 882Thr Arg Gly
Ala Pro Pro Tyr Pro Asp Val Asn Thr Phe Asp Ile Thr250 255 260
265gtt tac ttg ttg caa ggg aga aga ctc cta caa ccc gaa tac tgc cca
930Val Tyr Leu Leu Gln Gly Arg Arg Leu Leu Gln Pro Glu Tyr Cys Pro
270 275 280gac ccc tta tat gaa gta atg cta aaa tgc tgg cac cct aaa
gcc gaa 978Asp Pro Leu Tyr Glu Val Met Leu Lys Cys Trp His Pro Lys
Ala Glu 285 290 295atg cgc cca tcc ttt tct gaa ctg gtg tcc cgg ata
tca gcg atc ttc 1026Met Arg Pro Ser Phe Ser Glu Leu Val Ser Arg Ile
Ser Ala Ile Phe 300 305 310tct act ttc att ggg tagtcgac 1049Ser Thr
Phe Ile Gly 31595318PRTArtificial SequenceDescription of Artificial
Sequence Synthetic Plasmid 95Met Gly His His His His His His Met
Gly Asp Ser Asp Ile Ser Ser1 5 10 15Pro Leu Leu Gln Asn Thr Val His
Ile Asp Leu Ser Ala Leu Asn Pro 20 25 30Glu Leu Val Gln Ala Val Gln
His Val Val Ile Gly Pro Ser Ser Leu 35 40 45Ile Val His Phe Asn Glu
Val Ile Gly Arg Gly His Phe Gly Cys Val 50 55 60Tyr His Gly Thr Leu
Leu Asp Asn Asp Gly Lys Lys Ile His Cys Ala65 70 75 80Val Lys Ser
Leu Asn Arg Ile Thr Asp Ile Gly Glu Val Ser Gln Phe 85 90 95Leu Thr
Glu Gly Ile Ile Met Lys Asp Phe Ser His Pro Asn Val Leu 100 105
110Ser Leu Leu Gly Ile Cys Leu Arg Ser Glu Gly Ser Pro Leu Val Val
115 120 125Leu Pro Tyr Met Lys His Gly Asp Leu Arg Asn Phe Ile Arg
Asn Glu 130 135 140Thr His Asn Pro Thr Val Lys Asp Leu Ile Gly Phe
Gly Leu Gln Val145 150 155 160Ala Lys Gly Met Lys Tyr Leu Ala Ser
Lys Lys Phe Val His Arg Asp 165 170 175Leu Ala Ala Arg Asn Cys Met
Leu Asp Glu Lys Phe Thr Val Lys Val 180 185 190Ala Asp Phe Gly Leu
Ala Arg Asp Met Tyr Asp Lys Glu Tyr Tyr Ser 195 200 205Val His Asn
Lys Thr Gly Ala Lys Leu Pro Val Lys Trp Met Ala Leu 210 215 220Glu
Ser Leu Gln Thr Gln Lys Phe Thr Thr Lys Ser Asp Val Trp Ser225 230
235 240Phe Gly Val Leu Leu Trp Glu Leu Met Thr Arg Gly Ala Pro Pro
Tyr 245 250 255Pro Asp Val Asn Thr Phe Asp Ile Thr Val Tyr Leu Leu
Gln Gly Arg 260 265 270Arg Leu Leu Gln Pro Glu Tyr Cys Pro Asp Pro
Leu Tyr Glu Val Met 275 280 285Leu Lys Cys Trp His Pro Lys Ala Glu
Met Arg Pro Ser Phe Ser Glu 290 295 300Leu Val Ser Arg Ile Ser Ala
Ile Phe Ser Thr Phe Ile Gly305 310 3159630DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
96ccggatccat atgtctcagg agaggcccac 309733DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
97gaaaccctcg agtcaggact ccatctcttc ttg 33981236DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(88)..(1227) 98taatacgact cactataggg gaattgtgag
cggataacaa ttcccctcta gaaataattt 60tgtttaactt taagaaggag atatacc
atg ggc agc agc cat cat cat cat cat 114 Met Gly Ser Ser His His His
His His 1 5cac agc agc ggc ctg gtg ccg cgc ggc agc cat atg tct cag
gag agg 162His Ser Ser Gly Leu Val Pro Arg Gly Ser His Met Ser Gln
Glu Arg10 15 20 25ccc acg ttc tac cgg cag gag ctg aac aag aca atc
tgg gag gtg ccc 210Pro Thr Phe Tyr Arg Gln Glu Leu Asn Lys Thr Ile
Trp Glu Val Pro 30 35 40gag cgt tac cag aac ctg tct cca gtg ggc tct
ggc gcc tat ggc tct 258Glu Arg Tyr Gln Asn Leu Ser Pro Val Gly Ser
Gly Ala Tyr Gly Ser 45 50 55gtg tgt gct gct ttt gac aca aaa acg ggg
tta cgt gtg gca gtg aag 306Val Cys Ala Ala Phe Asp Thr Lys Thr Gly
Leu Arg Val Ala Val Lys 60 65 70aag ctc tcc aga cca ttt cag tcc atc
att cat gcg aaa aga acc tac 354Lys Leu Ser Arg Pro Phe Gln Ser Ile
Ile His Ala Lys Arg Thr Tyr 75 80 85aga gaa ctg cgg tta ctt aaa cac
atg aaa cat gaa aat gtg att ggt 402Arg Glu Leu Arg Leu Leu Lys His
Met Lys His Glu Asn Val Ile Gly90 95 100 105ctg ttg gac gtt ttt aca
cct gca agg tct ctg gag gaa ttc aat gat 450Leu Leu Asp Val Phe Thr
Pro Ala Arg Ser Leu Glu Glu Phe Asn Asp 110 115 120gtg tat ctg gtg
acc cat ctc atg ggg gca gat ctg aac aac att gtg 498Val Tyr Leu Val
Thr His Leu Met Gly Ala Asp Leu Asn Asn Ile Val 125 130 135aaa tgt
cag aag ctt aca gat gac cat gtt cag ttc ctt atc tac caa 546Lys Cys
Gln Lys Leu Thr Asp Asp His Val Gln Phe Leu Ile Tyr Gln 140 145
150att ctc cga ggt cta aag tat ata cat tca gct gac ata att cac agg
594Ile Leu Arg Gly Leu Lys Tyr Ile His Ser Ala Asp Ile Ile His Arg
155 160 165gac cta aaa cct agt aat cta gct gtg aat gaa gac tgt gag
ctg aag 642Asp Leu Lys Pro Ser Asn Leu Ala Val Asn Glu Asp Cys Glu
Leu Lys170 175 180 185att ctg gat ttt gga ctg gct cgg cac aca gat
gat gaa atg aca ggc 690Ile Leu Asp Phe Gly Leu Ala Arg His Thr Asp
Asp Glu Met Thr Gly 190 195 200tac gtg gcc act agg tgg tac agg gct
cct gag atc atg ctg aac tgg 738Tyr Val Ala Thr Arg Trp Tyr Arg Ala
Pro Glu Ile Met Leu Asn Trp 205 210 215atg cat tac aac cag aca gtt
gat att tgg tca gtg gga tgc ata atg 786Met His Tyr Asn Gln Thr Val
Asp Ile Trp Ser Val Gly Cys Ile Met 220 225 230gcc gag ctg ttg act
gga aga aca ttg ttt cct ggt aca gac cat att 834Ala Glu Leu Leu Thr
Gly Arg Thr Leu Phe Pro Gly Thr Asp His Ile 235 240 245gat cag ttg
aag ctc att tta aga ctc gtt gga acc cca ggg gct gag 882Asp Gln Leu
Lys Leu Ile Leu Arg Leu Val Gly Thr Pro Gly Ala Glu250 255 260
265ctt ttg aag aaa atc tcc tca gag tct gca aga aac tat att cag tct
930Leu Leu Lys Lys Ile Ser Ser Glu Ser Ala Arg Asn Tyr Ile Gln Ser
270 275 280ttg act cag atg ccg aag atg aac ttt gcg aat gta ttt att
ggt gcc 978Leu Thr Gln Met Pro Lys Met Asn Phe Ala Asn Val Phe Ile
Gly Ala 285 290 295aat ccc ctg gct gtc gac ttg ctg gag aag atg ctt
gta ttg gac tca 1026Asn Pro Leu Ala Val Asp Leu Leu Glu Lys Met Leu
Val Leu Asp Ser
300 305 310gat aag aga att aca gcg gcc caa gcc ctt gca cat gcc tac
ttt gct 1074Asp Lys Arg Ile Thr Ala Ala Gln Ala Leu Ala His Ala Tyr
Phe Ala 315 320 325cag tac cac gat cct gat gat gaa cca gtg gcc gat
cct tat gat cag 1122Gln Tyr His Asp Pro Asp Asp Glu Pro Val Ala Asp
Pro Tyr Asp Gln330 335 340 345tcc ttt gaa agc agg gac ctc ctt ata
gat gag tgg aaa agc ctg acc 1170Ser Phe Glu Ser Arg Asp Leu Leu Ile
Asp Glu Trp Lys Ser Leu Thr 350 355 360tat gat gaa gtc atc agc ttt
gtg cca cca ccc ctt gac caa gaa gag 1218Tyr Asp Glu Val Ile Ser Phe
Val Pro Pro Pro Leu Asp Gln Glu Glu 365 370 375atg gag tcc
tgactcgac 1236Met Glu Ser 38099380PRTArtificial SequenceDescription
of Artificial Sequence Synthetic Plasmid 99Met Gly Ser Ser His His
His His His His Ser Ser Gly Leu Val Pro1 5 10 15Arg Gly Ser His Met
Ser Gln Glu Arg Pro Thr Phe Tyr Arg Gln Glu 20 25 30Leu Asn Lys Thr
Ile Trp Glu Val Pro Glu Arg Tyr Gln Asn Leu Ser 35 40 45Pro Val Gly
Ser Gly Ala Tyr Gly Ser Val Cys Ala Ala Phe Asp Thr 50 55 60Lys Thr
Gly Leu Arg Val Ala Val Lys Lys Leu Ser Arg Pro Phe Gln65 70 75
80Ser Ile Ile His Ala Lys Arg Thr Tyr Arg Glu Leu Arg Leu Leu Lys
85 90 95His Met Lys His Glu Asn Val Ile Gly Leu Leu Asp Val Phe Thr
Pro 100 105 110Ala Arg Ser Leu Glu Glu Phe Asn Asp Val Tyr Leu Val
Thr His Leu 115 120 125Met Gly Ala Asp Leu Asn Asn Ile Val Lys Cys
Gln Lys Leu Thr Asp 130 135 140Asp His Val Gln Phe Leu Ile Tyr Gln
Ile Leu Arg Gly Leu Lys Tyr145 150 155 160Ile His Ser Ala Asp Ile
Ile His Arg Asp Leu Lys Pro Ser Asn Leu 165 170 175Ala Val Asn Glu
Asp Cys Glu Leu Lys Ile Leu Asp Phe Gly Leu Ala 180 185 190Arg His
Thr Asp Asp Glu Met Thr Gly Tyr Val Ala Thr Arg Trp Tyr 195 200
205Arg Ala Pro Glu Ile Met Leu Asn Trp Met His Tyr Asn Gln Thr Val
210 215 220Asp Ile Trp Ser Val Gly Cys Ile Met Ala Glu Leu Leu Thr
Gly Arg225 230 235 240Thr Leu Phe Pro Gly Thr Asp His Ile Asp Gln
Leu Lys Leu Ile Leu 245 250 255Arg Leu Val Gly Thr Pro Gly Ala Glu
Leu Leu Lys Lys Ile Ser Ser 260 265 270Glu Ser Ala Arg Asn Tyr Ile
Gln Ser Leu Thr Gln Met Pro Lys Met 275 280 285Asn Phe Ala Asn Val
Phe Ile Gly Ala Asn Pro Leu Ala Val Asp Leu 290 295 300Leu Glu Lys
Met Leu Val Leu Asp Ser Asp Lys Arg Ile Thr Ala Ala305 310 315
320Gln Ala Leu Ala His Ala Tyr Phe Ala Gln Tyr His Asp Pro Asp Asp
325 330 335Glu Pro Val Ala Asp Pro Tyr Asp Gln Ser Phe Glu Ser Arg
Asp Leu 340 345 350Leu Ile Asp Glu Trp Lys Ser Leu Thr Tyr Asp Glu
Val Ile Ser Phe 355 360 365Val Pro Pro Pro Leu Asp Gln Glu Glu Met
Glu Ser 370 375 38010034DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 100gctggcgcat atgaaggaga
aggagcccct ggag 3410133DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 101gaaagggtcg actttgctgg
gccccggcga cag 331021108DNAArtificial SequenceDescription of
Artificial Sequence Synthetic PlasmidCDS(113)..(994) 102agatcgatct
cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata 60acaattcccc
tctagaaata attttgttta actttaagaa ggagatatac at atg aag 118 Met Lys
1gag aag gag ccc ctg gag tcg cag tac cag gtg ggc ccg cta ctg ggc
166Glu Lys Glu Pro Leu Glu Ser Gln Tyr Gln Val Gly Pro Leu Leu Gly
5 10 15agc ggc ggc ttc ggc tcg gtc tac tca ggc atc cgc gtc tcc gac
aac 214Ser Gly Gly Phe Gly Ser Val Tyr Ser Gly Ile Arg Val Ser Asp
Asn 20 25 30ttg ccg gtg gcc atc aaa cac gtg gag aag gac cgg att tcc
gac tgg 262Leu Pro Val Ala Ile Lys His Val Glu Lys Asp Arg Ile Ser
Asp Trp35 40 45 50gga gag ctg cct aat ggc act cga gtg ccc atg gaa
gtg gtc ctg ctg 310Gly Glu Leu Pro Asn Gly Thr Arg Val Pro Met Glu
Val Val Leu Leu 55 60 65aag aag gtg agc tcg ggt ttc tcc ggc gtc att
agg ctc ctg gac tgg 358Lys Lys Val Ser Ser Gly Phe Ser Gly Val Ile
Arg Leu Leu Asp Trp 70 75 80ttc gag agg ccc gac agt ttc gtc ctg atc
ctg gag agg ccc gag ccg 406Phe Glu Arg Pro Asp Ser Phe Val Leu Ile
Leu Glu Arg Pro Glu Pro 85 90 95gtg caa gat ctc ttc gac ttc atc acg
gaa agg gga gcc ctg caa gag 454Val Gln Asp Leu Phe Asp Phe Ile Thr
Glu Arg Gly Ala Leu Gln Glu 100 105 110gag ctg gcc cgc agc ttc ttc
tgg cag gtg ctg gag gcc gtg cgg cac 502Glu Leu Ala Arg Ser Phe Phe
Trp Gln Val Leu Glu Ala Val Arg His115 120 125 130tgc cac aac tgc
ggg gtg ctc cac cgc gac atc aag gac gaa aac atc 550Cys His Asn Cys
Gly Val Leu His Arg Asp Ile Lys Asp Glu Asn Ile 135 140 145ctt atc
gac ctc aat cgc ggc gag ctc aag ctc atc gac ttc ggg tcg 598Leu Ile
Asp Leu Asn Arg Gly Glu Leu Lys Leu Ile Asp Phe Gly Ser 150 155
160ggg gcg ctg ctc aag gac acc gtc tac acg gac ttc gat ggg acc cga
646Gly Ala Leu Leu Lys Asp Thr Val Tyr Thr Asp Phe Asp Gly Thr Arg
165 170 175gtg tat agc cct cca gag tgg atc cgc tac cat cgc tac cat
ggc agg 694Val Tyr Ser Pro Pro Glu Trp Ile Arg Tyr His Arg Tyr His
Gly Arg 180 185 190tcg gcg gca gtc tgg tcc ctg ggg atc ctg ctg tat
gat atg gtg tgt 742Ser Ala Ala Val Trp Ser Leu Gly Ile Leu Leu Tyr
Asp Met Val Cys195 200 205 210gga gat att cct ttc gag cat gac gaa
gag atc atc agg ggc cag gtt 790Gly Asp Ile Pro Phe Glu His Asp Glu
Glu Ile Ile Arg Gly Gln Val 215 220 225ttc ttc agg cag agg gtc tct
tca gaa tgt cag cat ctc att aga tgg 838Phe Phe Arg Gln Arg Val Ser
Ser Glu Cys Gln His Leu Ile Arg Trp 230 235 240tgc ttg gcc ctg aga
cca tca gat agg cca acc ttc gaa gaa atc cag 886Cys Leu Ala Leu Arg
Pro Ser Asp Arg Pro Thr Phe Glu Glu Ile Gln 245 250 255aac cat cca
tgg atg caa gat gtt ctc ctg ccc cag gaa act gct gag 934Asn His Pro
Trp Met Gln Asp Val Leu Leu Pro Gln Glu Thr Ala Glu 260 265 270atc
cac ctc cac agc ctg tcg ccg ggg ccc agc aaa gtc gac cac cac 982Ile
His Leu His Ser Leu Ser Pro Gly Pro Ser Lys Val Asp His His275 280
285 290cac cac cac cac tgagatccgg ctgctaacaa agcccgaaag gaattcgagt
1034His His His Histggctgctgc caccgctgag caataactag cataacccct
tggggcctct aaacgggtct 1094tgaggggttt tttg 1108103294PRTArtificial
SequenceDescription of Artificial Sequence Synthetic Plasmid 103Met
Lys Glu Lys Glu Pro Leu Glu Ser Gln Tyr Gln Val Gly Pro Leu1 5 10
15Leu Gly Ser Gly Gly Phe Gly Ser Val Tyr Ser Gly Ile Arg Val Ser
20 25 30Asp Asn Leu Pro Val Ala Ile Lys His Val Glu Lys Asp Arg Ile
Ser 35 40 45Asp Trp Gly Glu Leu Pro Asn Gly Thr Arg Val Pro Met Glu
Val Val 50 55 60Leu Leu Lys Lys Val Ser Ser Gly Phe Ser Gly Val Ile
Arg Leu Leu65 70 75 80Asp Trp Phe Glu Arg Pro Asp Ser Phe Val Leu
Ile Leu Glu Arg Pro 85 90 95Glu Pro Val Gln Asp Leu Phe Asp Phe Ile
Thr Glu Arg Gly Ala Leu 100 105 110Gln Glu Glu Leu Ala Arg Ser Phe
Phe Trp Gln Val Leu Glu Ala Val 115 120 125Arg His Cys His Asn Cys
Gly Val Leu His Arg Asp Ile Lys Asp Glu 130 135 140Asn Ile Leu Ile
Asp Leu Asn Arg Gly Glu Leu Lys Leu Ile Asp Phe145 150 155 160Gly
Ser Gly Ala Leu Leu Lys Asp Thr Val Tyr Thr Asp Phe Asp Gly 165 170
175Thr Arg Val Tyr Ser Pro Pro Glu Trp Ile Arg Tyr His Arg Tyr His
180 185 190Gly Arg Ser Ala Ala Val Trp Ser Leu Gly Ile Leu Leu Tyr
Asp Met 195 200 205Val Cys Gly Asp Ile Pro Phe Glu His Asp Glu Glu
Ile Ile Arg Gly 210 215 220Gln Val Phe Phe Arg Gln Arg Val Ser Ser
Glu Cys Gln His Leu Ile225 230 235 240Arg Trp Cys Leu Ala Leu Arg
Pro Ser Asp Arg Pro Thr Phe Glu Glu 245 250 255Ile Gln Asn His Pro
Trp Met Gln Asp Val Leu Leu Pro Gln Glu Thr 260 265 270Ala Glu Ile
His Leu His Ser Leu Ser Pro Gly Pro Ser Lys Val Asp 275 280 285His
His His His His His 29010433DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 104gttcttcata tgcacaagtt
tgcccacaag cca 3310540DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 105gttcttgtcg accctcttaa
ccatcatctt ctccaggtct 401061285DNAArtificial SequenceDescription of
Artificial Sequence Synthetic PlasmidCDS(89)..(1171) 106taatacgact
cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 60tgtttaactt
taagaaggag atatacat atg cac aag ttt gcc cac aag cca 112 Met His Lys
Phe Ala His Lys Pro 1 5ccc atc tcc tca gct gag atg acc ttc cgg agg
ccc gcc cag gcc ttc 160Pro Ile Ser Ser Ala Glu Met Thr Phe Arg Arg
Pro Ala Gln Ala Phe 10 15 20ccg gtc agc tac tcc tct tcc ggt gcc cgc
cgg ccc tcg ctg gac tcc 208Pro Val Ser Tyr Ser Ser Ser Gly Ala Arg
Arg Pro Ser Leu Asp Ser25 30 35 40atg gag aac cag gtc tcc gtg gat
gcc ttc aag atc ctg gag gat cca 256Met Glu Asn Gln Val Ser Val Asp
Ala Phe Lys Ile Leu Glu Asp Pro 45 50 55aag tgg gaa ttc cct cgg aag
aac ttg gtt ctt gga aaa act cta gga 304Lys Trp Glu Phe Pro Arg Lys
Asn Leu Val Leu Gly Lys Thr Leu Gly 60 65 70gaa ggc gaa ttt gga aaa
gtg gtc aag gca acg gcc ttc cat ctg aaa 352Glu Gly Glu Phe Gly Lys
Val Val Lys Ala Thr Ala Phe His Leu Lys 75 80 85ggc aga gca ggg tac
acc acg gtg gcc gtg aag atg ctg aaa gag aac 400Gly Arg Ala Gly Tyr
Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn 90 95 100gcc tcc ccg
agt gag ctt cga gac ctg ctg tca gag ttc aac gtc ctg 448Ala Ser Pro
Ser Glu Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu105 110 115
120aag cag gtc aac cac cca cat gtc atc aaa ttg tat ggg gcc tgc agc
496Lys Gln Val Asn His Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser
125 130 135cag gat ggc ccg ctc ctc ctc atc gtg gag tac gcc aaa tac
ggc tcc 544Gln Asp Gly Pro Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr
Gly Ser 140 145 150ctg cgg ggc ttc ctc cgc gag agc cgc aaa gtg ggg
cct ggc tac ctg 592Leu Arg Gly Phe Leu Arg Glu Ser Arg Lys Val Gly
Pro Gly Tyr Leu 155 160 165ggc agt gga ggc agc cgc aac tcc agc tcc
ctg gac cac ccg gat gag 640Gly Ser Gly Gly Ser Arg Asn Ser Ser Ser
Leu Asp His Pro Asp Glu 170 175 180cgg gcc ctc acc atg ggc gac ctc
atc tca ttt gcc tgg cag atc tca 688Arg Ala Leu Thr Met Gly Asp Leu
Ile Ser Phe Ala Trp Gln Ile Ser185 190 195 200cag ggg atg cag tat
ctg gcc gag atg aag ctc gtt cat cgg gac ttg 736Gln Gly Met Gln Tyr
Leu Ala Glu Met Lys Leu Val His Arg Asp Leu 205 210 215gca gcc aga
aac atc ctg gta gct gag ggg cgg aag atg aag att tcg 784Ala Ala Arg
Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser 220 225 230gat
ttc ggc ttg tcc cga gat gtt tat gaa gag gat tcc tac gtg aag 832Asp
Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys 235 240
245agg agc cag ggt cgg att cca gtt aaa tgg atg gca att gaa tcc ctt
880Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu
250 255 260ttt gat cat atc tac acc acg caa agt gat gta tgg tct ttt
ggt gtc 928Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser Phe
Gly Val265 270 275 280ctg ctg tgg gag atc gtg acc cta ggg gga aac
ccc tat cct ggg att 976Leu Leu Trp Glu Ile Val Thr Leu Gly Gly Asn
Pro Tyr Pro Gly Ile 285 290 295cct cct gag cgg ctc ttc aac ctt ctg
aag acc ggc cac cgg atg gag 1024Pro Pro Glu Arg Leu Phe Asn Leu Leu
Lys Thr Gly His Arg Met Glu 300 305 310agg cca gac aac tgc agc gag
gag atg tac cgc ctg atg ctg caa tgc 1072Arg Pro Asp Asn Cys Ser Glu
Glu Met Tyr Arg Leu Met Leu Gln Cys 315 320 325tgg aag cag gag ccg
gac aaa agg ccg gtg ttt gcg gac atc agc aaa 1120Trp Lys Gln Glu Pro
Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys 330 335 340gac ctg gag
aag atg atg gtt aag agg gtc gac cac cac cac cac cac 1168Asp Leu Glu
Lys Met Met Val Lys Arg Val Asp His His His His His345 350 355
360cac tgagatccgg ctggccctac tggccgaaag gaattcgagg ccagcagggc
1221Hiscaccgctgag caataactag cataacccct tggggcctct aaacgggtct
tgaggggttt 1281tttg 1285107361PRTArtificial SequenceDescription of
Artificial Sequence Synthetic Plasmid 107Met His Lys Phe Ala His
Lys Pro Pro Ile Ser Ser Ala Glu Met Thr1 5 10 15Phe Arg Arg Pro Ala
Gln Ala Phe Pro Val Ser Tyr Ser Ser Ser Gly 20 25 30Ala Arg Arg Pro
Ser Leu Asp Ser Met Glu Asn Gln Val Ser Val Asp 35 40 45Ala Phe Lys
Ile Leu Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn 50 55 60Leu Val
Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val65 70 75
80Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr Val
85 90 95Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu Arg
Asp 100 105 110Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His
Pro His Val 115 120 125Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly
Pro Leu Leu Leu Ile 130 135 140Val Glu Tyr Ala Lys Tyr Gly Ser Leu
Arg Gly Phe Leu Arg Glu Ser145 150 155 160Arg Lys Val Gly Pro Gly
Tyr Leu Gly Ser Gly Gly Ser Arg Asn Ser 165 170 175Ser Ser Leu Asp
His Pro Asp Glu Arg Ala Leu Thr Met Gly Asp Leu 180 185 190Ile Ser
Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr Leu Ala Glu 195 200
205Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Ala
210 215 220Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser Arg
Asp Val225 230 235 240Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln
Gly Arg Ile Pro Val 245 250 255Lys Trp Met Ala Ile Glu Ser Leu Phe
Asp His Ile Tyr Thr Thr Gln 260 265 270Ser Asp Val Trp Ser Phe Gly
Val Leu Leu Trp Glu Ile Val Thr Leu 275 280 285Gly Gly Asn Pro Tyr
Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu 290 295 300Leu Lys Thr
Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu305 310 315
320Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys Arg
325 330 335Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met
Val Lys 340 345 350Arg Val Asp His His His
His His His 355 36010833DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 108gctggcccat atggtgacca
cctttgtggc cct 3310933DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 109gctactagtc gacctagagg
ttctccccgg gct 331101491DNAArtificial SequenceDescription of
Artificial Sequence Synthetic PlasmidCDS(88)..(1467) 110taatacgact
cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 60tgtttaactt
taagaaggag atatacc atg ggt cac cac cat cac cat cat atg 114 Met Gly
His His His His His His Met 1 5gtg acc acc ttt gtg gcc ctc tat gac
tat gag tct agg acg gag aca 162Val Thr Thr Phe Val Ala Leu Tyr Asp
Tyr Glu Ser Arg Thr Glu Thr10 15 20 25gac ctg tcc ttc aag aaa ggc
gag cgg ctc cag att gtc aac aac aca 210Asp Leu Ser Phe Lys Lys Gly
Glu Arg Leu Gln Ile Val Asn Asn Thr 30 35 40gag gga gac tgg tgg ctg
gcc cac tcg ctc agc aca gga cag aca ggc 258Glu Gly Asp Trp Trp Leu
Ala His Ser Leu Ser Thr Gly Gln Thr Gly 45 50 55tac atc ccc agc aac
tac gtg gcg ccc tcc gac tcc atc cag gct gag 306Tyr Ile Pro Ser Asn
Tyr Val Ala Pro Ser Asp Ser Ile Gln Ala Glu 60 65 70gag tgg tat ttt
ggc aag atc acc aga cgg gag tca gag cgg tta ctg 354Glu Trp Tyr Phe
Gly Lys Ile Thr Arg Arg Glu Ser Glu Arg Leu Leu 75 80 85ctc aat gca
gag aac ccg aga ggg acc ttc ctc gtg cga gaa agt gag 402Leu Asn Ala
Glu Asn Pro Arg Gly Thr Phe Leu Val Arg Glu Ser Glu90 95 100 105acc
acg aaa ggt gcc tac tgc ctc tca gtg tct gac ttc gac aac gcc 450Thr
Thr Lys Gly Ala Tyr Cys Leu Ser Val Ser Asp Phe Asp Asn Ala 110 115
120aag ggc ctc aac gtg aag cac tac aag atc cgc aag ctg gac agc ggc
498Lys Gly Leu Asn Val Lys His Tyr Lys Ile Arg Lys Leu Asp Ser Gly
125 130 135ggc ttc tac atc acc tcc cgc acc cag ttc aac agc ctg cag
cag ctg 546Gly Phe Tyr Ile Thr Ser Arg Thr Gln Phe Asn Ser Leu Gln
Gln Leu 140 145 150gtg gcc tac tac tcc aaa cac gcc gat ggc ctg tgc
cac cgc ctc acc 594Val Ala Tyr Tyr Ser Lys His Ala Asp Gly Leu Cys
His Arg Leu Thr 155 160 165acc gtg tgc ccc acg tcc aag ccg cag act
cag ggc ctg gcc aag gat 642Thr Val Cys Pro Thr Ser Lys Pro Gln Thr
Gln Gly Leu Ala Lys Asp170 175 180 185gcc tgg gag atc cct cgg gag
tcg ctg cgg ctg gag gtc aag ctg ggc 690Ala Trp Glu Ile Pro Arg Glu
Ser Leu Arg Leu Glu Val Lys Leu Gly 190 195 200cag ggc tgc ttt ggc
gag gtg tgg atg ggg acc tgg aac ggt acc acc 738Gln Gly Cys Phe Gly
Glu Val Trp Met Gly Thr Trp Asn Gly Thr Thr 205 210 215agg gtg gcc
atc aaa acc ctg aag cct ggc acg atg tct cca gag gcc 786Arg Val Ala
Ile Lys Thr Leu Lys Pro Gly Thr Met Ser Pro Glu Ala 220 225 230ttc
ctg cag gag gcc cag gtc atg aag aag ctg agg cat gag aag ctg 834Phe
Leu Gln Glu Ala Gln Val Met Lys Lys Leu Arg His Glu Lys Leu 235 240
245gtg cag ttg tat gct gtg gtt tca gag gag ccc att tac atc gtc acg
882Val Gln Leu Tyr Ala Val Val Ser Glu Glu Pro Ile Tyr Ile Val
Thr250 255 260 265gag tac atg agc aag ggg agt ttg ctg gac ttt ctc
aag ggg gag aca 930Glu Tyr Met Ser Lys Gly Ser Leu Leu Asp Phe Leu
Lys Gly Glu Thr 270 275 280ggc aag tac ctg cgg ctg cct cag ctg gtg
gac atg gct gct cag atc 978Gly Lys Tyr Leu Arg Leu Pro Gln Leu Val
Asp Met Ala Ala Gln Ile 285 290 295gcc tca ggc atg gcg tac gtg gag
cgg atg aac tac gtc cac cgg gac 1026Ala Ser Gly Met Ala Tyr Val Glu
Arg Met Asn Tyr Val His Arg Asp 300 305 310ctt cgt gca gcc aac atc
ctg gtg gga gag aac ctg gtg tgc aaa gtg 1074Leu Arg Ala Ala Asn Ile
Leu Val Gly Glu Asn Leu Val Cys Lys Val 315 320 325gcc gac ttt ggg
ctg gct cgg ctc att gaa gac aat gag tac acg gcg 1122Ala Asp Phe Gly
Leu Ala Arg Leu Ile Glu Asp Asn Glu Tyr Thr Ala330 335 340 345cgg
caa ggt gcc aaa ttc ccc atc aag tgg acg gct cca gaa gct gcc 1170Arg
Gln Gly Ala Lys Phe Pro Ile Lys Trp Thr Ala Pro Glu Ala Ala 350 355
360ctc tat ggc cgc ttc acc atc aag tcg gac gtg tgg tcc ttc ggg atc
1218Leu Tyr Gly Arg Phe Thr Ile Lys Ser Asp Val Trp Ser Phe Gly Ile
365 370 375ctg ctg act gag ctc acc aca aag gga cgg gtg ccc tac cct
ggg atg 1266Leu Leu Thr Glu Leu Thr Thr Lys Gly Arg Val Pro Tyr Pro
Gly Met 380 385 390gtg aac cgc gag gtg ctg gac cag gtg gag cgg ggc
tac cgg atg ccc 1314Val Asn Arg Glu Val Leu Asp Gln Val Glu Arg Gly
Tyr Arg Met Pro 395 400 405tgc ccg ccg gag tgt ccc gag tcc ctg cac
gac ctc atg tgc cag tgc 1362Cys Pro Pro Glu Cys Pro Glu Ser Leu His
Asp Leu Met Cys Gln Cys410 415 420 425tgg cgg aag gag cct gag gag
cgg ccc acc ttc gag tac ctg cag gcc 1410Trp Arg Lys Glu Pro Glu Glu
Arg Pro Thr Phe Glu Tyr Leu Gln Ala 430 435 440ttc ctg gag gac tac
ttc acg tcc acc gag ccc cag tac cag ccc ggg 1458Phe Leu Glu Asp Tyr
Phe Thr Ser Thr Glu Pro Gln Tyr Gln Pro Gly 445 450 455gag aac ctc
taggtcgacg aaggagatat atcc 1491Glu Asn Leu 460111460PRTArtificial
SequenceDescription of Artificial Sequence Synthetic Plasmid 111Met
Gly His His His His His His Met Val Thr Thr Phe Val Ala Leu1 5 10
15Tyr Asp Tyr Glu Ser Arg Thr Glu Thr Asp Leu Ser Phe Lys Lys Gly
20 25 30Glu Arg Leu Gln Ile Val Asn Asn Thr Glu Gly Asp Trp Trp Leu
Ala 35 40 45His Ser Leu Ser Thr Gly Gln Thr Gly Tyr Ile Pro Ser Asn
Tyr Val 50 55 60Ala Pro Ser Asp Ser Ile Gln Ala Glu Glu Trp Tyr Phe
Gly Lys Ile65 70 75 80Thr Arg Arg Glu Ser Glu Arg Leu Leu Leu Asn
Ala Glu Asn Pro Arg 85 90 95Gly Thr Phe Leu Val Arg Glu Ser Glu Thr
Thr Lys Gly Ala Tyr Cys 100 105 110Leu Ser Val Ser Asp Phe Asp Asn
Ala Lys Gly Leu Asn Val Lys His 115 120 125Tyr Lys Ile Arg Lys Leu
Asp Ser Gly Gly Phe Tyr Ile Thr Ser Arg 130 135 140Thr Gln Phe Asn
Ser Leu Gln Gln Leu Val Ala Tyr Tyr Ser Lys His145 150 155 160Ala
Asp Gly Leu Cys His Arg Leu Thr Thr Val Cys Pro Thr Ser Lys 165 170
175Pro Gln Thr Gln Gly Leu Ala Lys Asp Ala Trp Glu Ile Pro Arg Glu
180 185 190Ser Leu Arg Leu Glu Val Lys Leu Gly Gln Gly Cys Phe Gly
Glu Val 195 200 205Trp Met Gly Thr Trp Asn Gly Thr Thr Arg Val Ala
Ile Lys Thr Leu 210 215 220Lys Pro Gly Thr Met Ser Pro Glu Ala Phe
Leu Gln Glu Ala Gln Val225 230 235 240Met Lys Lys Leu Arg His Glu
Lys Leu Val Gln Leu Tyr Ala Val Val 245 250 255Ser Glu Glu Pro Ile
Tyr Ile Val Thr Glu Tyr Met Ser Lys Gly Ser 260 265 270Leu Leu Asp
Phe Leu Lys Gly Glu Thr Gly Lys Tyr Leu Arg Leu Pro 275 280 285Gln
Leu Val Asp Met Ala Ala Gln Ile Ala Ser Gly Met Ala Tyr Val 290 295
300Glu Arg Met Asn Tyr Val His Arg Asp Leu Arg Ala Ala Asn Ile
Leu305 310 315 320Val Gly Glu Asn Leu Val Cys Lys Val Ala Asp Phe
Gly Leu Ala Arg 325 330 335Leu Ile Glu Asp Asn Glu Tyr Thr Ala Arg
Gln Gly Ala Lys Phe Pro 340 345 350Ile Lys Trp Thr Ala Pro Glu Ala
Ala Leu Tyr Gly Arg Phe Thr Ile 355 360 365Lys Ser Asp Val Trp Ser
Phe Gly Ile Leu Leu Thr Glu Leu Thr Thr 370 375 380Lys Gly Arg Val
Pro Tyr Pro Gly Met Val Asn Arg Glu Val Leu Asp385 390 395 400Gln
Val Glu Arg Gly Tyr Arg Met Pro Cys Pro Pro Glu Cys Pro Glu 405 410
415Ser Leu His Asp Leu Met Cys Gln Cys Trp Arg Lys Glu Pro Glu Glu
420 425 430Arg Pro Thr Phe Glu Tyr Leu Gln Ala Phe Leu Glu Asp Tyr
Phe Thr 435 440 445Ser Thr Glu Pro Gln Tyr Gln Pro Gly Glu Asn Leu
450 455 46011239DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 112agagggatcc gccaccatgg acaagaagct
cttcctgaa 3911347DNAArtificial SequenceDescription of Artificial
Sequence Synthetic Primer 113acgaattcta gtggtggtgg tggtggtggt
gcccttccac cttgctg 471141014DNAArtificial SequenceDescription of
Artificial Sequence Synthetic PlasmidCDS(143)..(1006) 114agatcatgga
gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt 60tcgtaacagt
tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac
120catcgggcgc ggatccgcca cc atg gac aag aag ctc ttc ctg aag cgc gat
172 Met Asp Lys Lys Leu Phe Leu Lys Arg Asp 1 5 10aac ctc ctc ata
gct gac att gaa ctt ggc tgc ggc aac ttt ggc tca 220Asn Leu Leu Ile
Ala Asp Ile Glu Leu Gly Cys Gly Asn Phe Gly Ser 15 20 25gtg cgc cag
ggc gtg tac cgc atg cgc aag aag cag atc gac gtg gcc 268Val Arg Gln
Gly Val Tyr Arg Met Arg Lys Lys Gln Ile Asp Val Ala 30 35 40atc aag
gtg ctg aag cag ggc acg gag aag gca gac acg gaa gag atg 316Ile Lys
Val Leu Lys Gln Gly Thr Glu Lys Ala Asp Thr Glu Glu Met 45 50 55atg
cgc gag gcg cag atc atg cac cag ctg gac aac ccc tac atc gtg 364Met
Arg Glu Ala Gln Ile Met His Gln Leu Asp Asn Pro Tyr Ile Val 60 65
70cgg ctc att ggc gtc tgc cag gcc gag gcc ctc atg ctg gtc atg gag
412Arg Leu Ile Gly Val Cys Gln Ala Glu Ala Leu Met Leu Val Met
Glu75 80 85 90atg gct ggg ggc ggg ccg ctg cac aag ttc ctg gtc ggc
aag agg gag 460Met Ala Gly Gly Gly Pro Leu His Lys Phe Leu Val Gly
Lys Arg Glu 95 100 105gag atc cct gtg agc aat gtg gcc gag ctg ctg
cac cag gtg tcc atg 508Glu Ile Pro Val Ser Asn Val Ala Glu Leu Leu
His Gln Val Ser Met 110 115 120ggg atg aag tac ctg gag gag aag aac
ttt gtg cac cgt gac ctg gcg 556Gly Met Lys Tyr Leu Glu Glu Lys Asn
Phe Val His Arg Asp Leu Ala 125 130 135gcc cgc aac gtc ctg ctg gtt
aac cgg cac tac gcc aag atc agc gac 604Ala Arg Asn Val Leu Leu Val
Asn Arg His Tyr Ala Lys Ile Ser Asp 140 145 150ttt ggc ctc tcc aaa
gca ctg ggt gcc gac gac agc tac tac act gcc 652Phe Gly Leu Ser Lys
Ala Leu Gly Ala Asp Asp Ser Tyr Tyr Thr Ala155 160 165 170cgc tca
gca ggg aag tgg ccg ctc aag tgg tac gca ccc gaa tgc atc 700Arg Ser
Ala Gly Lys Trp Pro Leu Lys Trp Tyr Ala Pro Glu Cys Ile 175 180
185aac ttc cgc aag ttc tcc agc cgc agc gat gtc tgg agc tat ggg gtc
748Asn Phe Arg Lys Phe Ser Ser Arg Ser Asp Val Trp Ser Tyr Gly Val
190 195 200acc atg tgg gag gcc ttg tcc tac ggc cag aag ccc tac aag
aag atg 796Thr Met Trp Glu Ala Leu Ser Tyr Gly Gln Lys Pro Tyr Lys
Lys Met 205 210 215aaa ggg ccg gag gtc atg gcc ttc atc gag cag ggc
aag cgg atg gaa 844Lys Gly Pro Glu Val Met Ala Phe Ile Glu Gln Gly
Lys Arg Met Glu 220 225 230tgc cca cca gag tgt cca ccc gaa ctg tac
gca ctc atg agt gac tgc 892Cys Pro Pro Glu Cys Pro Pro Glu Leu Tyr
Ala Leu Met Ser Asp Cys235 240 245 250tgg atc tac aag tgg gag gat
cgc ccc gac ttc ctg acc gtg gag cag 940Trp Ile Tyr Lys Trp Glu Asp
Arg Pro Asp Phe Leu Thr Val Glu Gln 255 260 265cgc atg cga gcc tgt
tac tac agc ctg gcc agc aag gtg gaa ggg cac 988Arg Met Arg Ala Cys
Tyr Tyr Ser Leu Ala Ser Lys Val Glu Gly His 270 275 280cac cac cac
cac cac cac tagaattc 1014His His His His His His
285115288PRTArtificial SequenceDescription of Artificial Sequence
Synthetic Plasmid 115Met Asp Lys Lys Leu Phe Leu Lys Arg Asp Asn
Leu Leu Ile Ala Asp1 5 10 15Ile Glu Leu Gly Cys Gly Asn Phe Gly Ser
Val Arg Gln Gly Val Tyr 20 25 30Arg Met Arg Lys Lys Gln Ile Asp Val
Ala Ile Lys Val Leu Lys Gln 35 40 45Gly Thr Glu Lys Ala Asp Thr Glu
Glu Met Met Arg Glu Ala Gln Ile 50 55 60Met His Gln Leu Asp Asn Pro
Tyr Ile Val Arg Leu Ile Gly Val Cys65 70 75 80Gln Ala Glu Ala Leu
Met Leu Val Met Glu Met Ala Gly Gly Gly Pro 85 90 95Leu His Lys Phe
Leu Val Gly Lys Arg Glu Glu Ile Pro Val Ser Asn 100 105 110Val Ala
Glu Leu Leu His Gln Val Ser Met Gly Met Lys Tyr Leu Glu 115 120
125Glu Lys Asn Phe Val His Arg Asp Leu Ala Ala Arg Asn Val Leu Leu
130 135 140Val Asn Arg His Tyr Ala Lys Ile Ser Asp Phe Gly Leu Ser
Lys Ala145 150 155 160Leu Gly Ala Asp Asp Ser Tyr Tyr Thr Ala Arg
Ser Ala Gly Lys Trp 165 170 175Pro Leu Lys Trp Tyr Ala Pro Glu Cys
Ile Asn Phe Arg Lys Phe Ser 180 185 190Ser Arg Ser Asp Val Trp Ser
Tyr Gly Val Thr Met Trp Glu Ala Leu 195 200 205Ser Tyr Gly Gln Lys
Pro Tyr Lys Lys Met Lys Gly Pro Glu Val Met 210 215 220Ala Phe Ile
Glu Gln Gly Lys Arg Met Glu Cys Pro Pro Glu Cys Pro225 230 235
240Pro Glu Leu Tyr Ala Leu Met Ser Asp Cys Trp Ile Tyr Lys Trp Glu
245 250 255Asp Arg Pro Asp Phe Leu Thr Val Glu Gln Arg Met Arg Ala
Cys Tyr 260 265 270Tyr Ser Leu Ala Ser Lys Val Glu Gly His His His
His His His His 275 280 28511632DNAArtificial SequenceDescription
of Artificial Sequence Synthetic Primer 116gttgtgacat atgttccaga
tcccagagtt tg 3211731DNAArtificial SequenceDescription of
Artificial Sequence Synthetic Primer 117gttgtgagtc gactcactgg
gagggggcgg a 31118795DNAArtificial SequenceDescription of
Artificial Sequence Synthetic PlasmidCDS(85)..(654) 118tacgactcac
tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt 60ttaactttaa
gaaggagata tacc atg gct ggt tgc ctg aac gac atc ttc 111 Met Ala Gly
Cys Leu Asn Asp Ile Phe 1 5gaa gct cag aaa atc gaa tgg cac cat cac
cat cac cat atg ttc cag 159Glu Ala Gln Lys Ile Glu Trp His His His
His His His Met Phe Gln10 15 20 25atc cca gag ttt gag ccg agt gag
cag gaa gac tcc agc tct gca gag 207Ile Pro Glu Phe Glu Pro Ser Glu
Gln Glu Asp Ser Ser Ser Ala Glu 30 35 40agg ggc ctg ggc ccc agc ccc
gca ggg gac ggg ccc tca ggc tcc ggc 255Arg Gly Leu Gly Pro Ser Pro
Ala Gly Asp Gly Pro Ser Gly Ser Gly 45 50 55aag cat cat cgc cag gcc
cca ggc ctc ctg tgg gac gcc agt cac cag 303Lys His His Arg Gln Ala
Pro Gly Leu Leu Trp Asp Ala Ser His Gln 60 65 70cag gag cag cca acc
agc agc agc cat cat gga ggc gct ggg gct gtg 351Gln Glu Gln Pro Thr
Ser Ser Ser His His Gly Gly Ala Gly Ala Val 75 80 85gag atc cgg agt
cgc cac agc tcc tac ccc gcg ggg acg gag gac gac 399Glu Ile Arg Ser
Arg His Ser Ser Tyr Pro Ala Gly Thr Glu Asp Asp90 95 100 105gaa ggg
atg ggg gag gag ccc agc ccc ttt cgg ggc cgc tcg cgc tcg 447Glu Gly
Met Gly Glu Glu Pro Ser Pro Phe Arg Gly Arg Ser Arg Ser 110 115
120gcg ccc ccc aac ctc tgg gca gca cag cgc tat ggc cgc gag ctc cgg
495Ala Pro Pro Asn Leu Trp Ala Ala Gln Arg Tyr Gly Arg Glu Leu Arg
125 130 135agg atg agt gac gag ttt gtg gac tcc ttt aag aag gga ctt
cct cgc
543Arg Met Ser Asp Glu Phe Val Asp Ser Phe Lys Lys Gly Leu Pro Arg
140 145 150ccg aag agc gcg ggc aca gca acg cag atg cgg caa agc tcc
agc tgg 591Pro Lys Ser Ala Gly Thr Ala Thr Gln Met Arg Gln Ser Ser
Ser Trp 155 160 165acg cga gtc ttc cag tcc tgg tgg gat cgg aac ttg
ggc agg gga agc 639Thr Arg Val Phe Gln Ser Trp Trp Asp Arg Asn Leu
Gly Arg Gly Ser170 175 180 185tcc gcc ccc tcc cag tgagtcgacc
accaccacca ccaccactga gatccggctg 694Ser Ala Pro Ser Gln
190gccctactgg ccgaaaggaa ttcgaggcca gcagggccac cgctgagcaa
taactagcat 754aaccccttgg ggcctctaaa cgggtcttga ggggtttttt g
795119190PRTArtificial SequenceDescription of Artificial Sequence
Synthetic Plasmid 119Met Ala Gly Cys Leu Asn Asp Ile Phe Glu Ala
Gln Lys Ile Glu Trp1 5 10 15His His His His His His Met Phe Gln Ile
Pro Glu Phe Glu Pro Ser 20 25 30Glu Gln Glu Asp Ser Ser Ser Ala Glu
Arg Gly Leu Gly Pro Ser Pro 35 40 45Ala Gly Asp Gly Pro Ser Gly Ser
Gly Lys His His Arg Gln Ala Pro 50 55 60Gly Leu Leu Trp Asp Ala Ser
His Gln Gln Glu Gln Pro Thr Ser Ser65 70 75 80Ser His His Gly Gly
Ala Gly Ala Val Glu Ile Arg Ser Arg His Ser 85 90 95Ser Tyr Pro Ala
Gly Thr Glu Asp Asp Glu Gly Met Gly Glu Glu Pro 100 105 110Ser Pro
Phe Arg Gly Arg Ser Arg Ser Ala Pro Pro Asn Leu Trp Ala 115 120
125Ala Gln Arg Tyr Gly Arg Glu Leu Arg Arg Met Ser Asp Glu Phe Val
130 135 140Asp Ser Phe Lys Lys Gly Leu Pro Arg Pro Lys Ser Ala Gly
Thr Ala145 150 155 160Thr Gln Met Arg Gln Ser Ser Ser Trp Thr Arg
Val Phe Gln Ser Trp 165 170 175Trp Asp Arg Asn Leu Gly Arg Gly Ser
Ser Ala Pro Ser Gln 180 185 19012030DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
120cgggtcccat atgcccaaga agaagccgac 3012133DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
121gttcgttgtc gacgacgcca gcagcatggg ttg 3312233DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
122ctaattcatc tggagatcgc gcccgcaatc cgg 3312333DNAArtificial
SequenceDescription of Artificial Sequence Synthetic Primer
123ccggattgcg ggcgcgatct ccagatgaat tag 331241916DNAArtificial
SequenceDescription of Artificial Sequence Synthetic
PlasmidCDS(1)..(1908) 124atg tcc cct ata cta ggt tat tgg aaa att
aag ggc ctt gtg caa ccc 48Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile
Lys Gly Leu Val Gln Pro1 5 10 15act cga ctt ctt ttg gaa tat ctt gaa
gaa aaa tat gaa gag cat ttg 96Thr Arg Leu Leu Leu Glu Tyr Leu Glu
Glu Lys Tyr Glu Glu His Leu 20 25 30tat gag cgc gat gaa ggt gat aaa
tgg cga aac aaa aag ttt gaa ttg 144Tyr Glu Arg Asp Glu Gly Asp Lys
Trp Arg Asn Lys Lys Phe Glu Leu 35 40 45ggt ttg gag ttt ccc aat ctt
cct tat tat att gat ggt gat gtt aaa 192Gly Leu Glu Phe Pro Asn Leu
Pro Tyr Tyr Ile Asp Gly Asp Val Lys 50 55 60tta aca cag tct atg gcc
atc ata cgt tat ata gct gac aag cac aac 240Leu Thr Gln Ser Met Ala
Ile Ile Arg Tyr Ile Ala Asp Lys His Asn65 70 75 80atg ttg ggt ggt
tgt cca aaa gag cgt gca gag att tca atg ctt gaa 288Met Leu Gly Gly
Cys Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu 85 90 95gga gcg gtt
ttg gat att aga tac ggt gtt tcg aga att gca tat agt 336Gly Ala Val
Leu Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser 100 105 110aaa
gac ttt gaa act ctc aaa gtt gat ttt ctt agc aag cta cct gaa 384Lys
Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120
125atg ctg aaa atg ttc gaa gat cgt tta tgt cat aaa aca tat tta aat
432Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn
130 135 140ggt gat cat gta acc cat cct gac ttc atg ttg tat gac gct
ctt gat 480Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala
Leu Asp145 150 155 160gtt gtt tta tac atg gac cca atg tgc ctg gat
gcg ttc cca aaa tta 528Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp
Ala Phe Pro Lys Leu 165 170 175gtt tgt ttt aaa aaa cgt att gaa gct
atc cca caa att gat aag tac 576Val Cys Phe Lys Lys Arg Ile Glu Ala
Ile Pro Gln Ile Asp Lys Tyr 180 185 190ttg aaa tcc agc aag tat ata
gca tgg cct ttg cag ggc tgg caa gcc 624Leu Lys Ser Ser Lys Tyr Ile
Ala Trp Pro Leu Gln Gly Trp Gln Ala 195 200 205acg ttt ggt ggt ggc
gac cat cct cca aaa tcg gat ctg gtt ccg cgt 672Thr Phe Gly Gly Gly
Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220gga tct cat
atg ccc aag aag aag ccg acg ccc atc cag ctg aac ccg 720Gly Ser His
Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro225 230 235
240gcc ccc gac ggc tct gca gtt aac ggg acc agc tct gcg gag acc aac
768Ala Pro Asp Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn
245 250 255ttg gag gcc ttg cag aag aag ctg gag gag cta gag ctt gat
gag cag 816Leu Glu Ala Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp
Glu Gln 260 265 270cag cga aag cgc ctt gag gcc ttt ctt acc cag aag
cag aag gtg gga 864Gln Arg Lys Arg Leu Glu Ala Phe Leu Thr Gln Lys
Gln Lys Val Gly 275 280 285gaa ctg aag gat gac gac ttt gag aag atc
agt gag ctg ggg gct ggc 912Glu Leu Lys Asp Asp Asp Phe Glu Lys Ile
Ser Glu Leu Gly Ala Gly 290 295 300aat ggc ggt gtg gtg ttc aag gtc
tcc cac aag cct tct ggc ctg gtc 960Asn Gly Gly Val Val Phe Lys Val
Ser His Lys Pro Ser Gly Leu Val305 310 315 320atg gcc aga gcg cta
att cat ctg gag atc aaa ccc gca atc cgg aac 1008Met Ala Arg Ala Leu
Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn 325 330 335cag atc ata
agg gag ctg cag gtt ctg cat gag tgc aac tct ccg tac 1056Gln Ile Ile
Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr 340 345 350atc
gtg ggc ttc tat ggt gcg ttc tac agc gat ggc gag atc agt atc 1104Ile
Val Gly Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile 355 360
365tgc atg gag cac atg gat gga ggt tct ctg gat caa gtc ctg aag aaa
1152Cys Met Glu His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys
370 375 380gct gga aga att cct gaa caa att tta gga aaa gtt agc att
gct gta 1200Ala Gly Arg Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile
Ala Val385 390 395 400ata aaa ggc ctg aca tat ctg agg gag aag cac
aag atc atg cac aga 1248Ile Lys Gly Leu Thr Tyr Leu Arg Glu Lys His
Lys Ile Met His Arg 405 410 415gat gtc aag ccc tcc aac atc cta gtc
aac tcc cgt ggg gag atc aag 1296Asp Val Lys Pro Ser Asn Ile Leu Val
Asn Ser Arg Gly Glu Ile Lys 420 425 430ctc tgt gac ttt ggg gtc agc
ggg cag ctc atc gac tcc atg gcc aac 1344Leu Cys Asp Phe Gly Val Ser
Gly Gln Leu Ile Asp Ser Met Ala Asn 435 440 445tcc ttc gtg ggc aca
agg tcc tac atg tcg cca gaa aga ctc cag ggg 1392Ser Phe Val Gly Thr
Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly 450 455 460act cat tac
tct gtg cag tca gac atc tgg agc atg gga ctg tct ctg 1440Thr His Tyr
Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu465 470 475
480gta gag atg gcg gtt ggg agg tat ccc atc cct cct cca gat gcc aag
1488Val Glu Met Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys
485 490 495gag ctg gag ctg atg ttt ggg tgc cag gtg gaa gga gat gcg
gct gag 1536Glu Leu Glu Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala
Ala Glu 500 505 510acc cca ccc agg cca agg acc ccc ggg agg ccc ctt
agc tca tac gga 1584Thr Pro Pro Arg Pro Arg Thr Pro Gly Arg Pro Leu
Ser Ser Tyr Gly 515 520 525atg gac agc cga cct ccc atg gca att ttt
gag ttg ttg gat tac ata 1632Met Asp Ser Arg Pro Pro Met Ala Ile Phe
Glu Leu Leu Asp Tyr Ile 530 535 540gtc aac gag cct cct cca aaa ctg
ccc agt gga gtg ttc agt ctg gaa 1680Val Asn Glu Pro Pro Pro Lys Leu
Pro Ser Gly Val Phe Ser Leu Glu545 550 555 560ttt caa gat ttt gtg
aat aaa tgc tta ata aaa aac ccc gca gag aga 1728Phe Gln Asp Phe Val
Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg 565 570 575gca gat ttg
aag caa ctc atg gtt cat gct ttt atc aag aga tct gat 1776Ala Asp Leu
Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp 580 585 590gct
gag gaa gtg gat ttt gca ggt tgg ctc tgc tcc acc atc ggc ctt 1824Ala
Glu Glu Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu 595 600
605aac cag ccc agc aca cca acc cat gct gct ggc gtc gtc gac ctg aac
1872Asn Gln Pro Ser Thr Pro Thr His Ala Ala Gly Val Val Asp Leu Asn
610 615 620gac atc ttc gaa gct cag aaa atc gaa tgg cac cgt tagaattc
1916Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Arg625 630
635125636PRTArtificial SequenceDescription of Artificial Sequence
Synthetic Plasmid 125Met Ser Pro Ile Leu Gly Tyr Trp Lys Ile Lys
Gly Leu Val Gln Pro1 5 10 15Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu
Lys Tyr Glu Glu His Leu 20 25 30Tyr Glu Arg Asp Glu Gly Asp Lys Trp
Arg Asn Lys Lys Phe Glu Leu 35 40 45Gly Leu Glu Phe Pro Asn Leu Pro
Tyr Tyr Ile Asp Gly Asp Val Lys 50 55 60Leu Thr Gln Ser Met Ala Ile
Ile Arg Tyr Ile Ala Asp Lys His Asn65 70 75 80Met Leu Gly Gly Cys
Pro Lys Glu Arg Ala Glu Ile Ser Met Leu Glu 85 90 95Gly Ala Val Leu
Asp Ile Arg Tyr Gly Val Ser Arg Ile Ala Tyr Ser 100 105 110Lys Asp
Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120
125Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn
130 135 140Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala
Leu Asp145 150 155 160Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp
Ala Phe Pro Lys Leu 165 170 175Val Cys Phe Lys Lys Arg Ile Glu Ala
Ile Pro Gln Ile Asp Lys Tyr 180 185 190Leu Lys Ser Ser Lys Tyr Ile
Ala Trp Pro Leu Gln Gly Trp Gln Ala 195 200 205Thr Phe Gly Gly Gly
Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220Gly Ser His
Met Pro Lys Lys Lys Pro Thr Pro Ile Gln Leu Asn Pro225 230 235
240Ala Pro Asp Gly Ser Ala Val Asn Gly Thr Ser Ser Ala Glu Thr Asn
245 250 255Leu Glu Ala Leu Gln Lys Lys Leu Glu Glu Leu Glu Leu Asp
Glu Gln 260 265 270Gln Arg Lys Arg Leu Glu Ala Phe Leu Thr Gln Lys
Gln Lys Val Gly 275 280 285Glu Leu Lys Asp Asp Asp Phe Glu Lys Ile
Ser Glu Leu Gly Ala Gly 290 295 300Asn Gly Gly Val Val Phe Lys Val
Ser His Lys Pro Ser Gly Leu Val305 310 315 320Met Ala Arg Ala Leu
Ile His Leu Glu Ile Lys Pro Ala Ile Arg Asn 325 330 335Gln Ile Ile
Arg Glu Leu Gln Val Leu His Glu Cys Asn Ser Pro Tyr 340 345 350Ile
Val Gly Phe Tyr Gly Ala Phe Tyr Ser Asp Gly Glu Ile Ser Ile 355 360
365Cys Met Glu His Met Asp Gly Gly Ser Leu Asp Gln Val Leu Lys Lys
370 375 380Ala Gly Arg Ile Pro Glu Gln Ile Leu Gly Lys Val Ser Ile
Ala Val385 390 395 400Ile Lys Gly Leu Thr Tyr Leu Arg Glu Lys His
Lys Ile Met His Arg 405 410 415Asp Val Lys Pro Ser Asn Ile Leu Val
Asn Ser Arg Gly Glu Ile Lys 420 425 430Leu Cys Asp Phe Gly Val Ser
Gly Gln Leu Ile Asp Ser Met Ala Asn 435 440 445Ser Phe Val Gly Thr
Arg Ser Tyr Met Ser Pro Glu Arg Leu Gln Gly 450 455 460Thr His Tyr
Ser Val Gln Ser Asp Ile Trp Ser Met Gly Leu Ser Leu465 470 475
480Val Glu Met Ala Val Gly Arg Tyr Pro Ile Pro Pro Pro Asp Ala Lys
485 490 495Glu Leu Glu Leu Met Phe Gly Cys Gln Val Glu Gly Asp Ala
Ala Glu 500 505 510Thr Pro Pro Arg Pro Arg Thr Pro Gly Arg Pro Leu
Ser Ser Tyr Gly 515 520 525Met Asp Ser Arg Pro Pro Met Ala Ile Phe
Glu Leu Leu Asp Tyr Ile 530 535 540Val Asn Glu Pro Pro Pro Lys Leu
Pro Ser Gly Val Phe Ser Leu Glu545 550 555 560Phe Gln Asp Phe Val
Asn Lys Cys Leu Ile Lys Asn Pro Ala Glu Arg 565 570 575Ala Asp Leu
Lys Gln Leu Met Val His Ala Phe Ile Lys Arg Ser Asp 580 585 590Ala
Glu Glu Val Asp Phe Ala Gly Trp Leu Cys Ser Thr Ile Gly Leu 595 600
605Asn Gln Pro Ser Thr Pro Thr His Ala Ala Gly Val Val Asp Leu Asn
610 615 620Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Arg625 630
635
* * * * *
References