U.S. patent application number 14/170272 was filed with the patent office on 2016-06-02 for akt protein kinase inhibitors.
This patent application is currently assigned to Array BioPharma, Inc.. The applicant listed for this patent is Array BioPharma, Inc.. Invention is credited to April L. Bernacki, James Blake, Yongxin Han, John Josey, Nicolas C. Kallan, Scott Miller, Ian S. Mitchell, Mark Munson, Anthony Piscopio, Chang Rao, Keith L. Spencer, Peter Stengel, Guy P.A. Vigers, Bing Wang, Dengming Xiao, Rui Xu.
Application Number | 20160152576 14/170272 |
Document ID | / |
Family ID | 34632854 |
Filed Date | 2016-06-02 |
United States Patent
Application |
20160152576 |
Kind Code |
A9 |
Mitchell; Ian S. ; et
al. |
June 2, 2016 |
AKT PROTEIN KINASE INHIBITORS
Abstract
The present invention provides compounds, including resolved
enantiomers, diastereomers, solvates and pharmaceutically
acceptable salts thereof, comprising the Formula: A-L-CR where CR
is a cyclical core group, L is a linking group and A is as defined
herein. Also provided are methods of using the compounds of this
invention as AKT protein kinase inhibitors and for the treatment of
hyperproliferative diseases such as cancer.
Inventors: |
Mitchell; Ian S.; (Boulder,
CO) ; Spencer; Keith L.; (Boulder, CO) ;
Stengel; Peter; (Boulder, CO) ; Han; Yongxin;
(Boulder, CO) ; Kallan; Nicolas C.; (Boulder,
CO) ; Munson; Mark; (Boulder, CO) ; Vigers;
Guy P.A.; (Boulder, CO) ; Blake; James;
(Boulder, CO) ; Piscopio; Anthony; (Boulder,
CO) ; Josey; John; (Boulder, CO) ; Miller;
Scott; (Boulder, CO) ; Xiao; Dengming;
(Boulder, CO) ; Xu; Rui; (Boulder, CO) ;
Rao; Chang; (Boulder, CO) ; Wang; Bing;
(Boulder, CO) ; Bernacki; April L.; (Boulder,
CO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Array BioPharma, Inc. |
Boulder |
CO |
US |
|
|
Assignee: |
Array BioPharma, Inc.
Boulder
CO
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20140148436 A1 |
May 29, 2014 |
|
|
Family ID: |
34632854 |
Appl. No.: |
14/170272 |
Filed: |
January 31, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12567258 |
Sep 25, 2009 |
8680114 |
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14170272 |
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10993173 |
Nov 19, 2004 |
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12567258 |
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60524003 |
Nov 21, 2003 |
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Current U.S.
Class: |
514/210.21 ;
514/252.11; 514/252.14; 514/252.16; 514/252.17; 514/253.04;
514/253.05; 514/254.06; 514/266.22; 514/267; 544/250; 544/278;
544/280; 544/293; 544/295; 544/362; 544/363; 544/371 |
Current CPC
Class: |
Y10T 428/31786 20150401;
B44F 1/10 20130101; C07D 401/12 20130101; C07D 237/28 20130101;
A61P 35/00 20180101; C07D 491/048 20130101; C07D 409/12 20130101;
C07D 239/94 20130101; C07D 239/47 20130101; C07D 495/04 20130101;
Y10T 428/24802 20150115; C07D 403/12 20130101; C07D 403/04
20130101; C07D 409/04 20130101; Y10T 428/31855 20150401; C07D
487/04 20130101; C07D 471/04 20130101; C07D 239/42 20130101; A61P
29/00 20180101; G09F 13/16 20130101; C07D 231/56 20130101; Y10T
428/25 20150115; B41M 3/008 20130101; Y10T 428/31551 20150401; A61P
43/00 20180101; C07D 239/48 20130101; Y10T 428/31507 20150401; C07D
401/04 20130101; Y10T 428/24851 20150115 |
International
Class: |
C07D 239/94 20060101
C07D239/94; C07D 403/12 20060101 C07D403/12; C07D 495/04 20060101
C07D495/04; C07D 491/048 20060101 C07D491/048; C07D 231/56 20060101
C07D231/56; C07D 401/12 20060101 C07D401/12; C07D 409/04 20060101
C07D409/04; C07D 401/04 20060101 C07D401/04; C07D 409/12 20060101
C07D409/12; C07D 487/04 20060101 C07D487/04 |
Claims
1. A compound including resolved enantiomers, diastereomers,
solvates and pharmaceutically acceptable salts thereof, said
compound comprising Formula I: A-L-CR (I) where: CR is heteroaryl,
wherein said heteroaryl is optionally substituted with one or more
groups selected from halogen, hydroxyl, cyano, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl, heteroaryl, arylalkyl and
heteroarylalkyl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl
are further optionally substituted with one or more groups selected
from halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,
difluoromethyl, trifluoromethyl, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6
heterocycloalkyl, --SR.sup.21, --S(O)R.sup.24, --SO.sub.2R.sup.24,
--C(O)R.sup.21, C(O)OR.sup.21, --C(O)NR.sup.21R.sup.22,
--NR.sup.21R.sup.22 and --OR.sup.21; L is selected from:
##STR00124## ##STR00125## where R.sup.12 is hydrogen, halogen,
hydroxy, cyano, nitro, amino, azido, C.sub.1-C.sub.5 alkyl,
C.sub.2-C.sub.5 alkenyl, C.sub.2-C.sub.5 alkynyl, C.sub.1-C.sub.5
heteroalkyl, C.sub.2-C.sub.5 heteroalkenyl or C.sub.2-C.sub.5
heteroalkynyl, wherein any of said alkyl, alkenyl, alkynyl,
heteroalkyl, heteroalkenyl, and heteroalkynyl are optionally
substituted with one or more groups independently selected from
halogen, cyano, nitro, amino, azido, C.sub.1-C.sub.4 alkyl,
fluoromethyl, difluoromethyl, trifluoromethyl, methoxy,
fluoromethoxy, difluoromethoxy and trifluoromethoxy; ##STR00126## A
is W is N or CR.sup.15, provided that when L is a substituted or
unsubstituted piperazinylene, W must be CR.sup.15; G is hydrogen,
alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,
cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein any of
said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,
heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl is
optionally substituted with one or more groups selected from
halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl; B.sup.1 and B.sup.2 are independently absent or
C.sub.1-C.sub.4 alkylene, C.sub.1-C.sub.4 heteroalkylene,
C.sub.2-C.sub.4 alkenylene, C.sub.2-C.sub.4 heteroalkenylene,
C.sub.2-C.sub.4 alkynylene, C.sub.2-C.sub.4 heteroalkynylene,
C.sub.3-C.sub.6 cycloalkylene, and C.sub.3-C.sub.6
heterocycloalkylene, wherein any of said alkylene, heteroalkylene,
alkenylene, heteroalkenylene, alkynylene, heteroalkynylene,
cycloalkylene or heterocycloalkylene is optionally substituted with
one or more groups independently selected from halogen, hydroxyl,
cyano, nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4
heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21; R.sup.21, R.sup.22
and R.sup.23 independently are hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6
heterocycloalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl;
R.sup.24 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 heteroalkyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 heteroalkynyl,
C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl,
arylalkyl, heteroaryl or heteroarylalkyl; or any two of R.sup.21,
R.sup.22, R.sup.23 or R.sup.24 together with the atom(s) to which
they are attached form a 4 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl or heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,
aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, and
heterocyclylalkyl; R.sup.13 and R.sup.14 are independently
hydrogen, hydroxyl, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
heteroalkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6
heterocycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl,
--C(O)R.sup.21, C(O)OR.sup.21, C(.dbd.NR.sup.21)NR.sup.22R.sup.23
or --SO.sub.2R.sup.24, wherein any of said alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroalkyl,
heteroalkenyl, heteroalkynyl, arylalkyl or heteroarylalkyl is
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl; or R.sup.13 and R.sup.14 together with the atoms to
which they are attached form a 4 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21; or R.sup.13 and an atom of B.sup.2
together with N form a 4 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21; R.sup.15 is hydrogen,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl or
C.sub.2-C.sub.4 heteroalkynyl, wherein any of said alkyl,
heteroalkyl, alkenyl, heteroalkenyl, alkynyl or heteroalkynyl is
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, nitro, azido,
NR.sup.21R.sup.22 and OR.sup.21; or R.sup.13 and R.sup.15 together
with atoms to which they are attached form a 3 to 10 membered
carbocyclic, aryl, heteroaryl or heterocyclic ring, wherein any of
said carbocyclic, aryl, heteroaryl and heterocyclic rings are
optionally substituted with one or more groups independently
selected from halogen, cyano, nitro, azido, C.sub.1-C.sub.4 alkyl,
C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21; or,
when W is CR.sup.15, R.sup.15 and an atom of B.sup.1 or B.sup.2
together with C, form a 3 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21.
2. The compound of claim 1, where CR is selected from: ##STR00127##
where X is N or CR.sup.1; Y is CR.sup.2 or N, provided that when X
is N, Y must be CR.sup.2; Z is CR.sup.3R.sup.3a or NR.sup.2a,
provided that when X is N, Z must be CR.sup.3; D.sup.1, D.sup.2,
D.sup.3 and D.sup.4 are independently CR.sup.4 or N, provided that
no more than two of D.sup.1, D.sup.2, D.sup.3 or D.sup.4 are N; - -
- - - is an optional double bond; D.sup.5 is CR.sup.5R.sup.5a,
NR.sup.2a, O or S, provided that when D.sup.5 is O or S, D.sup.8
must be C, D.sup.7 must be CR.sup.7 or N, and either (i) Q must be
CR.sup.6 or CR.sup.6R.sup.6a or (ii) D.sup.7 must be CR.sup.7 or
CR.sup.7R.sup.7a; Q is CR.sup.6, N or C.dbd.O, provided that either
(w) when Q is N, one of D.sup.5, D.sup.7 and D.sup.8 must be C, or
(x) when Q is C.dbd.O, D.sup.5 must be CR.sup.5 or N, D.sup.7 must
be CR.sup.7 or N, and D.sup.8 must be C; D.sup.7 is CR.sup.7, N, O
or S, provided that when D.sup.7 is O or S, D.sup.8 must be C,
D.sup.5 must be CR.sup.5 or N, and either (y) Q must be CR.sup.6,
or (z) D.sup.5 must be CR.sup.5; D.sup.8 is C or N, provided that
when D.sup.8 is N, D.sup.5 must be CR.sup.5R.sup.5a and Q must be
CR.sup.6 or CR.sup.6R.sup.6a; either K or M is carbonyl, provided
that both K and M are not carbonyl; R.sup.1, R.sup.5, R.sup.5a and
R.sup.8 are independently hydrogen, halogen, hydroxyl, cyano,
nitro, azido, fluoromethyl, difluoromethyl, trifluoromethyl,
methoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, amino,
aminomethyl, dimethylamino, aminoethyl, diethylamino or ethoxy;
R.sup.2 is hydrogen, halogen, hydroxyl, cyano, nitro, amino, azido,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl,
C.sub.1-C.sub.6 heterocycloalkyl, C.sub.1-C.sub.6 aryl, or
C.sub.1-C.sub.6 heteroaryl, wherein any of said alkyl, heteroalkyl,
alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl are further optionally
substituted with one or more groups independently selected from
halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy, trifluoromethoxy, amino, aminomethyl,
dimethylamino, aminoethyl, diethylamino and ethoxy; R.sup.2a is
hydrogen, hydroxyl, cyano, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6
heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,
heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,
cycloalkyl, heterocycloalkyl, aryl or heteroaryl are optionally
substituted with one or more groups independently selected from
halogen, hydroxyl, cyano, nitro, azido fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy, trifluoromethoxy, amino, aminomethyl,
dimethylamino, aminoethyl, diethylamino or ethoxy; R.sup.3 and
R.sup.3a are independently hydrogen, halogen, hydroxyl, cyano,
nitro, amino azido, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
heteroalkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6
heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,
heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,
cycloalkyl, heterocycloalkyl, aryl or heteroaryl are further
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, nitro, azido, OR.sup.1,
NR.sup.1R.sup.2, and (C.dbd.O)R.sup.2; R.sup.4, R.sup.6, R.sup.6a,
R.sup.7, R.sup.7a and R.sup.10 are independently hydrogen,
hydroxyl, cyano, amino, nitro, azido, alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23 or --OR.sup.21, wherein any of
said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,
heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,
heteroaryl or heteroarylalkyl, is optionally substituted with one
or more groups independently selected from halogen, hydroxyl,
cyano, amino, nitro, azido, --NR.sup.21SO.sub.2R.sup.24,
--SO.sub.2NR.sup.21R.sup.22, --NR.sup.21S(O)R.sup.4,
--S(O)NR.sup.21R.sup.22, --C(O)R.sup.21, --C(O)OR.sup.21,
--OC(O)R.sup.21, --OC(O)OR.sup.21, --NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl
and heteroaryl, and wherein said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl may optionally be further
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl
and heteroaryl, --NR.sup.21R.sup.22, and --OR.sup.21; or R.sup.6
and R.sup.7 together with the atoms to which they are attached form
a 4 to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic
ring, wherein any of said carbocyclic, aryl, heteroaryl and
heterocyclic rings are optionally substituted with one or more
groups independently selected from halogen, cyano, nitro,
trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,
OR.sup.1, NR.sup.1R.sup.2, cycloalkyl, heterocycloalkyl, aryl,
heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, and
heterocyclylalkyl; and R.sup.9 is hydrogen, halogen, hydroxyl,
cyano, nitro, azido, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
heteroalkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6
heterocycloalkyl, aryl, heteroaryl, --NR.sup.21R.sup.22,
--OR.sup.21, --NR.sup.21SO.sub.2R.sup.24 and
--NR.sup.21C(O)R.sup.22, wherein any of said alkyl, heteroalkyl,
alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl are further optionally
substituted with one or more groups independently selected from
halogen, hydroxyl, cyano, nitro, azido, OR.sup.1, NR.sup.1R.sup.2,
and (C.dbd.O)R.sup.2.
3. The compound of claim 2, where CR is selected from:
##STR00128##
4. The compound of claim 3, where D.sup.5 is NH.
5. The compound of claim 2, where CR is selected from:
##STR00129##
6. The compound of claim 2, where L is selected from:
##STR00130##
7. The compound of claim 6, where L is: ##STR00131##
8. The compound of claim 2, where A is: ##STR00132##
9. The compound of claim 2, where A is: ##STR00133## where G is
hydrogen, alkyl, cycloalkyl, heterocycloaryl, aryl or heteroaryl,
wherein any of said alkyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl is optionally substituted with one or more groups
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
cycloalkyl, heterocycloalkyl aryl and heteroaryl.
10. The compound of claim 6, where A is: ##STR00134## where G is
hydrogen, alkyl, cycloalkyl, heterocycloaryl, aryl or heteroaryl,
wherein any of said alkyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl is optionally substituted with one or more groups
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 alkynyl,
cycloalkyl, heterocycloalkyl aryl and heteroaryl.
11. The compound of claim 6, wherein A comprises: ##STR00135##
where B.sup.1 and B.sup.2 are, independently, absent or
C.sub.1-C.sub.4 alkylene; R.sup.21a--R.sup.21c are independently H,
halogen, CH.sub.3, CF.sub.3, CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph,
OH, or OCH.sub.2Ph; R.sup.22a, R.sup.22b, and R.sup.24 are
independently H, CH.sub.3, or halogen; R.sup.23a is H; and
R.sup.23b is H, CH.sub.3, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.2,
CH.sub.2CH.sub.2NH.sub.2, CH.sub.2CH.sub.2NHCH.sub.2,
CH.sub.2CH.sub.2N(CH.sub.2).sub.2, --(C.dbd.O)CH.sub.2NH.sub.2 or
--(C.dbd.O)CH.sub.2CH.sub.2NH.sub.2; or R.sup.23a and R.sup.23b are
joined to complete a 5 or 6 membered heterocyclic ring.
12. The compound of claim 7, wherein A comprises: ##STR00136##
where B.sup.1 and B.sup.2 are, independently, absent or
C.sub.1-C.sub.4 alkylene; R.sup.21a--R.sup.21c are independently H,
halogen, CH.sub.3, CF.sub.3, CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph,
OH, or OCH.sub.2Ph; R.sup.22a, R.sup.22b, and R.sup.24 are
independently H, CH.sub.3, or halogen; R.sup.23a is H; and
R.sup.23b is H, CH.sub.3, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.2,
CH.sub.2CH.sub.2NH.sub.2, CH.sub.2CH.sub.2NHCH.sub.2,
CH.sub.2CH.sub.2N(CH.sub.2).sub.2, --(C.dbd.O)CH.sub.2NH.sub.2 or
--(C.dbd.O)CH.sub.2CH.sub.2NH.sub.2; or R.sup.23a and R.sup.23b are
joined to complete a 5 or 6 membered heterocyclic ring.
13. The compound of claim 6, wherein A comprises: ##STR00137##
wherein R.sup.25 and R.sup.26 are independently H or CH.sub.3; and
R.sup.27 is 1-naphthyl, 2-naphthyl, 3'-benzylthienyl, 2'-thienyl,
2'-pyridyl, 3'-pyridyl, 4'-pyridyl, 4'-thiazolyl, or
3,3-diphenyl.
14. The compound of claim 7, wherein A comprises: ##STR00138##
wherein R.sup.25 and R.sup.26 are independently H or CH.sub.3; and
R.sup.27 is 1-naphthyl, 2-naphthyl, 3'-benzylthienyl, 2'-thienyl,
2'-pyridyl, 3'-pyridyl, 4'-pyridyl, 4'-thiazolyl, or
3,3-diphenyl.
15. The compound of claim 6, where A is: ##STR00139##
16. The compound of claim 7, where A is: ##STR00140##
17. The compound of claim 7, where A is a D- or L-amino acid
selected from the naturally occurring amino acids,
4-hydroxyproline, hydroxylysine, demosine, isodemosine,
3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric
acid, cirtulline, homocysteine, homoserine, ornithine and
methionine sulfone.
18. The compound of claim 17, wherein the amino acid is alanine,
phenylalanine, histidine, or tryptophan.
19. The compound of claim 1, wherein the compound comprises:
##STR00141##
20. The compound of claim 1, wherein the compound comprises:
##STR00142## wherein R.sup.28 is s H, halogen, CH.sub.3, CF.sub.3,
CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph, OH, or OCH.sub.2Ph.
21. The compound of claim 1, wherein the compound comprises:
##STR00143## wherein D.sup.16 is O or N; and R.sup.29 is H,
halogen, CH.sub.3, CF.sub.3, CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph,
OH, or OCH.sub.2Ph.
22. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 1 to said mammal.
23. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 2 to said mammal.
24. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 3 to said mammal.
25. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 4 to said mammal.
26. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 5 to said mammal.
27. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 6 to said mammal.
28. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 7 to said mammal.
29. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 8 to said mammal.
30. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 9 to said mammal.
31. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 10 to said mammal.
32. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 11 to said mammal.
33. A method of treating hyperproliferative diseases in a mammal
comprising administering a therapeutically effective amount of the
compound defined in claim 12 to said mammal.
34. A composition comprising a compound of claim 1 and a
pharmaceutically acceptable carrier.
35. A composition comprising a compound of claim 2 and a
pharmaceutically acceptable carrier.
36. A composition comprising a compound of claim 3 and a
pharmaceutically acceptable carrier.
37. A composition comprising a compound of claim 4 and a
pharmaceutically acceptable carrier.
38. A composition comprising a compound of claim 5 and a
pharmaceutically acceptable carrier.
39. A composition comprising a compound of claim 7 and a
pharmaceutically acceptable carrier.
40. A composition comprising a compound of claim 9 and a
pharmaceutically acceptable carrier.
41. A compound according to any one of claims 1-21 for use as a
medicament.
42. A compound according to any one of claims 1-21 for use as a
medicament for the treatment of a hyperproliferative disorder.
43. The use of a compound according to any one of claims 1-21 in
the manufacture of a medicament for the treatment of a
hyperproliferative disorder or an inflammatory condition.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. application Ser.
No. 10/993,173 filed on Nov. 19, 2004, which claims the benefit of
U.S. Provisional Application Ser. No. 60/524,003, filed Nov. 21,
2003, which are incorporated herein by reference in their
entireties.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention relates to novel inhibitors of
serine/threonine protein kinases (e.g., AKT and related kinases),
pharmaceutical compositions containing the inhibitors, and methods
for preparing these inhibitors. The inhibitors are useful for the
treatment of hyperproliferative diseases, such as cancer and
inflammation, in mammals and especially in humans.
[0004] 2. Description of the State of the Art
[0005] Protein kinases are a class of enzymes that catalyze the
transfer of the .gamma.-phosphorate group from ATP to a recipient
protein, acting as a substrate. The specific target of the kinase
is the hydroxyl group of a serine, threonine or tyrosine residue.
As a result of this specific targeting, kinases are generally
referred to as serine/threonine protein kinases or tyrosine protein
kinases. The human genome is estimated to encode in excess of 500
distinct protein kinases.
[0006] The seemingly insignificant task of phosphorylation of a
serine, threonine or tyrosine residue belies the importance of
protein kinases in the processes of signal transduction and
regulation of cellular functions. Kinases are typically mediated by
transmembrane cellular receptors, such as G-protein coupled
receptors or growth factor receptors, which when activated by
extracellular ligands cause the phosphorylation of intracellular
proteins. Often, an interconnected series (or cascade) of protein
kinases is necessary to exert the overall effect of this initial
signal, which can ultimately result in effects as extreme as cell
death (apoptosis).
[0007] The ratio of phosphorylated to unphosphorylated protein is a
delicate equilibrium, with protein phosphatases acting as the
negative regulator of protein kinases, removing the phosphoryl
group as it is no longer required. As an example of this interplay,
the phosphorylation state of kinases can control whether a cell
undergoes division, arrests in the cell cycle or programmed cell
death. Should this kinase/phosphatase relationship become
disregulated, the potential consequences relating to disease are
enormous. For example, abnormal protein kinase activity or
expression may be correlated with numerous hyperproliferative
diseases, inflammation and tissue repair, and has been associated
with a large number of diseases ranging from the relatively
non-life threatening, such as psoriasis, to those which are almost
always fatal, such as glioblastoma multiforme, an aggressive brain
cancer.
[0008] Significantly, atypical protein phosphorylation and/or
expression is often reported to be one of the causative effects of
abnormal cellular proliferation, metastasis and cell survival in
cancer. The abnormal regulation and/or expression of various
kinases, including VEGF, ILK, AKT, ROCK, p70S6K, Bcl, PKA, PKC,
Raf, Src, PDK1, ErbB2, MEK, IKK, Cdk, EGFR, BAD, CHK1, CHK2 and
GSK3 amongst numerous others, has been specifically implicated in
cancer.
[0009] Recent data from the CDC indicate that cancer is the second
most common cause of death in the United States, with nearly a
quarter of all deaths reported being attributable to malignant
neoplasms (Anderson, National Vital Statistics Report, 2001, 49
(11):1). Despite recent advances in the understanding of the
genesis, progression and treatment of cancer, much still needs to
be done to improve the overall prognosis of cancer patients.
[0010] The phosphatidylinositol 3'-OH kinase (PI3K) pathway is one
of the signaling pathways that exerts its effect on numerous
cellular functions including cell cycle progression, proliferation,
motility, metabolism and survival. Activation of receptor protein
tyrosine kinases (RTKs) cause PI3K to phosphorylate
phosphatidylinositol (4,5)-diphosphate [PtdIns(4,5)P.sub.2],
generating the membrane-bound phosphatidylinositol
(3,4,5)-triphosphate [PtdIns(3,4,5)P.sub.3]. This in turn promotes
the recruitment of a variety of protein kinases from the cytoplasm
to the plasma membrane through the binding of PtdIns(3,4,5)P.sub.3
to the pleckstrin-homology (PH) domain of the kinase. Kinases
notable as key downstream targets of PI3K include
phosphoinositide-dependant kinase 1 (PDK1) and AKT (also known as
Protein Kinase B.) Phosphorylation of such kinases then permits the
activation or deactivation of numerous other pathways involving
mediators such as GSK3, mTOR, PRAS40, FKHD, NF-.kappa.B, BAD,
Caspase-9, etc.
[0011] An important negative feedback mechanism for the PI3K
pathway is PTEN, a phosphatase that catalyses the dephosphorylation
of PtdIns(3,4,5)P.sub.3 to PtdIns(4,5)P.sub.2 (Furnari, F. B., et
al, Cancer Res. 1998, 58:5002; Dahia, P. L. M., Hum. Molec. Genet.
1999, 8:185). It is of enormous significance that in greater than
60% of all solid tumors, PTEN is mutated into an inactive form,
permitting the constitutive-activation of the PI3K pathway. As the
majority of cancers are solid tumors, such an observation would
suggest that by specifically targeting either PI3K itself or the
individual downstream kinases in the PI3K pathway, one might able
to mitigate the effects of various cancers and restore normal
cellular function.
[0012] One of the best-characterized targets of the PI3K lipid
products is the AGC serine/threonine protein kinase AKT (Hemmings,
B. A., Science, 1997, 275:628). AKT is the human homologue of the
protooncogene v-akt of the acutely transforming retrovirus AKT8.
Its high sequence homology to protein kinases A and C has also
earned it the names Protein Kinase B (PKB) and Related to A and
C(RAC.) Three isoforms of AKT are known to exist, namely Akt1, Akt2
and Akt3, which exhibit an overall homology of 80% (Staal, S. P,
Proc. Natl. Acad. Sci., 1987, 84:5034; Nakatani, K, Biochem.
Biophys. Res. Commun., 1999, 257:906). In addition, both Akt2 and
Akt3 exhibit splice variants.
[0013] Upon recruitment to the cell membrane by
PtdInd(3,4,5)P.sub.3, AKT is phosphorylated (activated) by PDK1 at
T308, T309 and T305 for isoforms Akt1, 2 and 3, respectively, and
at S473, S474 and S472 for isoforms Akt1, 2 and 3, respectively.
Such phosphorylation occurs by an as yet unknown kinase (putatively
named PDK2), although PDK1 (Balendran, A., Curr. Biol., 1999,
9:393), autophosphorylation (Toker, A., J. Biol. Chem., 2000,
275:8271) and integrin-linked kinase (ILK) (Delcommenne, M., Proc.
Natl. Acad. Sci. USA, 1998, 95:11211) have been implicated in this
process. Although monophosphorylation of AKT activates the kinase,
bis(phosphorylation) is required for maximal kinase activity.
[0014] AKT is believed to assert its effect on cancer by
suppressing apoptosis and enhancing both angiogenesis and
proliferation. In addition, AKT has been shown to be overexpressed
in many forms of human cancer including, but not limited to, colon
(Zinda, et al, Clin. Cancer Res., 2001, 7:2475), ovarian (Cheng, J.
Q., et al., Proc. Natl. Acad. Sci. USA, 1992, 89:9267), brain (Haas
Kogan, D., et al, Curr. Biol., 1998, 8:1195), lung (Brognard, J.,
et al, Cancer Res., 2001, 61:3986), pancreatic (Cheng, J. Q., et
al., Proc. Natl. Acad. Sci., 1996, 93:3636), prostate (Graff, J.
R., et al, J. Biol. Chem., 2000, 275:24500) and gastric carcinomas
(Staal, S. P., et al., Proc. Natl. Acad. Sci. USA, 1987,
84:5034).
[0015] The development of kinase inhibitors that target abnormally
regulated pathways and ultimately result in disease is of enormous
ethical and commercial interest to the medical and pharmaceutical
community. As such, a compound that inhibits (1) recruitment of AKT
to the cell membrane, (2) activation by PDK1 or PDK2, (3) substrate
phosphorylation, or (4) one of the downstream targets of AKT would
therefore be a valid target as an anticancer agent, either as a
stand-alone therapy or in conjunction with other accepted
procedures.
SUMMARY OF THE INVENTION
[0016] This invention provides novel compounds that inhibit AKT
protein kinases, methods for producing these compounds, and
pharmaceutical compositions containing such compounds. The
compounds of the present invention have utility as therapeutic
agents for diseases and conditions that can be treated by the
inhibition of AKT protein kinases. More specifically, the present
invention includes compounds, including resolved enantiomers and
diastereomers, and pharmaceutically acceptable prodrugs,
metabolites, salts and solvates thereof, having the general Formula
I:
A-L-CR (I)
where:
[0017] CR is heteroaryl, wherein said heteroaryl is optionally
substituted with one or more groups selected from halogen,
hydroxyl, cyano, nitro, azido, --NR.sup.21SO.sub.2R.sup.24,
--SO.sub.2NR.sup.21R.sup.22, --NR.sup.21S(O)R.sup.4,
--S(O)NR.sup.21R.sup.22, --C(O)R.sup.21, --C(O)OR.sup.21,
--OC(O)R.sup.21, --OC(O)OR.sup.21, NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl, heteroaryl, arylalkyl and
heteroarylalkyl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl
are further optionally substituted with one or more groups selected
from halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,
difluoromethyl, trifluoromethyl, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6
heterocycloalkyl, --SR.sup.21, --S(O)R.sup.24, --SO.sub.2R.sup.24,
--C(O)R.sup.21, C(O)OR.sup.21, --C(O)NR.sup.21R.sup.22,
--NR.sup.21R.sup.22 and --OR.sup.21;
[0018] L is selected from:
##STR00001## ##STR00002##
[0019] R.sup.12 is hydrogen, halogen, hydroxy, cyano, nitro, amino,
azido, C.sub.1-C.sub.5 alkyl, C.sub.2-C.sub.5 alkenyl,
C.sub.2-C.sub.5 alkynyl, C.sub.1-C.sub.5 heteroalkyl,
C.sub.2-C.sub.5 heteroalkenyl or C.sub.2-C.sub.5 heteroalkynyl,
wherein any of said alkyl, alkenyl, alkynyl, heteroalkyl,
heteroalkenyl, and heteroalkynyl are optionally substituted with
one or more groups independently selected from halogen, cyano,
nitro, amino, azido, C.sub.1-C.sub.4 alkyl, fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy and trifluoromethoxy;
##STR00003##
[0020] A is
[0021] W is N or CR.sup.15, provided that when L is a substituted
or unsubstituted piperazinylene, W must be CR.sup.15;
[0022] G is hydrogen, alkyl, heteroalkyl, alkenyl, heteroalkenyl,
alkynyl, heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl is optionally substituted with
one or more groups selected from halogen, hydroxyl, cyano, amino,
nitro, azido, --NR.sup.21SO.sub.2R.sup.24,
--SO.sub.2NR.sup.21R.sup.22, --NR.sup.21S(O)R.sup.4,
--S(O)NR.sup.21R.sup.22, --C(O)R.sup.21, --C(O)OR.sup.21,
--OC(O)R.sup.21, --OC(O)OR.sup.21, --NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl;
[0023] B.sup.1 and B.sup.2 are independently absent or
C.sub.1-C.sub.4 alkylene, C.sub.1-C.sub.4 heteroalkylene,
C.sub.2-C.sub.4 alkenylene, C.sub.2-C.sub.4 heteroalkenylene,
C.sub.2-C.sub.4 alkynylene, C.sub.2-C.sub.4 heteroalkynylene,
C.sub.3-C.sub.6 cycloalkylene, and C.sub.3-C.sub.6
heterocycloalkylene, wherein any of said alkylene, heteroalkylene,
alkenylene, heteroalkenylene, alkynylene, heteroalkynylene,
cycloalkylene or heterocycloalkylene is optionally substituted with
one or more groups independently selected from halogen, hydroxyl,
cyano, nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4
heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21;
[0024] R.sup.21, R.sup.22 and R.sup.23 independently are hydrogen,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6
heteroalkenyl, C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl, arylalkyl,
heteroaryl or heteroarylalkyl;
[0025] R.sup.24 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 heteroalkyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 heteroalkynyl,
C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl,
arylalkyl, heteroaryl or heteroarylalkyl;
[0026] or any two of R.sup.21, R.sup.22, R.sup.23 or R.sup.24
together with the atom(s) to which they are attached form a 4 to 10
membered carbocyclic, aryl, heteroaryl or heterocyclic ring,
wherein any of said carbocyclic, aryl, heteroaryl or heterocyclic
rings are optionally substituted with one or more groups
independently selected from halogen, cyano, nitro, trifluoromethyl,
difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl,
arylalkyl, heteroarylalkyl, heterocyclyl, and
heterocyclylalkyl;
[0027] R.sup.13 and R.sup.14 are independently hydrogen, hydroxyl,
cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 heteroalkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.2-C.sub.6 heteroalkynyl,
C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6 heterocycloalkyl, aryl,
heteroaryl, arylalkyl, heteroarylalkyl, --C(O)R.sup.21,
C(O)OR.sup.21, C(.dbd.NR.sup.21)NR.sup.22R.sup.23 or
--SO.sub.2R.sup.24, wherein any of said alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroalkyl,
heteroalkenyl, heteroalkynyl, arylalkyl or heteroarylalkyl is
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl;
[0028] or R.sup.13 and R.sup.14 together with the atoms to which
they are attached form a 4 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21;
[0029] or R.sup.13 and an atom of B.sup.2 together with N form a 4
to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,
wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic
rings are optionally substituted with one or more groups
independently selected from halogen, cyano, nitro, azido,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21;
[0030] R.sup.15 is hydrogen, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl or C.sub.2-C.sub.4
heteroalkynyl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl or heteroalkynyl is optionally substituted
with one or more groups independently selected from halogen,
hydroxyl, cyano, nitro, azido, NR.sup.21R.sup.22 and OR.sup.21;
[0031] or R.sup.13 and R.sup.15 together with atoms to which they
are attached form a 3 to 10 membered carbocyclic, aryl, heteroaryl
or heterocyclic ring, wherein any of said carbocyclic, aryl,
heteroaryl and heterocyclic rings are optionally substituted with
one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21;
[0032] or, when W is CR.sup.15, R.sup.15 and an atom of B.sup.1 or
B.sup.2 together with C, form a 3 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21.
[0033] The invention also relates to pharmaceutical compositions
comprising an effective amount of an agent selected from compounds
of Formula I. Methods of making the compounds of Formula I are also
described.
[0034] In a further embodiment, the present invention provides
methods of inhibiting the activity of AKT protein kinases utilizing
compounds of Formula I.
[0035] In a further embodiment, the present invention provides a
method of treating diseases or medical conditions mediated by AKT
protein kinases. For example, this invention provides a method for
treatment of a hyperproliferative disorder in a warm-blooded animal
which comprises administering to such animal one or more compounds
of Formula I, or a pharmaceutically acceptable salt or in vivo
cleavable prodrug thereof in an amount effective to treat or
prevent said hyperproliferative disorder.
[0036] In a further embodiment, the present invention provides a
method of inhibiting the production of AKT protein kinases, which
comprises administering to a warm-blooded animal an effective
amount of a compound of Formula I, or a pharmaceutically acceptable
salt or in vivo cleavable prodrug thereof in an amount effective to
inhibit production of an AKT protein kinase.
[0037] In a further embodiment, the present invention provides a
method of providing AKT protein kinase inhibiting effect comprising
administering to a warm-blooded animal an effective amount of a
compound of Formula I, or a pharmaceutically acceptable salt or in
vivo cleavable prodrug thereof.
[0038] In a further embodiment, the present invention provides
treating or preventing an AKT protein kinase mediated condition,
comprising administering to a mammal a compound having Formula I or
a pharmaceutically-acceptable salt, in vivo cleavable prodrug or
pharmaceutical formulation thereof, in an amount effective to treat
or prevent said AKT protein kinase-mediated condition. AKT protein
kinase mediated conditions that can be treated according to the
methods of this invention include, but are not limited to, cancer,
inflammation and various proliferative, cardiovascular,
neurodegenerative, gynecological & dermatological diseases.
[0039] Hyperproliferative conditions that can be treated according
to the methods of this invention include, but are not limited to,
cancers of the head, neck, lung, breast, colon, ovary, bladder,
stomach, esophagus, uterus or prostate, among other kinds of
hyperproliferative disorders. In compounds and methods of this
invention can be used to treat diseases and conditions, including
rheumatoid arthritis, osteoarthritis, endometriosis,
atherosclerosis, vein graft stenosis, peri-anastomatic prothetic
graft stenosis, prostate hyperplasia, chronic obstructive pulmonary
disease, psoriasis, inhibition of neurological damage due to tissue
repair, scar tissue formation (and can aid in wound healing),
multiple sclerosis, inflammatory bowel disease, infections,
particularly bacterial, viral, retroviral or parasitic infections
(by increasing apoptosis), pulmonary disease, neoplasm, Parkinson's
disease, transplant rejection (as an immunosupressant), macular
degeneration and septic shock.
[0040] The compounds of Formula I may be used advantageously in
combination with other known therapeutic agents.
[0041] Additional advantages and novel features of this invention
shall be set forth in part in the description that follows, and in
part will become apparent to those skilled in the art upon
examination of the following specification, or may be learned by
the practice of the invention. The advantages of the invention may
be realized and attained by means of the instrumentalities,
combinations, compositions, and methods particularly pointed out in
the appended claims.
BRIEF DESCRIPTION OF THE FIGURES
[0042] The accompanying drawings, which are incorporated herein and
form a part of the specification, illustrate non-limiting
embodiments of the present invention, and together with the
description, serve to explain the principles of the invention.
[0043] In the Figures:
[0044] FIG. 1 shows a reaction scheme for the preparation of
compounds 8-11.
[0045] FIG. 2 shows a reaction scheme for the preparation of
compound 14.
[0046] FIG. 3 shows a reaction scheme for the preparation of
compound 18.
[0047] FIG. 4 shows a reaction scheme for the preparation of
compound 22.
[0048] FIG. 5 shows a reaction scheme for the preparation of
compound 28.
[0049] FIG. 6 shows a reaction scheme for the preparation of
compound 31.
[0050] FIG. 7 shows a reaction scheme for the preparation of
compounds 35 and 36.
[0051] FIG. 8 shows a reaction scheme for the preparation of
compounds 39 and 40.
[0052] FIG. 9 shows a reaction scheme for the preparation of
compound 40.
[0053] FIG. 10 shows a reaction scheme for the preparation of
compound 46.
[0054] FIG. 11 shows a reaction scheme for the preparation of
compound 50.
[0055] FIG. 12 shows a reaction scheme for the preparation of
compound 55.
[0056] FIG. 13 shows a reaction scheme for the preparation of
compounds 57 and 58.
[0057] FIG. 14 shows a reaction scheme for the preparation of
compounds 60 and 61.
[0058] FIG. 15 shows a reaction scheme for the preparation of
compounds 69-71.
[0059] FIG. 16 shows a reaction scheme for the preparation of
compounds 72-74.
[0060] FIG. 17 shows a reaction scheme for the preparation of
compound 78.
[0061] FIG. 18 shows a reaction scheme for the preparation of
compounds 80 and 81.
[0062] FIG. 19 shows a reaction scheme for the preparation of
compounds 85 and 86.
[0063] FIG. 20 shows a reaction scheme for the preparation of
compound 90.
[0064] FIG. 21 shows a reaction scheme for the preparation of
compounds 93-97.
[0065] FIG. 22 shows a reaction scheme for the preparation of
compounds 100 and 101.
[0066] FIG. 23 shows a reaction scheme for the preparation of
compounds 104-109.
[0067] FIG. 24 shows a reaction scheme for the preparation of
compounds 112-116.
[0068] FIG. 25 shows a reaction scheme for the preparation of
compounds 120-125.
[0069] FIG. 26 shows a reaction scheme for the preparation of
compounds 127 and 129.
[0070] FIG. 27 shows a reaction scheme for the preparation of
compounds 132 and 134.
[0071] FIG. 28 shows a reaction scheme for the preparation of
compounds 137 and 139.
[0072] FIG. 29 shows a reaction scheme for the preparation of
compounds 141-144.
[0073] FIG. 30 shows a reaction scheme for the preparation of
compound 148.
[0074] FIG. 31 shows a reaction scheme for the preparation of
compounds 151-153.
[0075] FIG. 32 shows a reaction scheme for the preparation of
compounds 155 and 156.
[0076] FIG. 33 shows a reaction scheme for the preparation of
compounds 161 and 162.
[0077] FIG. 34 shows a reaction scheme for the preparation of
compounds 171-175.
[0078] FIG. 35 shows a reaction scheme for the preparation of
compounds 178-182.
[0079] FIG. 36 shows a reaction scheme for the preparation of
compounds 179.
[0080] FIG. 37 shows a reaction scheme for the preparation of
compound 190.
[0081] FIG. 38 shows a reaction scheme for the preparation of
compounds 197-199.
[0082] FIG. 39 shows a reaction scheme for the preparation of
compounds 205-208.
[0083] FIG. 40 shows a reaction scheme for the preparation of
compounds 215 and 217.
[0084] FIG. 41 shows a reaction scheme for the preparation of
compounds 219, 221 and 223.
[0085] FIG. 42 shows a reaction scheme for the preparation of
compound 229.
[0086] FIG. 43 shows a reaction scheme for the preparation of
compounds 232 and 234.
[0087] FIG. 44 shows a reaction scheme for the preparation of
compounds 237-242.
[0088] FIG. 45 shows a reaction scheme for the preparation of
compounds 244 and 247.
[0089] FIG. 46 shows a reaction scheme for the preparation of
compounds 250, 251 and 254-256.
[0090] FIG. 47 shows a reaction scheme for the preparation of
compounds 263 and 265.
[0091] FIG. 48 shows a reaction scheme for the preparation of
compounds 269 and 271.
DETAILED DESCRIPTION OF THE INVENTION
[0092] The inventive compounds of Formula I are useful for
inhibiting AKT protein kinases. The compounds of Formula I may also
be useful as inhibitors of tyrosine kinases as well as serine and
threonine kinases in addition to AKT. Such compounds have utility
as therapeutic agents for diseases that can be treated by the
inhibition of the AKT protein kinase signaling pathway and tyrosine
and serine/threonine kinase receptor pathways. In general, the
invention includes compounds, including resolved enantiomers and
diastereomers, and pharmaceutically acceptable prodrugs,
metabolites, salts and solvates thereof, having the general Formula
I:
AA-L-CR I
[0093] where CR is heteroaryl, wherein said heteroaryl is
optionally substituted with one or more groups selected from
halogen, hydroxyl, cyano, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl, heteroaryl, arylalkyl and
heteroarylalkyl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl
are further optionally substituted with one or more groups selected
from halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,
difluoromethyl, trifluoromethyl, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6
heteroalkyl, C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6
heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6
heterocycloalkyl, --SR.sup.21, --S(O)R.sup.24, --SO.sub.2R.sup.24,
--C(O)R.sup.21, C(O)OR.sup.21, --C(O)NR.sup.21R.sup.22,
--NR.sup.21R.sup.22 and --OR.sup.21;
[0094] L is selected from:
##STR00004## ##STR00005##
[0095] R.sup.12 is hydrogen, halogen, hydroxy, cyano, nitro, amino,
azido, C.sub.1-C.sub.5 alkyl, C.sub.2-C.sub.5 alkenyl,
C.sub.2-C.sub.5 alkynyl, C.sub.1-C.sub.5 heteroalkyl,
C.sub.2-C.sub.5 heteroalkenyl or C.sub.2-C.sub.5 heteroalkynyl,
wherein any of said alkyl, alkenyl, alkynyl, heteroalkyl,
heteroalkenyl, and heteroalkynyl are optionally substituted with
one or more groups independently selected from halogen, cyano,
nitro, amino, azido, C.sub.1-C.sub.4 alkyl, fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy and trifluoromethoxy;
##STR00006##
[0096] A is
[0097] W is N or CR.sup.15, provided that when L is a substituted
or unsubstituted piperazinylene, W must be CR.sup.15;
[0098] G is hydrogen, alkyl, heteroalkyl, alkenyl, heteroalkenyl,
alkynyl, heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or
heteroaryl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl is optionally substituted with
one or more groups selected from halogen, hydroxyl, cyano, amino,
nitro, azido, --NR.sup.21SO.sub.2R.sup.24,
--SO.sub.2NR.sup.21R.sup.22, --NR.sup.21S(O)R.sup.4,
--S(O)NR.sup.21R.sup.22, --C(O)R.sup.21, --C(O)OR.sup.21,
--OC(O)R.sup.21, --OC(O)OR.sup.21, --NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --N.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.2C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl;
[0099] B.sup.1 and B.sup.2 are independently absent or
C.sub.1-C.sub.4 alkylene, C.sub.1-C.sub.4 heteroalkylene,
C.sub.2-C.sub.4 alkenylene, C.sub.2-C.sub.4 heteroalkenylene,
C.sub.2-C.sub.4 alkynylene, C.sub.2-C.sub.4 heteroalkynylene,
C.sub.3-C.sub.6 cycloalkylene, and C.sub.3-C.sub.6
heterocycloalkylene, wherein any of said alkylene, heteroalkylene,
alkenylene, heteroalkenylene, alkynylene, heteroalkynylene,
cycloalkylene or heterocycloalkylene is optionally substituted with
one or more groups independently selected from halogen, hydroxyl,
cyano, nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4
heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21;
[0100] R.sup.21, R.sup.22 and R.sup.23 independently are hydrogen,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6
heteroalkenyl, C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl, arylalkyl,
heteroaryl or heteroarylalkyl;
[0101] R.sup.24 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 heteroalkyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 heteroalkynyl,
C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl,
arylalkyl, heteroaryl or heteroarylalkyl;
[0102] or any two of R.sup.21, R.sup.22, R.sup.23 or R.sup.24
together with the atom(s) to which they are attached form a 4 to 10
membered carbocyclic, aryl, heteroaryl or heterocyclic ring,
wherein any of said carbocyclic, aryl, heteroaryl or heterocyclic
rings are optionally substituted with one or more groups
independently selected from halogen, cyano, nitro, trifluoromethyl,
difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl,
arylalkyl, heteroarylalkyl, heterocyclyl, and
heterocyclylalkyl;
[0103] R.sup.13 and R.sup.14 are independently hydrogen, hydroxyl,
cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 heteroalkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.2-C.sub.6 heteroalkynyl,
C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6 heterocycloalkyl, aryl,
heteroaryl, arylalkyl, heteroarylalkyl, --C(O)R.sup.21,
C(O)OR.sup.21, C(.dbd.NR.sup.21)NR.sup.22R.sup.23 or
--SO.sub.2R.sup.24, wherein any of said alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroalkyl,
heteroalkenyl, heteroalkynyl, arylalkyl or heteroarylalkyl is
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl aryl
and heteroaryl;
[0104] or R.sup.13 and R.sup.14 together with the atoms to which
they are attached form a 4 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21;
[0105] or R.sup.13 and an atom of B.sup.2 together with N form a 4
to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,
wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic
rings are optionally substituted with one or more groups
independently selected from halogen, cyano, nitro, azido,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, NR.sup.21R.sup.22 and OR.sup.21;
[0106] R.sup.15 is hydrogen, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4
heteroalkyl, C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4
heteroalkenyl, C.sub.2-C.sub.4 alkynyl or C.sub.2-C.sub.4
heteroalkynyl, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl or heteroalkynyl is optionally substituted
with one or more groups independently selected from halogen,
hydroxyl, cyano, nitro, azido, NR.sup.21R.sup.22 and OR.sup.21;
[0107] or R.sup.13 and R.sup.15 together with atoms to which they
are attached form a 3 to 10 membered carbocyclic, aryl, heteroaryl
or heterocyclic ring, wherein any of said carbocyclic, aryl,
heteroaryl and heterocyclic rings are optionally substituted with
one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21;
[0108] or, when W is CR.sup.15, R.sup.15 and an atom of B.sup.1 or
B.sup.2 together with C, form a 3 to 10 membered carbocyclic, aryl,
heteroaryl or heterocyclic ring, wherein any of said carbocyclic,
aryl, heteroaryl and heterocyclic rings are optionally substituted
with one or more groups independently selected from halogen, cyano,
nitro, azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
NR.sup.21R.sup.22 and OR.sup.21.
[0109] In one embodiment of the invention, CR is selected from:
##STR00007##
[0110] where X is N or CR.sup.1;
[0111] Y is CR.sup.2 or N, provided that when X is N, Y must be
CR.sup.2;
[0112] Z is CR.sup.3R.sup.3a or NR.sup.2a, provided that when X is
N, Z must be CR.sup.3;
[0113] D.sup.1, D.sup.2, D.sup.3 and D.sup.4 are independently
CR.sup.4 or N, provided that no more than two of D.sup.1, D.sup.2,
D.sup.3 or D.sup.4 are N;
[0114] - - - - - is an optional double bond;
[0115] D.sup.5 is CR.sup.5R.sup.5a, NR.sup.2a, O or S, provided
that when D.sup.5 is O or S, D.sup.8 must be C, D.sup.7 must be
CR.sup.7 or N, and either (i) Q must be CR.sup.6 or
CR.sup.6R.sup.6a or (ii) D.sup.7 must be CR.sup.7 or
CR.sup.7R.sup.7a;
[0116] Q is CR.sup.6, N or C.dbd.O, provided that either (w) when Q
is N, one of D.sup.5, D.sup.7 and D.sup.8 must be C, or (x) when Q
is C.dbd.O, D.sup.5 must be CR.sup.5 or N, D.sup.7 must be CR.sup.7
or N, and D.sup.8 must be C;
[0117] D.sup.7 is CR.sup.7, N, O or S, provided that when D.sup.7
is O or S, D.sup.8 must be C, D.sup.5 must be CR.sup.5 or N, and
either (y) Q must be CR.sup.6, or (z) D.sup.5 must be CR.sup.5;
[0118] D.sup.8 is C or N, provided that when D.sup.8 is N, D.sup.5
must be CR.sup.5R.sup.5a and Q must be CR.sup.6 or
CR.sup.6R.sup.6a;
[0119] either K or M is carbonyl, provided that both K and M are
not carbonyl;
[0120] R.sup.1, R.sup.5, R.sup.5a and R.sup.8 are independently
hydrogen, halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy, trifluoromethoxy, amino, aminomethyl,
dimethylamino, aminoethyl, diethylamino or ethoxy;
[0121] R.sup.2 is hydrogen, halogen, hydroxyl, cyano, nitro, amino,
azido, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.2-C.sub.4 heteroalkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.2-C.sub.4 heteroalkynyl,
C.sub.1-C.sub.6 cycloalkyl, C.sub.1-C.sub.6 heterocycloalkyl,
C.sub.1-C.sub.6 aryl, or C.sub.1-C.sub.6 heteroaryl, wherein any of
said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,
heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl are
further optionally substituted with one or more groups
independently selected from halogen, hydroxyl, cyano, nitro, azido,
fluoromethyl, difluoromethyl, trifluoromethyl, methoxy,
fluoromethoxy, difluoromethoxy, trifluoromethoxy, amino,
aminomethyl, dimethylamino, aminoethyl, diethylamino and
ethoxy;
[0122] R.sup.2a is hydrogen, hydroxyl, cyano, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl or heteroaryl, wherein any
of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,
heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl are
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, nitro, azido fluoromethyl,
difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,
difluoromethoxy, trifluoromethoxy, amino, aminomethyl,
dimethylamino, aminoethyl, diethylamino or ethoxy;
[0123] R.sup.3 and R.sup.3a are independently hydrogen, halogen,
hydroxyl, cyano, nitro, amino azido, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl,
C.sub.1-C.sub.6 heterocycloalkyl, aryl or heteroaryl, wherein any
of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,
heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl are
further optionally substituted with one or more groups
independently selected from halogen, hydroxyl, cyano, nitro, azido,
OR.sup.1, NR.sup.1R.sup.2, and (C.dbd.O)R.sup.2;
[0124] R.sup.4, R.sup.6, R.sup.6a, R.sup.7, R.sup.7a and R.sup.10
are independently hydrogen, hydroxyl, cyano, amino, nitro, azido,
alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,
cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, --NR.sup.21SO.sub.2R.sup.24,
--SO.sub.2NR.sup.21R.sup.22, --NR.sup.21S(O)R.sup.4,
--S(O)NR.sup.21R.sup.22, --C(O)R.sup.21, --C(O)OR.sup.21,
--OC(O)R.sup.21, --OC(O)OR.sup.21, --NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(NR.sup.21)NR.sup.22R.sup.23, --NR.sup.21C(O)R.sup.22,
--C(O)NR.sup.21R.sup.22, --SR.sup.21, --S(O)R.sup.24,
--SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
NR.sup.21C(O)NR.sup.22R.sup.23, --NR.sup.21C(NCN)NR.sup.22R.sup.23
or --OR.sup.21, wherein any of said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl,
is optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
--NR.sup.21SO.sub.2R.sup.24, --SO.sub.2NR.sup.21R.sup.22,
--NR.sup.21S(O)R.sup.4, --S(O)NR.sup.21R.sup.22, --C(O)R.sup.21,
--C(O)OR.sup.21, --OC(O)R.sup.21, --OC(O)OR.sup.21,
--NR.sup.21C(O)OR.sup.24,
--NR.sup.21C(.dbd.NR.sup.21)NR.sup.22R.sup.23,
--NR.sup.21C(O)R.sup.22, --C(O)NR.sup.21R.sup.22, --SR.sup.21,
--S(O)R.sup.24, --SO.sub.2R.sup.24, --NR.sup.21R.sup.22,
--NR.sup.21C(O)NR.sup.22R.sup.23,
--NR.sup.21C(NCN)NR.sup.22R.sup.23, --OR.sup.21, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl
and heteroaryl, and wherein said alkyl, heteroalkyl, alkenyl,
heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl may optionally be further
optionally substituted with one or more groups independently
selected from halogen, hydroxyl, cyano, amino, nitro, azido,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 heteroalkyl, C.sub.2-C.sub.4
alkenyl, C.sub.2-C.sub.4 heteroalkenyl, C.sub.2-C.sub.4 alkynyl,
C.sub.2-C.sub.4 heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl
and heteroaryl, --NR.sup.21R.sup.22 and --OR.sup.21;
[0125] or R.sup.6 and R.sup.7 together with the atoms to which they
are attached form a 4 to 10 membered carbocyclic, aryl, heteroaryl
or heterocyclic ring, wherein any of said carbocyclic, aryl,
heteroaryl and heterocyclic rings are optionally substituted with
one or more groups independently selected from halogen, cyano,
nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,
OR.sup.1, NR.sup.1R.sup.2, cycloalkyl, heterocycloalkyl, aryl,
heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, and
heterocyclylalkyl; and
[0126] R.sup.9 is hydrogen, halogen, hydroxyl, cyano, nitro, azido,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 heteroalkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 heteroalkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.6 heteroalkynyl, C.sub.1-C.sub.6 cycloalkyl,
C.sub.1-C.sub.6 heterocycloalkyl, aryl, heteroaryl,
--NR.sup.21R.sup.22, --OR.sup.21, --NR.sup.21SO.sub.2R.sup.24 and
--NR.sup.21C(O)R.sup.22, wherein any of said alkyl, heteroalkyl,
alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,
heterocycloalkyl, aryl or heteroaryl are further optionally
substituted with one or more groups independently selected from
halogen, hydroxyl, cyano, nitro, azido, OR.sup.1, NR.sup.1R.sup.2,
and (C.dbd.O)R.sup.2.
[0127] In one embodiment of the invention, CR is selected from
##STR00008##
[0128] where D.sup.5, D.sup.7, D.sup.8, X and Q are as defined
above.
[0129] According to another embodiment of the invention, A is:
##STR00009##
[0130] where B.sup.1 and B.sup.2 are, independently, absent or
C.sub.1-C.sub.4 alkylene;
[0131] R.sup.21a--R.sup.21c are independently H, halogen, CH.sub.3,
CF.sub.3, CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph, OH, or
OCH.sub.2Ph;
[0132] R.sup.22a, R.sup.22b, and R.sup.24 are independently H,
CH.sub.3, or halogen;
[0133] R.sup.23a is H; and
[0134] R.sup.23b is H, CH.sub.3, CH.sub.2NH.sub.2,
CH.sub.2NHCH.sub.2, CH.sub.2CH.sub.2NH.sub.2,
CH.sub.2CH.sub.2NHCH.sub.2, CH.sub.2CH.sub.2N(CH.sub.2).sub.2,
--(C.dbd.O)CH.sub.2NH.sub.2 or
--(C.dbd.O)CH.sub.2CH.sub.2NH.sub.2;
[0135] or R.sup.23a and R.sup.23b are joined to complete a 5 or 6
membered heterocyclic ring.
[0136] In yet another embodiment of this invention, A is
##STR00010##
[0137] where R.sup.25 and R.sup.26 are independently H or CH.sub.3;
and
[0138] R.sup.27 is 1-naphthyl, 2-naphthyl, 3'-benzylthienyl,
2'-thienyl, 2'-pyridyl, 3'-pyridyl, 4'-pyridyl, 4'-thiazolyl, or
3,3-diphenyl.
[0139] More specific examples of the A group of Formula I according
to this invention include, but are not limited to,
##STR00011##
[0140] where G, B.sup.1, R.sup.13 and R.sup.14 are as defined
above.
[0141] In an alternative embodiment, the A group of Formula I of
this invention is a D- or L-amino acid selected from the 20
naturally occurring amino acids commonly designated by three letter
symbols, and also includes unnatural amino acids including, but not
limited to, 4-hydroxyproline, hydroxylysine, demosine, isodemosine,
3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric
acid, cirtulline, homocysteine, homoserine, ornithine and
methionine sulfone. In one preferred embodiment, the A group of
Formula I is alanine, phenylalanine, histidine, or tryptophan.
[0142] A specific example of a compound based on Formula I is:
##STR00012##
[0143] Another example of a compound based on Formula I is:
##STR00013##
[0144] wherein R.sup.28 is H, halogen, CH.sub.3, CF.sub.3,
CH.sub.3O, CN, NO.sub.2, NH.sub.2, Ph, OH, or OCH.sub.2Ph.
[0145] Still another example of a compound based on Formula I
is:
##STR00014##
[0146] where D.sup.16 is O or N; and
[0147] R.sup.29 is H, halogen, CH.sub.3, CF.sub.3, CH.sub.3O, CN,
NO.sub.2, NH.sub.2, Ph, OH, or OCH.sub.2Ph.
[0148] The term "alkyl" as used herein refers to a saturated linear
or branched-chain monovalent hydrocarbon radical of one to twelve
carbon atoms, wherein the alkyl radical may be optionally
substituted independently with one or more substituents described
below. Examples of alkyl groups include, but are not limited to,
methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl,
tert-butyl, pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and
the like.
[0149] The term "alkylene" as used herein refers to a linear or
branched-chain saturated divalent hydrocarbon radical of one to
twelve carbon atoms, e.g., methylene, ethylene, propylene,
2-methylpropylene, pentylene, and the like. The alkylene radical
may be optionally substituted independently with one or more
substituents described herein.
[0150] The term "heteroalkyl" refers to saturated linear or
branched-chain monovalent hydrocarbon radical of one to twelve
carbon atoms, wherein at least one of the carbon atoms is replaced
with a heteroatom selected from N, O, or S, and wherein the radical
may be a carbon radical or heteroatom radical (i.e., the heteroatom
may appear in the middle or at the end of the radical). The
heteroalkyl radical may be optionally substituted independently
with one or more substituents described herein. The term
"heteroalkyl" encompasses alkoxy and heteroalkoxy radicals.
[0151] The term "heteroalkylene" as used herein refers to a linear
or branched-chain saturated divalent hydrocarbon radical of two to
twelve carbon atoms, wherein at least one of the carbon atoms is
replaced with a heteroatom selected from N, O, or S, and wherein
the radical may be a carbon radical or heteroatom radical (i.e.,
the heteroatom may appear in the middle or at the end of the
radical). The heteroalkylene radical may be optionally substituted
independently with one or more substituents described herein.
[0152] "Alkenyl" means a linear or branched-chain monovalent
hydrocarbon radical of two to twelve carbon atoms containing at
least one double bond, wherein the alkenyl radical may be
optionally substituted independently with one or more substituents
described below. Examples of alkenyl groups include, but are not
limited to: ethylene or vinyl (--CH.dbd.CH.sub.2), allyl
(--CH.sub.2CH.dbd.CH.sub.2), 1-cyclopent-1-enyl,
1-cyclopent-2-enyl, 1-cyclopent-3-enyl, 5-hexenyl (--CH.sub.2
CH.sub.2CH.sub.2CH.sub.2CH.dbd.CH.sub.2), 1-cyclohex-1-enyl,
1-cyclohex-2-enyl, and 1-cyclohex-3-enyl.
[0153] "Alkenylene" refers to an a linear or branched-chain
divalent hydrocarbon radical of one to twelve carbon atoms
containing at least one double bond, e.g., 1,2-ethylene
(--CH.dbd.CH--). The alkenylene radical may be optionally
substituted independently with one or more substituents described
herein.
[0154] The term "heteroalkenyl" refers to a linear or
branched-chain monovalent hydrocarbon radical of two to twelve
carbon atoms and at least one double bond, wherein at least one of
the carbon atoms is replaced with a heteroatom selected from N, O,
or S, and wherein the radical may be a carbon radical or heteroatom
radical (i.e., the heteroatom may appear in the middle or at the
end of the radical). The heteroalkenyl radical may be optionally
substituted independently with one or more substituents described
herein. The term "heteroalkenyl" encompasses alkenoxy and
heteroalkenoxy radicals.
[0155] "Heteroalkenylene" refers to an a linear or branched
saturated divalent hydrocarbon radical of one to twelve carbon
atoms containing at least one double bond, wherein at least one of
the carbon atoms is replaced with a heteroatom selected from N, O,
or S, and wherein the radical may be a carbon radical or heteroatom
radical (i.e., the heteroatom may appear in the middle or at the
end of the radical). The heteroalkenylene radical may be optionally
substituted independently with one or more substituents described
herein.
[0156] The term "allyl" refers to a radical having the formula
RC.dbd.CHCHR, wherein R is alkyl, alkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, or any substituent as defined
herein, wherein the allyl radical may be optionally substituted
independently with one or more substituents described herein.
[0157] The term "alkynyl" means a linear or branched-chain
monovalent hydrocarbon radical of two to twelve carbon atoms
containing at least one triple bond, wherein the alkynyl radical
may be optionally substituted independently with one or more
substituents described below. Examples of alkynyl groups include,
but are not limited to: acetylene (--C.ident.CH) and propargyl
(--CH.sub.2C.ident.CH).
[0158] "Alkynylene" refers to a linear or branched-chain divalent
hydrocarbon radical of one to twelve carbon atoms containing at
least one triple bond. The alkynylene radical may be optionally
substituted independently with one or more substituents described
herein. Typical alkynylene radicals include, but are not limited
to: acetylene (--C.ident.C--), propargyl (--CH.sub.2C.ident.C--),
and 4-pentynyl (--CH.sub.2CH.sub.2CH.sub.2C.ident.CH--).
[0159] The term "heteroalkynyl" refers to a linear or
branched-chain monovalent hydrocarbon radical of two to twelve
carbon atoms containing at least one triple bond, wherein at least
one of the carbon atoms is replaced with a heteroatom selected from
N, O, or S, and wherein the radical may be a carbon radical or
heteroatom radical (i.e., the heteroatom may appear in the middle
or at the end of the radical). The heteroalkynyl radical may be
optionally substituted independently with one or more substituents
described herein. The term "heteroalkynyl" encompasses alkynoxy and
heteroalkynoxy radicals.
[0160] The term "heteroalkynylene" refers to a linear or branched
divalent hydrocarbon radical of two to twelve carbons containing at
least one triple bond, wherein at least one of the carbon atoms is
replaced with a heteroatom selected from N, O, or S, and wherein
the radical may be a carbon radical or heteroatom radical (i.e.,
the heteroatom may appear in the middle or at the end of the
radical). The heteroalkynylene radical may be optionally
substituted independently with one or more substituents described
herein.
[0161] The terms "carbocycle," "carbocyclyl," or "cycloalkyl" refer
to saturated or partially unsaturated cyclic hydrocarbon radical
having from three to ten carbon atoms. The term "cycloalkyl"
includes monocyclic and polycyclic (e.g., bicyclic and tricyclic)
cycloalkyl structures, wherein the polycyclic structures optionally
include a saturated or partially unsaturated cycloalkyl fused to a
saturated or partially unsaturated cycloalkyl or heterocycloalkyl
ring or an aryl or heteroaryl ring. Examples of cycloalkyl groups
include, but are not limited to, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, and the like. The cycloalkyl
may be optionally substituted independently in one or more
substitutable positions with various groups. For example, such
cycloalkyl groups may be optionally substituted with, for example,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen, hydroxy,
cyano, nitro, amino, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0162] The terms "heterocycloalkyl," "heterocycle" or
"heterocyclyl" refer to a saturated or partially unsaturated
carbocyclic radical of 3 to 8 ring atoms in which at least one ring
atom is a heteroatom selected from nitrogen, oxygen and sulfur, the
remaining ring atoms being C, where one or more ring atoms may be
optionally substituted independently with one or more substituent
described below. The radical may be a carbon radical or heteroatom
radical. The term further includes bicyclic and tricyclic fused
ring systems which include a heterocycle fused one or more
carbocyclic or heterocyclic rings. "Heterocycloalkyl" also includes
radicals where heterocycle radicals are fused with aromatic or
heteroaromatic rings. Examples of heterocycloalkyl rings include,
but are not limited to, pyrrolidinyl, tetrahydrofuranyl,
dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,
dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,
thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl,
azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl,
thiepanyl, oxazepinyl, diazepinyl, thiazepinyl,
1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl,
2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl,
dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,
dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl,
3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,
azabicyclo[2.2.2]hexanyl, 3H-indolyl and quinolizinyl. Spiro
moieties are also included within the scope of this definition. The
foregoing groups, as derived from the groups listed above, may be
C-attached or N-attached where such is possible. For instance, a
group derived from pyrrole may be pyrrol-1-yl (N-attached) or
pyrrol-3-yl (C-attached). Further, a group derived from imidazole
may be imidazol-1-yl (N-attached) or imidazol-3-yl (C-attached). An
example of a heterocyclic group wherein 2 ring carbon atoms are
substituted with oxo (.dbd.O) moieties is
1,1-dioxo-thiomorpholinyl. The heterocycle groups herein are
unsubstituted or, as specified, substituted in one or more
substitutable positions with various groups. For example, such
heterocycle groups may be optionally substituted with, for example,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen, hydroxy,
cyano, nitro, amino, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0163] The terms "heterocycloalkylene" refers to a saturated or
partially unsaturated divalent carbocyclic radical of 3 to 8 ring
atoms in which at least one ring atom is a heteroatom selected from
nitrogen, oxygen and sulfur, the remaining ring atoms being C,
where one or more ring atoms may be optionally substituted
independently with one or more substituent described herein.
Examples include, but are not limited to, substituted and
unsubstituted piperidinylenes.
[0164] The term "aryl" refers to a monovalent aromatic carbocyclic
radical having a single ring (e.g., phenyl), multiple rings (e.g.,
biphenyl), or multiple condensed rings in which at least one is
aromatic, (e.g., 1,2,3,4-tetrahydronaphthyl, naphthyl), which is
optionally mono-, di-, or trisubstituted with, e.g., halogen, lower
alkyl, lower alkoxy, trifluoromethyl, aryl, heteroaryl, and
hydroxy.
[0165] The term "heteroaryl" refers to a monovalent aromatic
radical of 5-, 6-, or 7-membered rings which includes fused ring
systems (at least one of which is aromatic) of 5-10 atoms
containing at least one and up to four heteroatoms selected from
nitrogen, oxygen, or sulfur. Examples of heteroaryl groups are
pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl,
pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,
oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl,
indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl,
indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl,
pteridinyl, purinyl, oxadiazolyl, triazolyl, thiadiazolyl,
thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,
benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,
naphthyridinyl, and furopyridinyl. Spiro moieties are also included
within the scope of this definition. Heteroaryl groups are
optionally mono-, di-, or trisubstituted with, e.g., halogen, lower
alkyl, lower alkoxy, haloalkyl, aryl, heteroaryl, and hydroxy.
[0166] The term "halo" represents fluoro, chloro, bromo or iodo.
Likewise, the term "halogen" refers to a fluorine, chlorine,
bromine, or iodine substituent.
[0167] The term "arylalkyl" means an alkyl moiety (as defined
above) substituted with one or more aryl moiety (also as defined
above). More preferred arylalkyl radicals are
aryl-C.sub.1-3-alkyls. Examples include benzyl, phenylethyl, and
the like.
[0168] The term "heteroarylalkyl" means an alkyl moiety (as defined
above) substituted with a heteroaryl moiety (also as defined
above). More preferred heteroarylalkyl radicals are 5- or
6-membered heteroaryl-C.sub.1-3-alkyls. Examples include, but are
not limited to, oxazolylmethyl, pyridylethyl and the like.
[0169] The term "heterocyclylalkyl" means an alkyl moiety (as
defined above) substituted with a heterocyclyl moiety (also defined
above). More preferred heterocyclylalkyl radicals are 5- or
6-membered heterocyclyl-Cl_3-alkyls. An example includes, but is
not limited to, tetrahydropyranylmethyl.
[0170] The term "cycloalkylalkyl" means an alkyl moiety (as defined
above) substituted with a cycloalkyl moiety (also defined above).
More preferred heterocyclyl radicals are 5- or 6-membered
cycloalkyl-C.sub.1-3-alkyls. An example includes, but is not
limited to, cyclopropylmethyl.
[0171] The term "Me" means methyl, "Et" means ethyl, "Bu" means
butyl and "Ac" means acetyl.
[0172] In general, the various moieties or functional groups of the
compounds of Formula I may be optionally substituted by one or more
substituents. Examples of substituents suitable for purposes of
this invention include, but are not limited to, halo, alkyl, allyl,
alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,
heteroalkynyl, alkoxy, heteroalkoxy, G.sub.n-cycloalkyl,
G.sub.n-heterocycloalkyl, G.sub.n-OR, G.sub.n-NO.sub.2, G.sub.n-CN,
G.sub.n-CO.sub.2R, G.sub.n-(C.dbd.O)R, G.sub.n-O(C.dbd.O)R,
G.sub.n-O-alkyl, G.sub.n-OAr, G.sub.n-SH, G-SR, G.sub.n-SOR,
G.sub.n-SO.sup.2R, G.sub.n-S--ArG.sub.n-SOAr, G.sub.n-O.sub.2Ar,
aryl, heteroaryl, G.sub.n-Ar, G.sub.n-(C.dbd.O)NR.sup.2R.sup.3,
G.sub.n-NR.sup.2R.sup.3, G.sub.n-NR(C.dbd.O)R, G.sub.n-SO.sub.2
NR.sup.2R.sup.3, PO.sub.3H.sub.2, SO.sub.3H.sub.2, where G is
alkylene having from 1 to 4 carbons, or alkenylene or alkynylene
each having from 2 to 4 carbons; n is zero or 1; R.sub.1, R.sup.2,
and R.sup.3 are alkyl, allyl, alkenyl, alkynyl, heteroalkyl,
heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy,
G.sub.n-cycloalkyl, or G.sub.n-heterocycloalkyl; and Ar is aryl or
heteroaryl, wherein said alkyl, allyl, alkenyl, alkynyl,
heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,
heteroalkoxy, G.sub.n-cycloalkyl, G.sub.n-heterocycloalkyl,
alkylene, alkenylene, alkynylene, Ar, R.sup.1, R.sup.2, and R.sup.3
may be further substituted or unsubstituted.
[0173] It is to be understood that in instances where two or more
radicals are used in succession to define a substituent attached to
a structure, the first named radical is considered to be terminal
and the last named radical is considered to be attached to the
structure in question. Thus, for example, the radical arylalkyl is
attached to the structure in question by the alkyl group.
[0174] The compounds of this invention may possess one or more
asymmetric centers; such compounds can therefore be produced as
individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless
indicated otherwise, the description or naming of a particular
compound in the specification and claims is intended to include
both individual enantiomers and mixtures, racemic or otherwise,
thereof. Accordingly, this invention also includes racemates and
resolved enantiomers, and diastereomers compounds of the Formula I.
Methods for determining the stereochemistry and for the separation
of stereoisomers are well known in the art (see discussion in
Chapter 4 of "Advanced Organic Chemistry", 4th edition J. March,
John Wiley and Sons, New York, 1992).
[0175] In addition to compounds of the Formula I, the invention
also includes solvates, pharmaceutically active metabolites,
pharmaceutically acceptable prodrugs, and pharmaceutically
acceptable salts of such compounds.
[0176] The term "solvate" refers to an aggregate of a molecule with
one or more solvent molecules.
[0177] A "pharmaceutically active metabolite" is a
pharmacologically active product produced through metabolism in the
body of a specified compound or salt thereof. Metabolites of a
compound may be identified using routine techniques known in the
art and their activities determined using tests such as those
described herein.
[0178] Compounds of the present invention having functional groups
including, but not limited to, free amino, amido, hydroxy or
carboxylic groups can be converted into pharmaceutically acceptable
prodrugs. A "pharmaceutically acceptable prodrug" is a compound
that may be converted under physiological conditions or by
solvolysis to the specified compound or to a pharmaceutically
acceptable salt of such compound. Prodrugs include compounds
wherein an amino acid residue, or a polypeptide chain of two or
more (e.g., two, three or four) amino acid residues is covalently
joined through an amide or ester bond to a free amino, hydroxy or
carboxylic acid group of compounds of the present invention. The
amino acid residues include but are not limited to the 20 naturally
occurring amino acids commonly designated by three letter symbols
and also includes 4-hydroxyproline, hydroxylysine, demosine,
isodemosine, 3-methylhistidine, norvaline, beta-alanine,
gamma-aminobutyric acid, cirtulline, homocysteine, homoserine,
ornithine and methionine sulfone. One preferred prodrug of this
invention is a compound of Formula I covalently joined to a
phosphate residue. Another preferred prodrug of this invention is a
compound of Formula I covalently joined to a valine residue.
[0179] Additional types of prodrugs are also encompassed. For
instance, free carboxyl groups can be derivatized as amides or
alkyl esters. As another example, compounds of this invention
comprising free hydroxy groups may be derivatized as prodrugs by
converting the hydroxy group groups including to a phosphate ester,
hemisuccinate, dimethylaminoacetate, or
phosphoryloxymethyloxycarbonyl, as outlined in Advanced Drug
Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and
amino groups are also included, as are carbonate prodrugs,
sulfonate esters and sulfate esters of hydroxy groups.
Derivatization of hydroxy groups as (acyloxy)methyl and
(acyloxy)ethyl ethers wherein the acyl group may be an alkyl ester,
optionally substituted with groups including, but not limited to,
ether, amine and carboxylic acid functionalities, or where the acyl
group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.,
1996, 39, 10. More specific examples include replacement of the
hydrogen atom of the alcohol group with a group such as
(C.sub.1-C.sub.6)alkanoyloxymethyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxycarbonyloxymethyl,
N--(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl, where each .alpha.-aminoacyl
group is independently selected from the naturally occurring
L-amino acids, P(O)(OH).sub.2,
--P(O)(O(C.sub.1-C.sub.6)alkyl).sub.2 or glycosyl (the radical
resulting from the removal of a hydroxyl group of the hemiacetal
form of a carbohydrate).
[0180] Free amines can also be derivatized as amides, sulfonamides
or phosphonamides. All of these prodrug moieties may incorporate
groups including, but not limited to, ether, amine and carboxylic
acid functionalities. For example, a prodrug can be formed by the
replacement of a hydrogen atom in the amine group with a group such
as R-carbonyl, RO-carbonyl, NRR'-- carbonyl where R and R' are each
independently (C.sub.1-C.sub.10)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
benzyl, or R-carbonyl is a natural .alpha.-aminoacyl or natural
.alpha.-aminoacyl-natural .alpha.-aminoacyl, --C(OH)C(O)OY wherein
Y is H, (C.sub.1-C.sub.6)alkyl or benzyl, --C(OY.sub.0)Y.sub.1
wherein Y.sub.0 is (C.sub.1-C.sub.4) alkyl and Y.sub.1 is
(C.sub.1-C.sub.6)alkyl, carboxy(C.sub.1-C.sub.6)alkyl,
amino(C.sub.1-C.sub.4)alkyl or mono-N-- or
di-N,N--(C.sub.1-C.sub.6)alkylaminoalkyl, --C(Y.sub.2)Y.sub.3
wherein Y.sub.2 is H or methyl and Y.sub.3 is mono-N-- or
di-N,N--(C.sub.1-C.sub.6)alkylamino, morpholino, piperidin-1-yl or
pyrrolidin-1-yl.
[0181] A "pharmaceutically acceptable salt" is a salt that retains
the biological effectiveness of the free acids and bases of the
specified compound and that is not biologically or otherwise
undesirable. A compound of the invention may possess a sufficiently
acidic, a sufficiently basic, or both functional groups, and
accordingly react with any of a number of inorganic or organic
bases, and inorganic and organic acids, to form a pharmaceutically
acceptable sale. Examples of pharmaceutically acceptable salts
include those salts prepared by reaction of the compounds of the
present invention with a mineral or organic acid or an inorganic
base, such salts including sulfates, pyrosulfates, bisulfates,
sulfites, bisulfites, phosphates, monohydrogenphosphates,
dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides,
bromides, iodides, acetates, propionates, decanoates, caprylates,
acrylates, formates, isobutyrates, caproates, heptanoates,
propiolates, oxalates, malonates, succinates, suberates, sebacates,
fumarates, maleates, butyn-1,4-dioates, hexyne-1,6-dioates,
benzoates, chlorobenzoates, methylbenzoates, dinitromenzoates,
hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,
xylenesulfonates, pheylacetates, phenylpropionates,
phenylbutyrates, citrates, lactates, .gamma.-hydroxybutyrates,
glycollates, tartrates, methanesulfonates, propanesulfonates,
naphthalene-1-sulfonates, naphthalene-2-sulfonates, and
mandelates.
[0182] If the inventive 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 alphahydroxy 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.
[0183] If the inventive 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, but are not
limited to, organic salts derived from amino acids, such as glycine
and arginine, ammonia, primary, secondary, and tertiary amines, and
cyclic amines, such as piperidine, morpholine and piperazine, and
inorganic salts derived from sodium, calcium, potassium, magnesium,
manganese, iron, copper, zinc, aluminum and lithium.
[0184] The inventive compounds may be prepared using the reaction
routes and synthesis schemes as described herein, employing the
techniques available in the art using starting materials that are
readily available.
Therapeutic Aspects of the Invention
[0185] The invention also provides a pharmaceutical composition for
the treatment of a hyperproliferative disorder in a mammal which
comprises a therapeutically effective amount of a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite or hydrate thereof, and a pharmaceutically acceptable
carrier. In one embodiment, said pharmaceutical composition is for
the treatment of cancer such as skin, brain, lung, squamous cell,
bladder, gastric, pancreatic, breast, head, neck, renal, kidney,
ovarian, prostate, colorectal, esophageal, testicular,
gynecological, cardiac, liver, bone, meninges, spinal cord, blood,
skin, adrenal or thyroid cancer. In another embodiment, said
pharmaceutical composition is for the treatment of a non-cancerous
hyperproliferative disorder such as benign hyperplasia of the skin
(e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic
hypertrophy (BPH)).
[0186] The invention also relates to a method for the treatment of
a hyperproliferative disorder in a mammal that comprises
administering to said mammal a therapeutically effective amount of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug or hydrate thereof, in combination with an
anti-tumor agent selected from the group consisting of mitotic
inhibitors, alkylating agents, anti-metabolites, intercalating
antibiotics, growth factor inhibitors, cell cycle inhibitors,
enzyme inhibitors, topoisomerase inhibitors, biological response
modifiers, anti-hormones, angiogenesis inhibitors, and
anti-androgens.
[0187] The invention also relates to a method of treating
pancreatitis or kidney disease in a mammal that comprises
administering to said mammal a therapeutically effective amount of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug or hydrate thereof.
[0188] The invention also relates to a method of preventing
blastocyte implantation in a mammal that comprises administering to
said mammal a therapeutically effective amount of a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug
or hydrate thereof.
[0189] The invention also relates to a method of treating diseases
related to vasculogenesis or angiogenesis in a mammal that
comprises administering to said mammal a therapeutically effective
amount of a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug or hydrate thereof. In
one embodiment, said method is for treating a disease selected from
the group consisting of tumor angiogenesis, chronic inflammatory
disease such as rheumatoid arthritis, atherosclerosis, inflammatory
bowel disease, skin diseases such as psoriasis, excema, and
scleroderma, diabetes, diabetic retinopathy, retinopathy of
prematurity, age-related macular degeneration, hemangioma, glioma,
melanoma, Kaposi's sarcoma and ovarian, breast, lung, pancreatic,
prostate, colon and epidermoid cancer.
[0190] The invention also relates to a pharmaceutical composition
for treating a disease or condition related to inflammatory
disease, autoimmune disease, destructive bone disorders,
proliferative disorders, infectious disease, viral disease,
fibrotic disease or neurodegenerative disease in a mammal which
comprises a therapeutically effective amount of a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug
or hydrate thereof, and a pharmaceutically acceptable carrier.
Examples of the above diseases and/or conditions include but is not
limited to rheumatoid arthritis, atherosclerosis, inflammatory
bowel disease, skin diseases such as psoriasis, eczema, and
scleroderma, diabetes and diabetic complications, diabetic
retinopathy, retinopathy of prematurity, age-related macular
degeneration, hemangioma, chronic obstructive pulmonary disease,
idiopathic pulmonary fibrosis, allergic responses including asthma
allergic rhinitis and atopic dermatitis, renal disease and renal
failure, polycystic kidney disease, acute coronary syndrome,
congestive heart failure, osteoarthritis, neurofibromatosis, organ
transplant rejection, cachexia and pain.
[0191] Further provided is a compound of Formula I for use as a
medicament in the treatment of the diseases and conditions
described above in a warm-blooded animal, preferably a mammal, more
preferably a human, suffering from such disorder. Also provided is
the use of a compound of Formula I in the preparation of a
medicament for the treatment of the diseases and conditions
described above in a warm-blooded animal, preferably a mammal, more
preferably a human, suffering from such disorder.
[0192] Patients that can be treated with compounds of the present
invention, or pharmaceutically acceptable salts, prodrugs and
hydrates of said compounds, according to the methods of this
invention include, for example, patients that have been diagnosed
as having psoriasis, restenosis, atherosclerosis, BPH, lung cancer,
bone cancer, CMML, pancreatic cancer, skin cancer, cancer of the
head and neck, cutaneous or intraocular melanoma, uterine cancer,
ovarian cancer, rectal cancer, cancer of the anal region, stomach
cancer, colon cancer, breast cancer, testicular, gynecologic tumors
(e.g., uterine sarcomas, carcinoma of the fallopian tubes,
carcinoma of the endometrium, carcinoma of the cervix, carcinoma of
the vagina or carcinoma of the vulva), Hodgkin's disease, cancer of
the esophagus, cancer of the small intestine, cancer of the
endocrine system (e.g., cancer of the thyroid, parathyroid or
adrenal glands), sarcomas of soft tissues, cancer of the urethra,
cancer of the penis, prostate cancer, chronic or acute leukemia,
solid tumors of childhood, lymphocytic lymphomas, cancer of the
bladder, cancer of the kidney or ureter (e.g., renal cell
carcinoma, carcinoma of the renal pelvis), or neoplasms of the
central nervous system (e.g., primary CNS lymphona, spinal axis
tumors, brain stem gliomas or pituitary adenomas).
[0193] This invention also relates to a pharmaceutical composition
for inhibiting abnormal cell growth in a mammal which comprises an
amount of a compound of the present invention, or a
pharmaceutically acceptable salt or solvate or prodrug thereof, in
combination with an amount of a chemotherapeutic, wherein the
amounts of the compound, salt, solvate, or prodrug, and of the
chemotherapeutic are together effective in inhibiting abnormal cell
growth. Many chemotherapeutics are presently known in the art. In
one embodiment, the chemotherapeutic is selected from the group
consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, antitumor antibiotics, growth factor inhibitors,
cell cycle inhibitors, enzymes, topoisomerase inhibitors,
biological response modifiers, anti-hormones, angiogenesis
inhibitors, and anti-androgens.
[0194] This invention further relates to a method for inhibiting
abnormal cell growth in a mammal or treating a hyperproliferative
disorder which method comprises administering to the mammal an
amount of a compound of the present invention, or a
pharmaceutically acceptable salt or solvate or prodrug thereof, in
combination with radiation therapy, wherein the amounts of the
compound, salt, solvate, or prodrug, is in combination with the
radiation therapy effective in inhibiting abnormal cell growth or
treating the hyperproliferative disorder in the mammal. Techniques
for administering radiation therapy are known in the art, and these
techniques can be used in the combination therapy described herein.
The administration of the compound of the invention in this
combination therapy can be determined as described herein.
[0195] It is believed that the compounds of the present invention
can render abnormal cells more sensitive to treatment with
radiation for purposes of killing and/or inhibiting the growth of
such cells. Accordingly, this invention further relates to a method
for sensitizing abnormal cells in a mammal to treatment with
radiation which comprises administering to the mammal an amount of
a compound of the present invention or pharmaceutically acceptable
salt or solvate or prodrug thereof, which amount is effective is
sensitizing abnormal cells to treatment with radiation. The amount
of the compound, salt, or solvate in this method can be determined
according to the means for ascertaining effective amounts of such
compounds described herein.
[0196] Compounds and methods of this invention may also be used to
treat other diseases and conditions (e.g., inflammatory disease),
including rheumatoid arthritis, osteoarthritis, endometriosis,
atherosclerosis, vein graft stenosis, peri-anastomatic prosthetic
graft stenosis, prostate hyperplasia, chronic obstructive pulmonary
disease, psoriasis, inhibition of neurological damage due to tissue
repair, scar tissue formation (and can aid in wound healing),
multiple sclerosis, inflammatory bowel disease, infections,
particularly bacterial, viral, retroviral or parasitic infections
(by increasing apoptosis), pulmonary disease, neoplasm, Parkinson's
disease, transplant rejection (as an immunosuppressant), macular
degeneration and septic shock.
[0197] Therapeutically effective amounts of the compounds of the
invention may be used to treat diseases mediated by modulation or
regulation of AKT protein kinases, tyrosine kinases, additional
serine/threonine kinases, and/or dual specificity kinases. An
"effective amount" is intended to mean that amount of compound
that, when administered to a mammal in need of such treatment, is
sufficient to effect treatment for a disease mediated by the
activity of one or more AKT protein kinases, tyrosine kinases,
additional serine/threonine kinases, and/or dual specificity
kinases. Thus, for example, a therapeutically effective amount of a
compound selected from Formula I or a salt, active metabolite or
prodrug thereof, is a quantity sufficient to modulate, regulate, or
inhibit the activity of one or more AKT protein kinases, tyrosine
kinases, additional serine/threonine kinases, and/or dual
specificity kinases such that a disease condition which is mediated
by that activity is reduced or alleviated.
[0198] The terms "abnormal cell growth" and "hyperproliferative
disorder" are used interchangeably in this application.
[0199] "Abnormal cell growth," as used herein, unless otherwise
indicated, refers to cell growth that is independent of normal
regulatory mechanisms (e.g., loss of contact inhibition). This
includes, for example, the abnormal growth of: (1) tumor cells
(tumors) that proliferate by expressing a mutated tyrosine kinase
or overexpression of a receptor tyrosine kinase; (2) benign and
malignant cells of other proliferative diseases in which aberrant
tyrosine kinase activation occurs; (3) any tumors that proliferate
by receptor tyrosine kinases; (4) any tumors that proliferate by
aberrant serine/threonine kinase activation; and (5) benign and
malignant cells of other proliferative diseases in which aberrant
serine/theroine kinase activation occurs.
[0200] The amount of a given agent that will correspond to such an
amount will vary depending upon factors such as the particular
compound, disease condition and its severity, the identity (e.g.,
weight) of the mammal in need of treatment, but can nevertheless be
routinely determined by one skilled in the art. "Treating" is
intended to mean at least the mitigation of a disease condition in
a mammal, such as a human, that is affected, at least in part, by
the activity of one or more AKT protein kinases, tyrosine kinases,
additional serine/threonine kinases, and/or dual specificity
kinases, and includes, but is not limited to, preventing the
disease condition from occurring in a mammal, particularly when the
mammal is found to be predisposed to having the disease condition
but has not yet been diagnosed as having it; modulating and/or
inhibiting the disease condition; and/or alleviating the disease
condition.
[0201] In order to use a compound of the Formula I or a
pharmaceutically acceptable salt, solvate, metabolite or prodrug
thereof, for the therapeutic treatment (including prophylactic
treatment) of mammals including humans, it is normally formulated
in accordance with standard pharmaceutical practice as a
pharmaceutical composition. According to this aspect of the
invention there is provided a pharmaceutical composition that
comprises a compound of the Formula I, or a pharmaceutically
acceptable salt, solvate, metabolite or prodrug thereof, as defined
hereinbefore in association with a pharmaceutically acceptable
diluent or carrier.
[0202] To prepare the pharmaceutical compositions according to this
invention, a therapeutically or prophylactically effective amount
of a compound of Formula I or pharmaceutically acceptable salt,
solvate, metabolite or prodrug thereof (alone or together with an
additional therapeutic agent as disclosed herein) is preferably
intimately admixed with a pharmaceutically acceptable carrier
according to conventional pharmaceutical compounding techniques to
produce a dose. A carrier may take a wide variety of forms
depending on the form of preparation desired for administration,
e.g., oral or parenteral. Examples of suitable carriers include any
and all solvents, dispersion media, adjuvants, coatings,
antibacterial and antifungal agents, isotonic and absorption
delaying agents, sweeteners, stabilizers (to promote long term
storage), emulsifiers, binding agents, thickening agents, salts,
preservatives, solvents, dispersion media, coatings, antibacterial
and antifungal agents, isotonic and absorption delaying agents,
flavoring agents, and miscellaneous materials such as buffers and
absorbents that may be needed in order to prepare a particular
therapeutic composition. The use of such media and agents with
pharmaceutically active substances is well known in the art. Except
insofar as any conventional media or agent is incompatible with a
compound of Formula I, its use in the therapeutic compositions and
preparations is contemplated. Supplementary active ingredients can
also be incorporated into the compositions and preparations as
described herein.
[0203] The compositions of the invention may be in a form suitable
for oral use (for example as tablets, lozenges, hard or soft
capsules, aqueous or oily suspensions, emulsions, dispersible
powders or granules, syrups or elixirs), for topical use (for
example as creams, ointments, gels, or aqueous or oily solutions or
suspensions), for administration by inhalation (for example as a
finely divided powder or a liquid aerosol), for administration by
insuffiation (for example as a finely divided powder) or for
parenteral administration (for example as a sterile aqueous or oily
solution for intravenous, subcutaneous, or intramuscular dosing or
as a suppository for rectal dosing). For example, compositions
intended for oral use may contain one or more coloring, sweetening,
flavoring and/or preservative agents.
[0204] Suitable pharmaceutically-acceptable excipients for a tablet
formulation include, for example, inert diluents such as lactose,
sodium carbonate, calcium phosphate or calcium carbonate,
granulating and disintegrating agents such as corn starch or
algenic acid; binding agents such as starch; lubricating agents
such as magnesium stearate, stearic acid or talc; preservative
agents such as ethyl or propyl p-hydroxybenzoate, and
anti-oxidants, such as ascorbic acid. Tablet formulations may be
uncoated or coated either to modify their disintegration and the
subsequent absorption of the active ingredient within the
gastrointestinal tract, or to improve their stability and/or
appearance, in either case, using conventional coating agents and
procedures well known in the art.
[0205] Compositions for oral use may be in the form of hard gelatin
capsules in which the active ingredient is mixed with an inert
solid diluent, for example, calcium carbonate, calcium phosphate or
kaolin, or as soft gelatin capsules in which the active ingredient
is mixed with water or an oil such as peanut oil, liquid paraffin,
or olive oil.
[0206] Aqueous suspensions generally contain the active ingredient
in finely powdered form together with one or more suspending
agents, such as sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents such as lecithin or condensation products of an
alkylene oxide with fatty acids (for example polyoxethylene
stearate), or condensation products of ethylene oxide with long
chain aliphatic alcohols, for example heptadecaethyleneoxycetanol,
or condensation products of ethylene oxide with partial esters
derived from fatty acids and a hexitol such as polyoxyethylene
sorbitol monooleate, or condensation products of ethylene oxide
with partial esters derived from fatty acids and hexitol
anhydrides, for example polyethylene sorbitan monooleate. The
aqueous suspensions may also contain one or more preservatives
(such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as
ascorbic acid), coloring agents, flavoring agents, and/or
sweetening agents (such as sucrose, saccharine or aspartame).
[0207] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil (such as arachis oil, olive oil,
sesame oil or coconut oil) or in a mineral oil (such as liquid
paraffin). The oily suspensions may also contain a thickening agent
such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set out above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0208] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water generally contain
the active ingredient together with a dispersing or wetting agent,
suspending agent, and one or more preservatives. Suitable
dispersing or wetting agents and suspending agents are exemplified
by those already mentioned above. Additional excipients such as
sweetening, flavoring and coloring agents, may also be present.
[0209] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, such as olive oil or arachis oil, or a mineral oil
such as liquid paraffin, or a mixture of any of these. Suitable
emulsifying agents may be, for example, naturally-occurring gums
such as gum acacia or gum tragacanth, naturally-occurring
phosphatides such as soya bean, lecithin, an esters or partial
esters derived from fatty acids and hexitol anhydrides (for example
sorbitan monooleate) and condensation products of the said partial
esters with ethylene oxide such as polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening, flavoring
and preservative agents.
[0210] Syrups and elixirs may be formulated with sweetening agents
such as glycerol, propylene glycol, sorbitol, aspartame or sucrose,
and may also contain a demulcent, preservative, flavoring and/or
coloring agent.
[0211] The pharmaceutical compositions may also be in the form of a
sterile injectable aqueous or oily suspension, which may be
formulated according to known procedures using one or more of the
appropriate dispersing or wetting agents and suspending agents,
which have been mentioned above. A sterile injectable preparation
may also be a sterile injectable solution or suspension in a
non-toxic parenterally-acceptable diluent or solvent, for example a
solution in 1,3-butanediol.
[0212] Suppository formulations may be prepared by mixing the
active ingredient with a suitable non-irritating excipient, which
is solid at ordinary temperature s but liquid at the rectal
temperature and will therefore melt in the rectum to release the
drug. Suitable excipients include, for example, cocoa butter and
polyethylene glycols.
[0213] Topical formulations, such as creams, ointments, gels and
aqueous or oily solutions or suspensions, may generally be obtained
by formulating an active ingredient with a conventional, topically
acceptable, vehicle or diluent using conventional procedures well
known in the art.
[0214] Compositions for administration by insufflation may be in
the form of a finely divided powder containing particles of average
diameter of, for example, 30 .mu.m or much less, the powder itself
comprising either active ingredient alone or diluted with one or
more physiologically acceptable carriers such as lactose. The
powder for insufflation is then conveniently retained in a capsule
containing, for example, 1 to 50 mg of active ingredient for use
with a turbo-inhaler device, such as is used for insufflation of
the known agent sodium cromoglycate.
[0215] Compositions for administration by inhalation may be in the
form of a conventional pressurized aerosol arranged to dispense the
active ingredient either as an aerosol containing finely divided
solid or liquid droplets. Conventional aerosol propellants such as
volatile fluorinated hydrocarbons or hydrocarbons may be used and
the aerosol device is conveniently arranged to dispense a metered
quantity of active ingredient.
[0216] For further information on formulations, see Chapter 25.2 in
Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;
Chairman of Editorial Board), Pergamon Press 1990, which is
specifically incorporated herein by reference.
[0217] The amount of a compound of this invention that is combined
with one or more excipients to produce a single dosage form will
necessarily vary depending upon the host treated and the particular
route of administration. For example, a formulation intended for
oral administration to humans may contain, for example, from 0.5 mg
to 2 g of active agent compounded with an appropriate and
convenient amount of excipients, which may vary from about 5 to
about 98 percent by weight of the total composition. Dosage unit
forms will generally contain about 1 mg to about 500 mg of an
active ingredient. For further information on routes of
administration and dosage regimes, see Chapter 25.3 in Volume 5 of
Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of
Editorial Board), Pergamon Press 1990, which is specifically
incorporated herein by reference.
[0218] The size of the dose for therapeutic or prophylactic
purposes of a compound of Formula I will naturally vary according
to the nature and severity of the conditions, the age and sex of
the animal or patient and the route of administration, according to
well known principles of medicine.
[0219] The compounds of this invention may be used alone in
combination with other drugs and therapies used in the treatment of
disease states which would benefit from the inhibition of MEK. Such
treatment may involve, in addition to the compounds of the
invention, conventional surgery or radiotherapy or chemotherapy.
Such chemotherapy may include one or more of the following
categories of anti-tumor agents:
[0220] (i) antiproliferative/antineoplastic drugs and combinations
thereof, as used in medical oncology, such as alkylating agents
(for example, cis-platin, carboplatin, cyclophosphamide, nitrogen
mustard, melphalan, chlorambucil, busulphan and nitorsoureas);
anti-metabolites (for example, antifolates such as such as
fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed,
methotrexate, cytosine arabinside, hydroxyurea, or, one of the
preferred anti-metabolites disclosed in European Patent Application
No. 239362 such as
N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]--
2-thenoyl)-L-glutamic acid); antitumor antibiotics (for example,
anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin,
epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin);
antimitotic agents (for example, vinca alkaloids like vincristine,
vinblastine, vindesine and vinorelbine and taxoids like taxol and
taxotere); and topoisomerase inhibitors (for example
epipodophyllotoxins like eptoposide and teniposide, amsacrine,
topotecan and campothecin):
[0221] (ii) cytostatic agents such as antiestrogens (for example,
tamoxifen, toremifene, raloxifene, droloxifene and iodoxyfene),
estrogen receptor down regulators (for example, fulvestrant),
antiandrogens (for example, bicalutamide, flutamide, nilutamide,
cyproterone acetate and Casodex.TM.
(4'-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromet-
hyl)propionanilide)), LHRH antagonists or LHRH agonists (for
example, goserelin, leuporelin and buserelin), progestogens (for
example, megestrol acetate), aromatase inhibitors (for example,
anastrozole, letrozole, vorazole and exemestane) and inhibitors of
5.alpha.-reductase such as finasteride;
[0222] (iii) agents which inhibit cancer cell invasion (for
example, metalloproteinase inhibitors like marimastat and
inhibitors of urokinase plasminogne activator receptor
function);
[0223] (iv) inhibitors of growth factor function like growth factor
antibodies, growth factor receptor antibodies (for example, the
anti-erbB2 antibody trastumuzab [Herceptin.TM.] and the anti-erbB1
antibody cetuximab [C225]), farnesyl transferase inhibitors,
tyrosine kinase inhibitors and serine-threonine kinase inhibitors
(for example, inhibitors of the epidermal growth factor family
tyrosine kinases such as
N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-
-amine (gefitinib, AZD1839),
N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine
(erlotinib, OSI-774) and
6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazoli-
n-4-amine (CI 1033)), inhibitors of the platelet-derived growth
factor family and inhibitors of the hepatocyte growth factor
family;
[0224] (v) antiangiogenic agents such as those which inhibit the
effects of vascular endothelial growth factor (for example, the
anti-vascular endothelial cell growth factor antibody bevacizumab
[Avastin.TM.], compounds such as those disclosed in International
Patent Applications WO 97/22596, WO 97/30035, WO 97/32856, and WO
98/13354) and compounds that work by other mechanisms (for example,
linomide, inhibitors of integrin .alpha.v(33 function, MMP
inhibitors, COX-2 inhibitors and angiostatin);
[0225] (vi) vascular damaging agents such as Combretastatin A4 and
compounds disclosed in International Patent Applications WO
99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434, and
WO 02/08213;
[0226] (vii) antisense therapies (for example, those which are
directed to the targets listed above such as ISIS 2503, and
anti-ras antisense);
[0227] (viii) gene therapy approaches, including for example
GVAX.TM., approaches to replace aberrant genes such as aberrant p53
or aberrant BRCA1 or BRCA2, GDEPT (gene-directed enzyme prodrug
therapy) approaches such as those using cytosine deaminase,
thymidine kinase or a bacterial nitroreductase enzyme and
approaches to increase patient tolerance to chemotherapy or
radiotherapy such as multi-drug resistance gene therapy;
[0228] (ix) interferon; and
[0229] (x) immunotherapy approaches, including for example ex-vivo
and in-vivo approaches to increase the immunogenicity of patient
tumor cells, such as transfection with cytokines such as
interleukin 2, interleukin 4 or granulocyte-macrophage colony
stimulating factor, approaches to decrease T-cell anergy,
approaches to using transfected immune cells such as
cytokine-transfected dendritic cells, approaches using
cytokine-transfected tumor cell lines and approaches using
anti-idiotypic antibodies.
[0230] Such conjoint treatment may be achieved by way of the
simultaneous, sequential or separate dosing of the individual
components of treatment. Such combination products employ the
compounds of this invention within the dose range described
hereinbefore and the other pharmaceutically active agent within its
approved dose range.
[0231] According to this aspect of the invention there is provided
a pharmaceutical product comprising a compound of Formula I as
defined hereinbefore and an additional anit-tumor agent as definged
hereinbefore for the conjoint treatment of cancer.
[0232] Although the compounds of Formula I are primarily of value
as therapeutic agents for use in warm-blooded animals (including
man), they are also useful whenever it is required to control AKT
protein kinases, tyrosine kinases, additional serine/threonine
kinases, and/or dual specificity kinases. Thus, they are useful as
pharmacological standards for use in the development of new
biological tests and in the search for new pharmacological
agents.
[0233] The activity of the compounds of this invention may be
assayed for AKT protein kinases, tyrosine kinases, additional
serine/threonine kinases, and/or dual specificity kinases in vitro,
in vivo, or in a cell line. In vitro assays include assays that
determine inhibition of the kinase activity. Alternate in vitro
assays quantitate the ability of the inhibitor to bind to kinases
and may be measured either by radiolabelling the inhibitor prior to
binding, isolating the inhibitor/kinase complex and determining the
amount of radiolabel bound, or by running a competition experiment
where new inhibitors are incubated with known radioligands. These
and other useful in vitro and cell culture assays are well known to
those of skill in the art.
[0234] Although the invention has been described and illustrated
with a certain degree of particularity, it is understood that the
present disclosure has been made only by way of example, and that
numerous changes in the combination and arrangement of parts can be
resorted to by those skilled in the art without departing from the
spirit and scope of the invention, as hereinafter claimed.
[0235] The compounds of the present invention may be prepared in a
number of ways well known to one skilled in the art of organic
synthesis. The compounds of the present invention can be
synthesized using the methods described below, together with
synthetic methods known in the art of synthetic organic chemistry,
or variations thereon as appreciated by those skilled in the art.
Preferred methods include, but are not limited to, those described
below.
[0236] The novel compounds of the present invention may be prepared
using the reactions and techniques described in this section. The
reactions are performed in solvents appropriate to the reagents and
materials employed and are suitable for the transformations being
effected. Also, in the description of the synthetic methods
described below, it is to be understood that all proposed reaction
conditions, including choice of solvent, reaction atmosphere,
reaction temperature, duration of the experiment and workup
procedures, are chosen to be the conditions standard for that
reaction, which should be readily recognized by one skilled in the
art. It is understood by one skilled in the art of organic
synthesis that the functionality present on various portions of the
molecule must be compatible with the reagents and reactions
proposed. Such restrictions to the substituents that are compatible
with the reaction conditions will be readily apparent to one
skilled in the art and alternate methods must then be used.
[0237] The preparation of compounds of the present invention may be
carried out in a convergent or sequential synthetic manner. The
skills required in preparation and purification of such compounds
and the intermediates leading to these compounds are known to those
in the art. Purification procedures include, but are not limited
to, normal or reverse phase chromatography, crystallization, and
distillation.
[0238] An illustration of the preparation of compounds (8), (9),
(10) and (11) of the present invention is shown in FIG. 1. The
synthesis starts with the preparation of a substituted
quinazolinone (3) made by, for example, by the condensation of a
corresponding substituted aryl amino acid (1) and a corresponding
substituted amide (2) (see, for example, LeMahieu, et al, J. Med.
Chem., 1983, 26, 420-5 and references cited therein). Introduction
of a leaving group into quinazolinone (3) may be accomplished by
treatment with a halogenating agent (for example POCl.sub.3) to
give the chlorinated quinazoline (4.) The halogen leaving group is
then displaced with substituted and protected piperazine (5) (e.g.,
Boc, but any suitable protecting group may be used; see, T. W.
Greene et al., `Protective groups in organic synthesis`, John Wiley
and Sons, 1999, 3.sup.rd Ed., pp. 494-653). The piperazine (5) may
be introduced to chlorinated quinazoline (4) either neat or in the
presence of base. The piperazine protecting group may then be
removed by known methods (see, Greene et al, supra) (6).
[0239] Substitution of the piperazine secondary amine in
quinazoline intermediate (6) may be accomplished using a variety of
electrophiles and reaction conditions. For example, the piperazine
may be acylated by a suitably N-substituted or protected amino acid
(e.g., Boc, etc.) which may be introduced using a variety of
standard peptide coupling procedures under both solution phase and
solid phase conditions, to produce a product such as compound (8).
For representative examples, see Miklos Bodanszky, `Principles of
Peptide Synthesis,` Springer-Verlag, 1993, 2nd Ed., and C. Najera,
Synlett, 2002, 9, 1388-1403. As above (and if protected) the
N-protected amino acid unit may then be deprotected using
representative procedures (e.g., using acid on a Boc-group; Greene
et al., supra), and then manipulated as desired according to
procedures appreciated by those skilled in the art.
[0240] Compounds of the present invention similar to compound (8)
may be a prepared from quinazoline intermediate (6) by acylation
with a natural or an `unnatural` amino acid (7). The preparation of
`unnatural` amino acids is also well known to those skilled in the
art, and their use is included in the present invention (for
representative reviews, see, C. Najera, Synlett, 2002, 9,
1388-1403, and J.-A. Ma, Angew. Chemie, Int. Ed., 2003, 42,
4290-4299, and references therein).
[0241] Alternatively, the piperazine (6) may be acylated with an
acid or acid halide in the presence of base to generate a
substituted amine (10). Additionally, a substituted tertiary amine
(11) can be prepared by treating piperazine (6) with an appropriate
aldehyde (or surrogate) in the presence of a reducing agent (e.g.,
sodium cyanoborohydride). The piperazine (6) can also be treated
with an epoxide to give the amino alcohol (9.) All functional
groups may be further manipulated under standard conditions (e.g.,
reductions, alkylations, oxidations, palladium or nickel mediated
couplings, etc.) to further functionalize each compound.
[0242] The compounds described in FIG. 1 may be prepared either as
either the racemate, or as a single enantiomer (for example, using
an enantiomerically pure amino acid (7.)) If prepared as the
racemate, the corresponding enantiomers may be isolated by
separation of the racemic mixture of compound on a chiral
stationary phase column utilizing normal or reverse phase HPLC
techniques. Alternatively, a diastereomeric mixture of compound (8)
can be prepared by treatment of racemic compound (8) with an
appropriate chiral acid (or suitably activated derivative), for
example dibenzoyl tartrate or the like (see, for example, Kinbara,
K., et. al., J. Chem. Soc., Perkin Trans. 2, 1996, 2615; and
Tomori, H., et. al., Bull. Chem. Soc. Jpn., 1996, 3581). The
diastereomers would then be separated by traditional techniques
(i.e. silica chromatography, crystallization, HPLC, etc) followed
by removal of the chiral auxiliary to afford enantiomerically pure
compound (8.)
[0243] Other compounds of the present invention can be prepared
using the alternatively substituted and functionalised acids, amino
acids, hydroxy acids and variants thereof described in FIGS. 2-15.
For example, compound (14) of this invention may be prepared as
shown in FIG. 2. The hydroxyl group of substituted .alpha.-hydroxy
benzyl ester (12) is protected with an appropriate protecting group
(such as acetate) to give compound 13. The benzyl ester is then
converted to the corresponding carboxylic acid (for example by
hydrogenolysis) to give compound (14).
[0244] FIG. 3 shows the preparation of compound (18). Substituted
phenyl boronic acid (15), glyoxylic acid (16), and a chiral or
achiral mono-protected (using the Boc protecting group, for
example) diamine (17) (such as 3-Boc-aminopyrrolidine) are combined
in an appropriate solvent such as 1,2-dichloroethane and stirred at
elevated temperature to provide carboxylic acids (18).
[0245] FIG. 4 shows the preparation of compound (22).
2-(2-Aminoethoxy)ethanol (19) is protected with an appropriate
amine protecting group (such as Boc), and the hydroxyl group is
oxidized to the carboxylic acid to provide intermediate (20). The
acid in compound (20) is then converted to an ester using an
appropriate base (such as K.sub.2CO.sub.3) and alkyl halide to
furnish intermediate compound (21). Enolization of intermediate
compound (21) is accomplished with strong base (such as LDA or
LHMDS), followed by addition of a substituted benzyl halide yields
an alkylated ester, which is then converted by basic hydrolysis to
the corresponding acid (22).
[0246] Compound (28) may be prepared as shown in FIG. 5. The
substituted phenyl carboxylic acid (23) is transformed to the
appropriate ester (24) under acidic (mineral acid, R.sup.2OH) or
basic (K.sub.2CO.sub.3, R.sup.2X) conditions. Enolization of ester
(24) is accomplished with strong base (such as LDA), and addition
of a haloacetate ester (for example tert-butyl bromoacetate)
provides intermediate compound (25). Selective ester deprotection
is performed by treating compound (25) with acid (such as TFA) to
provide carboxylic acid (26). The carboxylic acid (26) is converted
to an acyl azide (using diphenylphosphoryl azide, for example),
which is then transformed to the corresponding carbamate-protected
amine by heating in an appropriate alcohol solvent (tert-butyl
alcohol, for example) in the presence or absence of a Lewis acid
(such as SnCL4) to provide compound (27). The carboxylic acid ester
is then converted to the corresponding acid by hydrolysis under
basic conditions (such as aq. LiOH in THF) to afford compound
(28).
[0247] FIG. 6 shows the preparation of carboxylic acid (31). Lactam
(29) is enolized with strong base (such as LDA/LiBr or LHMDS), and
addition of a substituted benzyl halide furnishes alkylated
intermediate compound (30). The lactam is then opened under basic
conditions (such as aq. LiOH, THF) to furnish carboxylic acid
(31).
[0248] Compound (36) may be prepared as shown in FIG. 7.
Condensation of an appropriately substituted benzaldehyde with
ethyl cyanoacetate provides compounds of structure (32). Treatment
with a reducing agent such as NaBH.sub.4 gives the saturated
compound (33), which is followed by cobalt-mediated hydride
reduction to give compound (34). The amine can then be protected
and the ester saponified to give compound (35). Coupling with a
piperazine can be accomplished using (for example) EDCI or PyBrop,
followed by deprotection to give final compound (36).
[0249] FIG. 8 shows the preparation of amino acid (40). Compound
(39) can be prepared by condensation of benzaldehydes with ethyl
cyanoacetate followed by catalytic hydrogenation according to the
procedures described by Lee, J. et al. (1999), 3060-3065. Compound
(39) can be converted to compound amino acid (40) by protection of
the primary amine followed by saponification under basic condition
(for example, aqueous LiOH solution).
[0250] An alternate approach to amino acid (40) is shown in FIG. 9.
Compound (41), where Pg is an appropriate protecting group (for
example, Boc), can be treated with a variety of organometallic
agents such as LDA in a suitable solvent such as THF or ether at
low temperature s to generate a dianion intermediate, which can be
quenched by suitable amount of benzyl halides to afford the
intermediate compound (42). Saponification under basic conditions
such as aqueous LiOH solution furnishes the desired product
(40).
[0251] FIG. 10 summarizes a synthesis of amino alcohols (46) from
compound (45). Compound (45) may be prepared from compound (44) by
a sequence of deprotonation, alkylation and saponification as
described in FIG. 9.
[0252] Preparation of compound (50) is shown in FIG. 11. Phenyl
acetic acid derivative (47) can be deprotonated by treatment with a
suitable organometallic agent such as LDA in a suitable solvent
such as THF or ether at low temperature s, and then reacted with
compound (48), where X is a suitable leaving group (for example Br,
Cl) and Pg is an appropriate protecting group (for example, Boc or
Ts), to yield intermediate compound (49) (Ho-sam A. et al. (1997)
J. Med. Chem., 40, 2196; Ohkanda et al. (2004), J. Med. Chem., 47,
432). Saponification of compound (49) under basic conditions (for
example, aqueous LiOH solution) gives acid (50).
[0253] Compound (55) may be prepared as shown in FIG. 12.
Esterification of the appropriately substituted and commercially
available acid (51) with an alcohol affords the desired ester (52).
Treatment of ester (52) with appropriate base and electrophile
(e.g., acrylate, etc. (53)) followed by ester cleavage with acid
(see, T. W. Greene et al., supra) affords intermediate compound
(54). Introduction of azide (affords acyl-azide) with activating
reagent followed by heating affects rearrangement of acid (54) to
the requisite N-protected amino-ester intermediate (for example
Boc, but any suitable protecting group may be used with the
appropriate alcohol solvent; see, Greene et al., supra. Treatment
of ester intermediate with hydroxide base affords the N-protected
amino acid (55).
[0254] Compound (58) may be prepared as shown in FIG. 13. Michael
addition of phenylacetic acid ethyl esters with tert-butyl acrylate
using catalytic base such as potassium tert-butoxide followed by
acid hydrolysis of the tert-butyl ester provides compound (56).
Curtius rearrangement using diphenylphosphorylazide followed by
saponification of the ester gives compound (57). Coupling with a
piperazine can be accomplished using EDCI or PyBrop, followed by
deprotection of the Boc group to give final compound (58).
[0255] FIG. 14 shows the preparation of compound (61). Alkylation
of phenylacetic acid ethyl esters with .alpha.-bromoacetate
tert-butyl ester using a base such as lithium
bis(trimethylsilyl)amide provides compound (59). The remainder of
the sequence is as that described in FIG. 13 to provide compound
(61).
[0256] Compound (71) may be prepared as shown in FIG. 15. The
displacement of 4-chloroquinazoline with ethyl isonipecotate
followed by saponification of the ester gives intermediate (68).
Treatment with a halogenating reagent such as thionyl chloride or
oxalyl chloride provides acid chloride (69). Reductive amination of
an appropriately substituted benzaldehyde with
N-Boc-ethylenediamine using NaCNBH.sub.3 or NaH(OAc).sub.3 in MeOH,
THF, or DCE as solvent gives the secondary amine (70). Reaction of
(69) with amine (70) followed by deprotection of the Boc group
provides compound (71).
[0257] Alternatively, compound of formula (74) may be prepared as
shown in FIG. 16. Reductive amination of an appropriately
substituted aniline with tert-butyl N-(2-oxoethyl)carbamate using
NaCNBH.sub.3 or NaH(OAc).sub.3 in MeOH, THF, or DCE as solvent
gives the secondary amine (72). Compound (72) can be purified by
removal of the Boc group followed by acid-base extraction and
chromatography to give compounds of structure (73) and then
converted back to compound (72) by treatment with Boc.sub.2O.
Reaction with intermediate (69) using DMAP as base followed by
deprotection of the Boc group with (for example) ethereal HCl and
substitution (if required) gives compound (74).
[0258] Compound (78) may be prepared as shown in FIG. 17. Compound
(75) can be prepared from 7-azaindole according to literature
procedures. Introduction of the piperazine can be accomplished by
melting N-benzylpiperidine with intermediate (75) to give
intermediate (76). Removal of the benzyl protecting group can be
accomplished using (for example) hydrogenation in the presence of
Pd--C in methanol. Coupling of a Boc-protected amino acid with
intermediate (77) can be accomplished using (for example) EDCI or
PyBrop, followed by deprotection of the Boc group to give compound
(78).
[0259] FIG. 18 shows preparation of compound (81). Compound (79)
can be prepared from 2-aminopyridine similar to literature
procedure (A. R. Katritzky et al., J. Org. Chem., 2003, 68,
4935-4937). Deprotection of the Boc group using (for example)
ethereal HCl gives intermediate (80). The piperazine can be coupled
to N-protected amino acids using (for example) EDCI or PyBrop
followed by deprotection to give final compound (81).
[0260] FIG. 19 illustrates the preparation of 5- and 6-substituted
indazole (86). Substituted nitro indazole (82), where R.sup.5 and
R.sup.7 are substituents which are suitable for use in the
subsequent reactions, may be reduced to amino indazole (83) using
standard conditions (for example catalytic hydrogenation,
zinc/acetic acid, Fe/HCl, SnCl.sub.2/MeOH or FeSO.sub.4 in water).
Amino indazole (83) can react with compound (84) (for example,
bis(2-chloroethyl)amine) in the presence of an acid scavenger (for
example, Na.sub.2CO.sub.3, K.sub.2CO.sub.3, or the like) to afford
the cyclized product (85). This reaction is performed in a suitable
solvent (for example, ethanol) by heating at about 50-150.degree.
C. The resulting piperazine compound (85) is then acylated by a
suitable acid (for example a protected amino acids) which may be
introduced using a variety of standard peptide coupling procedures
under solution phase or solid phase conditions. The coupling
product may require a separate deprotection step to remove any
protecting groups in R to afford the product (86). For example, a
Boc protecting group may be removed by treating with a strong acid
such as trifluoroacetic acid (TFA) or hydrochloric acid in the
presence of an inert solvent such as dichloromethane or methanol.
Removal of a Cbz group can be carried out by catalytic
hydrogenation with hydrogen in the presence of a palladium catalyst
or by transfer hydrogenation. An Fmoc group can be removed with a
low boiling point amine (for example piperidine or the like) in a
solvent such as DMF.
[0261] FIG. 20 describes the synthesis of 3-alkyl and 3-aryl
substituted indazole (90). The iodo intermediate (87) can be
prepared by the procedures shown in FIG. 20. Compound (87) is
protected using a suitable protecting group and treated with an
alkyl or aryl boronic acid or ester and a suitable Pd catalyst, for
example, Pd(PPh.sub.3).sub.4, to afford the desired 3-substituted
intermediate (89) which is then deprotected to give compound
(90).
[0262] The preparation of 3-amino substituted indazoles is outlined
in FIG. 21. Compound (92) can be prepared by reacting a suitably
monoprotected piperazine intermediate with a compound (91), where X
is a suitable leaving group (for example, bromo, iodo or OTf), via
Pd or Cu-mediated coupling (Buchwald et al. (2000), J. Org. Chem.,
65, 1144; Hartwig et al. (1998) Angew. Chem. Int. Ed. Eng., 37,
2046) to furnish intermediate (92). Removal of the protecting group
followed by amide coupling with an acid affords compound (93),
which is then treated with hydrazine to give 3-amino indazole
intermediate (94). Selective protection of the N-1 nitrogen affords
compound (95), which can be acylated with an acid halide in an
inert solvent (for example, dichloromethane, or the like) to afford
the amide (96). An organic base, such as diisopropylethylamine,
triethylamine, pyridine, or DMAP, may be added as an acid scavenger
to facilitate the coupling reaction. Transformation of compound
(96) into compound (97) can be accomplished by removal of the
protecting groups.
[0263] FIG. 22 describes a synthesis of a particular class of
compounds bearing an isoquinoline ring. Compound (100) can be
prepared by reacting suitably mono-protected piperazine (99) with
an isoquinoline compound substituted with a leaving group X, where
X is halide (for example, chloro, bromo, and iodo), or sulfonate
(for example OSO.sub.2CF.sub.3), in the presence of a base and a
palladium or a copper catalyst, according to known methods. Removal
of the protecting group Pg of compound (100) affords an amine
intermediate, which can be conveniently converted to compound (101)
by amide coupling with acids followed by optional removal of
protecting groups as described in FIG. 19.
[0264] FIG. 23 presents a synthesis of a particular class of
pyrimidines bearing a substituent at the 5-position. Compound (104)
can be prepared by S.sub.NAr reaction between a suitably
mono-protected piperazine compound (103) and a 4-chloro substituted
pyrimidine intermediate (102), where X is Br or I, in the presence
of an acid scavenger (for example, diisopropylethylamine or
triethylamine). Removal of the protecting group Pg followed by
amide coupling with an acid (106) affords intermediate (107).
Intermediate (107) can react with various coupling components (108)
via metal-mediated reactions to furnish product (109). For example,
compounds bearing an O- or S-linked substituent at the 5-position
of the pyrimidine ring can be prepared by reactions between
intermediate (107) and an alcohol or thiol in the presence of a
base (for example, Cs.sub.2CO.sub.3) and a Cu catalyst (for
example, CuCl, CuI, or the like) under modified Ullman coupling
conditions (Wolter, M. et. al. Org. Lett. 2002, 4, 973-976). In
some cases, an additive (for example,
2,2,6,6-tetramethyl-heptane-3,5-dione, pentane-2,4-dione,
1,10-phenethroline, or the like) is added to accelerate the
reaction. Alternatively, if sodium thiolates are available, base is
not required for the reaction. The coupling between compound (107)
and a thiol may also be accomplished by palladium-catalyzed
reactions (Kondo, T. et al. Chem. Rev., 2000, 100, 3205-3220;
Zheng, N. et al. J. Org. Chem., 1998, 63, 9606-9607). Compounds
bearing a N-linked substituent at the 5-position of the pyrimidine
ring can be prepared by a Pd or Cu mediated coupling between
intermediate (107) and amines (Buchwald et al. (2000), J. Org.
Chem., 65, 1144; Hartwig et al. (1998) Angew. Chem. Int. Ed. Eng.
37, 2046). Compounds bearing an alkyl or an aryl substituent at the
5-position of the pyrimidine ring can be prepared by Suzuki
coupling (Miyaura, N. Suzuki A. (1995), Chem. Rev. 95, 2457; Org.
React. (1997), 50, 1) between intermediates (107) and (108),
wherein Y is a boronic acid or boronic ester, in the presence of a
base (for example, Na.sub.2CO.sub.3 and Et.sub.3N), a catalytic
Pd(0) species (for example, Pd(PPh.sub.3).sub.4,
Pd(PPh.sub.3).sub.2Cl.sub.2, Pd.sub.2(dba).sub.3 and Pd(OAc).sub.2)
and a suitable ligand (such as PPh.sub.3 and AsPh.sub.3).
Alternatively, 5-alkyl and aryl substituted pyrimidines (109) may
also be prepared by Nigeshi or Kumada couplings between compounds
(107) and (108), wherein Y--R' is an organo zinc reagent, in the
presence of a Pd (for example, Pd(PPh.sub.3).sub.4) or Ni (for
example Ni(acac).sub.2) catalyst. Alternatively, 5-alkyl and aryl
substituted pyrimidines (109) may also be prepared by Stille
coupling between compounds (107) and (108), wherein Y--R' is an
organostannane reagent, in the presence of a Pd catalyst.
[0265] FIG. 24 describes an alternate synthesis of compounds
bearing an O-linked substituent at the 5-position. Compound (III)
can be prepared by Cu-catalyzed coupling of intermediate (110) with
benzyl alcohol. Removal of the benzyl group by hydrogenation
affords 5-hydroxylpyrimidine intermediate (112), which can be
converted to compound (115) by deprotection and amide coupling as
described in FIG. 19. Alkylation of compound (115) with alkyl
halides in the presence of a base (for example, K.sub.2CO.sub.3,
Cs.sub.2CO.sub.3, or the like) in an inert solvent (for example,
DMF) provides the desired compound (116).
[0266] Compound (125) can be synthesized as described in FIG. 25.
An appropriately substituted pyrimidine (119) may be prepared by
the condensation of a corresponding substituted malonic acid
diester (117) and a corresponding substituted formamidine (118) in
the presence of a base (for example, NaOEt). Treatment of
pyrimidine (119) with a halogenating agent (for example, POCl.sub.3
or POBr.sub.3) affords the dihalide (120). Displacing one of the
halogens with protected piperazine (121) gives the mono-substituted
compound (122), which can be converted to compound (123) by
reduction (for example, catalytic hydrogenation) of the second
halogen. Transformation of compound (123) into desired compound
(125) can be accomplished by the procedures described in FIG.
22.
[0267] FIG. 26 illustrates an approach to preparation of
5,6-disubstituted pyrimidines (129). Treatment of compound (126)
with a nucleophile (for example, an amine) either neat or in the
presence of a base can give the SNA, product (127). Alternatively,
compound (126) can be converted to compound (127) via various metal
mediated coupling reactions such as described in FIG. 23.
Transformation of compound (127) into desired compound (129) can be
accomplished by a sequence of deprotection, amide coupling and
optional deprotection as described in FIG. 22.
[0268] The preparation of compounds with an amino group at the
6-position of the pyrimidine ring is shown in FIG. 27. Compounds of
formula (131) can be prepared by palladium catalyzed coupling
reactions between intermediate (130) and an ammonia equivalent (for
example, benzophenone imine). For a review and leading references
for arylation of ammonia equivalents, see Muci, A. R., Buchwald, S.
L., Topics in Current Chemistry, 2002, 219, 131. Removal of the
protecting group Pg.sup.2 in compound (131) furnishes the amino
intermediate (132). Transformation of (132) into desired compound
(134) can be accomplished by the procedures described in FIG.
22.
[0269] FIG. 28 summarizes the preparation of compounds of the
invention bearing a cinnoline ring. Compound (137) can be prepared
by a one-pot process from 4-hydroxyl cinnoline (135) and protected
piperazine (136) through a triflate intermediate (Cacchi, S. et al.
Synlett, 1997, 1400). Sequential removal of the protecting group in
compound (137) followed by amide coupling and optional deprotection
affords compound (139).
[0270] FIG. 29 describes a synthesis of a compound containing a
diamino group. Protection of the amino group in compound (140)
gives a protected intermediate, which is subjected to Mitsunobu
reaction with pthalimide to furnish compound (141). The phthalimide
group can be selectively removed with a base (for example,
hydrazine and low boiling point amines). Acylation of compound
(142) with a acids using standard peptide coupling procedures
followed by removal of the protecting group affords the product
(144).
[0271] As shown in FIG. 30, the protecting group in compound (141)
can alternatively be first selectively removed under known
conditions to give compound (146), which can be coupled with an
acid to afford the amide (147). Removal of the phthalimide group
with a base (for example, hydrazine and a low boiling point amine)
leads to the product (148).
[0272] FIG. 31 summarizes a preparation of 1-substituted
quinolizinones (153). Treatment of compound (149) with an
organometallic base (for example, n-BuLi) followed by quenching
with 2-ethoxymethylenemalonic acid diethyl ester yields the Michael
addition product (150). Cyclization occurs when heating compound
(150) in an inert solvent (for example, xylene) to give
intermediate (151). The carboxylate group in intermediate (151) may
be removed by heating in an acidic solution (for example, aqueous
HCl or H.sub.2SO.sub.4 solution). Sequential removal of the
protecting group in compound (152) followed by amide coupling and
optional deprotection affords compound (153).
[0273] FIG. 32 describes a synthesis of compounds with a 4-hydroxyl
piperidine linker. Compound (155) can be prepared by S.sub.NAr
reaction between a suitably N-protected 4-hydroxyl piperidine
compound and a substituted quinazoline intermediate (154) where X
is leaving group (for example Cl or Br), in the presence of a base
(for example, NaH or triethylamine) in a suitable solvent such as
DMF, THF etc. Removal of the protecting group Pg in compound (155)
followed by amide coupling with an acid and optional deprotection
affords the desired compound (156).
[0274] Synthesis of compounds with a tetrahydropyridine linker is
described in FIG. 33. Compound (157), where Pg is an appropriate
protecting group, is treated with an organometallic agent (for
example, LDA) and N-phenyltrifluoromethanesulfonimide to give the
triflate (158) (Eastwood, P. R. (2000), Tetrahedron Lett., 3705).
Conversion of triflate (158) to the corresponding borinate ester
(159) is accomplished by reacting with a suitable diboron species,
such as dipinacolatodiboron, or other electrophilic source of
boron, with an appropriate palladium catalyst. Ester (159) is then
reacted with compound (160), where X is a leaving group (for
example, chloro, bromo, iodo or OTf), under palladium catalysis to
give intermediate (161). Alternatively, the cross=coupling reaction
also can be performed in the reverse direction by switching the
leaving group and boron species. For example, intermediate (161)
can be prepared by the reaction between compound (160), where X is
a boronic acid or ester, with the triflate (158). Sequential
removal of the protecting group in compound (161) followed by amide
coupling and optional deprotection affords compound (162).
[0275] FIG. 34 describes the preparation of 5,6-disubstituted
pyrrolopyrimidine (175). Compound (168) can be obtained from
commercial sources or can be prepared by literature methods (for
example, Eger, K. et al. (1987), J. Heterocycl. Chem. 24, 425-430;
Roth, H. J. et al. (1975), Arch. Pharm. 308, 179-185; Pichler, H.
et al. (1986), Liebigs Ann. Chem. 1986, 1485-1505). Condensation of
compound (168) with formic acid at elevated temperature affords
intermediate (169) (Traxler, P. M. et al. (1996), J. Med. Chem.,
39, 2285-2292). Treating compound (169) with a halogenating agent
(for example, POCl.sub.3) yields the halide (170). Removal of the
protecting group Pg followed by displacement of the halogen with
suitably protected piperazine (172), either neat or in the presence
of an acid scavenger (for example, diisopropylethylamine or
triethylamine), leads to intermediate (173). Transformation of
intermediate (173) into compound (175) can be accomplished by the
procedures described in FIG. 22.
[0276] The preparation of 3-substituted pyrazolopyrimidines (182)
is described in FIG. 35. Compound (176) can be obtained from
commercial sources or can be prepared by literature methods (for
example, Hamaguchi, M. et al. (1986), Heterocycles. 24, 2111-2115;
MaCall, M. A. et al. (1962), J. Org. Chem. 27, 2433-2439).
Condensation of compound (176) with hydrazine affords the cyano
intermediate (177), which can be converted to compound (178) by
condensing with formic acid at elevated temperature. Alternatively,
compound (96) can first be hydrolyzed to afford the primary amide,
which is then condensed with formamide at elevated temperature to
give the cyclized product (178). Treatment of (178) with a
halogenating agent (for example, POCl.sub.3) gives the halide
(179). Transformation of (179) into desired compound (182) can be
accomplished by the procedures described in FIG. 22.
[0277] An alternate route to the intermediate (179) for the
synthesis of 3-substituted pyrazolopyrimidines is shown in FIG. 36.
Regioselective deprotonation of 4,6-dichloropyrimidine at the C-5
position by treatment with an organometallic agent (for example,
LDA), followed by quenching with aldehyde (184) furnishes the
hydroxyl intermediate (185) (Radinov, R. et al. (1986), Synthesis,
11, 886-891; Radinov, R. et al. (1991), J. Org. Chem., 56,
4793-4796). Intermediate (185) can be oxidized with an oxidizing
agent (for example, CrO.sub.3 or MnO.sub.2) to give ketone (186).
Treatment with hydrazine in an inert solvent such as THF or DCM
yields the cyclized product (179). Transformation of (179) into
desired compound (182) can be accomplished by the procedures
described in FIG. 22.
[0278] As shown in FIG. 37, compound (190) may be prepared in two
ways. First, the substitution of heterocyclic core (188) with
aminoamido piperazine (189) followed by deprotection with acid
affords the desired product (190). Second, substitution of the
heterocyclic core (188) with N-protected piperazine (191) gives the
intermediate (192), which was subject to deprotection, coupling
with amino acid (193) and deprotection again with acid to provide
the final compound (190). The halide (188) may be obtained from
commercial sources or prepared by means known to those in the
art.
[0279] The tetrahydropyrado[2,3-d]pyrimidine derivative (199) may
be prepared as shown in FIG. 38. A 2-amino-3-pyradocarboxylic acid
(194) was heated with formamide to give the
4-hydroxypyradopyrimidine derivative (195), which was subject to
chlorination with (for example) POCl.sub.3 to afford the 4-chloro
pyridopyrimidine derivative (196). S.sub.NAr reactions of the
compounds (196) with 1-Boc-piperazine gave the intermediates (197).
Reduction of the intermediate (197) in the presence of catalytic
amount of (for example) PtO.sub.2 under hydrogen yielded the
tetrahydropyrido[2,3-d]pyrimidine derivatives (198). After
deprotection, the compounds (198) were subject to amide coupling
with N-protected amino acids and followed by deprotection with acid
to offer the product (199).
[0280] Dihydropyrrolo[2,3-d]pyrimidine derivatives may be prepared
as shown in FIG. 39. 2-Ethoxycarbonyl-succinic acid diethyl ester
(200) was heated with formamidine to provide
(4,6-Dihydroxypyrimidin-5-yl)-acetic acid methyl ester (201).
Halogenation of compound (201) with (for example) POCl.sub.3 gave
the dichloropyrimidine derivative (202). Treating compound (142)
with base (e.g., KH) in THF and an electrophile R.sup.3X affords
the intermediate (203). Reduction with an agent such as DIBAL-H
gives the alcohol, which was activated with (for example) MsCl in
the presence of base (e.g., TEA) to provide the mesylate
intermediate (204). Treating the mesylate (204) with (for example)
p-methoxybenzylamine yielded the dihydropyrrolo[2,3-d]pyrimidine
derivative (205). S.sub.NAr reaction of the compound (205) with
1-Boc-piperazine formed the intermediate (206). Deprotection of the
compound (206) with acid (e.g., TFA) gave the free amine (207).
Amide coupling of the free amine (207) with an appropriately
substituted/protected amino acid and followed by deprotection (if
necessary) gave the product (208).
[0281] Compound (217) of this invention may be prepared as shown in
FIG. 40. Thus palladium-catalyzed cross coupling of boronic acid
(213) and properly substituted aryl halide (214) affords the ester
intermediate, which is saponified by hydroxide base leading to acid
(215). The coupling of acid (215) and amine (216) under standard
conditions (e.g., EDCI, HOBt, etc.) gives the
N-protected/substituted advanced intermediate (for example Boc, but
any suitable protecting group may be used; see, Greene et al.,
supra. The N-protected/substituted intermediate (e.g., Boc) is
cleaved if necessary to afford product (217).
[0282] Compound (223) may be prepared as shown in FIG. 41.
Introduction of a leaving group into the appropriately substituted
and commercially available 4,3,0-heterocycle (218) may be
accomplished, for example, by treatment with a halogenating agent
(for example POCl.sub.3) to give the chloride (219). Displacement
of the leaving group with an appropriately substituted and
protected piperazine (220) (for example Boc, but any suitable
protecting group may be used; see, Greene et al., supra) either,
for example, neat or in the presence of base, followed by removal
of the piperazine protecting group (using references from the above
book) can give the advanced quinazoline intermediate (221).
Substitution of the piperazine secondary amine may then be
accomplished using a variety of electrophiles and reaction
conditions. The piperazine may be acylated by a suitably
N-substituted or protected amino acid (e.g., Boc, etc. (162)) which
may be introduced using a variety of standard peptide coupling
procedures under both solution phase and solid phase conditions to
yield compound (223). For representative examples, see Miklos
Bodanszky, `Principles of Peptide Synthesis,` Springer-Verlarg,
1993, 2nd Ed., and C. Najera, Synlett, 2002, 9, 1388-1403. As above
(and if protected) the N-protected amino acid unit may then be
deprotected using representative procedures (e.g., acid, for a
Boc-group) referenced in Greene et al., supra, and then manipulated
as desired according to procedures appreciated by those skilled in
the art. Alternatively (and as in FIG. 1), the amine (221) may be
reacted with any other electrophile, including (but not limited to)
epoxides, acid halides, aldehydes, etc., using procedures known to
those in the art of organic synthesis.
[0283] Compounds of formula (229) may be prepared as shown in FIG.
42. An S.sub.NAr reaction of compound (224) with a protected linker
(eg. Boc-piperazine) gives the piperazine (225). An
organometallic-mediated reaction may be used to install an
activated acetylene group (226) and treatment with base (for
example, KOtBu) gives the pyrrolopyrimidine (228). Deprotection of
the piperazine protecting group (with, for example in the case of a
Boc group, acid) and acylation (with for example a protected amino
acid, followed by deprotection if necessary) gives the desired
product (229).
[0284] Compounds of formula (234) may be prepared as shown in FIG.
43. Thus halogenation (eg. POCl.sub.3) of a suitably substituted
nitropyridone, (230), and subsequent treatment with a vinyl
Grignard reagent gives the appropriately halogenated
pyrrolopyridine (232). Subsequent S.sub.NAr displacement (or
alternatively a transition metal mediated reaction) with a suitably
substituted piperazine derivative (233) gives the desired product
(234).
[0285] FIG. 44 illustrates the general preparation of compounds of
the formula (242). Acylation of an appropriately substituted
aminothiophene (235) (using, for example, formic acid and ammonium
acetate under heat) and cyclisation using (for example) formamide
and ammonium formate at high temperature gives the appropriate
heterocycle. Halogenation, using (for example) oxalyl chloride then
gives the appropriately halogenated intermediate (237). This
intermediate may then be functionalised in multiple ways. For
example, displacement with an appropriately substituted piperazine
(using either heat or transition metal mediated reactions) will
give the desired product (242). Alternatively, the core may be
halogenated, using (for example) an organolithium base and a
halogen source (e.g., NCS, Br.sub.2, I.sub.2, etc.) to give
compound (238). In the case of bromination, both non-regioselective
and polybromination are observed, allowing an entry into more fully
substituted and functionalised derivatives, (239). These may then
be subjected to any number of anionic or transition metal-mediated
reactions (eg. Suzuki, Stille, Negishi, etc.) to provide further
functionality (e.g., (240) or (241)). In all cases, subsequent
displacement with an appropriately substituted piperazine (along
with subsequent functionalisation, if desired) gives rise to the
desired products (242).
[0286] Compounds (247) may be prepared as shown in FIG. 45.
4-Chloropyrrolo[2,3-d]pyrimidine (243) is oxidized with an
appropriate oxidizing agent (pyridinium tribromide, for example) in
an appropriate solvent (such as t-butanol), and the resulting
gem-dibromide is reduced under appropriate conditions (using
Zn/HOAc, for example) in an appropriate solvent (e.g., MeOH) to
give the lactam 244. Reaction of compound (244) with a
monoprotected diamine (245) (using the Boc protecting group, for
example) in an appropriate solvent (such as IPA or NMP) in the
presence or absence of base (such as triethylamine), followed by
deprotection with mineral acid (HCl, for example) furnishes
intermediate (246). Compound (246) is then combined with (for
example) a protected amino acid (using the Boc protecting group,
for example) in an appropriate solvent (such as DCM or DMF) with or
without base (such as triethylamine or DIEA) and treated with an
appropriate coupling agent (such as DCC, HBTU, or EDCI) to furnish
compound (247) after deprotection with acid (in the case of a
Boc-protecting group.)
[0287] FIG. 46 shows the general preparation of compounds (256). A
suitably substituted thiophenecarboxylic acid (248) may be
converted to the protected aminothiophene (249) by means of a
rearrangement, using (for example) diphenylphosphorylazide in the
presence of a suitable base and solvent (e.g., t-BuOH) at elevated
temperature. This can be treated with a suitable malonate
derivative (eg. 2-ethoxymethylene malonate) at high temperature to
give the pyridothiophene (250). Halogenation using (for example)
POCl.sub.3 in the presence of base (eg. NEt.sub.3) gives the
chloride (251) which may then be treated with an appropriate linker
(eg. Boc-piperazine) and appropriately deprotected to give the
piperazine intermediate (252). Acylation (using, for example, a
suitably protected amino acid under standard conditions) followed
by deprotection (if needed), gives the desired product (254).
Alternatively, in place of deprotection, the ester may be
saponified using aqueous basic conditions (e.g., LiOH in water and
methanol) to give the acid (255) which may then be removed by
decarboxylation by heating at high temperature in an appropriate
solvent (e.g., diphenyl ether.) After any additional and necessary
deprotection, the desired product is attained (256).
[0288] The general preparation of compounds (265) is shown in FIG.
47. Hence protection of an appropriately substituted halopyridone
(257) with (for example) an acetate group under standard
conditions) and a subsequent transition metal mediated introduction
of a functionalized actyylene (258) (for example, using
PdCl.sub.2(PPh.sub.3).sub.2 and CuI) gives the intermediate
acetylene (260). Treatment with base (e.g., aqueous
Na.sub.2CO.sub.3 and heat) effects cyclisation to give the
pyridylfuran core (261). Halogenation, via N-oxidation (eg. mcpba
oxidation) and treatment with a halogenating agent (eg. POCl.sub.3)
gives the halide (263), which is then displaced by (for example)
piperazine to give (264), and then subsequently further
functionalised by (for example) the introduction of a suitably
protected amino acid, followed by deprotection to give the desired
product (265).
[0289] FIG. 48 illustrates the general preparation of compound
(271). Coupling of the uracil derivative (266) with 3-aminopyrazole
(267) in the presence of base gives the pyrazolopyridone (268).
Halogenation (for example, using POCl.sub.3) gives the halide
(269), displacement with a suitably substituted linker (e.g.,
Boc-piperazine using heat) and removal of the protecting group
(e.g., TFA, for a Boc-group), followed by acylation using (for
example) a protected amino acid (followed by the appropriate
deprotection) gives the desired product (271).
[0290] In order to illustrate the invention, the following examples
are included. However, it is to be understood that these examples
do not limit the invention and are only meant to suggest a method
of practicing the invention. Persons skilled in the art will
recognize that the chemical reactions described may be readily
adapted to prepare a number of other analogs of the invention, and
alternative methods for preparing the compounds of this invention
are deemed to be within the scope of this invention. For example,
the synthesis of non-exemplified compounds according to the
invention may be successfully performed by modifications apparent
to those skilled in the art, e.g., by appropriately protecting
interfering groups, by utilizing other suitable reagents known in
the art other than those described, and/or by making routine
modifications of reaction conditions. Alternatively, other
reactions disclosed herein or known in the art will be recognized
as having applicability for preparing other compounds of the
invention.
BIOLOGICAL EXAMPLES
Example of AKT-1 Kinase Assay
[0291] The activity of the compounds described in the present
invention may be determined by the following procedure: This
procedure describes a kinase assay that measures the
phosphorylation of a fluorescently-labeled peptide by full-length
human recombinant active AKT-1 by fluorescent polarization using a
commercially available IMAP kit.
[0292] The assay materials come from an IMAP AKT Assay Bulk Kit,
product #R8059, from Molecular Devices, Sunnyvale, Calif. The kit
materials include an IMAP Reaction Buffer (5.times.): The diluted
1.times. IMAP Reaction Buffer contains 10 mM Tris-HCl, pH 7.2, 10
mM MgCl.sub.2, 0.1% BSA, 0.05% NaN3. DTT is routinely added to a
final concentration of 1 mM immediately prior to use.
[0293] Also included are IMAP Binding Buffer (5.times.), and IMAP
Binding Reagent. The Binding Solution is prepared as a 1:400
dilution of IMAP Binding Reagent into 1.times. IMAP Binding
Buffer.
[0294] Fluorescein-labeled AKT Substrate (Crosstide), having the
sequence (Fl)-GRPRTSSFAEG. A stock solution of 20 .mu.M is made up
in 1.times. IMAP Reaction Buffer.
[0295] The plates used include a Costar 3657 (382-well made of
polypropylene and having a white, v-bottom) that is used for
compound dilution and for preparing the compound-ATP mixture. The
assay plate was the Packard ProxyPlate.TM.-384 F.
[0296] The AKT-1 used was made from full-length, human recombinant
AKT-1 that is activated with PDK1 and MAP kinase 2.
[0297] The assay procedure starts the preparation of stock
solutions of compounds at 10 mM in DMSO. The stock solutions and
the control compound are serially diluted 1:2 nine times into DMSO
(10 .mu.L of compound+10 .mu.L of DMSO) to give 50.times. dilution
series over the desired dosing range. Then, 2.1-.mu.L aliquots of
the compounds in DMSO are transferred to a Costar 3657 plate
containing 50 .mu.L of 10.4 .mu.M ATP in 1.times. IMAP Reaction
Buffer containing 1 mM DTT. After thorough mixing, 2.5-.mu.L
aliquots are transferred to a ProxyPlate.TM.-384 F plate.
[0298] The assay is initiated by the addition of 2.5-.mu.L aliquots
of a solution containing 200 nM of fluorescently-labeled peptide
substrate and 4 nM AKT-1. The plate is centrifuged for 1 minute at
1000 g and incubated for 60 minute at ambient temperature. The
reaction is then quenched by the addition of 15 .mu.L of Binding
Solution, centrifuged again and incubated for an additional 30
minutes at ambient temperature prior to reading on a Victor 1420
Multilabel HTS Counter configured to measure fluorescence
polarization.
[0299] Several compounds of Formula I tested in the assay showed
activity for inhibiting AKT protein kinases, including
(2R)-2-amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-prop-
an-1-one,
(2R)-2-amino-3-(2-napthyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)--
propan-1-one, and
(2R)-2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2,b]pyridin-7-yl-piperazin--
1-yl)-propan-1-one.
PREPARATIVE EXAMPLES
[0300] The compounds of the present invention may be prepared
either as either the racemate or as a single enantiomer (for
example, using enantiomerically pure reagents. If prepared as the
racemate, the corresponding enantiomers may be isolated by
separation of the racemic mixture of on a chiral stationary phase
column utilizing normal or reverse phase HPLC techniques.
Alternatively, a diastereomeric mixture can be prepared by
treatment of the racemic mixture with an appropriate chiral acid
(or suitably activated derivative), for example dibenzoyl tartrate
or the like (see, for example, Kinbara, K., et. al., J. Chem. Soc.,
Perkin Trans. 2, 1996, 2615; and Tomori, H., et. al., Bull. Chem.
Soc. Jpn., 1996, 3581). The diastereomers would then be separated
by traditional techniques (i.e. silica chromatography,
crystallization, HPLC, etc) followed by removal of the chiral
auxiliary to afford enantiomerically pure material.
[0301] The examples below describe the synthesis of some of the
compounds of the invention. Unless otherwise indicated all
temperature s are set forth in degrees Celsius. Reagents were
purchased from commercial suppliers such as Aldrich Chemical
Company, Lancaster, TCI or Maybridge, and were used without further
purification unless otherwise indicated.
[0302] The reactions set forth below were done generally under a
positive pressure of nitrogen or argon or with a drying tube
(unless otherwise stated) in anhydrous solvents, and the reaction
flasks were typically fitted with rubber septa for the introduction
of substrates and reagents via syringe. Glassware was oven dried
and/or heat dried.
[0303] HPLC retention times (R.sub.t) are reported in minutes.
Unless stated otherwise, the following HPLC conditions were used to
obtain the reported retention times: column: Waters YMC ODS-AQ,
3.0.times.50 mm; 5-95% gradient MeCN in water (0.01% HFBA, 1% IPA);
flow rate: 1.00 mL/min; detected at 220 nm.
[0304] .sup.1H-NMR spectra were recorded on a Varian instrument
operating at 400 MHz. .sup.1H-NMR spectra were obtained as
CDCl.sub.3 solutions (reported in ppm), using chloroform as the
reference standard (7.25 ppm). Other NMR solvents were used as
needed. When peak multiplicities are reported, the following
abbreviations are used: s (singlet), d (doublet), t (triplet), m
(multiplet), br (broadened), dd (doublet of doublets), dt (doublet
of triplets).
Example 1A
##STR00015##
[0305] Preparation of 4-piperazinylquinazoline Amino Amides
[0306] Step 1:
[0307] To a solution of 4-chloroquinazoline (2.0 g, 12.2 mmol) (To
be, Masanori, et al., Bioorg. Med. Chem. 2003, 11(3), 383) and DIEA
(3.2 mL, 18.2 mmol) in 40 mL IPA was added Boc-piperazine (1.96 g,
12.81 mmol). The reaction mixture was heated to reflux and stirred
for 20 hours, after which it was cooled to room temperature and
concentrated by rotary evaporation. The residue was dissolved in
dichloromethane (DCM) and washed with 1N NaOH. The organic layer
was dried (Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation. The resulting oil was dissolved in 25 mL dioxane, and
4M HCl/dioxane (46 mL, 182 mmol) was added dropwise. The suspension
was sonicated for 2 minutes and stirred 13 hours at room
temperature, after which the reaction mixture was concentrated to
dryness by rotary evaporation. The resulting amine HCl salt was
dissolved in 2N NaOH and extracted with DCM. The organic layer was
dried (Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation. The resulting oil was purified on silica (9:1:0.02
DCM/MeOH/NH.sub.4OH) to give 4-piperazinylquinazoline as a yellow
oil (2.5 g, 96%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.74
(s, 1H), 7.92-7.86 (m, 2H), 7.76-7.70 (m, 1H), 7.48-7.42 (m, 1H),
3.75 (t, J=4.9 Hz, 4H), 3.09 (t, J=4.9 Hz, 4H), 1.89 (br s, 1H).
R.sub.t 0.70. MS (ESI+) [M+H].sup.+ 215.
[0308] Step 2:
[0309] To a Jones tube containing PS-CDI (Argonaut, 1.04 mmol/g,
2.2 equivalents) suspended in a solution of the
4-piperazinylquinazoline (1.0 equivalent) in CHCl.sub.3 was added a
solid Boc-protected amino acid (1.5 equivalents) (see Example 1B).
The reaction mixture was shaken for 15 hours at room temperature,
after which it was vacuum filtered, the resin rinsed with
CHCl.sub.3, and the filtrate concentrated by rotary evaporation. If
necessary, the crude coupled product was purified on silica
(DCM/EtOAc or DCM/MeOH). The resulting Boc-amino amide was
dissolved in minimal dioxane, and 4M HCl/dioxane (10 equivalents)
was added. The suspension was sonicated 5 minutes and stirred at
room temperature for 12 hours, after which it was concentrated by
rotary evaporation. The solids were dispersed in ether, isolated by
filtration with nitrogen pressure, and dried under reduced pressure
to give the corresponding 4-piperazinylquinazoline amino amide as
the hydrochloride salt. If necessary, the hydrochloride salts were
free-based with 1N NaOH, extracted with DCM, and the combined
organic layers were dried (Na.sub.2SO.sub.4), filtered,
concentrated by rotary evaporation, and dried under reduced
pressure.
Example 1B
[0310] The following amino acids were introduced as Boc-protected
amino acids to the 4-piperazinylquinazoline in Example 1, Step
2:
##STR00016## ##STR00017##
[0311] The compounds described in Examples 2-21 were prepared as
described in Example 1, Step 2, using 4-piperazinylquinazoline and
the appropriate amino acid shown in Example 1B.
Example 2
##STR00018##
[0312]
(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-
-one
[0313] R.sub.t 2.15. MS (ESI+) [M+H].sup.+ 362.
Example 3
##STR00019##
[0314]
(2R)-2-Amino-4-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1--
one
[0315] R.sub.t 2.26. MS (ESI+) [M+H].sup.+ 376.
Example 4
##STR00020##
[0316]
(2R)-2-Amino-3-(4-methoxphenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one
[0317] R.sub.t 2.15. MS (ESI+) [M+H].sup.+ 392.
Example 5
##STR00021##
[0318]
(2R)-2-Amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-3-(2-thienyl)-pro-
pan-1-one
[0319] R.sub.t 2.10. MS (ESI+) [M+H].sup.+ 368.
Example 6
##STR00022##
[0320]
(2R)-2-Amino-3-(3-indolyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-prop-
an-1-one. R.sub.t 2.24. MS (ESI+) [M+H].sup.+ 401.
Example 7
##STR00023##
[0321]
(2R)-2-Amino-3-(2-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one
[0322] R.sub.t 2.23. MS (ESI+) [M+H].sup.+ 396.
Example 8
##STR00024##
[0323]
(2R)-2-Amino-3-(3-chlorophenyl-(4-quinazolin-4-yl-piperazin-1-yl)-p-
ropan-1-one
[0324] R.sub.t 2.30. MS (ESI+) [M+H].sup.+ 396.
Example 9
##STR00025##
[0325]
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one
[0326] R.sub.t 2.36. MS (ESI+) [M+H].sup.+ 396. .sup.1H NMR (free
base; CDCl.sub.3, 400 MHz) .delta. 8.73 (s, 1H), 7.89 (d, J=8.4 Hz,
1H), 7.78 (d, J=8.4 Hz, 1H), 7.77-7.71 (m, 1H), 7.49-7.43 (m, 1H),
7.31-7.26 (m, 2H), 7.23-7.19 (m, 2H), 4.37 (br s, 1H), 3.92-3.82
(m, 1H), 3.73-3.57 (m, 6H), 3.39-3.30 (m, 1H), 3.29-3.19 (m, 1H),
3.12-3.02 (m, 2H).
Example 10
##STR00026##
[0327]
(2R)-2-Amino-3-(4-fluorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one (12)
[0328] R.sub.t 2.23. MS (ESI+) [M+H].sup.+ 380.
Example 11
##STR00027##
[0329]
(2R)-2-Amino-3-(2-naphthyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pr-
opan-1-one
[0330] R.sub.t 2.39. MS (ESI+) [M+H].sup.+ 412.
Example 12
##STR00028##
[0331]
(2R)-2-Amino-2-(2-indanyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-eth-
an-1-one
[0332] R.sub.t 2.33. MS (ESI+) [M+H].sup.+ 388.
Example 13
##STR00029##
[0333]
(.+-.)-2-(4-Fluorophenyl)-2-piperazinyl-1-(4-quinazolin-4-yl-pipera-
zin-1-yl)-ethan-1-one
[0334] R.sub.t 2.21 minutes. MS (ESI+) [M+H].sup.+ 435.
Example 14
##STR00030##
[0335]
(2R)-2-Amino-3-(2,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin--
1-yl)-propan-1-one
[0336] R.sub.t 2.39. MS (ESI+) [M+H].sup.+ 430.
Example 15
##STR00031##
[0337]
(2R)-2-Amino-3-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin--
1-yl)-propan-1-one
[0338] R.sub.t 2.43. MS (ESI+) [M+H].sup.+ 430.
Example 16
##STR00032##
[0339]
(2R)-2-(N-Methylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one
[0340] R.sub.t 2.15. MS (ESI+) [M+H].sup.+ 376.
Example 17
##STR00033##
[0341] Preparation of
(2R)-2-(N,N-Dimethylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)--
propan-1-one
[0342] To a solution of
(2R)-2-(N-Methylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-prop-
an-1-one (free-base, 14 mg, 0.037 mmol) in 1,2-dichloroethane was
added a solution of formaldehyde (37% w/w, 50 .mu.L, 0.61 mmol) and
then Na(OAc).sub.3BH (0.12 mmol). The reaction mixture was stirred
at room temperature for 3 hours, after which saturated NaHCO.sub.3
was added, and the reaction mixture was stirred 10 minutes. The
suspension was extracted with DCM, and the combined extracts were
dried (Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation. The residue was filtered through a Fluorosil plug with
ETOAC, and the filtrate was concentrated by rotary evaporation. The
resulting residue was dissolved in ether and excess 4M HCl/dioxane
was added. The yellow solids were isolated by filtration with
nitrogen pressure and dried under reduced pressure to afford the
desired product (7 mg, 41%) as the dihydrochloride salt. R.sub.t
2.20. MS (ESI+) [M+H].sup.+ 390.
Example 18
##STR00034##
[0343]
(4-Quinazolin-4-yl-piperazin-1-yl)-((3R)-1,2,3,4-tetrahydro-isoquin-
olin-3-yl)-methanone
[0344] R.sub.t 2.21. MS (ESI+) [M+H].sup.+ 374.
Example 19
##STR00035##
[0345] Preparation of
(2R)-2-(2-Aminoacetamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-p-
ropan-1-one
[0346]
(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-
-one (free-base) was acylated with Boc-glycine and deprotected
according to Step 2 of Example 1 to furnish
(2R)-2-(2-Aminoacetamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-p-
ropan-1-one. R.sub.t 2.23. MS (ESI+) [M+H].sup.+ 419.
Example 20
##STR00036##
[0347] Preparation of
(2R)-2-(3-aminopropionamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one
[0348]
(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-
-one (free-base) was acylated with Boc-homoalanine and deprotected
according to according to Step 2 of Example 1 to furnish
(2R)-2-(3-Aminopropionamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one. R.sub.t 2.24. MS (ESI+) [M+H].sup.+ 433.
[0349] For the compounds prepared below, the HPLC conditions used
to obtain the reported retention times (minutes) were: column:
Waters YMC ODS-AQ, 4.6.times.50 mm; 5-95% gradient MeCN in water
(0.01% HFBA, 1% IPA); flow rate: 2.00 mL/min; detected at 220
nm.
Example 21
##STR00037##
[0350] Preparation of
(2R)-2-[(2-Aminoethyl)amino]-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-pipe-
razin-1-yl)-propan-1-one
[0351]
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one (free-base, 41 mg, 0.104 mmol) was stirred at room
temperature with Boc-2-aminoacetaldehyde (16 mg, 0.104 mmol) in 0.7
mL methanol for 1 hour, after which NaBH.sub.4 (6 mg, 0.160 mmol)
was added. The reaction mixture was stirred for 3 hours and then
quenched with 1N NaOH. The reaction mixture was extracted with DCM,
and the combined extracts were dried (Na.sub.2SO.sub.4), filtered,
and concentrated by rotary evaporation. The crude residue was
purified on silica (15:1 DCM/MeOH). The resulting Boc-amine
intermediate was treated with 1.4 mL 4M HCl/dioxane and stirred at
room temperature for 14 hours, after which the reaction mixture was
diluted with ether. The solids were isolated by filtration with
nitrogen pressure and dried under reduced pressure to give the
desired product (29 mg, 51%) as the trihydrochloride salt. R.sub.t
1.69. MS (CI+) [M+H].sup.+ 439.
Example 22
##STR00038##
[0352] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-((2R)-2-methyl-4-quinazolin-4-yl-pipera-
zin-1-yl)-propan-1-one
[0353] 4-((3R)-3-methylpiperazinyl)quinazoline was prepared in a
similar fashion to Step 1 of Example 1, which was then acylated
with Boc-4-chloro-D-phenylalanine and deprotected according to Step
2 of Example 1 to furnish
(2R)-2-Amino-3-(4-chlorophenyl)-1-((2R)-2-methyl-4-quinazolin-4-yl-pipera-
zin-1-yl)-propan-1-one. R.sub.t 1.72. MS (CI+) [M+H].sup.+ 410.
Example 23
##STR00039##
[0354] Preparation of
2-Amino-3-(4-chlorophen)-1-[4-(6-phenylquinazolin-4-yl)-piperazin-1-yl]-p-
ropan-1-one
[0355] Step 1: A solution of 6-bromoquinazolin-4-ol (1.0 g, 4.44
mmol) in POCl.sub.3 (10 mL) was stirred and heated at 110.degree.
C. in a sealed tube overnight. The solution was cooled to room
temperature and poured onto ice (200 g.) The solution was extracted
with DCM (300 mL), washed with water (200 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to give the impure
6-bromo-4-chloroquinazoline as a brown solid that was not purified
further (1.5 g.) MS (APCI+) [M+H].sup.+ 243.1.
[0356] Step 2: A solution of the crude product from Step 1 (1.5 g),
piperazine-1-carboxylic acid tert-butyl ester (2.29 g, 12.3 mmol)
and triethylamine (2.15 mL, 15.4 mmol) in N-methylpyrrolidinone (50
mL) was stirred and heated at 80.degree. C. for 2 hours. The
solution was cooled to room temperature, diluted with EtOAc (200
mL), washed with water (3.times.200 mL) and dried over
Na.sub.2SO.sub.4. The mixture was purified by silica gel column
chromatography (50% EtOAc/hexanes) to give
4-(6-bromo-quinazolin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester as a colorless oil (1.5 g, 3.8 mmol, 85% from Step 1.) MS
(APCI+) [M+H].sup.+ 394.9 and 392.9. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.76 (1H, s), 8.02 (1H, s), 7.84-7.78 (2H, m), 3.74
(4H, s), 3.66 (4H, s), 1.51 (9H, s.)
[0357] Step 3: HCl (1.0 M in Et.sub.2O, 30 mL) was added to a
solution of 4-(6-bromo-quinazolin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester (1.50 g, 3.81 mmol) in MeOH (50 mL) and
stirred at room temperature overnight. The mixture was concentrated
in vacuo to give 6-bromo-4-piperazin-1-yl-quinazoline as the
bis-hydrochloride salt (1.3 g, 93%.) MS (APCI+) [M+H].sup.+
295.1.
[0358] Step 4: EDCI.HCl (230 mg, 1.2 mmol), HOBt (160 mg, 1.2 mmol)
and Boc-D-4-chlorophenylalanine (240 mg, 1.2 mmol) were added to a
stirred solution of 6-bromo-4-piperazin-1-yl-quinazoline
bis-hydrochloride (360 mg, 0.98 mmol) and triethylamine (0.30 mL,
1.2 mmol) in DMF (8 mL) at room temperature under nitrogen. Stirred
at room temperature overnight. Diluted with EtOAc (100 mL) and
washed with water (3.times.50 mL.) Dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The mixture was purified by silica gel
column chromatography (100% EtOAc) to give
[2-[4-(6-bromoquinazolin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-e-
thyl]-carbamic acid tert-butyl ester as a yellow solid (320 mg,
57%.) MS (APCI+) [M+H].sup.+ 575.9. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.75 (1H, s), 7.95 (1H, s), 7.85-7.79 (2H, m),
7.30-7.27 (2H, m), 7.17 (2H, d, J 7.1 Hz), 5.39 (1H, app. d, J 8.2
Hz), 4.86 (1H, app. d, J 7.3 Hz), 3.83-3.60 (6H, m), 3.36-3.26 (2H,
m), 3.04-2.96 (2H, m), 1.43 (9H, s.).
[0359] Step 5: THF (5 mL) was added to a stirred mixture of
Pd.sub.2 dba.sub.3 (8.0 mg, 0.0087 mmol) and triphenylarsine (11
mg, 0.035 mmol) at room temperature under nitrogen. The yellow
solution was stirred at room temperature for 2 minutes and then
transferred via cannula to a stirred solution of
[2-[4-(6-bromoquinazolin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-e-
thyl]-carbamic acid tert-butyl ester (50 mg, 0.087 mmol) and
phenylboronic acid (21 mg, 0.17 mmol) in ethylene glycol dimethyl
ether (5 mL) and aqueous sodium carbonate (2M, 5 mL) and stirred
and heated at 80.degree. C. under nitrogen overnight. The reaction
was cooled to room temperature, extracted into EtOAc (100 mL),
washed with water (50 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The mixture was purified by silica gel
column chromatography (100% EtOAc) to give
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-phenylquinazolin-4-yl)-piperazin-1-yl]--
ethyl}-carbamic acid tert-butyl ester as an oil (30 mg, 60%.) MS
(APCI+) [M+H].sup.+ 574.0 and 572.0.
[0360] Step 6: Trifluoroacetic acid (4 mL) was added to a stirred
solution of
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1--
yl]-ethyl}-carbamic acid tert-butyl ester (30 mg) in DCM (10 mL) at
room temperature. The solution was stirred at room temperature for
4 hours, quenched with aqueous NaOH (1N, 10 mL), diluted with EtOAc
(100 mL) and washed with aqueous NaOH (2.times.50 mL.) The organic
phase was dried over Na.sub.2SO.sub.4 and concentrated in vacuo to
give
2-amino-3-(4-chlorophenyl)-1-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1-yl-
]-propan-1-one as a colorless oil (23 mg.) .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 8.77 (1H, s), 8.02-8.00 (2H, m), 7.98 (1H, s),
7.64 (2H, d, J 7.4 Hz), 7.53 (2H, t, J 7.7 Hz), 7.44 (1H, t, J 7.3
Hz), 7.30 (2H, d, J 8.6 Hz), 7.17 (2H, d, J 8.6 Hz), 3.98 (1H, t, J
7.3 Hz), 3.87-3.60 (6H, m), 3.42-3.34 (2H, m), 2.97 (1H, dd, J 7.4
and 13.3 Hz), 2.84 (1H, dd, J 7.0 and 13.7 Hz.) The oil was taken
up into THF (10 mL) and treated with HCl (1.0M in Et.sub.2O, 10 mL)
and concentrated in vacuo to give the bis hydrochloride salt of
2-amino-3-(4-chlorophenyl)-1-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1-yl-
]-propan-1-one as a solid (21 mg.).
Example 24
##STR00040##
[0361] Preparation of
(2R)-2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2,b]pyridin-7-yl-piperazin--
1-yl)-propan-1-one dihydrochloride
[0362] Step 1: NaH (60% in mineral oil, 0.24 g) in DMF (15 mL) was
added Thieno[3,2-b]pyridin-7-ol (0.756 g) portionwise. The reaction
mixture was warmed at 40.degree. C. and stirred for 30 minutes.
After cooling, N-phenyltrifluoromethanesulfonimide (2.1 g) was
added, the reaction mixture was stirred at room temperature for 1
hour, and the Boc-piperazine (1.9 g) was added. The mixture was
stirred at 80.degree. C. for 2 hours. Ethyl acetate (100 mL) was
added and the resulting solution was washed with brine (2.times.50
mL), dried over sodium sulfate, concentrated under reduced pressure
and purified by chromatography (1:4 hexane/EtOAc) to give the
product as yellow oil 1.32 g (82.5%). R.sub.t 2.10 minutes. MS
(ESI+) [M+H].sup.+ 320.
[0363] Step 2: The 4-thieno[3,2-b]pyridin-7-piperazine-1-carboxylic
tert-butyl ester (1.32 g) in DCM (20 mL) was added the 4 N HCl in
dioxane (21 mL). The reaction was stirred at room temperature for
10 hours. The solvent was removed under reduced pressure and the
resulting amine HCl salt was dissolved in saturated sodium
bicarbonate (20 mL) and extracted with DCM (30 mL). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and concentrated by
rotary evaporation. to give 7-piperazin-1-yl-thieno[3,2-b]pyridine
as an off-white solid (0.85 g, 93.8%). R.sub.t 1.40 minutes. MS
(ESI+) [M+H].sup.+ 220.
[0364] Step 3: DIEA (0.07 mL) and HBTU (0.12 g) was added to the
solution of the
(2R)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(0.092 g) in THF (5 mL) at 0.degree. C. The mixture was stirred at
room temperature for 20 minutes, then
7-piperazin-1-yl-thieno[3,2-b]pyridine (0.056 g) was added. The
reaction was stirred at room temperature for 1 hr. 20 mL of EtOAc
was added and the organic layer was separated. The aqueous layer
extracted with EtOAc (20 mL). The combined organic layer was washed
with saturated sodium bicarbonate (20 mL) and dried over sodium
sulfate. After removal of solvent, the residue was purified by
flash chromatography (EtOAc) to give the product as white foam
solid (0.126 g, 98.5%). R.sub.t 2.43 minutes. MS (ESI+) [M+H].sup.+
501.
[0365] Step 4: The resulting Boc-amino amide (0.056 mg) was
dissolved in dioxane, and 4M HCl/dioxane (0.5 mL) was added. The
suspension was stirred at room temperature for 3 hours, after which
it was concentrated to give the corresponding amino amide as the
hydrochloride salt (0.53 g, 98%). R.sub.t 1.77 minutes. MS (ESI+)
[M+H].sup.+ 401.
Example 25
##STR00041##
[0366] Preparation of
2-Amino-3-(4-chlorophenyl)-1-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-y-
l)-propan-1-one dihydrochloride
[0367] Step 1: To a solution of Boc-D-Phe(4-Cl)--OH (3.65 g, 12.2
mmol), piperazine (10 g, 116 mmol) in DCM (200 mL) were added HOBT
(3.3 g, 24 mmol) and EDCI (4.7 g, 25 mmol). The mixture was stirred
at room temperature for 12 hours. The solution was washed with
water, brine and dried over magnesium sulfate. After filtration,
the organic solvent was evaporated and the residue was subject to
silica gel chromatography to afford the product
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester (0.58 g, 13%). .sup.1H NMR (CDCl.sub.3, 400 Hz)
.delta. 7.25 (d, J=8.4 Hz, 2H), 7.12 (d, J=8.0 Hz, 2H), 5.52 (d,
J=8.4 Hz, 1H), 4.83-4.77 (m, 1H), 3.63-3.45 (m, 2H), 3.37-3.32 (m,
1H), 3.08-3.04 (m, 1H), 2.99-2.90 (m, 2H), 2.81-2.70 (m, 3H),
2.42-2.38 (m, 1H), 1.41 (s, 9H). MS (ESI+) [M+H].sup.+ 369.
[0368] Step 2: To a solution of 3-aminothiophene-2-carboxylic
methyl ester (20 g, 127 mmol) in formic acid (100 mL) was added
ammonium acetate (13 g, 169 mmol). The mixture was refluxed for 3
hours. After cooling to room temperature, the precipitate was
filtered, washed with water and dried under vacuum to afford
3-formylaminothiophene-2-carboxylic acid methyl ester (20.5 g,
87%). .sup.1H NMR (DMSO 400 Hz) .delta. 10.38 (s, 1H), 8.42 (s,
1H), 8.00 (d, J=5.2 Hz, 1H), 7.90 (d, J=5.6 Hz, 1H), 3.84 (s, 3H).
MS (ESI+) [M+H].sup.+ 186.
[0369] Step 3: To a mixture of 3-formylaminothiophene-2-carboxylic
acid methyl ester (20.5 g, 111 mmol) and ammonium formate (21 g,
333 mmol) was added formamide (29.8 g, 662 mmol). The slurry was
heated to 140.degree. C. for 10 hours. After cooling, the solid was
filtered, washed with water and dried under vacuum to afford the
product 3H-Thieno[3,2-d]pyrimidin-4-one (12.5 g, 74%). .sup.1H NMR
(DMSO, 400 Hz) .delta. 12.31 (br, 1H), 8.18 (d, J=5.2 Hz, 1H), 8.16
(s, 1H), 7.41 (d, J=5.2 Hz, 1H). MS (ESI+) [M+H].sup.+ 153
[0370] Step 4: To a solution of DMF (13.2 mL, 170 mmol) in DCM (100
mL) at 0.degree. C. was added oxalyl chloride (22 mL, 252 mmol) in
DCM (100 mL) very slowly over 1 hour. To the resulting white gel
solution was added the 3H-thieno[3,2-d]pyrimidin-4-one (12 g, 79
mmol). The mixture was refluxed for 4 hours. After cooling, the
mixture was purred into water (500 mL) and extracted with DCM
(3.times.250 mL). Then the organic phase was dried over magnesium
sulfate, filtered and concentrated to afford
7-chloro-thieno[3,2-b]pyridine as white solid (13.4 g, 99%).
.sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 9.00 (s, 1H), 8.06 (d,
J=5.6 Hz, 1H), 7.61 (d, J=5.6 Hz, 1H). MS (ESI+) [M+H].sup.+
170
[0371] Step 5: The solution of
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester (60 mg, 0.163 mmol) and
4-Chlorothieno[3,2-d]pyrimidine 950 mg, 0.293 mmol) in Toluene (5
mL)/TEA (1 mL) was refluxed for 12 hours. After the solvent was
removed, the residue was subject to chromatography on silica gel to
afford
[1-(4-chlorobenzyl)-2-oxo-2-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl-
)-ethyl]-carbamic acid tert-butyl ester (71 mg, 86.7%). .sup.1H NMR
(CDCl.sub.3, 400 Hz) .delta. 8.60 (s, 1H), 7.76 (d, J=5.2 Hz, 1H),
7.45 (d, J=5.2 Hz, 1H), 7.26 (d, J=8.4 Hz, 1H), 7.15 (d, J=8.4 Hz,
1H), 5.39 (d, J=8.8 Hz, 1H), 4.88-4.82 (m, 1H), 3.94-3.76 (m, 4H),
3.69-3.58 (m, 3H), 3.25-3.22 (m, 1H), 2.98 (d, J=7.2 Hz, 2H), 1.43
(s, 9H). MS (ESI+) [M+H].sup.+ 503.
[0372] Step 6: To a solution of
[1-(4-chlorobenzyl)-2-oxo-2-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl-
)-ethyl]-carbamic acid tert-butyl ester in DCM (2 mL) was added HCl
in Dioxane (4M, 1 mL). The mixture was stirred at room temperature
for 4 hours. The solvent was removed to afford
2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-y-
l)-propan-1-one dihydrochloride quantitatively. MS (ESI+)
[M+H].sup.+ 402.
Example 26
##STR00042##
[0373] Preparation of
2-amino-3-(4-chlorophenyl)-1-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperaz-
in-1-yl]-propan-1-one dihydrochloride
[0374] Step 1: To a solution of 4,6-dichloro-5-aminopyrimidine (1
g, 6.1 mmol) in TEA (2 mL) and toluene (10 mL) was added
1-Boc-piperazine (2.3 g, 12.3 mmol). The mixture was refluxed for
12 hours. The solvent was removed and the residue was subject to
chromatography on silica gel to afford the product
4-(5-amino-6-chloropyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (1.9 g, 99%). .sup.1H NMR (CDCl.sub.3, 400 Hz)
.delta. 8.16 (s, 1H), 3.87 (s, 2H), 3.56 (m, 4H), 3.29 (m, 4H),
1.49 (s, 9H). MS (ESI+) [M+H].sup.+ 314.
[0375] Step 2: To a solution of
4-(5-amino-6-chloropyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (1 g, 3.19 mmol) and TMS-acetylene (1.5 g, 15
mmol) in TEA (10 mL) and THF (30 mL) were added
PdCl.sub.2(PPh.sub.3).sub.2 (0.33 g, 0.47 mmol) and CuI (0.1 g,
0.53 mmol) under N.sub.2. The mixture was heated to 80.degree. C.
for 20 hours. The solvent was removed and the residue was subject
to chromatography on silica gel to afford the product
4-(5-Amino-6-trimethylsilanylethynyl-pyrimidin-4-yl)-piperazine-1-carboxy-
lic acid tert-butyl ester (0.3 g, 25%). .sup.1H NMR (CDCl.sub.3,
400 Hz) .delta. 8.00 (s, 1H), 3.74 (s, 2H), 3.28-3.25 (m, 4H),
3.02-2.99 (m, 4H), 1.20 (s, 9H), 0.00 (s, 9H). MS (ESI+)
[M+H].sup.+ 376.
[0376] Step 3: To a solution of tBuOK (0.063 g, 0.56 mmol) in NMP
(4 mL) was added
4-(5-Amino-6-trimethylsilanylethynyl-pyrimidin-4-yl)-piperazine-
-1-carboxylic acid tert-butyl ester (0.1 g, 0.27 mmol) in NMP (1
mL) under N.sub.2. The mixture was vigorously stirred at room
temperature for 4 hours. The reaction was quenched with water (1
mL) and ethyl acetate (50 mL). The organic phase was washed brine
and water until NMP was gone, then dried over MgSO.sub.4, filtered
and concentrated. The residue was subject to chromatography on
silica gel to afford the product
4-(5H-Pyrrolo[3,2-d]pyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (43 mg, 53%). .sup.1H NMR (CDCl.sub.3, 400 Hz)
.delta. 9.47 (s, 1H), 8.49 (s, 1H), 7.37 (d, J=2.4 Hz, 1H), 6.62
(d, J=2.8 Hz, 1H), 3.85-3.82 (m, 4H), 3.61-3.58 (m, 4H). 1.49 (s,
9H). MS (ESI+) [M+H].sup.+ 304.
[0377] Step 4: To a solution of
4-(5H-Pyrrolo[3,2-d]pyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (43 mg, 0.14 mmol) in DCM (4 mL) was added HCl in
Dioxane (4M, 1 mL). The mixture was stirred at room temperature for
4 hours. The solvent was removed and the residue was treated with
TEA (2 mL), Boc-D-Phe(4-Cl)--OH (43 mg, 0.14 mmol), HOBT (30 mg,
0.222 mmol) and EDCI (41 mg, 0.214 mmol) in DCM (5 mL). The mixture
was stirred at room temperature for 12 hours. The solvent was
removed and the residue was subject to chromatography on silica gel
to give product
{1-(4-chlorobenzyl)-2-oxo-2-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazi-
n-1-yl]-ethyl}-carbamic acid tert-butyl ester (30 mg, 44%). .sup.1H
NMR (CDCl.sub.3, 400 Hz) .delta. 9.85 (s, 1H), 8.48 (s, 1H), 7.37
(s, 1H), 7.25-7.23 (d, J=8.4 Hz, 2H), 7.13-7.11 (d, J=8.4 Hz, 2H),
6.60 (s, 1H), 5.39-5.37 (d, J=8.8 Hz, 1H), 4.83-4.78 (m, 1H),
3.81-3.43 (m, 7H), 3.31-3.27 (m, 1H), 3.00-2.91 (m, 2H), 1.41 (s,
9H). MS (ESI+) [M+H].sup.+ 485.
[0378] Step 5: To a solution of
{1-(4-chlorobenzyl)-2-oxo-2-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazi-
n-1-yl]-ethyl}-carbamic acid tert-butyl ester in DCM (4 mL) was
added HCl in Dioxane (4M, 1 mL) and stirred for 4 hours. The
solvent was removed to afford
2-Amino-3-(4-chlorophenyl)-1-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)--
piperazin-1-yl]-propan-1-one dihydrochloride quantitatively. MS
(ESI+) [M+H].sup.+ 385.
Example 27
##STR00043##
[0379] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-
-1-yl]-propan-1-one
[0380] Step 1: To a solution of 4-hydroxy-3-nitro-pyridine (2 g, 14
mmol) in POCl.sub.3 (6 mL) was added PCl.sub.5 (2.5 g, 12 mmol).
The mixture was heated to reflux for 3 hours. The solvent was
evaporated and the residue was cooled with ice-water and vigorously
stirred with water (3 mL) and CHCl.sub.3 (6 mL). The aqueous was
extracted CHCl.sub.3 (5.times.5 mL). The organic phase was combined
and dried over MgSO.sub.4. After filtration, the solvent was
removed to afford the product 4-Chloro-3-nitro-pyridine (2.24 g,
99%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 9.13 (s, 1H), 8.70
(d, J=5.2 Hz, 1H), 7.55 (d, J=5.6 Hz, 1H). MS (ESI+) [M+H].sup.+
159.
[0381] Step 2: To a solution of 4-chloro-3-nitro-pyridine (2 g, 13
mmol) in dry THF (100 mL) under N.sub.2 at -78.degree. C. was added
excess vinyl magnesium bromide (1.0M, 40 mL, 40 mmol). The mixture
was stirred at -20.degree. C. for 8 hours before the reaction was
quenched with 20% NH.sub.4Cl (75 mL). The aqueous phase was
extracted with ethyl acetate (3.times.100 mL). The combined organic
layer was dried over MgSO.sub.4, filtered and concentrated. The
residue was subject to chromatography on silica gel to afford
7-Chloro-1H-pyrrolo[3,2-b]pyridine (0.3 g, 16%). .sup.1H NMR
(CD.sub.3OD, 400 Hz) .delta. 8.22 (d, J=5.2 Hz, 1H), 7.64 (d, J=3.2
Hz, 1H), 7.23 (d, J=5.2 Hz, 1H), 6.67 (d, J=3.2 Hz, 1H). MS (ESI+)
[M+H].sup.+ 153.
[0382] Step 3: To a solution of 7-Chloro-1H-pyrrolo[3,2-b]pyridine
(40 mg, 0.262 mmol) in xylene (4 mL) and TEA (1 mL) was added
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester (0.1 g, 0.27 mol). The mixture was refluxed for 6
days. The solvent was removed under vacuum and the residue was
subject to purification by HPLC to afford
{1-(4-chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin--
1-yl]-ethyl}-carbamic acid tert-butyl ester (10 mg, 8%). .sup.1H
NMR (CDCl.sub.3, 400 Hz) .delta. 11.81 (s, 1H), 7.95 (d, J=6.4 Hz,
1H), 7.40 (s, 1H), 7.15 (d, J=8.00 Hz, 2H), 6.46 (d, J=6.8 Hz, 2H),
5.42 (d, J=8.4 Hz, 1H), 4.82 (d, J=7.2 Hz, 1H), 3.87-3.28 (m, 8H),
2.98 (d, J=6.4 Hz, 2H), 1.41 (s, 9H). MS (ESI+) [M+H].sup.+
484.
[0383] Step 4: To a solution of
{1-(4-chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin--
1-yl]-ethyl}-carbamic acid tert-butyl ester in DCM (4 mL) was added
HCl in Dioxane (4M, 1 mL). The mixture was stirred at room
temperature for 4 hours. The solvent removed to afford the product
2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-
-1-yl]-propan-1-one quantitatively. MS (ESI+) [M+H].sup.+ 384.
Example 28
##STR00044##
[0384] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piper-
azin-1-yl]-propan-1-one
[0385] Step 1: To a solution of LDA (1.8M, 20.6 mL, 37.1 mmol) in
THF (65 mL) at -78.degree. C. was added
4-chloro-thieno[3,2-d]pyrimidine (5.26 g, 31 mmol) in THF (50 mL)
dropwise over 1 hour. After stirring for 20 minutes, I.sub.2 (12.7
g, 50 mmol) in THF (40 mL) was added to the mixture at -78.degree.
C. dropwise. The mixture was stirred at the same temperature for 20
minutes and then warmed up to room temperature for 2 hours. The
mixture was poured into water (100 mL) and stirred for 30 minutes.
The solid was filtered and washed with water and Hexane-Hexanes/DCM
(50:1) to afford the product 4-Chloro-6-iodothieno[3,2-d]pyrimidine
(6.86 g, 75%). .sup.1H NMR (DMSO, 400 Hz) .delta. 8.97 (s, 1H),
8.15 (s, 1H).
[0386] Step 2: To a solution of
4-chloro-6-iodothieno[3,2-d]pyrimidine (0.22 g, 0.742 mmol) in DCE
(5 mL)/TEA (2 mL) was added
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester (25 mg, 0.68 mmol). The mixture was refluxed for 2
hours. The solvent was removed and the residue was subject to
chromatography on silica gel to afford the product
{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1--
yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (44 mg, 95%).
.sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.47 (s, 1H), 7.64 (s,
1H), 7.27-7.25 (d, J=8.4 Hz, 2H), 7.17-7.15 (d, J=8.0 Hz, 2H), 5.44
(d, J=8.8 Hz, 1H), 4.85-4.81 (m, 1H), 3.86-3.50 (m, 7H), 3.24-3.19
(m, 1H), 2.99-2.97 (d, J=7.2 Hz, 2H), 1.43 (s, 9H). MS (ESI+)
[M+H].sup.+ 628.
[0387] Step 3: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1--
yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester in DCM (4 mL) was
added HCl in Dioxane (4M, 1 mL). The mixture was stirred at room
temperature for 4 hours. The solvent was removed to afford the
product
2-Amino-3-(4-chlorophenyl)-1-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piper-
azin-1-yl]-propan-1-one quantitatively. MS (ESI+) [M+H].sup.+
528.
Example 29
##STR00045##
[0388] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-y-
l)-piperazin-1-yl]-propan-1-one dihydrochloride
[0389] Step 1: To a solution of ZnBr.sub.2 (70 mg, 0.311 mmol) in
THF (2 mL) was added propargyl magnesium bromide (0.5M, 0.6 mL, 0.3
mmol) at room temperature. After stirring for 20 minutes, the
6-iodothieno[3,2-d]pyrimidine (50 mg, 0.08 mmol) was added. The
mixture was flushed with N.sub.2 and PdCl.sub.2(dppf) was added.
The mixture was stirred at room temperature under N.sub.2 for 12
hours. After filtration, the filtrate was concentrated and the
residue was subject to chromatography on silica gel to afford the
product
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl-
)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester (17 mg,
40%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.56 (s, 1H), 7.37
(s, 1H), 7.27-7.25 (d, J=8 Hz, 2H), 7.16-7.14 (d, J=8.4 Hz, 2H),
5.37-5.35 (d, J=8.4 Hz, 1H), 4.85-4.82 (m, 1H), 3.86-3.51 (m, 7H,
3.23-3.20 (m, 1H), 3.00-2.94 (m, 2H), 2.15 (s, 3H), 1.42 (s, 9H).
MS (ESI+) [M+H].sup.+ 540.
[0390] Step 2: To a solution of
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl-
)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester in DCM (4
mL) was added HCl in Dioxane (4M, 1 mL). The mixture was stirred
for 4 hours. The solvent was removed to afford the product
2-Amino-3-(4-chlorophenyl)-1-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-y-
l)-piperazin-1-yl]-propan-1-one dihydrochloride quantitatively. MS
(ESI+) [M+H].sup.+ 440.
Example 30
##STR00046##
[0391] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-
-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0392] Step 1: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1--
yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (50 mg, 0.080 mmol)
in DMF (3 mL) were added 2M Na.sub.2CO.sub.3 (0.1 mL) and
3-thiophenyl boronic acid (15 mg, 0.117 mmol). The mixture was
bubbled N.sub.2 for 20 minutes and then Pd(PPh.sub.3).sub.4 (10 mg,
0.012 mmol) was added. The mixture was heated to 90.degree. C. for
12 hours. The solvent was removed under vacuum and the residue was
subject to chromatography on silica gel to afford the product
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4--
yl)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester (16 mg,
34.4%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.57 (s, 1H),
7.70-7.42 (m, 4H), 7.27 (d, J=8.4, 2H), 7.16 (d, J=8.4 Hz, 2H),
5.36 (d, J=8.8 Hz, 1H), 4.87-4.84 (m, 1H), 3.91-3.59 (m, 7H),
3.27-3.24 (d, J=11.2 Hz, 1H), 3.00-2.99 (d, J=7.2 Hz, 2H), 1.43 (s,
9H). MS (ESI+) [M+H].sup.+ 584.
[0393] Step 2: To a solution of product
{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4--
yl)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester in DCM (4
mE) was added HCl in Dioxane (4M, 1 mL). The mixture was stirred at
room temperature for 4 hours. The solvent was removed to afford the
product
2-Amino-3-(4-chlorophenyl)-1-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-
-yl)-piperazin-1-yl]-propan-1-one dihydrochloride quantitatively.
MS (ESI+) [M+H].sup.+ 484.
Example 31
##STR00047##
[0394] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin--
4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0395] Step 1: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1--
yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (50 mg, 0.080
mmol), sodium methylthiolate (12 mg, 2.15 mmol) and
1,3-di-tert-butyl-propane-dione 940 mg, 0.22 mmol) was purged with
N.sub.2. NMP (2 mL) and CuCl (5 mg, 0.05 mmol) were added. The
mixture was heated to 130.degree. C. for 3 hours. After cooling
down, the reaction was diluted with ethyl acetate and filtered. The
filtrate was washed with water, brine and dried over MgSO.sub.4.
After filtration, the solvent was removed and the residue was
subject to chromatography on silica gel to afford the product
{1-(4-chlorobenzyl)-2-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-p-
iperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester. (20 mg,
46%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.61 (s, 1H), 7.57
(s, 1H), 7.28-7.26 (d, J=6.4 Hz, 2H), 7.18-7.16 (d, J=8.4 Hz, 2H),
5.33-5.31 (d, J=8.8 Hz, 1H), 4.81-4.79 (m, 1H), 4.01-3.71 (m, 7H),
3.24-3.19 (m, 1H), 3.03-2.98 (m, 2H), 2.74 (s, 3H), 1.43 (s, 9H).
MS (ESI+) [M+H].sup.+ 548.
[0396] Step 2: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-p-
iperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester in DCM
(4 mL) was added HCl in Dioxane (4M, 1 mL). The mixture was stirred
at room temperature for 4 hours. The solvent was removed to afford
the product
2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin--
4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride quantitatively.
MS (ESI+) [M+H].sup.+ 448.
Example 32
##STR00048##
[0397] Preparation of
4-{4-[2-Amino-3-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-thieno[3,2-d]-
pyrimidine-6-carbonitrile dihydrochloride
[0398] Step 1: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1--
yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (50 mg, 0.080 mmol)
in Pyridine (5 mL) was added CuCN (20 mg, 0.223 mmol). The mixture
was refluxed under N.sub.2 for 12 hours. The solvent was removed
and the residue was subject to chromatography on silica gel to
afford
{1-(4-chlorobenzyl)-2-[4-(6-cyanothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-
-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (29 mg, 69%).
.sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.65 (s, 1H), 7.95 (s,
1H), 7.28-7.26 (d, J=7.6 Hz, 2H), 7.18-7.16 (d, J=8.4 Hz, 2H).
5.35-5.33 (d, J=8.4 Hz, 1H), 4.85-4.80 (m, 1H), 3.91-3.56 (m, 7H,
3.24-3.19 (m, 1H), 3.01-2.99 (d, J=8.0 Hz, 2H), 1.44 (s, 9H). MS
(ESI+) [M+H].sup.+ 527.
[0399] Step 2: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-cyanothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-
-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester in DCM (4 mL) was
added HCl in Dioxane (4M, 1 mL). The mixture was stirred at room
temperature for 10 hours. The solvent was removed and the residue
was subject to purification by HPLC to afford the product
4-{4-[2-Amino-3-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-thieno[3,2-d]-
pyrimidine-6-carbonitrile dihydrochloride (12 mg, 44%). MS (ESI+)
[M+H].sup.+ 427.
Example 33
##STR00049##
[0400] Preparation of
2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride
[0401] Step 1: To a solution of
4-chloro-6-iodothieno[3,2-d]pyrimidine (0.5 g, 1.7 mmol) in DCE (5
mL)/TEA (1 mL) was added 1-Benzyl-piperazine (0.3 g, 1.69 mmol).
The mixture was refluxed for 1 hour. The solvent was removed and
the residue was subject to chromatography on silica gel to afford
(4-Benzyl-piperazin-1-yl)-6-iodothieno[3,2-d]pyrimidine (0.65 g,
88%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.47 (s, 1H) 7.60
(s, 1H), 7.35-7.23 (m, 5H), 3.95-3.92 (m, 4H), 3.51 (s, 2H),
2.59-2.54 (m, 4H). MS (ESI+) [M+H].sup.+ 437.
[0402] Step 2: To a suspension of ZnBr.sub.2 (0.5 g, 2.2 mmol)
dried under vacuum in THF (10 mL) was added MeMgBr (3M, 0.6 mL, 1.8
mmol) at room temperature dropwise. After addition, the mixture was
stirred for 1 hour, then
(4-Benzyl-piperazin-1-yl)-6-iodothieno[3,2-d]pyrimidine (0.4 g,
0.92 mmol) was added followed by PdCl.sub.2(dppf) (30 mg) under
N.sub.2. The mixture was heated to 60.degree. C. for 2 hours. The
reaction was quenched with water. The organic phase was separated
and dried over MgSO.sub.4. After filtration, the solvent was
removed and the residue was subject to chromatography on silica gel
to give the product
4-(4-Benzyl-piperazin-1-yl)-6-methylthieno[3,2-d]pyrimidine (0.16
g, 54%). .sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.52 (s, 1H),
7.35-7.26 (m, 5H), 7.07 (s, 1H), 3.98-3.95 (m, 4H), 3.56 (s, 2H),
2.59-2.57 (m, 7H). MS (ESI+) [M+H].sup.+ 325.
[0403] Step 3: To a solution of
4-(4-Benzyl-piperazin-1-yl)-6-methylthieno[3,2-d]pyrimidine (65 mg,
0.20 mmol) in MeOH (10 mL) was added Pd/C (10%, 20 mg) and two drop
of TFA. The mixture was stirred under H.sub.2 balloon for 4 hours.
The catalyst was filtered off and the filtrate was concentrated.
The residue was dissolved in DCM (6 mL) and TEA (2 mL), then
Boc-D-Phe(4-Cl)--OH (59 mg, 0.20 mmol) was added, followed by HOBT
(50 mg, 0.37 mmol) and EDCI (74 mg, 0.39 mmol). The mixture was
stirred at room temperature for 12 hours. The solvent was removed
and the residue was subject to chromatography on silica gel to give
the product
{1-(4-chlorobenzyl)-2-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin--
1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (60 mg, 59%).
.sup.1H NMR (CDCl.sub.3, 400 Hz) .delta. 8.54 (s, 1H), 7.28-7.26
(d, J=7.2 Hz, 2H), 7.17-7.15 (d, J=8.4 Hz, 2H), 7.08 (s, 1H),
5.37-5.35 9d, J=8.8 Hz, 1H), 4.86-4.84 (m, 1H), 3.87-3.57 (m, 7H),
3.24-3.22 (m, 1H), 3.00-2.98 (d, J=7.2 Hz, 2H), 2.63 (s, 3H), 1.43
(s, 9H). MS (ESI+) [M+H].sup.+ 516.
[0404] Step 4: To a solution of
{1-(4-chlorobenzyl)-2-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin--
1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester in DCM (4 mL) was
added HCl in Dioxane (4M, 1 mL). The mixture was stirred for 4
hours. The solvent was removed to afford the product
2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride quantitatively. MS (ESI+)
[M+H].sup.+ 416.
Example 34
##STR00050##
[0405] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride
[0406] Step 1: A mixture of 5-aminoindazole (2.53 g, 19.0 mmol),
bis(2-chloroethyl)amine hydrochloride (3.60 g, 20.1 mmol) and
ethanol (30 mL) was heated at reflux overnight. The mixture was
allowed to cool to room temperature. Na.sub.2CO.sub.3 (2.14 g, 20.2
mmol) was added and the reaction mixture heated at reflux for 8
hours. After cooling, the mixture was filtered and the filtrate
evaporated in vacuo. The residue was dissolved in 1 N HCl (100 mL)
and extracted with DCM (2.times.50 mL). The aqueous phase was made
basic with 4 N NaOH (30 mL) and extracted with EtOAc (2.times.100
mL). The combined organic layers were washed with brine, dried and
concentrated. The residue was purified by column chromatography
(20:1 DCM/MeOH to 20:1:0.5 DCM/MeOH/Et.sub.3N) to
5-piperazin-1-yl-1H-indazole (1.26 g, 33%) as a brown solid.
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 12.80 (s, 1H), 7.89 (s,
1H), 7.40 (d, J=8.8 Hz, 1H), 7.16 (dd, J=8.8 Hz, J=2.0 Hz, 1H),
7.07 (s, 1H), 3.17 (s, 1H), 2.99 (m, 4H), 2.89 (m, 4H). LCMS
(APCI+) m/z 203 [M+H].sup.+; Rt=1.33 minutes.
[0407] Step 2: To a solution of (D)-Boc-4-chlorophenylalanine
(0.119 g, 0.396 mmol) and 5-piperazin-1-yl-1H-indazole (0.100 g,
0.494 mmol) in DMF (5 mL) was added EDCI (0.152 g, 0.791 mmol),
HOBt (0.121 g, 0.791 mmol) and triethylamine (0.110 mL, 0.791
mmol). The reaction mixture was stirred at room temperature
overnight. The mixture was partitioned between water and EtOAc. The
aqueous phase was extracted with EtOAc. The combined organic layers
were washed with aqueous NaHCO.sub.3, brine, dried and
concentrated. The residue was purified by column chromatography
(80:1 to 50:1 DCM/MeOH) to give
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester (0.176 g, 92%) as a white solid.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 10.12 (s, 1H), 7.98 (s,
1H), 7.41 (d, J=8.8 Hz, 1H), 7.28 (d, J=8.4 Hz, 2H), 7.16 (d, J=8.4
Hz, 2H), 7.12 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.07 (s, 1H), 5.46 (m,
1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m, 1H), 3.31 (m, 1H), 3.07
(m, 1H), 2.99 (d, J=6.8 Hz, 2H), 2.91 (m, 2H), 2.49 (m, 1H), 1.43
(s, 9H). LCMS (APCI+) m/z 484, 486 [M+H].sup.+; Rt=3.01
minutes.
[0408] Step 3: To a solution of
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester (0.176 g, 0.364 mmol) in DCM (10
mL) was added 4 N HCl in dioxane (1 mL). The mixture was stirred at
room temperature overnight and then evaporated. The resulting solid
was suspended in isopropyl alcohol-ether (1:5) and stirred for 30
minutes. The mixture was filtered to give
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride (0.151 g, 91%) as a white solid. .sup.1H
NMR (CD.sub.3OD, 400 MHz) .delta. 8.24 (s, 1H), 8.03 (s, 1H), 7.75
(d, J=9.2 Hz, 1H), 7.65 (d, J=9.2 Hz, 1H), 7.47 (d, J=8.0 Hz, 2H),
7.37 (d, J=8.0 Hz, 2H), 4.82 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H),
3.73 (m, 2H), 3.51 (m, 2H), 3.22 (dd, J=13.2 Hz, J=6.0 Hz, 1H),
3.15 (dd, J=13.2 Hz, J=9.2 Hz, 1H), 2.87 (m, 1H). LCMS (APCI+) m/z
384, 386 [M+H].sup.+; Rt=1.92 minutes.
Example 35
##STR00051##
[0409] Preparation of
(2S)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride
[0410] Step 1:
(2S)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester was prepared by the procedures
described in Example 34, Step 2, substituting
(D)-Boc-4-chlorophenylalanine with (L)-Boc-4-chlorophenylalanine.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 10.03 (s, 1H), 7.98 (s,
1H), 7.41 (d, J=8.8 Hz, 1H), 7.28 (d, J=8.4 Hz, 2H), 7.16 (d, J=8.4
Hz, 2H), 7.12 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.07 (s, 1H), 5.46 (m,
1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m, 1H), 3.31 (m, 1H), 3.07
(m, 1H), 2.99 (d, J=6.8 Hz, 2H), 2.91 (m, 2H), 2.49 (m, 1H), 1.43
(s, 9H).
[0411] Step 2:
(2S)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride was prepared by the procedures described
in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2S)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.24 (s, 1H), 8.03 (s, 1H), 7.75 (d, J=9.2 Hz, 1H),
7.65 (d, J=9.2 Hz, 1H), 7.47 (d, J=8.0 Hz, 2H), 7.37 (d, J=8.0 Hz,
2H), 4.82 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H), 3.73 (m, 2H), 3.51
(m, 2H), 3.22 (dd, J=13.2 Hz, J=6.0 Hz, 1H), 3.15 (dd, J=13.2 Hz,
J=9.2 Hz, 1H), 2.87 (m, 1H). LCMS (APCI+) m/z 384, 386 [M+H].sup.+;
Rt=1.92 minutes.
Example 36
##STR00052##
[0412] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-6-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride
[0413] Step 1: 6-piperazin-1-yl-1H-indazole was prepared by the
procedures described in Example 34, Step 1, substituting
5-aminoindazole with 6-aminoindazole. LCMS (APCI+) m/z 203
[M+H].sup.+; Rt=1.53 minutes.
[0414] Step 2:
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-6-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester was prepared by the procedures
described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 6-piperazin-1-yl-1H-indazole.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.91 (s, 1H), 7.94 (d,
J=2.0 Hz, 1H), 7.61 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.28 (d, J=8.4
Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 6.86 (dd, J=8.8 Hz, J=2.0 Hz, 1H),
6.73 (s, 1H), 5.42 (m, 1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m,
1H), 3.29 (m, 1H), 3.19 (m, 1H), 3.00 (m, 4H), 2.59 (m, 1H), 1.43
(s, 9H). LCMS (APCI+) m/z 484, 486 [M+H].sup.+; Rt=3.40
minutes.
[0415] Step 3:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-6-yl)-piperazin-1-yl]-pr-
opan-1-one dihydrochloride was prepared by the procedures described
in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-6-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.30 (s, 1H), 7.78 (d, J=9.2 Hz, 1H), 7.40 (d, J=8.4
Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 7.15 (d, J=9.2 Hz, 1H), 6.97 (s,
1H), 4.76 (m, 1H), 3.80 (m, 2H), 3.62 (m, 1H), 3.40 (m, 1H),
3.10-3.30 (m, 5H), 2.72 (m, 1H). LCMS (APCI+) m/z 384, 386
[M+H].sup.+; Rt=2.02 minutes.
Example 37
##STR00053##
[0416] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1-methyl-1H-indazol-5-yl)-piperazin-
-1-yl]-propan-1-one dihydrochloride
[0417] Step 1: To a solution of 5-piperazin-1-yl-1H-indazole (0.34
g, 1.3 mmol) in 1,4-dioxane (5 mL) was added 3N NaOH (0.42 mL, 1.3
mmol). After cooling to 0.degree. C., a solution of
tert-butylcarbonate (0.25 g, 1.3 mmol) in 1,4-dioxane (1 mL) was
added dropwise. The reaction mixture was stirred at room
temperature overnight and then poured into water and extracted with
EtOAc. The combined organic layers were washed with saturated
aqueous NaHCO.sub.3, water, brine, dried and concentrated. The
residue was purified by column chromatography (EtOAc:hexanes, 1:1)
to give 4-(1H-Indazol-5-yl)-piperazine-1-carboxylic acid tert-butyl
ester (0.31 g, 82%) as a white solid. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 10.01 (s, 1H), 7.80 (s, 1H), 7.42 (d, J=9.2 Hz, 1H),
7.21 (dd, J=9.2 Hz, J=2.0 Hz, 1H), 7.16 (d, J=2.0 Hz, 1H), 3.62 (m,
4H), 3.09 (m, 4H), 1.50 (s, 9H). LCMS (APCI+) m/z 303 [M+H].sup.+;
Rt=2.50 minutes.
[0418] Step 2: To a stirred suspension of NaH (60%, 4 mg, 0.1 mmol)
in DMF (0.5 mL) was added dropwise a solution of
4-(1H-Indazol-5-yl)-piperazine-1-carboxylic acid tert-butyl ester
(0.100 g, 0.33 mmol) in DMF (1 mL). After stirring for 30 minutes,
methyl iodide (0.026 g, 0.18 mmol) was added dropwise. The mixture
was stirred at room temperature for 2 hours and then partitioned
between EtOAc and water. The aqueous phase was extracted with
EtOAc. The combined organic layers were washed with brine, dried
and concentrated. The residue was purified by column chromatography
(hexanes:EtOAc, 2:1) to give
4-(1-Methyl-1H-indazol-5-yl)-piperazine-1-carboxylic acid
tert-butyl ester (0.023 g, 22%) as a white solid. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.87 (s, 1H), 7.32 (d, J=9.2 Hz, 1H),
7.20 (d, J=9.2 Hz, 1H), 7.12 (s, 1H), 4.04 (s, 3H), 3.62 (m, 4H),
3.09 (m, 4H), 1.50 (s, 9H). LCMS (APCI+) m/z 317 [M+H].sup.+;
Rt=3.31 minutes.
[0419] Step 3: 1-Methyl-5-piperazin-1-yl-1H-indazole
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(1-Methyl-1H-indazol-5-yl)-piperazine-1-carboxylic acid
tert-butyl ester. LCMS (APCI+) m/z 217 [M+H].sup.+; Rt=1.15
minutes.
[0420] Step 4:
(2R)-{1-(4-chlorobenzyl)-2-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]--
2-oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with
1-Methyl-5-piperazin-1-yl-1H-indazole dihydrochloride. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.87 (s, 1H), 7.31 (d, J=9.2 Hz, 1H),
7.28 (d, J=8.0 Hz, 2H), 7.17 (d, J=8.0 Hz, 2H), 7.12 (d, J=9.2 Hz,
1H), 7.04 (s, 1H), 5.41 (m, 1H), 4.87 (m, 1H), 4.04 (s, 3H), 3.73
(m, 2H), 3.50 (m, 1H), 3.31 (m, 1H), 3.07 (m, 1H), 2.99 (d, J=7.2
Hz, 2H), 2.94 (m, 2H), 2.48 (m, 1H), 1.43 (s, 9H). LCMS (APCI+) m/z
498, 500 [M+H].sup.+; Rt=3.27 minutes.
[0421] Step 5:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1-methyl-1H-indazol-5-yl)-piperazin-
-1-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]--
2-oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.13 (s, 1H), 7.96 (s, 1H), 7.76 (d,
J=9.2 Hz, 1H), 7.62 (dd, J=9.2 Hz, J=2.0 Hz, 1H), 7.47 (d, J=8.4
Hz, 2H), 7.37 (d, J=8.4 Hz, 2H), 4.81 (m, 1H), 4.11 (m, 4H), 3.98
(m, 1H), 3.90 (m, 1H), 3.69 (m, 2H), 3.49 (m, 2H), 3.16 (m, 2H),
2.84 (m, 1H). LCMS (APCI+) m/z 398, 400 [M+H].sup.+; Rt=2.05
minutes.
Example 38
##STR00054##
[0422] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-pro-
pan-1-one dihydrochloride
[0423] Step 1: A round bottom flask charged with 6-bromo
isoquinoline (prepared from 4-bromobenzaldehyde according to the
literature: Neiko Nerenz, et al. (1998) J. Chem. Soc. Perkin Trans.
2, 437-447, 0.200 g, 0.961 mmol), 1-Boc piperazine (0.215 g, 1.15
mmol), K.sub.3PO.sub.4 (0.286 g, 1.35 mmol),
(2'-dicyclohexylphosphanyl-biphenyl-2-yl)-dimethylamine (0.028 g,
0.072 mmol) and Pd.sub.2 dba.sub.3 (0.022 g, 0.024 mmol) in dry DME
(2 mL) was purged under N.sub.2 and heated at reflux for 5 hours.
After cooling, the mixture was partitioned between EtOAc and
H.sub.2O. The organic layer was washed with brine, dried and
concentrated. The residue was purified by column chromatography
(1:1 hexanes/EtOAc,) to give
4-Isoquinolin-6-yl-piperazine-1-carboxylic acid tert-butyl ester
(0.210 g, 70%) as a white solid. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 9.04 (s, 1H), 8.39 (dd, J=6.8 Hz, J=2.8 Hz, 1H), 7.83 (dd,
J=9.2 Hz, J=2.8 Hz, 1H), 7.45 (d, J=6.8 Hz, 1H), 7.32 (dd, J=9.2
Hz, J=2.4 Hz, 1H), 6.98 (s, 1H), 3.64 (m, 4H), 3.35 (m, 4H), 1.50
(s, 9H). LCMS (APCI+) m/z 314 [M+H].sup.+; Rt=2.14 minutes.
[0424] Step 2: 6-piperazin-1-yl-isoquinoline dihydrochloride was
prepared by the procedures described in Example 34, Step 3,
substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-Isoquinolin-6-yl-piperazine-1-carboxylic acid tert-butyl ester.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 9.28 (s, 1H), 8.30 (d,
J=9.2 Hz, 1H), 8.24 (d, J=7.2 Hz, 1H), 8.04 (d, J=6.8 Hz, 1H), 7.83
(dd, J=9.2 Hz, J=2.4 Hz, 1H), 7.49 (s, 1H), 3.98 (m, 4H), 3.45 (m,
4H). LCMS (APCI+) m/z 214 [M+H].sup.+; Rt=1.76 minutes.
[0425] Step 3:
(2R)-[1-(4-chlorobenzyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethy-
l]-carbamic acid tert-butyl ester was prepared by the procedures
described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 6-piperazin-1-yl-isoquinoline
dihydrochloride. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.04 (s,
1H), 8.40 (dd, J=5.6 Hz, J=3.2 Hz, 1H), 7.83 (dd, J=9.2 Hz, J=2.4
Hz, 1H), 7.46 (d, J=6.0 Hz, 1H), 7.27 (m, 3H), 7.17 (d, J=8.0 Hz,
2H), 6.90 (s, 1H), 5.41 (m, 1H), 4.87 (m, 1H), 3.76 (m, 2H), 3.58
(m, 1H), 3.31 (m, 2H), 3.20 (m, 2H), 2.99 (d, J=6.8 Hz, 2H), 2.78
(m, 1H), 1.43 (s, 9H). LCMS (APCI+) m/z 495, 497 [M+H].sup.+;
Rt=2.50 minutes.
[0426] Step 4:
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-pro-
pan-1-one dihydrochloride was prepared by the procedures described
in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[1-(4-chlorobenzyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethy-
l]-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 9.18 (s, 1H), 8.22 (d, J=9.2 Hz, 1H), 8.16 (d, J=6.8
Hz, 1H), 7.95 (d, J=6.8 Hz, 1H), 7.69 (dd, J=9.2 Hz, J=2.4 Hz, 1H),
7.38 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 7.27 (s, 1H), 4.75
(m, 1H), 3.58-3.84 (m, 6H), 3.10-3.30 (m, 4H). LCMS (APCI+) m/z
395, 397 [M+H].sup.+; Rt=2.68 minutes.
Example 39
##STR00055##
[0427] Preparation of
(2R)-2-Amino-3-(1H-indol-3-yl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-prop-
an-1-one dihydrochloride
[0428] Step 1:
(2R)-[1-(1H-Indol-3-ylmethyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-
-ethyl]-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 6-piperazin-1-yl-isoquinoline
dihydrochloride and substituting (D)-Boc-4-chlorophenylalanine with
(D)-Boc-tryptophan. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.00
(s, 1H), 8.37 (d, J=6.4 Hz, 1H), 8.19 (s, 1H), 7.76 (d, J=9.2 Hz,
1H), 7.72 (d, J=7.6 Hz, 1H), 7.41 (d, J=6.0 Hz, 1H), 7.30 (d, J=7.6
Hz, 1H), 7.18 (m, 2H), 7.12 (d, J=8.8 Hz, 1H), 7.08 (s, 1H), 6.70
(s, 1H), 5.53 (d, J=8.4 Hz, 1H), 5.03 (m, 1H), 3.79 (m, 1H), 3.47
(m, 1H), 3.05-3.40 (m, 6H), 2.73 (m, 1H), 1.97 (m, 1H), 1.46 (s,
9H). LCMS (APCI+) m/z 500 [M+H].sup.+; Rt=2.75 minutes.
[0429] Step 2:
(2R)-2-Amino-3-(1H-indol-3-yl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-prop-
an-1-one dihydrochloride was prepared by the procedures described
in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[1-(1H-Indol-3-ylmethyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-
-ethyl]-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 10.65 (s, 1H), 9.14 (s, 1H), 8.15 (m, 2H), 7.89
(d, J=6.8 Hz, 1H), 7.56 (d, J=5.6 Hz, 1H), 7.51 (d, J=9.2 Hz, 1H),
7.30 (m, 2H), 7.07 (m, 3H), 4.70 (m, 1H), 3.74 (m, 1H), 3.64 (m,
1H), 3.39 (m, 5H), 3.01 (m, 1H), 2.86 (m, 1H), 2.36 (m, 1H). LCMS
(APCI+) m/z 400 [M+H].sup.+; Rt=1.73 minutes.
Example 40
##STR00056##
[0430] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylsulfanylpyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride
[0431] Step 1: A mixture of 4-chloro-5-iodopyrimidine (3.00 g, 12.5
mmol) (prepared from 4(3H)-pyrimidinone according to the
literature: Takao Sakamoto, et al. (1986) Chem. Pharm. Bull.,
2719-2724), Et.sub.3N (5.22 mL, 37.4 mmol), 1-Boc piperazine (2.79
g, 15.0 mmol) and NMP (30 mL) was heated at 75.degree. C. for 6
hours. After cooling, the reaction mixture was partitioned between
EtOAc and water. The aqueous phase was extracted with EtOAc. The
combined organic layers were washed with brine, dried and
concentrated. The residue was purified by column chromatography
(2:1 hexanes/EtOAc) to give
4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester (4.81 g, 99%) as a white solid. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.70 (s, 1H), 8.62 (s, 1H), 3.57 (s, 8H), 1.49 (s,
9H). LCMS (APCI+) m/z 391 [M+H].sup.+; Rt=2.96 minutes.
[0432] Step 2: 5-Iodo-4-piperazin-1-yl-pyrimidine dihydrochloride
was prepared by the procedures described in Example 34, Step 3,
substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.87 (s, 1H), 8.84
(s, 1H), 4.36 (t, J=4.2 Hz, 4H), 3.47 (t, J=4.2 Hz, 4H). LCMS
(APCI+) m/z 291 [M+H].sup.+; Rt=1.45 minutes.
[0433] Step 3:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with
5-Iodo-4-piperazin-1-yl-pyrimidine dihydrochloride. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.70 (s, 1H), 8.62 (s, 1H), 7.27 (d,
J=8.0 Hz, 2H), 7.15 (d, J=8.0 Hz, 2H), 5.38 (m, 1H), 4.84 (m, 1H),
3.73 (m, 2H), 3.65 (m, 1H), 3.53 (m, 2H), 3.45 (m, 2H), 3.24 (m,
1H), 3.12 (m, 1H), 2.98 (m, 2H), 1.42 (s, 9H). LCMS (APCI+) m/z
572, 574 [M+H].sup.+; Rt=3.25 minutes.
[0434] Step 4: A round bottom flask was charged with sodium
methanethiolate (17 mg, 0.25 mmol),
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester (71 mg, 0.12 mmol) and
2,2,6,6-tetramethyl-heptane-3,5-dione (6 mg, 0.25 equivalents).
After vacuum purging and refilling of N.sub.2, NMP (3 mL) and CuCl
(6 mg, 0.06 mmol) was added to this mixture. The reaction was
stirred at 130.degree. C. for 1 hour. After cooling, the reaction
mixture was diluted with EtOAc and filtered through Celite. The
filtrate was washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
flash column chromatography (1:1 hexanes/EtOAc) to give
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin--
1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (24 mg, 39%) as a
colorless gum. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.56 (s,
1H), 8.29 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H),
5.37 (m, 1H), 4.84 (m, 1H), 3.70 (m, 1H), 3.61 (m, 2H), 3.51 (m,
4H), 3.20 (m, 2H), 2.97 (m, 2H), 2.42 (s, 3H), 1.42 (s, 9H). LCMS
(APCI+) m/z 492, 494 [M+H].sup.+; Rt=3.57 minutes.
[0435] Step 5:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylsulfanylpyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride was prepared by the
procedures described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin--
1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.64 (s, 1H), 8.30 (s, 1H), 7.39 (d,
J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.09 (m, 2H),
4.00 (m, 1H), 3.83 (m, 1H), 3.68 (m, 3H), 3.17 (m, 3H), 2.55 (s,
3H). LCMS (APCI+) m/z 392, 394 [M+H].sup.+; Rt=1.85 minutes.
Example 41
##STR00057##
[0436] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride
[0437] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester (0.150 g, 0.262 mmol),
phenylboronic acid (0.042 g, 0.341 mmol) and 2N sodium carbonate
solution (0.34 mL, 0.68 mmol) were stirred in DME (3 mL) and the
mixture was degassed with N.sub.2 for 15 minutes.
Tetrakis(triphenylphosphine) palladium (0) (0.015 g, 0.013 mmol)
was added and the mixture heated at 80.degree. C. for 24 hours. The
mixture was cooled to room temperature and partitioned between DCM
and water. The aqueous layer was extracted with DCM. The combined
organic layers were washed with saturated aqueous NaHCO.sub.3 and
brine, dried and concentrated. The residue was purified by flash
chromatography on silica gel, eluting with hexanes:EtOAc (3:1 to
1:1) to give
(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-phenylpyrimidin-4-yl)-piperazin-1--
yl]-ethyl}-carbamic acid tert-butyl ester (0.074 g, 54%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.65 (s,
1H), 8.21 (s, 1H), 7.46 (m, 2H), 7.38 (m, 2H), 7.25 (m, 3H), 7.09
(d, J=8.4 Hz, 2H), 5.30 (m, 1H), 4.73 (m, 1H), 3.50 (m, 1H), 3.43
(m, 1H), 3.30 (m, 2H), 3.16 (m, 2H), 2.97 (m, 1H), 2.91 (d, J=7.6
Hz, 2H), 2.85 (m, 1H), 1.39 (s, 9H). LCMS (APCI+) m/z 522, 524
[M+H].sup.+; Rt=2.72 minutes.
[0438] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-phenylpyrimidin-4-yl)-piperazin-1--
yl]-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.75 (s, 1H), 8.20 (s, 1H), 7.56 (m, 3H), 7.44 (d,
J=6.8 Hz, 2H), 7.37 (d, J=8.0 Hz, 2H), 7.25 (d, J=8.0 Hz, 2H), 4.62
(m, 1H), 3.47-3.54 (m, 6H), 3.38-3.44 (m, 4H). LCMS (APCI+) m/z
422, 424 [M+H].sup.+; Rt=2.40 minutes.
Example 42
##STR00058##
[0439] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride
[0440] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-pipe-
razin-1-yl]-ethyl}-carbamic acid tert-butyl ester was prepared by
the procedures described in Example 41, Step 1, substituting
phenylboronic acid with 3-thiopheneboronic acid. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.64 (s, 1H), 8.27 (s, 1H), 7.46 (d,
J=4.8 Hz, 1H), 7.33 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.16 (d, J=4.8
Hz, 1H), 7.11 (d, J=8.4 Hz, 2H), 5.33 (m, 1H), 4.76 (m, 1H), 3.55
(m, 1H), 3.48 (m, 1H), 3.27 (m, 2H), 3.16 (m, 2H), 2.98 (m, 1H),
2.92 (d, J=6.8 Hz, 2H), 2.82 (m, 1H), 1.40 (s, 9H). LCMS (APCI+)
m/z 528, 530 [M+H].sup.+; Rt=2.75 minutes.
[0441] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-pip-
erazin-1-yl]-propan-1-one dihydrochloride was prepared by the
procedures described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-pipe-
razin-1-yl]-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.74 (s, 1H), 8.24 (s, 1H), 7.70 (s,
1H), 7.65 (s, 1H), 7.39 (d, J=8.0 Hz, 2H), 7.27 (d, J=8.0 Hz, 2H),
7.22 (d, J=4.8 Hz, 1H), 4.62 (m, 1H), 3.48-3.55 (m, 6H), 3.38-3.44
(m, 4H). LCMS (APCI+) m/z 428, 430 [M+H].sup.+; Rt=2.76
minutes.
Example 43
##STR00059##
[0442] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenoxypyrimidin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride
[0443] A round bottom flask was charged with phenol (33 mg, 0.35
mmol),
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester (100 mg, 0.175 mmol),
Cs.sub.2CO.sub.3 (114 mg, 0.350 mmol) and
2,2,6,6-tetramethyl-heptane-3,5-dione (8 mg, 0.25 equivalents).
After vacuum purging and refilling of N.sub.2, NMP (3 mL) and CuCl
(9 mg, 0.09 mmol) was added to this mixture. The reaction was
stirred at 130.degree. C. for 2 hours. After cooling, the reaction
was diluted with EtOAc and filtered. The filtrate was washed with
water, brine and dried. Evaporation of the solvent followed by
flash column chromatography (10:1 DCM/MeOH) gave the title compound
as a free base, which was converted to dihydrochloride salt (36 mg,
38%) by treatment with 4 N HCl in dioxane. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.64 (s, 1H), 7.96 (s, 1H), 7.70 (s, 1H), 7.65 (s,
1H), 7.49 (m, 2H), 7.37 (m, 2H), 7.29 (m, 3H), 7.14 (d, J=8.0 Hz,
2H), 4.70 (m, 1H), 4.11 (m, 2H), 4.00 (m, 1H), 3.76 (m, 2H), 3.57
(m, 2H), 3.10 (m, 3H). LCMS (APCI+) m/z 438, 440 [M+H].sup.+;
Rt=2.13 minutes.
Example 44
##STR00060##
[0444] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-iodopyrimidin-4-yl)-piperazin-1-y-
l]-propan-1-one dihydrochloride
[0445] The title compound was prepared by the procedures described
in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.77 (s, 1H), 8.73 (s, 1H), 7.41 (d, J=8.4 Hz,
2H), 7.33 (d, J=8.4 Hz, 2H), 4.74 (m, 1H), 4.05 (m, 2H), 3.95 (m,
1H), 3.84 (m, 1H), 3.69 (m, 3H), 3.22 (m, 1H), 3.13 (m, 2H). LCMS
(APCI+) m/z 472, 474 [M+H].sup.+; Rt=2.06 minutes.
Example 45
##STR00061##
[0446] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-chloropyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride
[0447] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 40, Step 1, substituting
4-chloro-5-iodopyrimidine with 4,5-dichloropyrimidine (prepared
from 5-chloropyrimidin-4-ol according to the literature: Chestfield
J. et al. (1955) J. Chem. Soc. Abstracts, 3478-3481), and
substituting 1-Boc piperazine with
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.57
(s, 1H), 8.32 (s, 1H), 7.27 (d, J=8.0 Hz, 2H), 7.15 (d, J=8.0 Hz,
2H), 5.36 (m, 1H), 4.85 (m, 1H), 3.50-3.80 (m, 6H), 3.24 (m, 2H),
2.98 (m, 2H), 1.43 (s, 9H). LCMS (APCI+) m/z 480, 482, 484
[M+H].sup.+; Rt=3.24 minutes.
[0448] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-chloropyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.74 (s, 1H), 8.57 (s, 1H), 7.40 (d, J=7.6 Hz,
2H), 7.33 (d, J=7.6 Hz, 2H), 4.74 (m, 1H), 4.13 (m, 2H), 4.03 (m,
1H), 3.85 (m, 1H), 3.72 (m, 3H), 3.22 (m, 1H), 3.16 (m, 2H). LCMS
(APCI+) m/z 380, 382, 384 [M+H].sup.+; Rt=2.01 minutes.
Example 46
##STR00062##
[0449] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride
[0450] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 40, Step 1, substituting
4-chloro-5-iodopyrimidine with 4-chloro-5-fluoropyrimidine
(prepared from 5-fluoropyrimidin-4-ol according to the literature
(Kheifets, G. M. et al., 2000, Russian. Org. Chem., 1373-1387), and
substituting 1-Boc piperazine with
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.41
(d, J=2.8 Hz, 1H), 8.12 (d, J=6.4 Hz, 1H), 7.27 (d, J=8.0 Hz, 2H),
7.15 (d, J=8.0 Hz, 2H), 5.35 (m, 1H), 4.83 (m, 1H), 3.45-3.75 (m,
6H), 3.35 (m, 1H), 3.18 (m, 1H), 2.98 (d, J=7.2 Hz, 2H), 1.43 (s,
9H). LCMS (APCI+) m/z 464, 466 [M+H].sup.+; Rt=2.93 minutes.
[0451] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.66 (s, 1H), 8.50 (s, 1H), 7.39 (d, J=8.0 Hz,
2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.05 (m, 2H), 3.97 (m,
1H), 3.88 (m, 1H), 3.70 (m, 3H), 3.23 (m, 1H), 3.15 (m, 2H). LCMS
(APCI+) m/z 364, 366 [M+H].sup.+; Rt=1.88 minutes.
Example 47
##STR00063##
[0452] Preparation of
(2R)-2-Amino-1-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-3-(4-chloro-
phenyl)-propan-1-one dihydrochloride
[0453] Step 1: A sealed tube charged with
4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester (780 mg, 2.00 mmol), CuI (38 mg, 0.20 mmol),
1,10-phenathroline (72 mg, 0.4 mmol), Cs.sub.2CO.sub.3 (912 mg, 2.8
mmol), benzyl alcohol (0.62 mL, 6.0 mmol) and toluene (2 mL) was
heated at 110.degree. C. for 40 hours. The resulting suspension was
cooled to room temperature and filtered through a silica gel pad,
eluting with EtOAc. Evaporation of the solvent followed by flash
chromatography on silica gel (10:1 hexanes/EtOAc) provided
4-(5-benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (0.640 g, 86%) as a white solid. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.36 (s, 1H), 7.99 (s, 1H), 7.39 (m,
5H), 5.08 (s, 2H), 3.73 (m, 4H), 3.48 (m, 4H), 1.47 (s, 9H). LCMS
(APCI+) m/z 371 [M+H].sup.+; Rt=2.52 minutes.
[0454] Step 2: 5-Benzyloxy-4-piperazin-1-yl-pyrimidine
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(5-Benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester. LCMS (APCI+) m/z 271 [M+H].sup.+; Rt=1.64
minutes.
[0455] Step 3:
(2R)-[2-[4-(5-Benzyloxypyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-
-2-oxo-ethyl]-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with
5-Benzyloxy-4-piperazin-1-yl-pyrimidine dihydrochloride. .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.34 (s, 1H), 8.00 (s, 1H), 7.38
(m, 5H), 7.24 (d, J=8.0 Hz, 2H), 7.12 (d, J=8.0 Hz, 2H), 5.37 (d,
J=8.8 Hz, 1H), 5.06 (s, 2H), 4.81 (m, 1H), 3.64 (m, 5H), 3.44 (m,
1H), 3.34 (m, 1H), 3.13 (m, 1H), 2.96 (m, 2H), 1.41 (s, 9H). LCMS
(APCI+) m/z 552, 554 [M+H].sup.+; Rt=2.79 minutes.
[0456] Step 4:
(2R)-2-Amino-1-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-3-(4-chloro-
phenyl)-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[2-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-
-2-oxo-ethyl]-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.49 (s, 1H), 8.11 (s, 1H), 7.48 (m,
5H), 7.36 (d, J=8.4 Hz, 2H), 7.28 (d, J=8.4 Hz, 2H), 5.20 (s, 2H),
4.68 (m, 1H), 4.07 (m, 2H), 3.97 (m, 1H), 3.69 (m, 2H), 3.59 (m,
2H), 3.12 (m, 3H). LCMS (APCI+) m/z 452, 454 [M+H].sup.+; Rt=2.02
minutes.
Example 48
##STR00064##
[0457] Preparation of
(2R)-2-Amino-1-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophen-
yl)-propan-1-one dihydrochloride
[0458] Step 1: 4-piperazin-1-yl-pyrimidin-5-ylamine dihydrochloride
was prepared by the procedures described in Example 34, Step 3,
substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(5-aminopyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester (prepared from 4,6-dichloro-nitropyrimidine according to the
procedures described in U.S. Pat. No. 5,563,142). LCMS (APCI+) m/z
180 [M+H].sup.+; Rt=1.12 minutes.
[0459] Step 2:
(2R)-[2-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-o-
xo-ethyl]-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with
4-piperazin-1-yl-pyrimidin-5-ylamine dihydrochloride. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.38 (s, 1H), 7.97 (s, 1H), 7.27 (d,
J=8.0 Hz, 2H), 7.14 (d, J=8.0 Hz, 2H), 5.38 (d, J=8.4 Hz, 1H), 4.85
(m, 1H), 3.66 (m, 2H), 3.52 (m, 2H), 3.44 (s, 2H), 3.23 (m, 4H),
2.93 (m, 2H), 1.42 (s, 9H). LCMS (APCI+) m/z 461, 463 [M+H].sup.+;
Rt=2.38 minutes.
[0460] Step 3:
(2R)-2-Amino-1-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophen-
yl)-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[2-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-o-
xo-ethyl]-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.41 (s, 1H), 7.80 (s, 1H), 7.40 (d, J=8.4 Hz,
2H), 7.32 (d, J=8.4 Hz, 2H), 4.77 (m, 1H), 3.84 (m, 4H), 3.63 (m,
2H), 3.50 (m, 1H), 3.18 (m, 3H). LCMS (APCI+) m/z 361, 363
[M+H].sup.+; Rt=1.68 minutes.
Example 49
##STR00065##
[0461] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methoxypyrimidin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride
[0462] Step 1: To a stirred solution of
4-(5-benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (0.540 g, 1.46 mmol) in MeOH (20 mL) under N.sub.2
was cautiously added 10% Pd on carbon (40 mg). The reaction vessel
was evacuated under vacuum and then put under an atmosphere of
hydrogen using a balloon. The mixture was stirred for 2 hours at
room temperature. At this time the hydrogen gas was evacuated and
the catalyst was removed by filtration. The filtrate was
concentrated. The residue was purified by flash chromatography (1:1
hexanes/EtOAc) to give
4-(5-Hydroxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester (0.390 g, 95%) as a white solid. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.18 (s, 1H), 7.75 (s, 1H), 3.93 (m, 4H), 3.54 (m,
4H), 1.41 (s, 9H). LCMS (APCI+) m/z 281 [M+H].sup.+; Rt=2.01
minutes.
[0463] Step 2: 4-piperazin-1-yl-pyrimidin-5-ol dihydrochloride was
prepared by the procedures described in Example 34, Step 3,
substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(5-Hydroxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl
ester. LCMS (APCI+) m/z 181 [M+H].sup.+; Rt=1.15 minutes.
[0464] Step 3:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-hydroxypyrimidin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 4-piperazin-1-yl-pyrimidin-5-ol
dihydrochloride. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.18 (s,
1H), 7.76 (s, 1H), 7.25 (d, J=8.0 Hz, 2H), 7.14 (d, J=8.0 Hz, 2H),
5.39 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.55-3.95 (m, 5H), 3.51 (m,
2H), 3.21 (m, 1H), 2.96 (m, 2H), 1.42 (s, 9H). LCMS (APCI+) m/z
462, 464 [M+H].sup.+; Rt=2.41 minutes.
[0465] A mixture of methyl iodide (18 mg, 0.13 mmol),
K.sub.2CO.sub.3 (18 mg, 0.13 mmol) and
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-hydroxypyrimidin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester (50 mg, 0.11 mmol) in
DMF (2 mL) was stirred at room temperature for 1 hour. The reaction
mixture was diluted with water and extracted with EtOAc. The
combined organic layers were washed with brine, dried and
concentrated. The residue was purified by flash chromatography
(hexanes:EtOAc, 1:1) to give
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester (20 mg, 39%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.34 (s,
1H), 7.94 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H),
5.37 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.87 (s, 3H), 3.65 (m, 5H),
3.48 (m, 1H), 3.33 (m, 1H), 3.20 (m, 1H), 2.96 (m, 2H), 1.42 (s,
9H). LCMS (APCI+) m/z 476, 478 [M+H].sup.+; Rt=2.44 minutes.
[0466]
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methoxypyrimidin-4-yl)-pipe-
razin-1-yl]-propan-1-one dihydrochloride was prepared by the
procedures described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.50 (s, 1H), 8.01 (s, 1H), 7.38 (d, J=8.0 Hz,
2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.12 (m, 2H), 4.04 (m,
1H), 3.97 (s, 3H), 3.81 (m, 2H), 3.67 (m, 2H), 3.16 (m, 3H). LCMS
(APCI+) m/z 376, 378 [M+H].sup.+; Rt=1.77 minutes.
Example 50
##STR00066##
[0467] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-isopropoxypyrimidin-4-yl)-piperaz-
in-1-yl]-propan-1-one dihydrochloride
[0468] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl-
]-2-oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 49, Step 2, substituting methyl
iodide with isopropyl bromide. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.33 (s, 1H), 7.94 (s, 1H), 7.26 (d, J=8.0 Hz, 2H), 7.14
(d, J=8.0 Hz, 2H), 5.39 (d, J=8.8 Hz, 1H), 4.84 (m, 1H), 4.48 (m,
1H), 3.63 (m, 5H), 3.48 (m, 1H), 3.33 (m, 1H), 3.14 (m, 1H), 2.97
(m, 2H), 1.42 (s, 9H), 1.34 (d, J=6.4 Hz, 6H). LCMS (APCI+) m/z
504, 506 [M+H].sup.+; Rt=2.61 minutes.
[0469] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-isopropoxypyrimidin-4-yl)-piperaz-
in-1-yl]-propan-1-one dihydrochloride was prepared by the
procedures described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl-
]-2-oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.47 (s, 1H), 8.03 (s, 1H), 7.39 (d,
J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.73 (m, 2H), 4.13 (m, 2H),
4.02 (m, 1H), 3.81 (m, 2H), 3.66 (m, 2H), 3.17 (m, 3H), 1.42 (d,
J=6.4 Hz, 6H). LCMS (APCI+) m/z 404, 406 [M+H].sup.+; Rt=1.86
minutes.
Example 51
##STR00067##
[0470] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylpyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride
[0471] Step 1: A mixture of
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-ox-
o-ethyl}-carbamic acid tert-butyl ester (0.070 g, 0.12 mmol),
methylboronic acid (0.022 g, 0.37 mmol), K.sub.2CO.sub.3 (0.085 g,
0.61 mmol) and PdCl.sub.2(PPh.sub.3).sub.2 (0.0086 g, 0.012 mmol)
in DMF (2 mL) was heated at 100.degree. C. for 16 hours under
nitrogen. The mixture was cooled to room temperature and
partitioned between EtOAc and water. The organic layer was washed
with saturated aqueous NaHCO.sub.3 and brine, dried and
concentrated. The residue was purified by flash chromatography on
silica gel, eluting with DCM/MeOH (70:1) to give
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester (0.022 g, 39%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.62 (s,
1H), 8.20 (s, 1H), 7.26 (d, J=8.0 Hz, 2H), 7.14 (d, J=8.0 Hz, 2H),
5.37 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.70 (m, 1H), 3.60 (m, 1H),
3.42 (m, 1H), 3.32 (m, 4H), 3.02 (m, 3H), 2.20 (s, 3H), 1.42 (s,
9H). LCMS (APCI+) m/z 460, 462 [M+H].sup.+; Rt=2.38 minutes.
[0472] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylpyrimidin-4-yl)-piperazin-1-
-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-2--
oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 8.65 (s, 1H), 8.13 (s, 1H), 7.39 (d, J=8.4 Hz,
2H), 7.32 (d, J=8.4 Hz, 2H), 4.73 (m, 1H), 3.40 (m, 2H), 3.96 (m,
1H), 3.80 (m, 1H), 3.67 (m, 3H), 3.21 (m, 1H), 3.14 (m, 2H). LCMS
(APCI+) m/z 360, 362 [M+H].sup.+; Rt=1.70 minutes.
Example 52
##STR00068##
[0473] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-cinnolin-4-yl-piperazin-1-yl)-propan-
-1-one dihydrochloride
[0474] Step 1: To a suspension of NaH (60% in mineral oil, 0.099 g,
2.46 mmol) in DMF (5 mL) was added cinnolin-4-ol (prepared from
2-aminoacetophenone according to the procedures described in U.S.
Pat. No. 4,620,000), 0.300 g, 2.05 mmol) in DMF (2 mL) dropwise.
The reaction mixture was warmed at 40.degree. C. and stirred for 30
minutes. After cooling, N-phenyltrifluoromethanesulfonimide (0.880
g, 2.46 mmol) in DMF (2 mL) was added, and the reaction mixture was
stirred at room temperature for 1 hour. 1-Boc piperazine (0.765 g,
4.11 mmol) was added to the mixture. The reaction was stirred at
80.degree. C. for 4 hours. After cooling, the mixture was
partitioned between EtOAc and water. The aqueous phase was
extracted with EtOAc. The combined organic layers were washed with
water, brine, dried and concentrated. The residue was purified by
column chromatography (1:1 to 1:3 hexanes/EtOAc) to give
4-Cinnolin-4-yl-piperazine-1-carboxylic acid tert-butyl ester
(0.246 g, 38%) as a yellow oil. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.91 (s, 1H), 8.47 (d, J=8.4 Hz, 1H), 7.97 (d, J=8.4 Hz,
1H), 7.80 (t, J=7.2 Hz, 1H), 7.69 (t, J=7.2 Hz, 1H), 3.74 (m, 4H),
3.34 (m, 4H), 1.51 (s, 9H). LCMS (APCI+) m/z 315 [M+H].sup.+;
Rt=2.14 minutes.
[0475] Step 2: 4-piperazin-1-yl-cinnoline dihydrochloride was
prepared by the procedures described in Example 34, Step 3,
substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-Cinnolin-4-yl-piperazine-1-carboxylic acid tert-butyl ester.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.96 (s, 1H), 8.33 (d,
J=8.8 Hz, 1H), 8.15 (t, J=7.2 Hz, 1H), 8.07 (d, J=8.4 Hz, 1H), 7.88
(t, J=7.2 Hz, 1H), 4.42 (m, 4H), 3.64 (m, 4H). LCMS (APCI+) m/z 215
[M+H].sup.+; Rt=1.46 minutes.
[0476] Step 3:
(2R)-[1-(4-chlorobenzyl)-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]--
carbamic acid tert-butyl ester was prepared by the procedures
described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 4-piperazin-1-yl-cinnoline
dihydrochloride. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.82 (s,
1H), 8.48 (d, J=8.4 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.80 (t, J=7.2
Hz, 1H), 7.70 (t, J=7.2 Hz, 1H), 7.31 (d, J=8.4 Hz, 2H), 7.18 (d,
J=8.4 Hz, 2H), 5.36 (m, 1H), 4.88 (m, 1H), 3.85 (m, 2H), 3.69 (m,
1H), 3.40 (m, 1H), 3.28 (m, 1H), 3.17 (m, 2H), 3.01 (d, J=7.2 Hz,
2H), 2.78 (m, 1H), 1.43 (s, 9H). LCMS (APCI+) m/z 496, 498
[M+H].sup.+; Rt=2.44 minutes.
[0477] Step 4:
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-cinnolin-4-yl-piperazin-1-yl)-propan-
-1-one dihydrochloride was prepared by the procedures described in
Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[1-(4-chlorobenzyl)-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]--
carbamic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.75 (s, 1H), 8.27 (d, J=8.4 Hz, 1H), 8.10 (t, J=7.2 Hz,
1H), 7.99 (d, J=8.4 Hz, 1H), 7.82 (t, J=7.2 Hz, 1H), 7.42 (d, J=8.0
Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 4.74 (m, 1H), 4.19 (m, 3H), 3.89
(m, 4H), 3.41 (m, 1H), 3.18 (d, J=7.6 Hz, 2H). LCMS (APCI+) m/z
396, 398 [M+H].sup.+; Rt=2.46 minutes.
Example 53
##STR00069##
[0478] Preparation of
(2R)-2-Amino-1-(4-cinnolin-4-yl-piperazin-1-yl)-3-phenylpropan-1-one
dihydrochloride
[0479] Step 1:
(2R)-[1-Benzyl-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamic
acid tert-butyl ester was prepared by the procedures described in
Example 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with
4-piperazin-1-yl-cinnoline dihydrochloride, and substituting
(D)-Boc-4-chlorophenylalanine with (D)-Boc-phenylalanine. .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.73 (s, 1H), 8.46 (d, J=8.4 Hz,
1H), 7.85 (d, J=8.4 Hz, 1H), 7.79 (t, J=8.0 Hz, 1H), 7.68 (t, J=8.0
Hz, 1H), 7.29 (m, 5H), 5.42 (d, J=8.4 Hz, 1H), 4.91 (m, 1H), 3.89
(m, 1H), 3.67 (m, 1H), 3.57 (m, 1H), 3.32 (m, 2H), 3.08 (m, 2H),
2.99 (m, 2H), 2.32 (m, 1H), 1.45 (s, 9H). LCMS (APCI+) m/z 462
[M+H].sup.+; Rt=2.30 minutes.
[0480] Step 2:
(2R)-2-Amino-1-(4-cinnolin-4-yl-piperazin-1-yl)-3-phenylpropan-1-one
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[1-Benzyl-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamic
acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.72 (s, 1H), 8.25 (d, J=8.4 Hz, 1H), 8.09 (t, J=8.0 Hz, 1H), 7.99
(d, J=8.4 Hz, 1H), 7.80 (t, J=8.0 Hz, 1H), 7.37 (m, 2H), 4.75 (m,
1H), 4.24 (m, 1H), 4.12 (m, 2H), 3.89 (m, 3H), 3.65 (m, 1H), 3.23
(m, 3H). LCMS (APCI+) m/z 362 [M+H].sup.+; Rt=2.38 minutes.
Example 54
##STR00070##
[0481] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methylquinazolin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride
[0482] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 40, Step 1, substituting
4-chloro-5-iodopyrimidine with 4-chloro-2-methylquinazoline, and
substituting 1-Boc piperazine with
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.84
(d, J=8.4 Hz, 1H), 7.77 (d, J=8.4 Hz, 1H), 7.71 (t, J=8.4 Hz, 1H),
7.41 (t, J=8.4 Hz, 1H), 7.28 (d, J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz,
2H), 5.37 (d, J=8.4 Hz, 1H), 4.87 (m, 1H), 3.73 (m, 3H), 3.60 (m,
4H), 3.29 (m, 2H), 2.98 (m, 2H), 2.84 (s, 3H), 1.43 (s, 9H). LCMS
(APCI+) m/z 510, 512 [M+H].sup.+; Rt=2.50 minutes.
[0483] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methylquinazolin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.19 (d, J=8.0 Hz, 1H), 8.01 (t,
J=8.0 Hz, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.72 (t, J=8.0 Hz, 1H), 7.40
(d, J=8.4 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 4.76 (m, 1H), 4.27 (m,
2H), 4.19 (m, 1H), 3.70-3.95 (m, 4H), 3.17 (m, 3H). LCMS (APCI+)
m/z 410, 412 [M+H].sup.+; Rt=1.80 minutes.
Example 55
##STR00071##
[0484] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-chloroquinazolin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride
[0485] Step 1:
(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester was prepared by the
procedures described in Example 40, Step 1, substituting
4-chloro-5-iodopyrimidine with 2,4-dichloroquinazoline, and
substituting 1-Boc piperazine with
[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acid
tert-butyl ester. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.84
(d, J=8.0 Hz, 1H), 7.77 (m, 2H), 7.47 (t, J=8.0 Hz, 1H), 7.29 (d,
J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 5.35 (d, J=8.8 Hz, 1H), 4.84
(m, 1H), 3.74 (m, 5H), 3.40 (m, 1H), 3.31 (m, 1H), 2.99 (d, J=8.0
Hz, 2H), 1.43 (s, 9H). LCMS (APCI+) m/z 530, 532, 534 [M+H].sup.+;
Rt=3.74 minutes.
[0486] Step 2:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-chloroquinazolin-4-yl)-piperazin--
1-yl]-propan-1-one dihydrochloride was prepared by the procedures
described in Example 34, Step 3, substituting
(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.16 (d, J=8.4 Hz, 1H), 7.97 (t,
J=8.4 Hz, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.69 (t, J=8.4 Hz, 1H), 7.40
(d, J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.74 (m, 1H), 4.14 (m,
2H), 4.06 (m, 1H), 3.91 (m, 1H), 3.78 (m, 3H), 3.33 (m, 1H), 3.17
(m, 2H). LCMS (APCI+) m/z 430, 432, 434 [M+H].sup.+; Rt=2.24
minutes.
Example 56
##STR00072##
[0487] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methoxyquinazolin-4-yl)-piperazin-
-1-yl]-propan-1-one dihydrochloride
[0488] To a solution of
(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-
-oxo-ethyl}-carbamic acid tert-butyl ester (23 mg, 0.043 mmol) in
MeOH (1 mL) was added 4 N HCl in dioxane (1 mL). The mixture was
stirred at room temperature for 2 days and then evaporated to give
the title compound (14 mg, 65%) as a white solid. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.17 (d, J=7.2 Hz, 1H), 7.94 (t,
J=7.2 Hz, 1H), 7.60 (d, J=8.0 Hz, 2H), 7.39 (m, 4H), 4.75 (m, 1H),
4.10-4.35 (m, 6H), 3.86 (m, 4H), 3.17 (m, 3H). LCMS (APCI+) m/z
426, 428 [M+H].sup.+; Rt=1.86 minutes.
Example 57
##STR00073##
[0489] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinolin-4-yl-piperazin-1-yl)-propan-
-1-one
[0490] Step 1: To a solution of 4-chloroquinoline (2.0 g, 12.2
mmol) in toluene (100 mL) was added piperazine (7.98 g, 92.7 mmol).
The reaction mixture was heated to reflux and stirred for 96 hours,
after which it was cooled to room temperature and then further
cooled to 0.degree. C. The resulting mixture was filtered to remove
the hydrochloride salts that had precipitated. After washing the
salts with toluene, the combined filtrate was washed with 10%
aqueous acetic acid (2.times.25 mL). The combined aqueous extracts
were washed with diethyl ether (25 mL) and then basified to pH 8-10
by adding 1M NaOH. The resulting aqueous mixture was extracted with
dichloromethane (3.times.25 mL). The combined organic extracts were
dried (Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation. The crude residue, 4-piperazinylquinoline, was
obtained as a yellow solid (2.54 g, 97%) and used without further
purification. .sup.1H NMR was consistent with that reported in the
literature (Abel, M. D., et al., Journal of Heterocyclic Chemistry
(1996), 33(2), 415-420).
[0491] Step 2: To a Jones tube containing PS-CDI (Argonaut, 1.04
mmol/g, 2.2 equivalents) suspended in a solution of
4-piperazinylquinoline (1.0 equivalent) in CHCl.sub.3 was added the
solid Boc-protected amino acid (1.5 equivalents.). The reaction
mixture was shaken for 15 hours at room temperature, after which it
was vacuum filtered, the resin rinsed with CHCl.sub.3, and the
filtrate concentrated by rotary evaporation. If necessary, the
crude coupled product was purified on silica (DCM/EtOAc or
DCM/MeOH). The resulting Boc-amino amide was dissolved in minimal
dioxane, and 4M HCl/dioxane (10 equivalents) was added. The
suspension was sonicated 5 minutes and stirred at room temperature
for 12 hours, after which it was concentrated by rotary
evaporation. The solids were dispersed in ether, isolated by
filtration with nitrogen pressure, and dried under reduced pressure
to give the corresponding amino amide as the hydrochloride salt,
which was 90% pure by HPLC analysis. R.sub.t 2.36. MS (APCI+)
[M+H].sup.+ 395. R.sub.t 2.36. MS (APCI+) [M+H].sup.+ 395.
Example 58
##STR00074##
[0492] Preparation of 1-Quinazolin-4-yl-piperidine-4-carboxylic
acid (2-amino-ethyl)-(4-chlorobenzyl)-amide dihydrochloride
[0493] Step 1: Triethylamine (12.7 mL, 91.1 mmol) was added to a
solution of 4-chloro-quinazoline (5.00 g, 30.4 mmol) and ethyl
isonipecotate (4.78 g, 30.4 mmol) in THF (80 mL) at ambient
temperature. After being refluxed for 12 hours, the mixture was
concentrated in vacuo, and the resulting residue was partitioned
between DCM and aqueous 0.1 M NaOH. The separated DCM layer was
dried (Na.sub.2SO.sub.4), filtered, and concentrated in vacuo. The
resulting solids were suspended in Et.sub.2O and filtered to give
1-quinazolin-4-yl-piperidine-4-carboxylic acid ethyl ester (8.0 g,
28.0 mmol). This material was dissolved in EtOH (50 mL) and THF (50
mL) followed by the addition of NaOH (3.73 g, 93.4 mmol) in
H.sub.2O (50 mL). After being stirred for 12 hours, the mixture was
neutralized with 1.0 N HCl (93.5 mL) and concentrated in vacuo.
After the volatile organics are removed, a white precipitate forms
in the resulting aqueous solution. The solid was filtered off and
air-dried to give 1-quinazolin-4-yl-piperidine-4-carboxylic acid
(6.5 g). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 12.28 (bs,
1H), 8.57 (s, 1H), 7.91 (m, 1H), 7.75 (m, 2H), 7.49 (m, 1H), 4.13
(m, 2H), 3.22 (m, 2H), 2.57 (m, 1H), 1.93 (m, 2H), 1.72 (m,
2H).
[0494] Step 2: Thionyl chloride (1.37 mL, 18.8 mmol) was added to a
suspension of 1-quinazolin-4-yl-piperidine-4-carboxylic acid (2.20
g, 8.55 mmol) in DCM (40 mL), which results in a clear solution.
After being stirred for 2 hours, a precipitate forms and was
filtered off to give 1-quinazolin-4-yl-piperidine-4-carbonyl
chloride hydrochloride as a white solid (2.0 g).
[0495] Step 3: A solution of tert-butyl N-(2-aminoethyl)carbamate
(1.00 g, 6.24 mmol) and 4-chlorobenzaldehyde (0.90 g, 6.37 mmol) in
DCE (10 mL) was stirred for 30 minutes, followed by the addition of
NaBH(OAc).sub.3 (1.98 g, 9.36 mmol) in a single portion. After
being stirred for 12 hours, the mixture was acidified to pH 2 with
0.2 N HCl, and extracted with DCM (3 times, each discarded). The
acidic aqueous layer was basified to pH 10 with 2.0 M NaOH, and
extracted with DCM. The DCM extracts were dried (Na.sub.2SO.sub.4),
filtered, and concentrated in vacuo to give
[2-(4-chlorobenzylamino)-ethyl]-carbamic acid tert-butyl ester as
an oil (1.0 g). LCMS (APCI+) m/z 285, 287 [M+H].sup.+.
[0496] Step 4: 1-Quinazolin-4-yl-piperidine-4-carbonyl chloride
hydrochloride (197 mg, 0.63 mmol) was added to a solution of
[2-(4-chlorobenzylamino)-ethyl]-carbamic acid tert-butyl ester (180
mg, 0.63 mmol) and DMAP (154 mg, 1.26 mmol) in DCM (6.5 mL) cooled
in an ice bath. After being stirred for 12 hours, the mixture was
partitioned between DCM (50 mL) and H.sub.2O (80 mL) containing 1
mL of 1.0 M HCl. The DCM layer was drained off and the acidic
aqueous layer was extracted 3 more times with DCM. The combined DCM
extracts were dried (Na.sub.2SO.sub.4), filtered, and concentrated
in vacuo. The crude material was chromatographed (SiO.sub.2) using
EtOAc as eluent to give
{2-[(4-chlorobenzyl)-(1-quinazolin-4-yl-piperidine-4-carbonyl)-amino]-eth-
yl}-carbamic acid tert-butyl ester (190 mg).
[0497] Step 5:
1{2-[(4-Chlorobenzyl)-(1-quinazolin-4-yl-piperidine-4-carbonyl)-amino]-et-
hyl}-carbamic acid tert-butyl ester (190 mg, 0.36 mmol) was
dissolved in DCM (5 mL) followed by the addition of 2.0 M HCl in
Et.sub.2O (2 mL). After being stirred for 12 hours, the mixture was
diluted with DCE and concentrated in vacuo. The resulting white
solid was then suspended in MeCN and concentrated in vacuo
(repeated twice) to give 1-quinazolin-4-yl-piperidine-4-carboxylic
acid (2-amino-ethyl)-(4-chlorobenzyl)-amide dihydrochloride as a
white powder (120 mg). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
8.83 (m, 1H), 8.51 (bs, 2H), 8.20 (m, 2H), 8.01 (m, 2H), 7.71 (m,
1H), 7.48 (d, J=8.3 Hz, 1H), 7.39 (d, J=8.3 Hz, 1H), 7.31 (d, J=8.3
Hz, 1H), 7.25 (d, J=8.3 Hz, 1H), 4.79 (m, 2H), 4.55 (s, 1H), 3.59
(m, 7H), 3.03 (m, 1H), 2.90 (m, 1H), 1.92 (m, 4H). LCMS (APCI+) m/z
424, 426 [M+H].
Example 59
##STR00075##
[0498] Preparation of
2-(R)-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-piper-
azin-1-yl]-propan-1-one dihydrochloride
[0499] Step 1: A mixture of 4-chloro-1H-pyrrolo[2,3-b]pyridine
(2.50 g, 16.4 mmol) and N-benzyl piperazine (3.18 g, 18.0 mmol)
were melted at 175.degree. C. for 3 hours in a sealed tube,
resulting in the formation of a crystalline solid mass. A solution
of 0.1 M aqueous NaOH (10 mL) was added and the solid was broken up
to give a suspension. Filtration gave
4-(4-benzyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]pyridine as a white
solid (3.90 g). LCMS (APCI+) m/z 293 [M+H].sup.+.
[0500] Step 2: A solution of
4-(4-benzyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]pyridine (3.90 g, 13.3
mmol) and Pd(OH).sub.2/C (937 mg, 1.33 mmol) in MeOH (60 mL) was
stirred under 1 atmosphere of H.sub.2 for 2 d. The mixture was
diluted with MeOH, filtered through diatomaceous earth, and the
filtrate was concentrated in vacuo to give
4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine as a solid (100 mg kept
as free base). The remaining material was suspended in MeOH and
treated with 2.0 M HCl in Et.sub.2O. This mixture was concentrated
in vacuo to give 4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine
dihydrochloride (2.30 g). .sup.1H NMR (DMSO-d.sub.6, 400 MHz)
.delta. 11.5 (bs, 1H), 9.43 (bs, 2H), 7.96 (d, J=5.4 Hz, 1H), 7.25
(d, J=3.3 Hz, 1H), 6.49 (d, J=3.4 Hz, 1H), 6.45 (d, J=5.4 Hz, 1H),
3.56 (bs, 4H), 3.21 (bs, 4H).
[0501] Step 3: PyBrop (407 mg, 0.87 mmol) was added in a single
portion to a solution of (R)--N-Boc-4-chlorophenylalanine (458 mg,
1.53 mmol) and 4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine (200 mg,
0.73 mmol) in DCM (5 mL) cooled in an ice bath. DIEA (0.66 mL, 3.78
mmol) was then dropped in, the ice bath was removed, and the
mixture was stirred for 12 hours at ambient temperature. The
mixture was diluted with DCM and washed with 0.1 N HCl. The
separated DCM layer was dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo. This material was dissolved in MeOH (4 mL)
followed by the addition of LiOH monohydrate (122 mg, 2.91 mmol) in
H.sub.2O (2 mL) and stirred for 12 hours. The mixture was
concentrated in vacuo and chromatographed (SiO.sub.2) using 2%
MeOH/DCM followed by 5% MeOH/DCM as eluent. The resulting material
was dissolved in DCM (4 mL) and 2.0 M HCl in Et.sub.2O (2 mL), and
then stirred for 12 hours. The mixture was concentrated in vacuo
and chromatographed (SiO.sub.2) using 10% MeOH/DCM followed by 10%
(7 N NH.sub.3 in MeOH)/DCM as eluent. The purified material was
dissolved in MeOH followed by the addition of 2.0 N HCl in
Et.sub.2O, and then concentrated in vacuo. The resulting glass was
suspended in Et.sub.2OO and concentrated in vacuo (repeat twice) to
give
2-(R)-amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-piper-
azin-1-yl]-propan-1-one dihydrochloride as an off-white powder (50
mg). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 12.6 (s, 1H), 8.56
(bs, 3H), 8.03 (d, J=7.1 Hz, 1H), 7.38 (m, 3H), 7.32 (m, 2H), 6.84
(s, 1H), 6.69 (d, J=7.1 Hz, 1H), 4.63 (bs, 1H), 3.88-3.58 (m, 7H),
3.33 (m, 1H), 3.14 (m, 1H), 3.05 (m, 1H). LCMS (APCI+) m/z 384, 386
[M+H].sup.+.
Example 60
##STR00076##
[0502] Preparation of
3-Amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-one
dihydrochloride
[0503] Step 1: n-BuLi (1.60M in hexanes, 40.7 mL, 65.1 mmol) was
added to a 0.degree. C. solution of diisopropylamine (9.4 mL, 67.0)
in 280 mL THF. The mixture was allowed to stir at 0.degree. C. for
30 minutes, then cooled to -78.degree. C. A solution of
p-tolyl-acetic acid methyl ester (10.48 g, 63.8 mmol; prepared from
p-tolyl-acetic acid) in 10 mL of THF was added to the -78.degree.
C. LDA solution by syringe, which was then stirred for 45 minutes.
Neat tert-butyl bromoacetate (28 mL) was added by syringe, and the
reaction was stirred 15 minutes at -78.degree. C. The bath was
removed, and the reaction was allowed to warm to room temperature.
After stirring an additional 5 hours, the reaction mixture was
quenched with saturated NH.sub.4Cl solution, and the organics were
removed in vacuo. The oily mixture was extracted with ethyl
acetate, and the organics were combined. The organic was dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The crude oil was
purified by on silica gel (25:1 hexanes:EtOAc) to afford the
2-p-tolyl-succinic acid 4-tert-butyl ester 1-methyl ester as a pale
yellow oil (15.3 g, 86%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.16 (d, J=8.0 Hz, 2H), 7.12 (d, J=8.0 Hz, 2H), 3.99 (dd, J=10.4,
5.6 Hz, 1H), 3.66 (s, 3H), 3.09 (dd, J=16.8, 10.4 Hz, 1H), 2.57
(dd, J=16.8, 5.6 Hz, 1H), 2.32 (s, 3H), 1.41 (m, 1H). HPLC
R.sub.t=3.71 min.
[0504] Step 2: A solution of 2-p-tolyl-succinic acid 4-tert-butyl
ester 1-methyl ester (15.3 g, 54.8 mmol) in 110 mL of DCM was
treated with neat TFA (63 mL) at room temperature. The mixture was
stirred for five hours to completion, after which the reaction
mixture was concentrated and dried in vacuo overnight to afford a
white solid. The solid was suspended in 190 mL of toluene, cooled
to 0.degree. C., and treated successively with diphenylphosphoryl
azide (13.4 mL, 62.1 mmol) and triethyl amine (19.7 mL, 141 mmol).
The reaction mixture (homogeneous) was allowed to warm to room
temperature and stirred for four hours to completion. The solution
was quenched with 1% citric acid solution and extracted with EtOAc.
The combined organic was washed with brine, dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to give a
light brown oil. The crude azide was dissolved in 190 mL of
tert-butanol, treated with neat SnCl.sub.4 (0.25 mL, 2.82 mmol),
and carefully heated to 90.degree. C. with evolution of nitrogen.
The mixture was stirred at 90.degree. C. for 2.5 hours and cooled
to room temperature. The solution was quenched with saturated
NaHCO.sub.3 solution and then concentrated. The oily mixture was
extracted with EtOAc, and the combined organic was washed with
brine, dried over MgSO.sub.4, filtered, and concentrated in vacuo.
The residue was purified by on silica gel (9:1 hexanes:EtOAc) to
afford the 3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid
methyl ester as a pale yellow oil (12.3 g, 74%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.14 (s, 4H), 4.86 (br s, 1H), 3.85
(m, 1H), 3.68 (s, 3H), 3.58 (m, 1H), 3.49 (m, 1H), 2.33 (s, 3H),
1.42 (s, 9H). HPLC R.sub.t=3.31 min.
[0505] Step 3: The 3-tert-butoxycarbonylamino-2-p-tolyl-propionic
acid methyl ester (12.3 g, 41.9 mmol) was dissolved in 200 mL 1:1
THF:water and treated with lithium hydroxide monohydrate (2.64 g,
62.9 mmol) at room temperature. The reaction was stirred at room
temperature overnight to completion and concentrated in vacuo. The
oily mixture was partitioned with water and washed with EtOAc
(discarded). The aqueous was treated with solid KHSO.sub.4 until
pH<2, then extracted with EtOAc. The combined organic was dried
over Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to
afford the 3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid as a
white solid (10.95 g, 93%). .sup.1H NMR (ca. 1:1 mixture of
rotamers) (CDCl.sub.3, 400 MHz) .delta. 10.40-8.40 (br s, 1H), 7.16
(d, J=8.4 Hz, 2H), 7.13 (d, J=8.4 Hz, 2H), 6.80 or 4.91 (br s, 1H),
3.86 and 3.75 (m, 1H), 3.55 (m, 1H), 3.47 (m, 2H), 2.31 (s, 3H),
1.44 and 1.41 (s, 9H). LCMS (APCI--) m/z 557 [2M-H].sup.-. HPLC
R.sub.t=2.80 min.
[0506] Step 4: The 4-piperazin-1-yl-quinazoline dihydrochloride (50
mg, 0.174 mmol, free-based with 2N NaOH and extracted with DCM),
HOBt monohydrate (27 mg, 0.174 mmol), and
3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid (58 mg, 0.209
mmol) were dissolved in 1.3 mL of DCM/3-5 drops of THF. The
reaction mixture was treated with DCC (43 mg, 0.209 mmol) and
allowed to stir at room temperature for 2.5 hours to completion.
The mixture was diluted with DCM, vacuum filtered through
compressed Celite, and rinsed with DCM. The filtrate was stirred
with 2N sodium hydroxide solution for five minutes, transferred to
seperatory funnel, and extracted with DCM. The combined organic was
dried over Na.sub.2SO.sub.4, filtered, and concentrated. The
residue was purified on silica gel (1:19 DCM:EtOAc) to afford
3-Boc-amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-one.
The material was dissolved in 1.0 mL of 1,4-dioxane and treated
with 1.0 mL of 4M HCl in 1,4-dioxane (precipitation). The mixture
was stirred at room temperature overnight to completion, then
concentrated to dryness. The solid was dissolved in a minimal
amount of MeOH, then triturated with diethyl ether. The resulting
solid was isolated by filtration through a fritted funnel with
nitrogen pressure, rinsed with ether, and dried in vacuo to afford
the
3-amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-one
dihydrochloride as a pale yellow powder (60 mg, 77%). .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 8.84 (s, 1H), 8.16 (d, J=8.4 Hz,
1H), 8.00 (m, 4H), 7.91 (d, J=8.0 Hz, 1H), 7.69 (t, J=8.0 Hz, 1H),
7.20 (s, 4H), 4.40 (m, 1H), 4.14 (m, 3H), 3.81 (m, 4H), 3.45 (m,
2H), 2.94 (m, 1H), 2.28 (s, 3H). LCMS (APCI+) m/z 376 [M+H].sup.+.
HPLC R.sub.t=1.67 min.
Example 61
##STR00077##
[0507] Preparation of
4-Amino-2-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-o-
ne
[0508] Step 1: The (4-chlorophenyl)-acetic acid (20.0 g, 106 mmol)
was dissolved in 220 mL of ethanol at ambient temperature. A
catalytic amount of sulfuric acid (10 drops) was added to afford a
light yellow solution. The reaction was allowed to stir overnight
to completion and was concentrated to 30 mL. The concentrate was
partitioned between ethyl acetate and half-saturated NaHCO.sub.3
solution. The aqueous was extracted with ethyl acetate, and the
organics were combined. The organic was washed with water, brine,
separated, dried over MgSO.sub.4, filtered, and concentrated in
vacuo to afford the desired pure (4-chlorophenyl)-acetic acid ethyl
ester as a pale yellow oil (21.0 g, 99%). .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 7.29 (d, J=8.4 Hz, 2H), 7.22 (d, J=8.4 Hz, 2H),
4.15 (q, J=7.2 Hz, 2H), 3.58 (s, 2H), 1.25 (t, J=7.2 Hz, 3H).
[0509] Step 2: The (4-chlorophenyl)-acetic acid ethyl ester (9.52
g, 47.9 mmol) was dissolved in 80 mL of THF, cooled to 0.degree.
C., and treated with potassium tert-butoxide (538 mg, 4.79 mmol).
The resulting orange solution was allowed to stir for 15 minutes at
0.degree. C., then cooled to -78.degree. C. The tert-butyl acrylate
(7.72 mL, 52.7 mmol) was added in three equal portions over ten
minutes. The solution was allowed to stir overnight warming slowly
to room temperature. The reaction solution was concentrated in
vacuo, and the residue was partitioned between ethyl acetate and
saturated NH.sub.4Cl solution. The aqueous was extracted with ethyl
acetate, and the organics were combined. The organic was washed
with brine, separated, dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by chromatography
(silica gel eluted with 9:1 hexanes:EtOAc) to afford the
2-(4-chlorophenyl)-pentanedioic acid 5-tert-butyl ester 1-ethyl
ester in greater than 80% purity (9.00 g, 57%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.31-7.21 (m, 4H), 4.13 (q, J=7.6 Hz,
2H), 3.58 (t, J=8.0 Hz, 1H), 2.27 (m, 1H), 2.16 (t, J=7.2 Hz, 2H),
2.04 (m, 1H), 1.43 (s, 9H), 1.21 (t, J=7.6 Hz, 3H).
[0510] Step 3: The 2-(4-chlorophenyl)-pentanedioic acid
5-tert-butyl ester 1-ethyl ester (9.00 g, 27.5 mmol) was dissolved
in 40 mL of DCM at room temperature and treated slowly with 40 mL
of TFA. The solution was allowed to stir for three hours to
completion, then concentrated in vacuo. The residue was stored
under vacuum overnight then dissolved in 80 mL of toluene. The
solution was degassed under nitrogen, cooled to 0.degree. C.,
treated with triethyl amine (8.44 mL, 60.6 mmol), and treated with
diphenylphosphoryl azide (6.53 mL, 30.3 mmol), respectively. The
reaction was allowed to warm to room temperature and stir for three
hours, then concentrated in vacuo. The residue was re-dissolved in
ethyl acetate and washed with 1 w/w % citric acid solution. The
organic was dried over MgSO.sub.4, filtered, and concentrated
(<30.degree. C.) to afford the intermediate azide as a yellow
oil. The material was immediately dissolved in 80 mL of
tert-butanol and treated with SnCl.sub.4 (1.65 mL of a 1.0M sol'n
in DCM, 1.65 mmol). The solution was heated to 80.degree. C. for
one hour to give evolution of nitrogen gas. The reaction mixture
was treated with saturated NaHCO.sub.3 (20 mL), and concentrated in
vacuo to give a gel. The residue was partitioned between ethyl
acetate and water, and the aqueous was extracted with ethyl
acetate. The organic was washed with brine, separated, dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
purified by chromatography (silica gel eluted with 4:1
hexanes:EtOAc, R.sub.f=0.20) to give the pure
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid ethyl
ester as a colorless oil (5.61 g, 60%). .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 7.29 (d, J=8.4 Hz, 2H), 7.23 (d, J=8.4 Hz, 2H),
4.51 (brs, 1H), 4.12 (m, 2H), 3.57 (t, J=7.6 Hz, 1H), 3.09 (m, 2H),
2.25 (m, 1H), 1.93 (m, 1H), 1.43 (s, 9H), 1.20 (t, J=7.2 Hz,
3H).
[0511] Step 4: The
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid ethyl
ester (5.61 g, 16.4 mmol) was dissolved in 40 mL of THF and 10 mL
of water, then cooled to 0.degree. C. The mixture was treated with
lithium hydroxide mono-hydrate (1.38 g, 32.8 mmol) to afford a
yellow solution. The ice-bath was removed, and the mixture was
allowed to stir overnight to room temperature. The reaction mixture
was concentrated to approximately 15 mL and diluted with water. The
aqueous was washed with ethyl acetate (discarded) then treated with
3M HCl solution until acidic (pH=2-3). The resulting white
precipitate was extracted with ethyl acetate, and the organics were
combined. The organic was washed with brine, separated, dried over
MgSO.sub.4, filtered, and concentrated in vacuo to afford the
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid as a
colorless oil. The material was re-dissolved in a minimal amount of
warm (60.degree. C.) hexanes and re-concentrated to afford the pure
desired product as a white foam. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 12.05-9.51 (brs, 1H), 7.31-7.25 (m, 4H), 4.59 (brs, 1H),
3.61 (d, J=7.6 Hz, 1H), 3.14 (brs, 2H), 2.28 (m, 1H), 1.92 (m, 1H),
1.43 (s, 9H).
[0512] Step 5: The 4-piperazin-1-yl-quinazoline (60 mg, 0.21 mmol)
and 4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid (62
mg. 0.21 mmol) were dissolved in 1.5 mL of DCM and cooled to
0.degree. C. The solution was treated with PyBrop (98 mg, 0.21
mmol) and DIEA (74 .mu.L, 0.42 mmol), respectively. The mixture was
allowed to warm to room temperature overnight, and the contents
were partitioned between ethyl acetate and saturated NH.sub.4Cl
solution. The aqueous was extracted with ethyl acetate, and the
organics were combined. The organic was washed with NaHCO.sub.3
solution, separated, dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by chromatography
(silica gel eluted with hexanes/EtOAc gradients) to afford the pure
Boc-intermediate as a colorless oil. The material was dissolved in
1.0 mL of 1,4-dioxane and treated with 1.0 mL of 4M HCl in dioxane
(2.79 mmol). The solution was allowed to stir at room temperature
overnight to completion. The resulting suspension was diluted with
diethyl ether and isolated by vacuum filtration. The pad of solid
was allowed to dry under a stream of dry nitrogen to afford the
pure
4-amino-2-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-o-
ne bis-hydrochloride as a white solid (32 mg, 37%). A small amount
of the product was free-based for analytical purposes. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.73 (s, 1H), 7.91 (d, J=8.4 Hz, 1H),
7.81 (d, J=8.4 Hz, 1H), 7.75 (t, J=7.6 Hz, 1H), 7.47 (t, J=7.6 Hz,
1H), 7.32 (d, J=8.8 Hz, 2H), 7.25 (d, J=8.8 Hz, 2H), 4.01 (dd,
J=7.2 Hz, 1H), 3.94 (m, 1H), 3.81 (m, 1H), 3.71 (m, 3H), 3.62 (m,
2H), 3.25 (m, 1H), 2.69 (t, J=6.8 Hz, 2H), 2.26 (m, 1H), 1.84 (m,
1H), 1.40 (brs, 2H). LCMS (APCI+) m/z 410 [M+H].sup.+; Rt=1.64
min.
Example 62
##STR00078##
[0513] Preparation of
(3E)-4-phenyl-2-piperazin-1-yl-1-(4-quinazolin-4-yl-piperazin-1-yl)-but-3-
-en-1-one trihydrochloride
[0514]
(3E)-4-Phenyl-2-piperazin-1-yl-1-(4-quinazolin-4-yl-piperazin-1-yl)-
-but-3-en-1-one trihydrochloride (17 mg, 47%) was prepared by the
procedures described for the preparation of Example 1A using
(3E)-2-(4-Boc-piperazinyl)-4-phenylbut-3-enoic acid. LCMS (APCI+)
m/z 443 [M+H].sup.+. HPLC Rt 2.31 min.
Example 63
##STR00079##
[0515] Preparation
3-amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-yl)-butyramide
dihydrochloride
[0516] Step 1: To a solution of 4-chloroquinazoline (2.0 g, 12.2
mmol) in 45 mL IPA was added Boc-4-aminopiperidine (2.56 g, 12.8
mmol) and DIEA (3.2 mL, 18.2 mmol). The reaction mixture was heated
to reflux and stirred 16 hours, after which the reaction mixture
was cooled to room temperature and concentrated. The residue was
dissolved in EtOAc and washed with water, 1N NaOH, brine, dried
(Na.sub.2SO.sub.4), filtered, and concentrated to provide
4-(4-Boc-aminopiperidin-1-yl)quinazoline, which was used directly
in the next step.
[0517] Step 2: To a solution of crude
4-(4-Boc-aminopiperidin-1-yl)quinazoline in 40 mL 1:1 dioxane:DCM
was added 20 mL 4M HCl/dioxane. The resulting suspension was
stirred at room temperature for 14 hours, after which it was
concentrated to dryness. The residue was stirred in DCM and 1M
NaOH, the phases were separated, and the aqueous phase was
extracted with DCM. The combined organic phases were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
purified on silica gel (8:1 EtOAc:MeOH to furnish
4-(4-aminopiperidin-1-yl)quinazoline (2.7 g, 96%) as a yellow oil.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.73 (s, 1H), 7.92-7.85
(m, 2H), 7.76-7.70 (m, 1H), 7.48-7.42 (m, 1H), 4.35-4.26 (m, 2H),
3.26-3.17 (m, 2H), 3.09-2.99 (m, 1H), 2.05-1.96 (m, 2H), 1.66-1.52
(m, 2H), 1.47 (br s, 2H). LCMS (APCI+) m/z 329 [M+H].sup.+. HPLC Rt
1.57 min.
[0518] Step 3:
3-Amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-yl)-butyramide
dihydrochloride (10 mg, 33%) was prepared from
4-(4-aminopiperidin-1-yl)quinazoline according to the procedure
employed for Example 1A, Step 2, using
Boc-.beta.-homophenylalanine. LCMS (APCI+) m/z 390 [M+H].sup.+.
HPLC Rt 1.94 min.
Example 64
##STR00080##
[0519] Preparation of
3-amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-ylmethyl)-butyramide
dihydrochloride
[0520]
3-Amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-ylmethyl)-butyram-
ide dihydrochloride (13 mg, 42%) was prepared by the procedures
described for the preparation of Example 63 using
4-Boc-aminomethylpiperidine in the S.sub.NAr step. LCMS (APCI+) m/z
404 [M+H].sup.+. HPLC Rt 1.98 min.
Example 65
##STR00081##
[0521] Preparation of
3-amino-4-phenyl-1-[4-(quinazolin-4-ylamino)-piperidin-1-yl]-butan-1-one
dihydrochloride
[0522]
3-Amino-4-phenyl-1-[4-(quinazolin-4-ylamino)-piperidin-1-yl]-butan--
1-one dihydrochloride (19 mg, 59%) was prepared by the procedures
described for the preparation of Example 63 using
4-amino-1-Boc-piperidine in the S.sub.NAr step. LCMS (APCI+) m/z
390 [M+H].sup.+. HPLC Rt 2.24 min.
Example 66
##STR00082##
[0523] Preparation of
3-amino-4-phenyl-1-[3-(quinazolin-4-ylamino)-pyrrolidin-1-yl]-butan-1-one
dihydrochloride
[0524]
3-Amino-4-phenyl-1-[3-(quinazolin-4-ylamino)-pyrrolidin-1-yl]-butan-
-1-one dihydrochloride (16 mg, 51%) was prepared by the procedures
described for the preparation of Example 63 using
3-amino-Boc-pyrrolidine in the S.sub.NAr step. LCMS (APCI+) m/z 376
[M+H].sup.+. HPLC Rt 2.17 min.
Example 67
##STR00083##
[0525] Preparation of
3-amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-propyl]-butyramide
dihydrochloride
[0526]
3-Amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-propyl]-butyramide
dihydrochloride (14 mg, 46%) was prepared by the procedures
described for the preparation of Example 63 using
Boc-propylenediamine in the S.sub.NAr step. LCMS (APCI+) m/z 364
[M+H].sup.+. HPLC Rt 2.14 min.
Example 68
##STR00084##
[0527] Preparation of
3-amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-ethyl]-butyramide
dihydrochloride
[0528]
3-Amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-ethyl]-butyramide
dihydrochloride (11 mg, 37%) was prepared by the procedures
described for the preparation of Example 63 using
Boc-ethylenediamine in the S.sub.NAr step. LCMS (APCI+) m/z 350
[M+H].sup.+. HPLC Rt 2.07 min.
Example 69
##STR00085##
[0529] Preparation of
3-Amino-4-phenyl-N-(3-quinazolin-4-yl-3-aza-bicyclo[3.1.0]hex-6-yl)-butyr-
amide dihydrochloride
[0530]
3-Amino-4-phenyl-N-(3-quinazolin-4-yl-3-aza-bicyclo[3.1.0]hex-6-yl)-
-butyramide dihydrochloride (19 mg, 59%, mixture of diastereomers)
was prepared by the procedures described for the preparation of
Example 63 using
(1.alpha.,5.alpha.,6.alpha.)-6-Boc-amino-3-azabicyclo[3.1.0]hexane
(prepared according to the literature: Tamim F. Braish et al. 1996,
1100-1102) in the S.sub.NAr step. LCMS (APCI+) m/z 388 [M+H].sup.+.
HPLC Rt 2.13, 2.23 min.
Example 70
##STR00086##
[0531] Preparation of
(2R)-2-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pi-
perazin-1-yl]-propan-1-one dihydrochloride
[0532] Step 1: A solution containing
4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.0 g, 32.6 mmol),
Boc-piperazine (15 g, 81 mmol), and DIEA (19.8 mL, 114 mmol) in 130
mL IPA was stirred at 80 C for 18 hours, after which the reaction
was concentrated. The crude was flashed on silica gel (20:1
DCM:MeOH) to give a yellow powder, which was recrystallized from
MeOH/minimal DCM to give
4-Boc-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine as a white
crystalline solid (3 crops). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 11.37 (br s, 1H), 8.34 (s, 1H), 7.12 (d, J=3.3 Hz, 1H),
6.49 (d, J=3.5 Hz, 1H), 4.01-3.94 (m, 4H), 3.64-3.56 (m, 4H), 1.48
(s, 9H). HPLC Rt 2.05 min.
[0533] Step 2: To a solution of
4-Boc-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine in 225 mL DCM was
added dropwise by addition funnel 120 mL 4M HCl/dioxane, and the
resulting suspension was stirred at room temperature 18 hours. The
reaction mixture was then diluted with ether, and the solids were
isolated by filtration through a fritted funnel with nitrogen
pressure, rinsed with ether, and dried in vacuo to give
4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (8.44
g, 94%) as a white powder. .sup.1H NMR (DMSO-d6, 400 MHz) .delta.
12.75 (1H, s), 9.63 (2H, s), 8.44 (1H, s), 7.50 (1H, s), 6.95 (1H,
s), 4.26-4.21 (4H, m), 3.33-3.26 (4H, m). LC/MS (APCI+) m/z 204
[M+H].sup.+.
[0534] Step 3: To a solution of
4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (30
mg, 0.11 mmol), HOBt.H.sub.2O (17 mg, 0.11 mmol), TEA (45 .mu.L,
0.33 mmol), and (D)-Boc-4-chlorophenylalanine (39 mg, 0.13 mmol) in
1.6 mL DMF was added DCC (27 mg, 0.13 mmol.) The reaction mixture
was stirred at room temperature for 4 hours, after which it was
concentrated. The residue was suspended in DCM, and the solids were
removed by vacuum filtration through cotton plug and rinsed with
DCM. The filtrate was concentrated, and the crude purified on
silica gel (1:1 to 1:4 DCM:EtOAc) to afford
(2R)-2-Boc-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one, which was used in the next
step.
[0535] Step 4: To a solution of
(2R)-2-Boc-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one in 1 mL dioxane was added 1 mL 4M
HCl/dioxane. The resulting suspension was stirred at room
temperature overnight, after which it was concentrated to dryness.
The solids were dissolved in minimal MeOH and then triturated with
ether. The resulting solids were isolated by filtration through a
fritted funnel with nitrogen pressure, rinsed with ether, and dried
in vacuo to give
(2R)-2-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pi-
perazin-1-yl]-propan-1-one dihydrochloride (19 mg, 38%) as a pink
powder. .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 8.16 (1H, s), 7.27
(1H, s), 7.18-7.11 (4H, m), 6.68 (1H, s), 4.60-4.56 (1H, m),
3.98-3.69 (4H, m), 3.61-3.52 (2H, m), 3.45-3.37 (1H, m), 3.20-3.12
(1H, m), 3.00-2.91 (2H, m). LCMS (APCI+) m/z 385 [M+H].sup.+. HPLC
Rt 1.68 min.
Example 71
##STR00087##
[0536] Preparation of
(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-y-
l)-piperazin-1-yl]-propan-1-one dihydrochloride
[0537] Step 1: nBuLi (1.6M in hexanes, 20 mL, 32 mmol) was added to
a stirred solution of (4R,5S)-4-Methyl-5-phenyl-oxazolidin-2-one
(5.2 g, 29 mmol) in THF (60 mL) at -78 C under N2. The solution was
stirred at -78.degree. C. for 10 mL and then
3-(4-Chlorophenyl)-propionyl chloride (6.0 g, 29 mmol) was added
and the solution allowed to warm to room temperature over 1 hour.
The solution was quenched with saturated aqueous NH4Cl, extracted
into DCM (2.times.200 mL), dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The product was purified by column
chromatography on silica (50% EtOAc/hexanes) to give
(4R,5S)-3-[3-(4-Chlorophenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-o-
ne (4.8 g, 48%.) .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.44-7.35 (3H, m), 7.30-7.24 (4H, m), 7.19 (2H, d, J 8.0 Hz), 5.64
(1H, d, J 7.4 Hz), 4.77-4.71 (1H, m), 3.34-3.17 (2H, m), 2.98 (2H,
t, J 7.7 Hz), 0.88 (3H, d, J 6.7 Hz.)
[0538] Step 2: NaHMDS (1.0M, 17 mL, 17 mmol) was added to a stirred
solution of
(4R,5S)-3-[3-(4-Chlorophenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-o-
ne (4.8 g, 14 mmol) in THF (200 mL) at -78.degree. C. under
N.sub.2. Stirred at -78.degree. C. for 45 minutes and then
Bromo-acetic acid tert-butyl ester (2.5 mL, 17 mmol) was added
dropwise over 10 minutes. The solution was allowed to warm to
-20.degree. C. over 4 hours and then quenched with saturated
aqueous NH.sub.4Cl. The product was extracted into EtOAc
(2.times.300 mL), dried over Na.sub.2SO.sub.4, concentrated in
vacuo and purified by column chromatography on silica (20%
EtOAc/hexanes) to give
(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazol-
idin-3-yl)-4-oxo-butyric acid tert-butyl ester (5.1 g, 80%.)
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.44-7.21 (9H, m), 5.45
(1H, d, J 7.3 Hz), 4.68-4.62 (1H, m), 4.51-4.42 (1H, m), 3.01 (1H,
dd, J 13.0 and 6.2 Hz), 2.78 (1H, dd, J 16.7 and 10.6 Hz), 2.63
(1H, dd, J 13.2 and 9.0 Hz), 2.32 (1H, dd, J 6.7 and 4.3 Hz), 1.38
(9H, s), 0.89 (3H, d, J 6.6 Hz.)
[0539] Step 3: A solution of
(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-y-
l)-4-oxo-butyric acid tert-butyl ester (5.1 g, 11 mmol) in DCM (100
mL) was treated with TFA (50 mL) and stirred at room temperature
for 1 hour. The solution was concentrated in vacuo, taken up into
toluene and then conc. in vacuo. Placed on high vacuum for 6 hours
to give
(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-y-
l)-4-oxo-butyric acid (4.5 g, 100%.) .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 7.44-7.37 (3H, m), 7.31-7.21 (6H, m), 5.48 (1H, d, J
6.9 Hz), 4.70-4.63 (1H, m), 4.50-4.42 (1H, m), 3.05 (1H, dd, J 13.3
and 6.2 Hz), 2.89 (1H, dd, J 17.7 and 10.6 Hz), 2.63 (1H, dd, J
13.2 and 9.0 Hz), 2.43 (1H, dd, J 17.6 and 4.4 Hz), 0.86 (3H, d, J
6.6 Hz.)
[0540] Step 4: NEt3 (700 uL, 5.0 mmol) was added to a stirred
solution of
(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-y-
l)-4-oxo-butyric acid (1.0 g, 2.5 mmol) in PhMe (50 mL) at
0.degree. C. under N.sub.2. This was followed by the addition of
the diphenylphosphoryl azide (650 .mu.L, 3.0 mmol.) The solution
was stirred at 0.degree. C. for 15 minutes and then stirred at room
temperature overnight. The solution was washed with 1% citric acid,
extracted into EtOAc and concentrated in vacuo. Taken up into tBuOH
(50 mL), SnCl4 (1.0M in DCM, 0.1 mL) added and stirred and heated
at 85.degree. C. for 5 hours. Cooled to room temperature and
quenched with saturated aqueous bicarbonate. Stirred at RT for 10
minutes and then concentrated in vacuo. The product was taken up
into water (100 mL) and extracted into EtOAc (2.times.200 mL.)
Dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The product
was purified by column chromatography on silica (10% EtOAc/hexanes)
and then on the Biotage to give
[(2S)-2-(4-Chlorobenzyl)-3-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3--
yl)-3-oxo-propyl]-carbamic acid tert-butyl ester (300 mg, 25%.)
LCMS (APCI+) m/z 373 [M-Boc+H].sup.+; Rt: 3.92 min.
[0541] Step 5: To a solution of
[(2S)-2-(4-Chlorobenzyl)-3-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3--
yl)-3-oxo-propyl]-carbamic acid tert-butyl ester (300 mg, 0.63
mmol) in THF/H.sub.2O (30/10 mL) at 0.degree. C. was added LiOH (80
mg, 1.9 mmol) and H.sub.2O.sub.2 (30% by volume, 3.0 mL, 0.63 mmol)
and stirred at 0.degree. C. for 30 minutes. Then Na.sub.2SO.sub.3
(saturated solution, 10 mL) was added slowly & cautiously.
Diluted with EtOAc (100 mL) and extracted into water (2.times.100
mL.) The aq. layer was acidified (1N HCl) and extracted into EtOAc
(3.times.100 mL.) Dried over Na.sub.2SO.sub.4 and concentrated in
vacuo to give
(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionic
acid (150 mg, 75%.) LCMS (APCI--) m/z 322 [M-Boc+H].sup.-; Rt: 2.23
min.
[0542] Step 6: NEt.sub.3 (150 .mu.L, 1.1 mmol) was added to a
stirred suspension of 4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine
dihydrochloride (100 mg, 0.36 mmol),
(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionic
acid (130 mg, 0.40 mmol), EDCI (83 mg, 0.44 mmol) and HOBt (59 mg,
0.44 mmol) in DMF (15 mL) at RT. Stirred at RT overnight. Poured
into EtOAc (100 mL), washed with water (100 mL), 1N NaOH (50 mL),
dried over Na2SO4, concentrated in vacuo, purified by column
chromatography on silica (100% EtOAc) to give
(S)-{2-(4-Chlorobenzyl)-3-oxo-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pipe-
razin-1-yl]-propyl}-carbamic acid tert-butyl ester. LCMS (APCI+)
m/z 499 [M+H].sup.+; Rt: 2.70 minutes. This was taken up into DCM
(50 mL) and stirred with TFA (5 mL) overnight. Poured into EtOAc
(100 mL) and washed with 1N NaOH (2.times.100 mL), dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. Formed HCl salt
(Et.sub.2O.HCl, 2M in diethyl ether) to give
(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-y-
l)-piperazin-1-yl]-propan-1-one dihydrochloride (100 mg, 59%.) LCMS
(APCI+) m/z 399 [M+H].sup.+; Rt: 1.82 min. [Free base: .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 9.97 (1H, br.s), 8.32 (1H, s), 7.24
(2H, d, J 7.4 Hz), 7.13 (2H, d, J 7.4 Hz), 7.09 (1H, d, J 2.4 Hz),
6.45 (1H, d, J 2.3 Hz), 4.00-3.68 (5H, m), 3.60-3.53 (1H, m),
3.46-3.40 (1H, m), 3.33-3.27 (1H, m), 3.16-3.05 (2H, m), 2.92-2.83
(2H, m), 2.77 (1H, dd, J 13.3 and 5.5 Hz.)]
Example 72
##STR00088##
[0543] Preparation of
(R)-2-Amino-1-[4-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-y-
l]-3-(4-chlorophenyl)-propan-1-one, dihydrochloride
[0544] Step 1: The 4-hydroxypyrazolopyrimidine (2.5 g, 18 mmol) was
dissolved in POCl.sub.3 (34 mL, 0.37 mol) and N,N-dimethyl aniline
(4.7 mL, 37 mmol.) This mixture was heated to reflux (120.degree.
C.) for 1.5 hours to afford a dark red solution. The mixture was
concentrated to a viscous oil and cooled to 0.degree. C. in an ice
bath. The oil was poured into a mixture of ice-water and was
stirred for 5 minutes. The acidic melt was extracted with ether
(4.times.100 mL), and the organics were combined. The organic was
washed with cold water, then cold half saturated NaHCO3 solution,
then brine, separated, dried over MgSO.sub.4, filtered, and
concentrated in vacuo to afford
4-Chloro-1H-pyrazolo[3,4-d]pyrimidine (1.1 g, 39%) as a light
yellow powder. .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 8.79 (1H, s),
8.41 (1H, s.)
[0545] Step 2: To a suspension of
4-Chloro-1H-pyrazolo[3,4-d]pyrimidine (1.1 g, 7.1 mmol) in
CHCl.sub.3 (50 mL) was added NBS (1.49 g, 8.4 mmol.) The mixture
was stirred at room temperature for 5 hours, cooled to 0 C and the
solids were isolated by vacuum filtration, rinsed with cold
CHCl.sub.3, and air dried. The solid was purified by column
chromatography on silica (50% EtOAc/hexanes) to give
3-Bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.3, 77%.)
[0546] Step 3: To a solution of
3-Bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.3 g, 5.5 mmol) in
DMF (42 mL) at 0.degree. C. was added NaH (180 mg, 7.7 mmol) in
portions. The reaction mixture was stirred at 0.degree. C. for 5
minutes, then stirred at room temperature for 1.5 hours, after
which it was cooled back to 0.degree. C. Neat PhSO.sub.2Cl (0.7 mL,
5.6 mmol) was added and the reaction mixture was warmed to room
temperature and stirred overnight. The reaction was quenched with
saturated aqueous NH.sub.4Cl and diluted further with H.sub.2O. The
resulting precipitate was isolated by vacuum filtration to give
1-Benzenesulfonyl-3-bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine
(1.8 g, 88%.) .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 9.11 (1H, s),
8.11 (2H, d, J 8.1 Hz), 7.84 (1H, t, J 7.5 Hz), 7.70 (2H, t, J 8.0
Hz.)
[0547] Step 4: A solution of
1-Benzenesulfonyl-3-bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine
(1.8 g, 4.8 mmol), Boc-piperazine (1.4 g, 7.2 mmol) and DIPEA (2.1
mL, 12 mmol) in IPA (40 mL) was stirred and heated at reflux
overnight. The reaction mixture was cooled to -10.degree. C., the
solids isolated by vacuum filtration, rinsed with cold IPA and
dried further on high vacuum line to give
4-(1-Benzenesulfonyl-3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piper-
azine-1-carboxylic acid tert-butyl ester (1.7 g, 67%) as a white
powder. LCMS (APCI+) m/z 523 and 525 [M+H].sup.+; Rt: 3.57 min.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.53 (1H, s), 8.23 (2H,
d, J 8.5 Hz), 7.65 (1H, t, J 6.9 Hz), 7.54 (2H, t, J 7.8 Hz),
3.82-3.79 (4H, m), 3.60-3.57 (4H, m), 1.48 (9H, s.)
[0548] Step 5: Anhydrous HCl (4N in dioxane, 10 mL) was added to a
stirred solution of
1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
(210 mg, 0.40 mmol) in MeOH (20 mL) and stirred at room temperature
overnight. The suspension was concentrated in vacuo to give
1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
dihydrochloride (200 mg, 100%.) LCMS (APCI+) m/z 423 and 425
[M+H].sup.+; Rt: 1.98 min.
[0549] Step 6: DIPEA (84 ul, 0.48 mmol) was added to a suspension
of
1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
dihydrochloride (40 mg, 0.081 mmol) and
(D)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(27 mg, 0.089 mmol) in DCM (10 mL) at room temperature. Then, HBTU
(34 mg, 0.089 mmol) was added and the reaction stirred at room
temperature overnight. MeOH (5 mL) and 3M LiOH (3 mL) were added
and the mixture stirred and heated at 50.degree. C. for 2 hours.
The mixture was diluted with saturated aqueous NaHCO.sub.3 (10 mL),
extracted into DCM, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The crude mixture was purified by column chromatography on
silica (50% EtOAc/hexanes) to give
(R)-[2-[4-(3-Bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-1-(4-
-chlorobenzyl)-2-oxo-ethyl]-carbamic acid tert-butyl ester. LCMS
(APCI+) m/z 564 and 566 [M+H].sup.+; Rt: 3.04 min. This was taken
up into MeOH (10 mL) and treated with anhydrous HCl (4M in dioxane,
20 mL.) The solution was stirred at room temperature overnight and
concentrated in vacuo to give
(R)-2-Amino-1-[4-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-y-
l]-3-(4-chlorophenyl)-propan-1-one, dihydrochloride (13 mg, 30%.)
LCMS (APCI+) m/z 464 and 466 [M+H].sup.+; Rt: 2.00 min.
Example 73
##STR00089##
[0550] Preparation of
(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyri-
midin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0551] Step 1: To a suspension of
4-Chloro-7H-pyrrolo[2,3-d]pyrimidine (2.5 g, 16 mmol) in CDCl.sub.3
(65 mL) was added NBS (2.9 g, 16 mmol) and the reaction mixture
stirred and heated at reflux for 2.5 hours. The mixture was cooled
to room temperature, the solids isolated by vacuum filtration,
rinsed with cold CHCl.sub.3 and air dried to give
5-Bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (3.0 g, 79%.) LCMS
(APCI+) m/z 232 and 234 [M+H].sup.+; Rt: 2.32 min. .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 12.98 (1H, br. s), 8.63 (1H, s), 7.96
(1H, s.)
[0552] Step 2: To a solution of
5-Bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (3.0 g, 13 mmol) in
DMF (40 mL) at 0.degree. C. was added NaH (60% w/w in mineral oil,
720 mg, 18.1 mmol) and the mixture stirred at 0.degree. C. under
N.sub.2 for 30 minutes. Then PhSO.sub.2Cl (1.7 g, 13 mmol) was
added and the reaction stirred at room temperature for 2 hours,
after which H.sub.2O (200 mL) was added, causing precipitation. The
precipitate was collected by filtration and dried under vacuum to
give 7-Benzenesulfonyl-5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine
(4.9 g, 100%.) .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 8.85 (1H, s),
8.45 (1H, s), 8.19 (2H, d, J 8.7 Hz), 7.81 (1H, t, J 7.4 Hz), 7.70
(2H, t, J 7.8 Hz.)
[0553] Step 3: A suspension of
7-Benzenesulfonyl-5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (4.9
g, 13 mmol), piperazine-1-carboxylic acid tert-butyl ester (3.7 g,
20 mmol), and DIPEA (5.7 mL, 33 mmol) in IPA (30 mL) was stirred
and heated at reflux for 6 hours. The mixture was cooled to
-10.degree. C., the solids collected by vacuum filtration, rinsed
with cold IPA and dried under vacuum to give
4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine--
1-carboxylic acid tert-butyl ester (5.9 g, 86%.) LCMS (APCI+) m/z
522 and 524 [M+H].sup.+; Rt: 3.92 min. .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 8.48 (1H, s), 8.21 (2H, d, J 8.2 Hz), 7.66-7.62 (2H,
m), 7.54 (2H, t, J 7.8 Hz), 3.62-3.55 (8H, m), 1.48 (9H, s.)
[0554] Step 4: MeZnCl (2.0M in THF, 720 uL, 1.4 mmol) was added to
a stirred solution
4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine--
1-carboxylic acid tert-butyl ester (250 mg, 0.48 mmol) and
tetrakis(triphenylphosphine)palladium(0) (140 mg, 0.12 mmol) in THF
(10 mL) at room temperature under N.sub.2. The solution was stirred
and heated at reflux for 2 hours, cooled to room temperature,
quenched with saturated aqueous NH.sub.4Cl, extracted into EtOAc
(2.times.100 mL), dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The product was purified on a Biotage (silica, 40%
EtOAc/hexanes) to give
4-(7-Benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-
-1-carboxylic acid tert-butyl ester (210 mg, 94%.) LCMS (APCI+) m/z
458 [M+H].sup.+; Rt: 3.73. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.49 (1H, s), 8.18 (2H, d, J 8.2 Hz), 7.59 (1H, t, J 6.9
Hz), 7.50 (2H, t, J 7.8 Hz), 7.34 (1H, s), 3.59-3.54 (4H, m),
3.48-3.44 (4H, m), 2.35 (3H, s), 1.48 (9H, s.)
[0555] Step 5: A solution of
4-(7-Benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-
-1-carboxylic acid tert-butyl ester (4.0 g, 13 mmol) in DCM (100
mL) was treated with anhydrous HCl (4M in dioxane, 100 mL) and
stirred at room temperature overnight. The suspension was
concentrated in vacuo to give
7-Benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine
dihydrochloride (5.4 g, 100%.) LCMS (APCI+) m/z 358 [M+H].sup.+;
Rt: 1.79.
[0556] Step 6: DIPEA (120 L, 0.70 mmol) was added to a suspension
of
7-Benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine
dihydrochloride (50 mg, 0.12 mmol) and
(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionic
acid (40 mg, 0.13 mmol) in DCM (10 mL) at room temperature. Then,
HBTU (48 mg, 0.13 mmol) was added and the reaction stirred at room
temperature overnight. MeOH (5 mL) and 3M LiOH (1.2 mL) were added
and the mixture stirred and heated at 50.degree. C. for 2 hours.
The mixture was diluted with saturated aqueous NaHCO.sub.3 (10 mL),
extracted into DCM, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The crude mixture was purified by column chromatography on
silica (50% EtOAc/hexanes) to give
(S)-{2-(4-Chlorobenzyl)-3-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-p-
iperazin-1-yl]-3-oxo-propyl}-carbamic acid tert-butyl ester. LCMS
(APCI+) m/z 253 [M+H].sup.+; Rt: 2.85 min. This was taken up into
MeOH (10 mL) and treated with anhydrous HCl (4M in dioxane, 20 mL.)
The solution was stirred at room temperature overnight and
concentrated in vacuo to give
(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyri-
midin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride (21 mg,
37%.) LCMS (APCI+) m/z 413 [M+H].sup.+; Rt: 1.82 min.
Example 73
##STR00090##
[0557] The Preparation of
(2S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,-
3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one
dihydrochloride
[0558] Step 1: To a solution of
4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine--
1-carboxylic acid tert-butyl ester (150 mg, 0.29 mmol) in 4 mL DME
(degassed with nitrogen prior to use) was added 0.94M aqueous
Na.sub.2CO.sub.3 (0.61 mL, 0.57 mmol) and Pd(PPh.sub.3).sub.4 (66
mg, 0.057 mmol). The reaction mixture was stirred 5 minutes, then
2-thiophene boronic acid (55 mg, 0.43 mmol) was added. The reaction
mixture heated to reflux and stirred 16 hours, after which it was
cooled to room temperature and DME was removed by rotary
evaporation. The reaction mixture was diluted with H.sub.2O and
extracted with DCM, and the combined extracts were dried
(Na.sub.2SO.sub.4), filtered, and concentrated. The crude was
purified on silica gel (8:1 to 4:1 hexanes:EtOAc) to furnish
4-(7-Benzenesulfonyl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pip-
erazine-1-carboxylic acid tert-butyl ester (120 mg, 79%) as a beige
powder. LCMS (APCI+) m/z 526 [M+H].sup.+; Rt: 3.25 min.
[0559] Step 2: To a solution of
4-(7-Benzenesulfonyl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pip-
erazine-1-carboxylic acid tert-butyl ester (120 mg, 0.23 mmol) in 2
mL dioxane was added 1.5 mL 4M HCl/dioxane. The reaction mixture
was stirred at room temperature 6 hours, after which it was diluted
with ether, and the solids were isolated by filtration through a
fritted funnel with nitrogen pressure, rinsed with ether, and dried
in vacuo to give
7-Benzenesulfonyl-4-piperazin-1-yl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrim-
idine dihydrochloride (110 mg, 95%) as a yellow powder. LCMS
(APCI+) m/z 426 [M+H].sup.+; Rt: 1.95 min.
[0560] Step 3: To a solution of
7-Benzenesulfonyl-4-piperazin-1-yl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrim-
idine dihydrochloride (50 mg, 0.10 mmol), DIEA (0.10 mL, 0.60
mmol), and (2S)-2-(Boc-aminomethyl)-3-(4-chlorophenyl)-propionic
acid (38 mg, 0.12 mmol) in 2 mL DCM was added HBTU (44 mg, 0.12
mmol). The reaction mixture was stirred at room temperature 2
hours, after which 2 mL MeOH and 0.5 mL 3M LiOH were added. The
reaction mixture was heated to 35.degree. C. and stirred 2 hours,
after which saturated NaHCO.sub.3 was added, and the mixture was
extracted with DCM. The combined extracts were dried
(Na.sub.2SO.sub.4), filtered, and concentrated. The crude was
purified on silica gel (flushed with 2:1 DCM:EtOAc, then gradient
to 1:4 DCM:EtOAc) to give
(2S)-2-Boc-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7-
H-pyrrolo[2,3-d]pyrimidin-4-yl)-pip-erazin-1-yl]-propan-1-one,
which was used in the next step.
[0561] Step 4: To a solution of
(2S)-2-Boc-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrol-
o[2,3-d]pyrimidin-4-yl)-pip-erazin-1-yl]-propan-1-one in 1.5 mL
dioxane was added 1.5 mL 4M HCl/dioxane. The resulting suspension
was stirred at room temperature 16 hours, after which it was
concentrated to dryness. The solids were dissolved in minimal MeOH,
and the product was triturated by the addition of ether. The solids
were isolated by filtration through a fritted funnel with nitrogen
pressure, rinsed with ether, and dried in vacuo to give
(2S)-2-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,-
3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
(37 mg, 67%) a pale yellow powder. .sup.1H NMR (D.sub.2O, 400 MHz)
.delta. 8.18 (1H, s), 7.41 (1H, d, J 5.4 Hz), 7.30 (1H, s), 7.19
(2H, d, J 8.6 Hz), 7.06 (1H, dd, J 5.0 and 3.5 Hz), 7.01 (2H, d, J
8.0 Hz), 6.88 (1H, d, J 3.5 Hz), 3.47-3.14 (5H, m), 3.08-3.00 (3H,
m), 2.86-2.77 (3H, m), 2.59 (1H, t, J 12.0 Hz), 2.13-2.06 (1H, m).
LCMS (APCI+) m/z 481 [M+H].sup.+; Rt: 1.87 min.
Example 75
##STR00091##
[0562] Preparation of
2-Amino-3-(4-chloro-2-methyl-phenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-
-propan-1-one, dihydrochloride
[0563] Step 1: A mixture containing 4-Chloro-2-methyl-benzoic acid
(4 g, 23 mmol) and LiAlH4 (890 mg, 23.5 mmol) in 250 mL of THF
under a nitrogen atmosphere was allowed to stir at room temperature
for 2 hours. The reaction was quenched with sodium sulfate
decahydrate. The mixture was filtered through a pad of Celite and
the filter cake washed with THF. The filtrate was concentrated
under reduced pressure. Purification of the residue via biotage
eluting with 30% ethyl acetate/hexanes gave
(4-Chloro-2-methyl-phenyl)-methanol (3.70 g, 100%) as a colorless
oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.30-7.25 (1H, m),
7.18-7.14 (2H, m), 4.66 (2H, d, J 5.8 Hz), 2.32 (3H, s.)
[0564] Step 2: A solution containing gave
(4-Chloro-2-methyl-phenyl)-methanol (2 g, 13 mmol) and PBr3 (1.3
mL, 14 mmol) in 150 mL of diethyl ether was allowed to stir at room
temperature overnight. The reaction was diluted with ether and
washed with water. The organic phase was dried over magnesium
sulfate. Filtration, removal of solvent and purification of the
residue via biotage eluting with 20% ethyl acetate/hexanes gave
1-Bromomethyl-4-chloro-2-methyl-benzene (1.89 g, 67%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.26-7.11
(3H, m), 4.67 (2H, s), 2.39 (3H, s.)
[0565] Step 3: To a solution containing
(Benzhydrylidene-amino)-acetic acid ethyl ester (2.3 g, 8.6 mmol)
in 50 mL of DMSO under a nitrogen atmosphere was added potassium
t-butoxide (1.2 g, 11 mmol) After stirring for 20 minutes,
1-Bromomethyl-4-chloro-2-methyl-benzene (1.89 g, 8.6 mmol) was
added and the reaction allowed to stir at room temperature
overnight. The reaction was diluted with ethyl acetate and washed
with brine. The organic phase was dried over magnesium sulfate.
Filtration, removal of solvent and purification of the residue via
biotage eluting with 20% ethyl acetate/hexanes gave
2-(Benzhydrylidene-amino)-3-(4-chloro-2-methyl-phenyl)-propionic
acid ethyl ester (1.75 g, 50%) as a yellow oil. LCMS (APCI+) m/z
406 [M+H].sup.+; Rt: 4.16 min.
[0566] Step 4: A mixture containing
2-(Benzhydrylidene-amino)-3-(4-chloro-2-methyl-phenyl)-propionic
acid ethyl ester (1.7 g, 4.2 mmol) and 90 mL of 3N HCl was heated
at 75 C overnight. The reaction was cooled to room temperature and
washed with ethyl acetate. The aqueous phase was concentrated under
reduced pressure to afford
2-Amino-3-(4-chloro-2-methyl-phenyl)-propionic acid (640 mg, 72%)
as white solid. LCMS (APCI+) m/z 214 [M+H].sup.+; Rt: 1.83 min.
.sup.1H NMR (D.sub.2O, 400 MHz) .delta. 7.18 (1H, s), 7.11 (1H, d,
J 8.7 Hz), 7.05 (1H, d, J 8.3 Hz), 4.05-4.00 (1H, m), 3.23 (1H, dd,
J 14.3 and 6.1 Hz), 2.98 (1H, dd, J 14.4 and 8.5 Hz), 2.19 (3H,
s.)
[0567] Step 5: To a solution containing
2-Amino-3-(4-chloro-2-methyl-phenyl)-propionic acid (640 mg, 3.0
mmol), 25 mL of dioxane and 9 mL of 1N sodium hydroxide was added
boc anhydride (0.73 g, 3.3 mmol.) The reaction was allowed to stir
at room temperature for 3 hours. The reaction was diluted with
water and washed with DCM. The aqueous phase was acidified with 1N
HCl and extracted with ethyl acetate. The organic phase was dried
over magnesium sulfate. Filtration and removal of solvent gave
2-tert-Butoxycarbonylamino-3-(4-chloro-2-methylphenyl)-propionic
acid (640 mg, 80%) as a white solid. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 7.13 (1H, s), 7.07 (2H, s), 5.14-5.08 (1H, m),
4.55-4.48 (1H, m), 3.24-3.16 (1H, m), 2.96-2.88 (1H, m), 2.33 (3H,
s), 1.38 (9H, s.)
[0568] Step 6: To a solution containing
2-tert-Butoxycarbonylamino-3-(4-chloro-2-methyl-phenyl)-propionic
acid (200 mg, 0.64 mmol) in 30 mL of DMF was added HOBT (0.12 g,
0.76 mmol), EDCI (0.15 g, 0.76 mmol) and NMM (0.19 g, 1.9 mmol)
under a nitrogen atmosphere. After stirring for 10 minutes,
4-piperazin-1-yl-quinazoline (200 mg, 0.93 mmol) was added and
stirring continued overnight. The reaction was diluted with ethyl
acetate and washed with water. The organic phase was dried over
magnesium sulfate. Filtration, removal of solvent and purification
of the residue via biotage eluting with 10% MeOH/DCM gave
[1-(4-Chloro-2-methylbenzyl)-2-oxo-2-(4-quinazolin-4-yl-piperazin-1-yl)-e-
thyl]-carbamic acid tert-butyl ester (0.31 g, 95%.) LCMS (APCI+)
m/z 510 [M+H].sup.+; Rt: 2.54 min.
[0569] Step 7: To a solution containing
[1-(4-Chloro-2-methylbenzyl)-2-oxo-2-(4-quinazolin-4-yl-piperazin-1-yl)-e-
thyl]-carbamic acid tert-butyl ester (0.30 g, 0.59 mmol) in 30 mL
of DCM under a nitrogen atmosphere was added TFA (1.4 mL.) After
stirring at room temperature overnight, the reaction was
concentrated under reduced pressure. The residue was dissolved in
DCM and 2N HCl in ether added. The solids were filtered and dried
to afford
2-Amino-3-(4-chloro-2-methyl-phenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-
-propan-1-one dihydrochloride (0.217 g) as an off-white solid. LCMS
(APCI+) m/z 410 [M+H].sup.+; Rt: 1.87 min. .sup.1H NMR (D2O, 400
MHz) .delta. 8.48 (1H, s), 7.91-7.86 (2H, m), 7.66-7.59 (2H, m),
7.08-7.02 (2H, m), 4.60-4.54 (1H, m), 4.20-4.12 (1H, m), 3.86-3.72
(3H, m), 3.61-3.38 (2H, m), 3.28-3.16 (2H, m), 3.01-2.94 (1H, m),
2.76-2.68 (1H, m), 2.19 (3H, s.)
Example 76
##STR00092##
[0570] Preparation of
4-{4-[3-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropy-
rrolo[2,3-b]pyrimidin-2-one dihydrochloride
[0571] Step 1: A solution of
4-chloro-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one (prepared
according to the literature: Li Sun et al. Bioorg. and Med. Chem.
Lett. 2002, 12, 2153-2157; 690 mg, 3.7 mmol), Boc-piperazine (630
mg, 3.7 mmol), and DIEA (0.96 mL, 5.5 mmol) in 20 mL IPA was heated
to reflux and stirred 14 hours, after which the reaction mixture
was concentrated. The crude was purified on silica gel (1:2
DCM:EtOAc to 1:4 DCM:EtOAc gradient) to give
4-Boc-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one,
which was used in the next step.
[0572] Step 2: To a solution of
4-Boc-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one in 25
mL dioxane, was added 15 mL 4M HCl/dioxane. The resulting
suspension was stirred at room temperature 15 hours, after which it
was diluted with ether. The solids were isolated by filtration
through a fritted funnel with nitrogen pressure, rinsed with ether,
and dried in vacuo to furnish
4-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one
dihydrochloride (350 mg, 100%) as a red powder. .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 11.12 (1H, s), 9.26 (2H, br. s), 8.25
(1H, s), 3.89-3.84 (4H, m), 3.77 (2H, s), 3.16-3.09 (4H, m). LCMS
(APCI+) m/z 220 [M+H].sup.+; Rt: 0.68 min.
[0573] Step 3: To a solution of give
4-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one
dihydrochloride (40 mg, 0.14 mmol), HOBt.H.sub.2O (21 mg, 0.14
mmol), TEA (57 .mu.L, 0.41 mmol), and
3-Boc-amino-2-(4-chlorophenyl)-propionic acid (prepared from
4-chlorophenylacetic acid methyl ester using the procedures
described for the preparation of A109; 49 mg, 0.16 mmol) in 1.2 mL
5:1 DCM:THF was added DCC (34 mg, 0.16 mmol). The reaction mixture
was stirred at room temperature 4 hours, after which it was
concentrated. The residue was suspended in DCM, and solids were
removed by vacuum filtration through a cotton plug and rinsed with
DCM. The filtrate was concentrated, and the crude was purified on
silica gel (1:1 to 1:5 DCM:EtOAc gradient) to give
4-{4-[3-Boc-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihyd-
ropyrrolo[2,3-b]pyrimidin-2-one, which was used in the next step.
LCMS (APCI+) m/z 401 [M-Boc+H].sup.+; Rt: 2.16 min.
[0574] Step 4: To a solution of
4-{4-[3-Boc-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihyd-
ropyrrolo[2,3-b]pyrimidin-2-one in 1.2 mL dioxane was added 1.2 mL
4M HCl/dioxane. The resulting suspension was stirred at room
temperature 15 hours, after which it was concentrated to dryness.
The solids were dissolved in minimal MeOH, and the product was
triturated by the addition of ether. The resulting solids were
isolated by filtration through a fritted funnel with nitrogen
pressure, rinsed with ether, and dried in vacuo to give
4-{4-[3-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropy-
rrolo[2,3-b]pyridin-2-one dihydrochloride (28 mg, 43%) as a dark
pink powder. .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 8.10 (1H, s),
7.31-7.14 (4H, m), 4.28-4.22 (1H, m), 3.92-3.82 (1H, m), 3.73-2.94
(11H, m). LCMS (APCI+) m/z 401 [M+H].sup.+; Rt: 1.98 min.
Example 77
##STR00093##
[0575] Preparation of
4-Amino-2-(4-chlorophenyl)-2-fluoro-1-(4-quinazolin-4-yl-piperazin-1-l)-b-
utan-1-one dihydrochloride
[0576] Step 1: A solution of 4-chloromandelic acid (12.3 g, 65.9
mmol) in toluene (50 mL), EtOH (16 mL), and concentrated
H.sub.2SO.sub.4 (0.1 mL) was refluxed for 12 hours while removing
water using a Dean-Stark trap. The mixture was concentrated in
vacuo, diluted with DCM, and washed with dilute aqueous
NaHCO.sub.3. The separated DCM layer was dried (MgSO.sub.4),
filtered, and concentrated in vacuo to give
(4-chlorophenyl)-hydroxy-acetic acid ethyl ester as a colorless oil
(10.0 g) that crystallized upon standing.
[0577] Step 2: (4-Chlorophenyl)-hydroxy-acetic acid ethyl ester
(10.0 g, 46.6 mmol) in DCM (35 mL) was cannulated into a solution
of [bis(2-methoxyethyl)amino]sulfur trifluoride (9.45 mL, 51.3
mmol) in DCM (35 mL) cooled at -78.degree. C. After being stirred
for 12 hours and allowed to warm to ambient temperature, the
mixture was poured into saturated aqueous NaHCO.sub.3. The mixture
was extracted with DCM and the organic extracts were dried
(MgSO.sub.4), filtered, and concentrated in vacuo. The crude
material was chromatographed (SiO.sub.2) using DCM as eluent to
give (4-chlorophenyl)-fluoroacetic acid ethyl ester as a colorless
oil (7.0 g).
[0578] Step 3: Potassium tert-butoxide (155 mg, 1.38 mmol) was
added to a solution of (4-chlorophenyl)-fluoroacetic acid ethyl
ester (3.00 g, 13.8 mmol) in THF (25 mL) at 0.degree. C. to give an
orange-red color. After 15 minutes, the mixture was cooled to
-78.degree. C. and t-butyl acrylate (2.23 mL, 15.2 mmol) was added
neat. After being stirred and allowed to warm to ambient
temperature for 12 hours, the mixture was quenched with saturated
NH.sub.4Cl, concentrated in vacuo, diluted with H.sub.2O, and
extracted with DCM. The DCM extracts were dried (MgSO.sub.4),
filtered, and concentrated in vacuo. The crude material was
chromatographed (SiO.sub.2) using DCM as eluent to give a colorless
oil (1.20 g). A solution of the oil in DCM (6 mL) and TFA (4 mL)
was stirred overnight. The mixture was diluted with toluene (40 mL)
and concentrated in vacuo. The crude product was dissolved in
dilute aqueous NaHCO.sub.3 and extracted with DCM (twice,
discarded). The aqueous layer was acidified to pH 1.0 with 1.0 N
HCl and extracted with DCM (twice). The organic extracts were dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo to give
2-(4-chlorophenyl)-2-fluoro-pentanedioic acid 1-ethyl ester as an
oil (1.0 g).
[0579] Step 4: Triethylamine (0.53 mL, 3.81 mmol) was added to a
solution of 2-(4-chlorophenyl)-2-fluoro-pentanedioic acid 1-ethyl
ester (1.00 g, 3.46 mmol) in t-BuOH (20 mL) followed by the
addition of diphenylphosphoryl azide (0.82 mL, 3.81 mmol). The
mixture was heated at 95.degree. C. for 3 hours, concentrated in
vacuo, and partitioned between dilute aqueous NaHCO.sub.3 and DCM.
The separated DCM layer was dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo. The crude product was chromatographed
(SiO.sub.2) using DCM as eluent to give
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid
ethyl ester (600 mg). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.42 (m, 2H), 7.35 (m, 2H), 5.25 (m, 0.4H), 4.61 (m, 0.6H), 4.19
(m, 2H), 3.35 (m, 0.6H), 3.23 (m, 1.4H), 2.59 (m, 1H), 2.34 (m,
1H), 1.40 (m, 9H), 1.24 (m, 3H). LCMS (APCI+) m/z 260 [M+H].sup.+
(loss of Boc group).
[0580] Step 5: Lithium hydroxide monohydrate (0.27 g, 6.45 mmol) in
H.sub.2O (5 mL) was added to a solution of
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid
ethyl ester (580 mg, 1.61 mmol) in THF (5 mL) and MeOH (5 mL).
After being stirred for 12 hours, the mixture was concentrated in
vacuo, diluted with H.sub.2O, and extracted with DCM (3 times,
discarded). The aqueous phase was then acidified to pH 1 and
extracted with DCM (2 times). The organic extracts were dried
(Na.sub.2SO.sub.4), filtered, and concentrated in vacuo to give
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid
(400 mg). The material was used in the following step without
further purification.
[0581] Step 6: PyBrop (562 mg, 1.21 mmol) was added in a single
portion to a solution of
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid
(400 mg, 1.21 mmol) and 4-piperazin-1-yl-quinazoline
dihydrochloride (346 mg, 1.21 mmol) in DCM (12 mL) cooled in an ice
bath. DIEA (0.84 mL, 4.82 mmol) was added and the mixture was
allowed to warm to ambient temperature and stirred for 12 hours.
The mixture was diluted with DCM and washed with 0.1 N HCl. The
separated DCM layer was dried (Na.sub.2SO.sub.4), filtered, and
concentrated in vacuo. The crude material was chromatographed
(SiO.sub.2) using 2% MeOH/DCM followed by 5% MeOH/DCM as eluent to
give
[3-(4-chlorophenyl)-3-fluoro-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-b-
utyl]-carbamic acid tert-butyl ester (230 mg). LCMS (APCI+) m/z
528, 530 [M+H].sup.+.
[0582] Step 7: A solution of
[3-(4-chlorophenyl)-3-fluoro-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-b-
utyl]-carbamic acid tert-butyl ester (226 mg, 0.43 mmol) in DCM (2
mL) and 2.0 M HCl in Et.sub.2O (1 mL) was stirred for 12 hours. The
mixture was concentrated in vacuo and chromatographed (SiO.sub.2)
using 10% MeOH/DCM followed by 10% (7 N NH.sub.3 in MeOH)/DCM as
eluent. The purified material was dissolved in MeOH followed by the
addition of 2.0 N HCl in Et.sub.2O, and then concentrated in vacuo.
The resulting glass was heated at reflux in isopropyl alcohol and
concentrated in vacuo. The resulting solid was suspended in
acetonitrile and concentrated in vacuo (repeated twice) to give
4-amino-2-(4-chlorophenyl)-2-fluoro-1-(4-quinazolin-4-yl-piperazin-1-yl)--
butan-1-one dihydrochloride as a white powder. .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 8.86 (s, 1H), 8.18 (m, 4H), 8.01
(m, 2H), 7.69 (m, 1H), 7.58 (d, J=8.4 Hz, 2H), 7.40 (d, J=8.6 Hz,
2H), 4.23 (m, 2H), 4.09 (m, 1H), 3.80 (m, 4H), 3.47 (m, 1H), 2.86
(m, 1H), 2.67 (m, 2H), 2.41 (m, 1H). LCMS (APCI+) m/z 428, 430
[M+H].sup.+.
Example 78
##STR00094##
[0583] Preparation of
3-Amino-2-(4-bromo-2-fluoro-benzyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one dihydrochloride
[0584] Step 1: To a stirred solution of diisopropyl amine (1.3 mL,
9.0 mmol) in THF (20 mL) was added n-BuLi (1.6 M solution in
hexanes, 5.6 mL, 9.0 mmol) at 0.degree. C. The reaction was stirred
at 0.degree. C. for 15 minutes and then cooled to -78.degree. C. A
solution of 3-tert-butoxycarbonylamino-propionic acid tert-butyl
ester (1.0 g, 4.1 mmol) in THF (5 mL) was added dropwise. The
mixture was stirred at -78.degree. C. for 2 hours. A solution of
4-Bromo-1-bromomethyl-2-fluoro-benzene (1.3 g, 4.9 mmol) in THF (4
mL) was added dropwise. After completion, the dry-ice bath was
removed and the reaction was warmed to 0.degree. C. in an ice bath.
After stirring at 0.degree. C. for 30 minutes, the reaction was
poured into saturated NH.sub.4Cl aqueous solution. The organic
layer was separated. The aqueous layer was extracted with EtOAc.
The combined organic layers were washed with brine, dried and
concentrated. The residue was purified by column chromatography
(hexane:EtOAc, 20:1 to 5:1) to give
2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic
acid tert-butyl ester (1.35 g, 77%) as a colorless oil. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.20 (d, J=8.0 Hz, 2H), 7.08 (t,
J=8.0 Hz, 1H), 4.86 (m, 1H), 3.28 (m, 2H), 2.92 (m, 3H), 1.43 (s,
9H), 1.36 (s, 9H).
[0585] Step 2:
2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic
acid tert-butyl ester (1.30 g, 3.01 mmol) was dissolved in THF (12
mL) and MeOH (12 mL). A solution of LiOH monohydrate (0.50 g, 12.0
mmol) in H.sub.2O (12 mL) was added. The mixture was heated at
reflux overnight. After cooling, the solvents were evaporated in
vacuo. The residue was dissolved in water and extracted with ether
(2.times.). The aqueous phase was acidified with 1N HCl and
extracted with EtOAc. The combined organic layers were washed with
brine, dried and concentrated to give
2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic
acid (1.00 g, 88%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.26 (m, 2H), 7.19 (m, 1H), 3.27 (m, 2H), 2.86 (m,
3H), 1.43 (s, 9H).
[0586] Step 3:
3-Amino-2-(4-bromo-2-fluoro-benzyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one dihydrochloride was prepared by
substituting 5-piperazin-1-yl-1H-indazole with
4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride and
substituting (D)-Boc-4-chlorophenylalanine with
2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic
acid in Example 34, Step 2, then removing the Boc protecting group
as described in Example 34, Step 3. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.37 (s, 1H), 7.40 (d, J=3.6 Hz, 1H), 7.36 (m, 1H),
7.31 (m, 1H), 7.24 (t, J=8.0 Hz, 1H), 6.95 (d, J=3.6 Hz, 1H), 4.14
(m, 2H), 4.02 (m, 1H), 3.87 (m, 4H), 3.55 (m, 2H), 3.34 (m, 1H),
3.11 (m, 1H), 2.98 (m, 2H). LCMS (APCI+) m/z 461, 463 [M+H].sup.+;
Rt=1.79 min.
Example 79
##STR00095##
[0587] Preparation of
2-(R)-Amino-3-(4-chlorophenyl)-1-(4-imidazo[1,2-a]pyridin-3-yl-piperazin--
1-yl)-propan-1-one dihydrochloride
[0588] Step 1: A mixture of tert-butyl 1-piperazinecarboxylate
(7.45 g, 40.0 mmol) and benzotriazole (4.76 g, 40.0 mmol) in
H.sub.2O (200 mL) was stirred for 1 hour. Glyoxal (40 wt. % in
water, 2.90 g, 20 mmol) was then added and the mixture was stirred
for 12 hours to produce a white precipitate. The precipitate was
filtered off and washed with H.sub.2O. The solids were dissolved in
DCM, dried (Na.sub.2SO.sub.4), filtered, and concentrated in vacuo
to give 1,2-(benzotriazol-1-yl)-1,2-(4-piperazine-1-carboxylic acid
tert-butyl ester)ethane off-white solid (10.0 g).
[0589] Step 2: A mixture of 2-aminopyridine (282 mg, 3.00 mmol) and
1,2-(benzotriazol-1-yl)-1,2-(4-piperazine-1-carboxylic acid
tert-butyl ester)ethane (1.90 g, 3.00 mmol) in dichloroethane (30
mL) was refluxed for 3 hours. Powdered KOH (555 mg, 9.90 mmol) was
then added, and the mixture was stirred for 12 hours. The mixture
was filtered and the filtrate was concentrated in vacuo. The
resulting material was chromatographed (SiO.sub.2) using DCM
followed by 5% MeOH/DCM as eluent to give
4-imidazo[1,2-a]pyridin-3-yl-piperazine-1-carboxylic acid
tert-butyl ester (900 mg). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 7.96 (m, 1H), 7.52 (m, 1H), 7.25 (m, 1H), 7.10 (m, 1H),
6.78 (m, 1H), 3.61 (m, 4H), 2.98 (m, 4H), 1.47 (s, 9H).
[0590] Step 3: A solution of
4-imidazo[1,2-a]pyridin-3-yl-piperazine-1-carboxylic acid
tert-butyl ester (950 mg, 3.14 mmol) in DCM (10 mL) and 2.0 N HCl
in Et.sub.2O (5 mL) was stirred for 12 hours. A precipitate formed
and was filtered off to give
3-piperazin-1-yl-imidazo[1,2-a]pyridine dihydrochloride as a red
solid (800 mg).
[0591] Step 4: Triethylamine (0.30 mL, 2.18 mmol) was added to a
solution of (R)--N-Boc-4-chlorophenylalanine (392 mg, 1.31 mmol),
3-piperazin-1-yl-imidazo[1,2-a]pyridine dihydrochloride (300 mg,
1.09 mmol), and 1-hydroxybenzotriazole (177 mg, 1.31 mmol) in DMF
(5 mL) followed by the addition of
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (251
mg, 1.31 mmol) in a single portion. After being stirred for 12
hours, the mixture was diluted with H.sub.2O, basified to pH 12
with 1.0 M NaOH, and extracted with DCM. The DCM extracts were
dried (Na.sub.2SO.sub.4), filtered, and concentrated in vacuo. The
residue was chromatographed (SiO.sub.2) using DCM:Et.sub.2O (1:1)
followed by 10% MeOH/DCM as eluent to give
(R)-[1-(4-chlorobenzyl)-2-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-
-1-yl)-2-oxo-ethyl]-carbamic acid tert-butyl ester (350 mg).
[0592] Step 5: A solution of
(R)-[1-(4-chlorobenzyl)-2-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-1-yl)-2-
-oxo-ethyl]-carbamic acid tert-butyl ester (350 mg, 0.72 mmol) in
DCM (3 mL) and 2.0 N HCl in Et.sub.2O (2 mL) was stirred for 12
hours. The mixture was concentrated in vacuo and the resulting
material was chromatographed (SiO.sub.2) using 10% MeOH/DCM
followed by 10% (7 N NH.sub.3 in MeOH)/DCM as eluent. This material
was dissolved in MeOH followed by the addition of 2.0 N HCl in
Et.sub.2O, and then concentrated in vacuo. The resulting gum was
stripped from isopropyl alcohol, and then from acetonitrile to give
2-(R)-amino-3-(4-chlorophenyl)-1-(4-imidazo[1,2-a]pyridin-3-yl-piperazin--
1-yl)-propan-1-one dihydrochloride as a white solid (100 mg).
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 8.66 (d, J=6.8 Hz, 1H),
8.49 (bs, 3H), 7.89 (m, 3H), 7.45 (m, 1H), 7.38 (d, J=8.3 Hz, 2H),
7.26 (d, J=8.3 Hz, 2H), 4.69 (bs, 1H), 3.49 (m, 6H), 3.08 (m, 1H),
2.94 (m, 3H), 2.85 (m, 1H). LCMS (APCI+) m/z 384, 386
[M+H].sup.+.
Example 80
##STR00096##
[0593] Preparation of
2(R)-Amino-3-(4-chlorophenyl)-3-methyl-1-(4-quinazolin-4-yl-piperazin-1-y-
l)-butan-1-one
[0594] Step 1: The 2-(4-chlorophenyl)-2-methyl-propionic acid (6.10
g, 30.7 mmol) was dissolved in 120 mL of dry THF at room
temperature. A 70% w/w solution of Red-Al (28.25 mL, 0.101 mol) was
added dropwise via syringe over 5 minutes (vigrous bubbling). The
mixture was heated to reflux for three hours. The solution was
cooled to 0 C and carefully quenched with the addition of saturated
sodium tartrate solution (100 mL, violent hydrogen evolution) and
100 mL of water. The aqueous mixture was extracted with ethyl
acetate, and the combined organic was washed with diluted
NaHCO.sub.3 solution, then brine, separated, dried over MgSO.sub.4,
filtered, and concentrated in vacuo to afford
2-(4-chlorophenyl)-2-methyl-propan-1-ol as a colorless oil (5.70 g,
99%). The material was used without purification. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.32 (s, 5H), 3.62 (d, J=4.4 Hz, 2H),
1.33 (s, 6H), 1.23 (t, J=4.4 Hz, 1H).
[0595] Step 2: The DMSO (436 mL, 61.4 mmol) was dissolved in 100 mL
of DCM and treated with oxalyl chloride (4.02 .mu.L, 46.6 mmol) at
-78.degree. C. The solution stirred for 30 minutes at -78.degree.
C. before the 2-(4-chlorophenyl)-2-methyl-propan-1-ol (5.67 g, 30.7
mmol) was added dropwise as a solution in 10 mL of DCM. After
addition was complete, the solution was stirred for two hours at
-78.degree. C., and then treated with triethyl amine (25.7 mL, 184
mmol). The solution was allowed to warm to ambient temperature and
stir for three hours. The solution was quenched with the addition
of water and partitioned with more DCM. The aqueous was extracted
with DCM, and the combined organics were dried over MgSO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
chromatography (silica, hexanes/ethyl acetate gradients) to afford
2-(4-chlorophenyl)-2-methylpropionaldehyde as a yellow oil (5.60 g,
99%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.48 (s, 1H), 7.35
(d, J=8.8 Hz, 2H), 7.21 (d, J=8.8 Hz, 1H), 1.45 (s, 6H).
[0596] Step 3: The 2-(4-chlorophenyl)-2-methylpropionaldehyde (5.60
g, 30.7 mmol) and (S)-4-methyl-benzenesulfinic acid amide (5.00 g,
32.2 mmol) were dissolved in 300 mL of DCM and treated with
Ti(OEt).sub.4 (32.1 mL, 153 mmol). The mixture was heated to reflux
under nitrogen for four hours. The solution was cooled in an ice
bath and quenched with the dropwise-addition of 200 mL of water.
The resulting precipitate (Ti salts) were removed by filtration
through a plug of celite and washed with DCM. The resulting
filtrate was separated, and the aqueous was extracted with more
DCM. The combined organic was dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by filtration
through a plug of silica gel (hexanes:ethyl acetate, 1:1) to afford
the (R)-4-methylbenzenesulfinic acid
[2-(4-chlorophenyl)-2-methylpropylidene]-amide as a colorless oil,
which solidified upon standing to give a white solid (9.28 g, 95%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.14 (s, 1H), 7.54 (d,
J=8.4 Hz, 2H), 7.31 (d, J=8.8 Hz, 2H), 7.27 (d, J=8.8 Hz, 2H), 7.14
(m, 2H), 2.42 (s, 3H), 1.52 (s, 3H), 1.47 (s, 3H).
[0597] Step 4: The diethyl aluminum cyanide (43.5 mL of a 1.0M
solution in toluene, 43.5 mmol) was added to isopropanol (28.9 mL,
377 mmol) and stirred at 10.degree. C. for 15 minutes. This
solution was cannulated into the (R)-4-methylbenzenesulfinic acid
[2-(4-chlorophenyl)-2-methyl-propylidene]-amide (9.28 g, 29.0 mmol)
as a solution in 290 mL of THF at -78.degree. C. This solution was
allowed to stir for 15 minutes at -78.degree. C. then allowed to
warm slowly to room temperature overnight. The solution was
quenched with the addition of diluted NaHCO.sub.3 solution and
extracted with ethyl acetate. The combined organic was washed with
brine, separated, dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo to afford the 4-methyl-benzenesulfinic acid
[2-(4-chlorophenyl)-1-cyano-2-methyl-propyl]-amide as a colorless
oil (9.55 g, 95% yield). The material was heated to 110.degree. C.
(reflux) in concentrated HCl solution over the weekend. The
solution was cooled to room temperature, washed with ether, then
concentrated in vacuo to give the
(S)-2-amino-3-(4-chlorophenyl)-3-methyl-butyric acid hydrochloride
salt a white solid (1.61 g, 21%). .sup.1H NMR (DMSO-d.sub.6, 400
MHz) .delta. 8.19 (brs, 1H), 7.48 (d, J=8.0 Hz, 2H), 7.41 (d, J=8.0
Hz, 2H), 7.13 (s, 3H), 4.15 (s, 1H), 1.42 (s, 3H), 1.40 (s, 3H).
LCMS (APCI+) m/z 228 [M+H].sup.+; Rt=1.81 min.
[0598] Step 5: The (S)-2-amino-3-(4-chlorophenyl)-3-methyl-butyric
acid hydrochloride salt (1.00 g, 3.79 mmol) was dissolved in 6 mL
of 2M NaOH solution at room temperature and treated with
di-tert-butyl di-carbonate (957 L, 4.16 mmol). The solution was
allowed to stir for four hours to completion, and the aqueous
solution was acidified with the addition of 1M HCl solution
(pH=2-3). The aqueous was extracted with ethyl acetate, and the
organics were combined. The organic was washed with brine,
separated, dried over MgSO.sub.4, filtered, and concentrated in
vacuo to give a colorless oil. The residue was purified by
chromatography (silica, hexanes/ethyl acetate gradients) to afford
the
(S)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-3-methyl-butyric
acid (636 mg, 51%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
12.67-10.55 (brs, 1H), 7.29 (m, 4H), 4.97 (d, J=8.4 Hz, 1H), 4.55
(d, J=8.4 Hz, 1H), 1.45 (s, 9H), 1.38 (s, 6H). LCMS (APCI+) m/z 228
[M-Boc+H].sup.+; Rt=3.20 min.
[0599] Step 6: The 4-piperazin-1-yl-quinazoline bis-hydrochloride
(220 mg, 0.766 mmol),
(S)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-3-methyl-butyric
acid (251 mg, 0.766 mmol, 1.0 equiv), 1-hydroxybenzotriazole (109
mg, 0.804 mmol, 1.05 equiv), and EDCI (154 mg, 0.804 mmol, 1.05
equiv) were dissolved/suspended in 6.0 mL of DMF. The mixture was
treated with triethylamine (427 .mu.L, 3.06 mmol) and allowed to
stir overnight to completion. The reaction was partitioned between
ethyl acetate and diluted NaHCO.sub.3 solution. The aqueous was
extracted with ethyl acetate, and the organics were combined. The
organic was washed with water, then brine, separated, dried over
MgSO.sub.4, filtered, and concentrated in vacuo. The residue was
eluted through a small plug of silica gel with ethyl acetate and
concentrated in vacuo. The protected intermediate was immediately
dissolved in 1 mL of dioxane and treated with 4M HCl in dioxane
(1.92 mL, 7.66 mmol) at room temperature for four hours. The
resulting precipitate was triturated with ether, then filtered to
afford the
(R)-2-amino-3-(4-chlorophenyl)-3-methyl-1-(4-quinazolin-4-yl-piperazin-1--
yl)-butan-1-one bis-hydrochloride as a light-yellow solid (277 mg,
73%) upon drying under vacuum. .sup.1H NMR (DMSO-d.sub.6, 400 MHz)
.delta. 8.89 (s, 1H), 8.46 (brs, 3H), 8.17 (d, J=8.4 Hz, 1H), 8.06
(t, J=7.6 Hz, 1H), 7.97 (d, J=8.4 Hz, 1H), 7.74 (d, J=8.0 Hz, 1H),
7.50 (d, J=8.0 Hz, 2H), 7.42 (d, J=8.0 Hz, 2H), 4.53 (brs, 1H),
4.09 (m, 3H), 3.90 (m, 3H), 3.32 (m, 1H), 3.04 (t, J=9.2 Hz, 1H),
1.51 (s, 3H), 1.43 (s, 3H). LCMS (APCI+) m/z 424 [M+H].sup.+;
Rt=1.90 min.
Example 81
##STR00097##
[0600] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza--
fluoren-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0601] Step 1:
4-(6,7,8,9-Tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazine-1-carboxyl-
ic acid tert-butyl ester was prepared by the procedures described
in Example 40, Step 1, substituting 4-chloro-5-iodopyrimidine with
4-Chloro-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorene (prepared from
2-Amino-1-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-1H-indole-3-carbonitrile
according to the literature: Traxler, P. M. et. al. (1996), J. Med.
Chem., 39, 2285-2292). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
10.24 (s, 1H), 8.32 (s, 1H), 3.59 (m, 4H), 3.55 (m, 4H), 2.78 (m,
4H), 1.92 (m, 2H), 1.82 (m, 2H), 1.43 (s, 9H). LCMS (APCI+) m/z 358
[M+H].sup.+; Rt=3.12 min.
[0602] Step 2:
4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorene
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(6,7,8,9-Tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazine-1-carboxyl-
ic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.42 (s, 1H), 4.14 (m, 4H), 3.48 (m, 4H), 2.83 (m, 4H), 1.96 (m,
2H), 1.86 (m, 2H). LCMS (APCI+) m/z 258 [M+H].sup.+; Rt=1.52
min.
[0603] Step 3: To a suspension of
4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorene
dihydrochloride (20 mg, 0.061 mmol) and
(D)-Boc-4-chlorophenylalanine (20 mg, 0.067 mmol) were added DIEA
(63 .mu.L, 0.36 mmol) and HBTU (25 mg, 0.067 mmol). The reaction
was stirred at room temperature for 2 hours. The mixture was
partitioned between water and EtOAc. The organic layer was washed
with aqueous NaHCO.sub.3 and brine, dried and concentrated. The
residue was purified by column chromatography (DCM:MeOH, 40:1 to
20:1) to give
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza-f-
luoren-4-yl)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester
as a colorless oil. Removal of the Boc protecting group by
procedures described in Example 34, Step 3 afforded
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza--
fluoren-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride (28 mg,
90%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.29 (s, 1H), 7.42 (d, J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.74
(m, 1H), 3.75 (m, 6H), 3.38 (m, 1H), 3.16 (m, 3H), 2.77 (m, 4H),
1.92 (m, 4H) LCMS (APCI+) m/z 439, 441 [M+H].sup.+; Rt=2.16
min.
Example 82
##STR00098##
[0604] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-
-propan-1-one
[0605] Step 1: To a stirred suspension of NaH (60%, 0.146 g, 3.65
mmol) in DMF (15 mL) was added dropwise a solution of
4-Hydroxypiperidine-1-carboxylic acid tert-butyl ester (0.611 g,
3.04 mmol) in DMF (5 mL) at 0.degree. C. The reaction was stirred
for 1 hour and then 4-chloroquinazoline (0.500 g, 3.04 mmol) was
added. The mixture was allowed to warm to room temperature and
stirred overnight. The mixture was partitioned between H.sub.2O and
EtOAc. The aqueous phase was extracted with EtOAc. The combined
organic layers were washed with brine, dried and concentrated. The
residue was purified by column chromatography (hexanes:EtOAc, 2:1)
to give 4-(Quinazolin-4-yloxy)-piperidine-1-carboxylic acid
tert-butyl ester (0.76 g, 76%) as a colorless oil. Removal of the
Boc group by the procedures described in Example 34, Step 3
afforded 4-(Piperidin-4-yloxy)-quinazoline dihydrochloride as a
white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 9.35 (s,
1H), 8.59 (d, J=8.4 Hz, 1H), 8.31 (td, J=7.2 Hz, J=1.2 Hz 1H), 8.11
(d, J=8.4 Hz, 1H), 8.04 (t, J=7.2 Hz, 1H), 6.00 (m, 1H), 3.55 (m,
2H), 3.40 (m, 2H), 2.42 (m, 4H). LCMS (APCI+) m/z 230 [M+H].sup.+;
Rt=1.67 min.
[0606] Step 2:
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(quinazolin-4-yloxy)-piperidin-1-yl]--
ethyl}-carbamic acid 9H-fluoren-9-ylmethyl ester was prepared by
the procedures described in Example 34, Step 2, substituting
5-piperazin-1-yl-1H-indazole with 4-(Piperidin-4-yloxy)-quinazoline
dihydrochloride and substituting (D)-Boc-4-chlorophenylalanine with
(D)-Fmoc-4-chlorophenylalanine. .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.77 (s, 1H), 8.12 (m, 1H), 8.02 (s, 1H), 7.94 (m, 1H),
7.84 (m, 1H), 7.77 (m, 2H), 7.59 (m, 4H), 7.41 (m, 2H), 7.30 (m,
4H), 7.15 (m, 2H), 5.71 (d, J=8.4 Hz, 1H), 5.55 (m, 1H), 4.94 (m,
1H), 4.40 (m, 2H), 4.21 (m, 1H), 3.85 (m, 1H), 3.61 (m, 1H), 3.38
(m, 1H), 3.02 (m, 2H), 1.94 (m, 5H). LCMS (APCI+) m/z 633, 635
[M+H].sup.+; Rt=3.98 min.
[0607] Step 3: To a stirred solution of
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(quinazolin-4-yloxy)-piperidin-1-yl]--
ethyl}-carbamic acid 9H-fluoren-9-ylmethyl ester (0.166 g, 0.262
mmol) in DCM (5 mL) was added piperidine (1 mL). The reaction was
stirred at room temperature for 4 hours. The volatiles were
evaporated. The residue was purified by column chromatography
(DCM:MeOH, 50:1 to 10:1) to give
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-
-propan-1-one (0.099 g, 92%) as a colorless oil. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.77 (s, 1H), 8.15 (m, 1H), 7.94 (d,
J=8.4 Hz, 1H), 7.85 (m, 1H), 7.59 (m, 1H), 7.30 (d, J=8.0 Hz, 2H),
7.17 (d, J=8.0 Hz, 2H), 5.57 (m, 1H), 3.95 (m, 2H), 3.10-3.70 (m,
3H), 2.95 (m, 1H), 2.82 (m, 1H), 1.30-2.10 (m, 4H). LCMS (APCI+)
m/z 411, 413 [M+H].sup.+; Rt=2.19 min.
Example 83
##STR00099##
[0608] Preparation of
4-Amino-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-
-1-one dihydrochloride
[0609] Step 1: An analogous reaction to that described in example
61 steps 1-3, but starting with (3,4-dichlorophenyl)-acetic acid
methyl ester yielded
4-tert-butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.41 (m, 2H), 7.17 (d,
J=8 Hz, 1H), 4.61 (brs, 1H), 3.58 (m, 1H), 3.16 (m, 2H), 2.28 (m,
1H), 1.92 (m, 1H), 1.44 (s, 9H).
[0610] Step 2: The 4-piperazin-1-yl-quinazoline (20 mg, 0.070 mmol)
was dissolved in 1 mL CHCl.sub.3 and
4-tert-butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid (36
mg, 0.10 mmol) was added. PS-carbodiimide resin (0.21 mmol) was
added and the mixture was shaken overnight. The reaction mixture
was filtered and the filtrate was concentrated in vacuo. The
residue was purified by column chromatography (silica gel eluted
with 1:4 DCM/EtOAc) to afford the pure Boc-protected intermediate.
The material was dissolved in 1.0 mL of 1,4-dioxane and treated
with 1.0 mL of 4M HCl in dioxane (4 mmol). The solution was allowed
to stir at room temperature overnight to completion. The reaction
mixture was concentrated to dryness and the residue was triturated
with 2 mL diethyl ether. The solid was filtered under a nitrogen
atmosphere and was allowed to dry under vacuum for 2 hours to
afford
4-amino-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-butan-1-one dihydrochloride (15 mg, 42%). .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.69 (s, 1H), 8.22 (d, J=9 Hz, 1H),
8.04 (t, J=8 Hz, 1H), 7.77 (m, 2H), 7.54 (m, 2H), 7.31 (d, J=8.6
MHz, 1H), 4.30 (m, 4H), 3.98 (m, 4H), 3.60 (m, 1H), 2.98 (m, 1H),
2.86 (m, 1H), 2.36 (m, 1H), 2.02 (m, 1H).
Example 84
##STR00100##
[0611] Preparation of
2-(3,4-Dichlorophenyl)-4-methylamino-1-(4-quinazolin-4-yl-piperazin-1-yl)-
-butan-1-one dihydrochloride
[0612]
2-(3,4-dichlorophenyl)-4-Boc-amino-1-(4-quinazolin-4-yl-piperazin-1-
-yl)-butan-1-one was N-methylated following a literature procedure
(Mahavir Prashad et al. Org. Lett. 2003, 5(2), 125-128) to give
[3-(3,4-Dichlorophenyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-
-methyl-carbamic acid tert-butyl ester, which was treated with
excess HCl/dioxane to furnish
2-(3,4-dichlorophenyl)-4-methylamino-1-(4-quinazolin-4-yl-piperazin-1-yl)-
-butan-1-one dihydrochloride (7 mg, 31%). .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 8.85 (br s, 2H), 8.84 (s, 1H), 8.17
(d, J=7.6 Hz, 1H), 8.01 (m, 1H), 7.89 (d, J=7.6 Hz, 1H), 7.69-7.61
(m, 3H), 7.33 (d, J=7.6 Hz, 1H), 4.40 (m, 1H), 4.11 (br s, 3H),
3.95 (m, 2H), 3.78 (m, 1H), 3.73 (m, 2H), 3.45 (m, 3H), 2.80 (m,
1H), 2.71 (m, 1H), 2.25 (m, 1H), 1.97 (m, 1H). LCMS (APCI+) m/z 458
[M+H].sup.+. HPLC R.sub.t=1.78 min.
Example 85
##STR00101##
[0613] Preparation of
1-{4-[4-Amino-2-(3,4-dichlorophenyl)-butyryl]-piperazin-1-yl}-quinolizin--
4-one hydrochloride
[0614] Step 1: To a solution of
4-Pyridin-2-ylmethyl-piperazine-1-carboxylic acid tert-butyl ester
(prepared from 1-Pyridin-2-ylmethyl-piperazine according to the
literature: J. Med. Chem. (1993), 36, 2984) (2.00 g, 7.21 mmol) in
THF (15 mL) was added n-BuLi (1.6 M in hexanes, 5.0 mL, 7.9 mmol)
at -78.degree. C. The mixture was allowed to warm to room
temperature and stirred for 30 minutes. The solution was then
cooled to -78.degree. C. and a solution of diethyl
ethoxymethylenemalonate (1.72 g, 7.93 mmol) in THF (2 mL) was added
dropwise. The reaction mixture was allowed to warm to 0.degree. C.
over 1 hour and stirred at 0.degree. C. for 1 hour. The reaction
was poured into water and extracted with EtOAc. The combined
organic layers were washed with brine, dried and concentrated. The
residue was purified by column chromatography (hexanes:EtOAc, 8:1
to 1;1) to give
2-[2-(4-tert-Butoxycarbonyl-piperazin-1-yl)-1-ethoxy-2-pyridin-2--
yl-ethyl]-malonic acid diethyl ester (2.40 g, 67%) as a mixture of
diastereomers. Diastereomer 1: .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.55 (d, J=4.4 Hz, 1H), 7.63 (td, J=7.6 Hz, J=1.6 Hz, 1H),
7.21 (d, J=7.6 Hz, 1H), 7.16 (dd, J=7.6 Hz, J=1.6 Hz, 1H), 4.74 (m,
1H), 4.16 (m, 5H), 3.75 (q, J=7.2 Hz, 2H), 3.52 (d, J=4.4 Hz, 1H),
3.38 (m, 4H), 2.67 (m, 2H), 2.44 (m, 2H), 1.40 (s, 9H), 1.22 (m,
6H), 1.44 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 494 [M+H].sup.+;
Rt=3.61 min. Diastereomer 2: .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.58 (d, J=4.8 Hz, 1H), 7.59 (td, J=7.6 Hz, J=1.6 Hz, 1H),
7.15 (dd, J=7.6 Hz, J=1.6 Hz, 1H), 7.08 (d, J=8.0 Hz, 1H), 4.91 (m,
1H), 4.16 (m, 5H), 3.53 (m, 1H), 3.22 (m, 6H), 2.50 (m, 2H), 2.19
(m, 2H), 1.34 (s, 9H), 1.24 (m, 6H), 0.69 (t, J=7.2 Hz, 3H). LCMS
(APCI+) m/z 494 [M+H].sup.+; Rt=3.62 min.
[0615] Step 2:
2-[2-(4-tert-Butoxycarbonyl-piperazin-1-yl)-1-ethoxy-2-pyridin-2-yl-ethyl-
]-malonic acid diethyl ester (2.40 g, 4.86 mmol) was dissolved in
xylene (20 mL) and heated at 140.degree. C. for 12 hours. After
cooling, the volatiles were evaporated and the residue was purified
by column chromatography (EtOAc) to give
1-(4-tert-Butoxycarbonyl-piperazin-1-yl)-4-oxo-1,9a-dihydro-4H-quinolizin-
e-3-carboxylic acid ethyl ester (1.39 g, 71%) as an orange solid.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.48 (d, J=7.2 Hz, 1H),
8.34 (s, 1H), 7.69 (td, J=7.2 Hz, J=1.2 Hz, 1H), 7.23 (m, 1H), 4.44
(q, J=7.2 Hz, 2H), 4.14 (m, 2H), 3.10 (m, 2H), 2.88 (m, 4H), 1.51
(s, 9H), 1.43 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 402 [M+H].sup.+;
Rt=2.85 min.
[0616] Step 3: A mixture of
1-(4-tert-Butoxycarbonyl-piperazin-1-yl)-4-oxo-1,9a-dihydro-4H-quinolizin-
e-3-carboxylic acid ethyl ester (0.320 g, 0.797 mmol) in
concentrated HCl (5 mL) was refluxed for 30 minutes. After cooling,
the reaction was basified with aqueous NaHCO.sub.3 solution and
thoroughly extracted with DCM. The combined organic layers were
washed with brine, dried and concentrated to give
1-piperazin-1-yl-1,9a-dihydro-quinolizin-4-one (0.075 g, 41%) as a
yellow oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.16 (d,
J=7.2 Hz, 1H), 8.20 (d, J=9.2 Hz, 1H), 7.71 (d, J=9.6 Hz, 1H), 7.37
(t, J=7.2 Hz, 1H), 7.03 (t, J=6.4 Hz, 1H), 6.61 (d, J=9.2 Hz, 1H),
3.07 (m, 4H), 2.87 (m, 4H). LCMS (APCI+) m/z 230 [M+H].sup.+;
Rt=0.29 min.
[0617] Step 4:
1-{4-[4-Amino-2-(3,4-dichlorophenyl)-butyryl]-piperazin-1-yl}-quinolizin--
4-one hydrochloride was prepared by substituting
5-piperazin-1-yl-1H-indazole with
1-piperazin-1-yl-1,9a-dihydro-quinolizin-4-one and substituting
(D)-Boc-4-chlorophenylalanine with
4-tert-Butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid in
Example 34, Step 2, then removing the Boc protecting group as
described in Example 34, Step 3. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 9.25 (d, J=7.2 Hz, 1H), 9.57 (d, J=8.8 Hz, 1H), 8.00 (m,
2H), 7.71 (d, J=7.2 Hz, 1H), 7.58 (m, 2H), 7.33 (d, J=8.0 Hz, 1H),
7.02 (d, J=8.4 Hz, 1H), 4.31 (m, 1H), 3.82 (m, 5H), 3.65 (m, 2H),
3.34 (m, 1H), 2.98 (m, 1H), 2.86 (m, 1H), 2.36 (m, 1H), 2.03 (m,
1H). LCMS (APCI+) m/z 459, 461, 463 [M+H].sup.+; Rt=2.02 min.
Example 86
##STR00102##
[0618] Preparation of
(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperidin-1-yl)-propan-1-one
dihydrochloride
[0619] Step 1: To a 25 mL flask was charged
4-Trifluoromethanesulfonyloxy-3,6-dihydro-2H-pyridine-1-carboxylic
acid benzyl ester (prepared from 4-Oxo-piperidine-1-carboxylic acid
benzyl ester according to the literature: Wustrow, D. J. et. al.
(1991), Synthesis, 993-995. 1.14 g, 3.12 mmol), 4-chloroquinazoline
(0.512 g, 3.12 mmol), lithium chloride (0.397 g, 9.36 mmol),
Pd(PPh.sub.3).sub.4 (0.180 g, 0.156 mmol) and hexamethyl ditin
(1.02 g, 3.12 mmol). 1,4-Dioxane (20 mL) was added and the reaction
was degassed with N.sub.2 for 15 minutes. The mixture was stirred
at reflux overnight. After cooling, the black suspension was poured
into saturated aqueous potassium fluoride solution. The mixture was
diluted with EtOAc and stirred for 2 hours. The organic phase was
separated. The aqueous phase was extracted with EtOAc. The combined
organic layers were washed with brine, dried and concentrated. The
residue was purified by column chromatography (hexanes:EtOAc, 2:3)
to give 4-Quinazolin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid
benzyl ester (0.790 g, 73%) as a colorless oil. .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 9.27 (s, 1H), 8.19 (d, J=7.2 Hz, 1H),
8.07 (d, J=8.8 Hz, 1H), 7.91 (m, 1H), 7.62 (t, J=7.2 Hz, 1H), 7.41
(m, 6H), 6.18 (m, 1H), 5.23 (s, 2H), 4.32 (s, 2H), 3.85 (t, J=5.6
Hz, 1H), 2.80 (br s, 2H). LCMS (APCI+) m/z 346 [M+H].sup.+; Rt=3.16
min.
[0620] Step 2: To a stirred solution of
4-Quinazolin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid benzyl
ester (0.907 g, 2.63 mmol) in MeOH (30 mL) under N.sub.2 was
cautiously added 10% Pd on carbon (100 mg). The reaction was
hydrogenated at 50 psi using a parr shaker for 3 days. The catalyst
was removed by filtration. The filtrate was evaporated under
vacuum. The resulting residue was purified by column chromatography
(DCM:MeOH, 20:1 to DCM:MeOH:Et.sub.3N, 100:10:1) to give
4-(1,2,3,6-Tetrahydro-pyridin-4-yl)-quinazoline (0.358 g, 64%) as a
white waxy solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.27
(s, 1H), 8.19 (d, J=8.4 Hz, 1H), 8.06 (d, J=8.4 Hz, 1H), 7.88 (t,
J=8.4 Hz, 1H), 7.65 (t, J=8.4 Hz, 1H), 3.71 (m, 1H), 3.31 (m, 2H),
2.90 (m, 2H), 1.99 (m, 4H). LCMS (APCI+) m/z 214 [M+H].sup.+;
Rt=1.60 min.
[0621] Step 3:
(2R)-[1-Benzyl-2-oxo-2-(4-quinazolin-4-yl-piperidin-1-yl)-ethyl]-carbamic
acid tert-butyl ester was prepared by substituting
5-piperazin-1-yl-1H-indazole with
4-(1,2,3,6-Tetrahydro-pyridin-4-yl)-quinazoline and substituting
(D)-Boc-4-chlorophenylalanine with (D)-Boc-phenylalanine in Example
34, Step 2. .sup.1H NMR (CDCl.sub.3, 400 MHz) (1:1 mixture of
rotamers) .delta. 9.26, 9.21 (2s, 1H, rotamers), 8.08 (m, 2H), 7.89
(m, 1H), 7.65 (m, 1H), 7.28 (m, 5H), 5.51, 5.43 (2d, J=8.4 Hz, 1H,
rotamers), 4.92 (m, 1H), 4.72 (m, 1H), 3.95, 3.83 (2d, J=13.2 Hz,
1H, rotamers), 3.65 (m, 1H), 2.20-3.30 (m, 4H), 1.20-2.10 (m, 4H),
1.44, 1.42 (2s, 9H, rotamers). LCMS (APCI+) m/z 461 [M+H].sup.+;
Rt=3.23 min.
[0622] Step 4:
(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperidin-1-yl)-propan-1-one
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
(2R)-[1-Benzyl-2-oxo-2-(4-quinazolin-4-yl-piperidin-1-yl)-ethyl]-carbamic
acid tert-butyl ester. LCMS (APCI+) m/z 361 [M+H].sup.+; Rt=2.38
min.
Example 87
##STR00103##
[0623] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6--
dihydro-2H-pyridin-1-yl]-propan-1-one dihydrochloride
[0624] Step 1: To a nitrogen flushed flask containing
4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine--
1-carboxylic acid tert-butyl ester (prepared from
tert-butyl-4-oxopiperidine-1-carboxylate according to the
literature: Eastwood, P. R. (2000), Tetrahedron Lett., 3705-3708.
127 mg, 0.410 mmol), K.sub.2CO.sub.3 (142 mg, 1.02 mmol) and
dichloro[1,1'-bis(diphenylphosphino)-ferrocene]palladium(II)
dichloromethane adduct (17 mg, 0.020 mmol) was added a solution of
1-Benzenesulfonyl-4-chloro-1H-pyrrolo[2,3-b]pyridine (100 mg, 0.342
mmol) in DMF (3 mL). The mixture was heated at 80.degree. C. for 36
hours. The mixture was cooled to room temperature and partitioned
between EtOAc and water. The combined organic layers were washed
with saturated aqueous NaHCO.sub.3 and brine, dried and
concentrated. The residue was purified by flash chromatography on
silica gel, eluting with hexanes:EtOAc (2:1) to give
4-(1-Benzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2-
H-pyridine-1-carboxylic acid tert-butyl ester (0.068 g, 45%) as a
colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.37 (d,
J=5.2 Hz, 1H), 8.20 (d, J=8.0 Hz, 2H), 7.73 (d, J=4.0 Hz, 1H), 7.56
(m, 1H), 7.49 (m, 2H), 7.03 (d, J=5.2 Hz, 1H), 6.73 (d, J=4.0 Hz,
1H), 6.14 (br s, 1H), 4.12 (m, 2H), 3.65 (m, 2H), 2.55 (m, 2H),
1.50 (s, 9H). LCMS (APCI+) m/z 440 [M+H].sup.+; Rt=3.84 min.
[0625] Step 2:
1-Benzenesulfonyl-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-pyrrolo[2,3-b]py-
ridine dihydrochloride was prepared by the procedures described in
Example 34, Step 3, substituting
(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(1-Benzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridi-
ne-1-carboxylic acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.36 (d, J=5.2 Hz, 1H), 8.15 (d, J=8.0 Hz, 2H), 7.93
(d, J=4.0 Hz, 1H), 7.76 (m, 1H), 7.56 (m, 2H), 7.29 (d, J=5.2 Hz,
1H), 6.97 (d, J=4.0 Hz, 1H), 6.29 (br s, 1H), 3.93 (m, 2H), 3.60
(m, 2H), 2.85 (m, 2H). LCMS (APCI+) m/z 340 [M+H].sup.+; Rt=1.87
min.
[0626] Step 3: To a solution of
1-Benzenesulfonyl-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-pyrrolo[2,3-b]py-
ridine dihydrochloride (25 mg, 0.061 mmol) and
(D)-Boc-4-chlorophenylalanine (20 mg, 0.067 mmol) in DMF (2 mL)
were added DIEA (63 .mu.L, 0.36 mmol) and HBTU (25 mg, 0.067 mmol).
The reaction was stirred at room temperature for 2 hours. The
mixture was partitioned between water and EtOAc. The organic layer
was washed with aqueous NaHCO.sub.3 and brine, dried and
concentrated. The residue was dissolved in THF (0.3 mL) and MeOH
(0.3 mL). A solution of lithium hydroxide monohydrate (10 mg, 0.24
mmol) in H.sub.2O (0.3 mL) was added. The mixture was heated at
50.degree. C. overnight. After cooling, the reaction was
partitioned between EtOAc and water. The organic layer was washed
with brine, dried and concentrated. The residue was purified by
column chromatography (hexanes:EtOAc, 1:1 to 3:1) to give
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-d-
ihydro-2H-pyridin-1-yl]-ethyl}-carbamic acid tert-butyl ester as a
colorless oil. Removal of the Boc group by the procedures described
in Example 34, Step 3 afforded
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6--
dihydro-2H-pyridin-1-yl]-propan-1-one dihydrochloride (9 mg, 33%)
as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) (1:1 mixture of
rotamers) .delta. 8.37 (m, 1H), 7.73 (s, 1H), 7.47, 7.40 (2d, J=6.0
Hz, rotamers), 7.33 (m, 3H), 7.23 (d, J=8.0 Hz, 2H), 6.99 (m, 1H),
6.33, 6.31 (2s, 1H, rotamers), 4.74 (m, 1H), 4.36 (m, 1H), 4.21 (m,
1H), 3.45-3.90 (m, 2H), 3.05-3.30 (m, 2H), 1.90-2.70 (m, 2H). LCMS
(APCI+) m/z 381, 383 [M+H].sup.+; Rt=1.95 min.
Example 88
##STR00104##
[0627] Preparation of
2(R)-Amino-3-(4-chlorophenyl)-N-(1-quinazolin-4-yl-azetidin-3-yl)-propion-
amide
[0628] Step 1: The (1-benzhydrylazetidin-3-yl)-carbamic acid
tert-butyl ester (500 mg, 1.48 mmol), Pd/C (10% w/w, 157 mg, 0.07
mmol), and ammonium formate (932 mg, 14.8 mmol) were weighed into a
25 RBF equipped with a condenser, degassed 3 times, and
suspended/dissolved in 6 mL of methanol. The mixture was heated to
60 C for 4 hours to completion and was allowed to cool to room
temperature. The mixture was filtered through a plug of celite
washed with ethanol, and the filtrate was concentrated in vacuo.
The residue was re-dissolved in 30 mL of DCM, dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to afford the
crude intermediate. The intermediate and 4-chloroquinazoline (268
mg, 1.63 mmol) were dissolved in 6 mL of NMP, then treated with
diisopropylethyl amine (515 .mu.L, 2.96 mmol). The solution was
heated to 80.degree. C. overnight to completion affording an orange
mixture. After cooling to room temperature, the solution was
diluted with ethyl acetate and poured into diluted NaHCO.sub.3
solution. The aqueous was extracted with ethyl acetate, and the
organics were combined. The organic was washed with water, brine,
separated, dried over MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified by chromatography (silica gel
eluted with 9:1 MeOH:EtOAc) to afford the pure
(1-quinazolin-4-yl-azetidin-3-yl)-carbamic acid tert-butyl ester as
a tan solid (390 mg, 88%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz)
.delta. 8.47 (s, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.78 (t, J=7.6 Hz,
1H), 7.72 (d, J=8.4 Hz, 1H), 7.67 (m, 1H), 7.48 (t, J=7.6 Hz, 1H),
4.73 (m, 2H), 4.50 (m, 1H), 4.30 (m, 2H), 1.41 (s, 9H). LCMS
(APCI+) m/z 301 [M+H].sup.+; Rt=2.26 min.
[0629] Step 2: The (1-quinazolin-4-yl-azetidin-3-yl)-carbamic acid
tert-butyl ester (390 mg, 1.30 mmol) was dissolved in 7 mL of 4M
HCl and allowed to stir at 80.degree. C. to completion after three
hours. The aqueous solution was washed with ether (discarded), and
the aqueous layer was concentrated in vacuo to afford the
de-protected intermediate as a white solid. The flask containing
this solid was charged with HOBt (193 mg, 1.43 mmol), EDCI (274 mg,
1.43 mmol), and the
2(S)-tert-butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(389 mg. 1.30 mmol). The mixture was suspended/dissolved in 12.0 mL
of DMF and treated with TEA (905 L, 6.49 mmol). The mixture was
allowed to stir for four hours to completion. The contents were
partitioned between ethyl acetate and diluted NaHCO.sub.3 solution.
The aqueous was extracted with ethyl acetate, and the organics were
combined. The organic was washed with water, brine, separated,
dried over MgSO.sub.4, filtered, and concentrated in vacuo to
afford the crude Boc-intermediate as a white solid. The material
was dissolved in 7 mL of DCM and treated with 4.0 mL of TFA. After
two hours, the reaction solution was concentrated in vacuo to
afford a pale yellow oil. The contents were partitioned between
ethyl acetate and diluted NaHCO.sub.3 solution. The aqueous was
extracted with ethyl acetate, and the organics were combined. The
organic was washed with brine, separated, dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The residue
was purified by chromatography (silica gel eluted with 9:1
MeOH:DCM) to afford the pure
2(R)-amino-3-(4-chlorophenyl)-N-(1-quinazolin-4-yl-azetidin-3-yl)-propion-
amide as a colorless oil (177 mg, 30%). .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 8.60 (s, 1H), 8.02 (brs, 1H), 7.82 (d, J=8.4 Hz,
1H), 7.75 (d, J=8.8 Hz, 1H), 7.72 (t, J=8.4 Hz, 1H), 7.40 (t, J=7.6
Hz, 1H), 7.29 (d, J=8.4 Hz, 1H), 7.15 (d, J=8.4 Hz, 1H), 4.85 (m,
3H), 4.29 (dd, J=15.6, 5.6 Hz, 2H), 3.64 (dd, J=8.8, 4.0 Hz, 1H),
3.22 (dd, J=13.6, 4.0 Hz, 1H), 2.78 (dd, J=14.0, 8.8 Hz, 1H). LCMS
(APCI+) m/z 382 [M+H].sup.+; Rt=0.76 min.
Example 89
##STR00105##
[0630] Preparation of
4-Amino-1-[4-(6-amino-5-cyclopropyl-pyrimidin-4-yl)-piperazin-1-yl]-2-(4--
dichlorophenyl)-butan-1-one dihydrochloride
[0631] Step 1: To a stirred solution of
5-cyclopropyl-pyrimidine-4,6-diol (1.35 g, 8.87 mmol) in DCE (35
mL) was added slowly POCl.sub.3 (4.14 mL, 44.4 mmol) followed by
DIEA (1.72 g, 13.3 mmol). The reaction mixture was heated to reflux
for 2 days. After cooling, the solvent was evaporated in vacuo. The
residue was partitioned between 5% NaHCO.sub.3 and EtOAc. The
organic phase was washed with brine, dried, and passed through a
Silica gel pad to give 4,6-dichloro-5-cyclopropyl-pyrimidine (1.30
g, 78%) as a yellow oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
(s, 1H), 1.65 (m, 1H), 1.03 (m, 2H), 1.58 (m, 2H).
[0632] Step 2:
4-(5-Cyclopropyl-6-chloro-pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester was prepared by the procedures described in
Example 40, Step 1, substituting 4-chloro-5-iodopyrimidine with
5-cyclopropyl-4,6-dichloro-pyrimidine. .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.31 (s, 1H), 3.64 (m, 4H), 3.54 (m, 4H), 1.73 (m,
1H), 1.46 (s, 9H), 1.14 (m, 2H), 0.65 (m, 2H). LCMS (APCI+) m/z
339, 341 [M+H].sup.+; Rt=2.38 min.
[0633] Step 3: A round bottom flask was charged with Pd(OAc).sub.2
(84 mg, 0.37 mmol) and rac-BINAP (234 mg, 0.37 mmol) and purged
with N.sub.2. To the flask was added
4-(5-cyclopropyl-6-chloro-pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester (1.27 g, 3.75 mmol), benzophenone imine (815
mg, 4.50 mmol), NaOBu.sup.t (793 mg, 8.25 mmol) and toluene (24
mL). The mixture was heated to 95.degree. C. for 1 hour. After
cooling to room temperature, the reaction was diluted with EtOAc,
filtered through Celite, and concentrated. The crude product was
purified by column chromatography (hexanes:EtOAc, 2:1) to give
4-[6-(Benzhydrylidene-amino)-5-cyclopropyl-pyrimidin-4-yl]-piperazine-1-c-
arboxylic acid tert-butyl ester (1.62 g, 89%) as a colorless oil.
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.38 (s, 1H), 7.41 (m,
10H), 3.40 (m, 4H), 3.32 (m, 4H), 1.46 (s, 9H), 0.93 (m, 1H), 0.86
(m, 2H), 0.59 (m, 2H). LCMS (APCI+) m/z 484 [M+H].sup.+; Rt=3.94
min.
[0634] Step 4: To a stirred solution of
4-[6-(Benzhydrylidene-amino)-5-cyclopropyl-pyrimidin-4-yl]-piperazine-1-c-
arboxylic acid tert-butyl ester (1.60 g, 3.31 mmol) in MeOH (70 mL)
was added hydroxylamine hydrochloride (0.41 g, 6.0 mmol) and NaOAc
(0.65 g, 7.9 mmol). After stirring at room temperature overnight,
the reaction mixture was partitioned between 0.1 N NaOH and DCM.
The organic layer was dried and concentrated. The residue was
purified by column (DCM:MeOH, 20:1) to give
4-(6-Amino-5-cyclopropyl-pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester (1.00 g, 95%) as a colorless syrup. .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.12 (s, 1H), 4.97 (s, 2H), 3.52
(m, 4H), 3.50 (m, 4H), 1.48 (s, 9H), 1.44 (m, 1H), 1.02 (m, 2H),
0.64 (m, 2H). LCMS (APCI+) m/z 320 [M+H].sup.+; Rt=2.51 min.
[0635] Step 5: 5-Cyclopropyl-6-piperazin-1-yl-pyrimidin-4-ylamine
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(6-Amino-5-cyclopropyl-pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.18 (s, 1H), 4.12 (m, 4H), 3.31 (m, 4H), 1.66 (m, 1H), 1.18 (m,
2H), 0.58 (m, 2H). LCMS (APCI+) m/z 220 [M+H].sup.+; Rt=0.74
min.
[0636] Step 6:
4-Amino-1-[4-(6-amino-5-cyclopropyl-pyrimidin-4-yl)-piperazin-1-yl]-2-(4--
chlorophenyl)-butan-1-one dihydrochloride was prepared by
substituting 5-piperazin-1-yl-1H-indazole with
5-Cyclopropyl-6-piperazin-1-yl-pyrimidin-4-ylamine dihydrochloride
and substituting (D)-Boc-4-chlorophenylalanine with
4-tert-Butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid in
Example 34, Step 2, then removing the Boc protecting group as
described in Example 34, Step 3. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.04 (s, 1H), 7.41 (d, J=8.4 Hz, 2H), 7.33 (d, J=8.4 Hz,
2H), 4.19 (m, 1H), 3.95 (m, 1H), 3.85 (m, 2H), 3.61 (m, 3H), 3.22
(m, 2H), 2.95 (m, 1H), 2.82 (m, 1H), 2.31 (m, 1H), 2.01 (m, 1H),
1.56 (m, 1H), 1.08 (m, 2H), 0.47 (m, 2H). LCMS (APCI+) m/z 415, 417
[M+H].sup.+; Rt=1.87 min.
Example 90
##STR00106##
[0637] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-furo[3,2-b]pyridin-7-yl-piperazin-1--
yl)-propan-1-one dihydrochloride
[0638] Step 1: A solution of 3-hydroxy-2-bromopyridine (21.4 g, 123
mmol) in acetic anhydride (25 g, 245 mmol) was refluxed for 1 hour.
After cooling, the mixture was poured into ice water, neutralized
with Na.sub.2CO.sub.3 and extracted with ether. The organic phase
was dried and concentrated. The residue was subject to column
chromatography, eluted by ether to afford Acetic acid
2-bromo-pyridin-3-yl ester (26.2 g, 99%). .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 8.28 (m, 1H), 7.46 (m, 1H), 7.30 (m, 1H), 2.38 (s,
3H).
[0639] Step 2: To a solution of PdCl.sub.2(PPh.sub.3).sub.2 (1.4 g,
2.0 mmol) and CuI (0.4 g, 2.1 mmol) in TEA (100 mL) and THF (200
mL) under nitrogen was added a mixture of
2-bromo-3-acetoxy-pyridine (13.1 g, 60.6 mmol) and TMS-acetylene
(7.0 g, 71 mmol) in THF (100 mL) in one portion. The mixture was
stirred at room temperature for 1 hour. Then quenched with
saturated NaHCO.sub.3 (50 mL) and MeOH (50 mL). The mixture was
stirred at 80.degree. C. for 2 hours. After cooling, the mixture
was extracted with ether. The organic phase was dried and
concentrated. The residue was subject to column chromatography to
afford Furo[3,2-b]pyridine (1.54 g, 21%). .sup.1H NMR (CDCl.sub.3
400 MHz) .delta. 8.55 (m, 1H), 7.84 (m, 1H), 7.75 (m, 1H), 7.72 (m,
1H), 6.99 (m, 1H).
[0640] Step 3: To a solution of Furo[3,2-b]pyridine (1.5 g, 13.0
mmol) in CHCl.sub.3 (30 mL) was added MCPBA (2.9 g, 17.0 mmol). The
mixture was stirred at room temperature for 16 hours. Then the
mixture was filtered through a alumina (140 g, basic) and washed
with ethyl acetate/hexane (1:1) and DCM/MeOH (20:1) to give
Furo[3,2-b]pyridine 4-oxide (1.49 g, 88%). .sup.1H NMR
(CDCl.sub.3,400 MHz) .delta. 8.25 (m, 1H), 7.81 (m, 1H), 7.51 (m,
1H), 7.23 (m, 1H).
[0641] Step 4: To a solution of Furo[3,2-b]pyridine 4-oxide in
CHCl.sub.3 (5 mL) was added POCl.sub.3 (5 g, 33 mmol). The mixture
was refluxed for 2 hours. After cooling, the mixture was quenched
with ice water and neutralized with NaHCO.sub.3. Extracted with
CHCl.sub.3 (3.times.100 mL). The organic phase was dried and
concentrated. The residue was subject to column chromatography to
afford 7-Chloro-furo[3,2-b]pyridine (0.74 g, 50%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.46 (m, 1H), 7.92 (m, 1H), 7.28 (m,
1H), 7.04 (m, 1H).
[0642] Step 5: A mixture of 7-Chloro-furo[3,2-b]pyridine (0.73 g,
4.75 mmol) and piperazine (1.2 g, 14 mmol) in a sealed tube was
heated to 130.degree. C. for 4 hours. After cooling, the solid was
dissolved in MeOH and DCM, concentrated and subject to column
chromatography, eluted by DCM/MeOH (10:1-1:1) to give
7-piperazin-1-yl-furo[3,2-b]pyridine (0.21 g, 22%). MS (APCI+)
[M+H].sup.+ 204.
[0643] Step 6: To a solution of
7-piperazin-1-yl-furo[3,2-b]pyridine (0.21 g, 1.03 mmol) and
(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(0.6 g, 2.0 mmol) in DMF (10 mL) and TEA (2 mL) was added HOBT (0.3
g, 2.2 mmol) and EDCI (0.42 g, 2.2 mmol). The mixture was stirred
at room temperature for 4 hours. The solvent was removed and the
residue was subject to column chromatography to afford
(2R)-[1-(4-Chlorobenzyl)-2-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-2-o-
xo-ethyl]-carbamic acid tert-butyl ester (7 mg, 1.4%). MS (APCI+)
[M+H].sup.+ 486.
[0644] Step 7: To a solution of
(2R)-[1-(4-Chlorobenzyl)-2-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-2-o-
xo-ethyl]-carbamic acid tert-butyl ester (7 mg, 0.014 mmol) in DCM
(4 mL) was added HCl in dioxane (4M, 1 mL). The mixture was stirred
at room temperature for 4 hours. The solvent was removed to give
(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-furo[3,2-b]pyridin-7-yl-piperazin-1--
yl)-propan-1-one as HCl salt (6 mg, 99%). MS (APCI+) [M+H].sup.+
386.
Example 91
##STR00107##
[0645] Preparation of
(7S,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyri-
do[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one
dihydrochloride and
(7R,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyri-
do[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one
dihydrochloride
[0646] Step 1: A solution of 2-Chloro-6-methyl-nicotinic acid (3.00
g, 17.5 mmol) in ammonia in MeOH (7M, 60 mL) in a bomb was heated
to 120.degree. C. overnight. After cooling, the solvent was removed
and the residue was neutralized with 2N HCl. The precipitate was
filtered, washed with water and dried to afford
2-Amino-6-methyl-nicotinic acid (1.44 g, 54%). .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.21 (d, J=7.6 Hz, 1H), 6.60 (d,
J=7.6 Hz, 1H), 2.41 (s, 3H). MS (APCI+) [M+H].sup.+ 153.
[0647] Step 2: A mixture of 2-Amino-6-methyl-nicotinic acid (1.44
g, 9.46 mmol) and formamide (8.0 g, 178 mmol) was stirred at
170.degree. C. for 2 hours. After cooling, the mixture was quenched
with water (4 mL). The precipitate was filtered, washed with water
and dried to afford 7-Methyl-pyrido[2,3-d]pyrimidin-4-ol (0.79 g,
51%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.49 (d, J=8.4 Hz,
1H), 8.22 (s, 1H), 7.35 (d, J=8.0 Hz, 1H), 2.75 (s, 3H). MS (APCI+)
[M+H].sup.+ 162.
[0648] Step 3: To a solution of
7-Methyl-pyrido[2,3-d]pyrimidin-4-ol (0.78 g, 4.84 mmol) in DCE (30
mL) was added DIEA (1.0 mL, 1.19 mmol), followed by POCl.sub.3 (2.4
mL, 26.1 mmol). The mixture was refluxed overnight. After cooling,
the solvent was removed and the residue was dissolved in water (50
mL) and extracted with ethyl acetate (3.times.100 mL). The organic
phase was dried and concentrated. The residue was subject to column
chromatography, eluted by hexane/ethyl acetate (2:1) to give
4-Chloro-7-methyl-pyrido[2,3-d]pyrimidine (0.66 g, 76%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 9.22 (s, 1H), 8.49 (d, J=8.8 Hz,
1H), 7.57 (d, J=8.4 Hz, 1H), 2.88 (s, 3H). MS (APCI+) [M+H].sup.+
180.
[0649] Step 4: The mixture of
4-Chloro-7-methyl-pyrido[2,3-d]pyrimidine (0.66 g, 3.67 mmol) and
1-Boc-piperazine (0.75 g, 4.03 mmol) in DCE (40 mL) and TEA (5 mL)
was refluxed for 1 hour. After cooling, the solvent was removed and
the residue was subject to column chromatography, eluted by ethyl
acetate-DCM/MeOH (10:1) to give
4-(7-Methyl-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester (1.2 g, 99%). .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.82 (s, 1H), 8.11 (d, J=8.4 Hz, 1H), 7.28 (d, J=8.8
Hz, 1H), 3.79 (m, 4H), 3.65 (m, 4H), 2.75 (s, 3H), 1.50 (s, 9H). MS
(APCI+) [M+H].sup.+ 330.
[0650] Step 5: A solution of
4-(7-Methyl-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylic
acid tert-butyl ester (1.2 g, 3.64 mmol), PtO.sub.2 (42 mg, 0.18
mmol) in MeOH (40 mL) and TFA (2 mL) was stirred under H.sub.2 (1
atm) at room temperature for 4 hours. The catalyst was filtered and
the solvent was removed. The residue was subject to column
chromatography, eluted by DCM/MeOH (20:1) to give
4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1--
carboxylic acid tert-butyl ester as TFA salt (0.43 g, 27%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 9.75 (s, 1H), 7.97 (s, 1H), 3.60
(m, 10H), 2.60 (m, 2H), 2.00 (m, 1H), 1.48 (s, 9H), 1.35 (d, J=6.4
Hz, 3H). MS (APCI+) [M+H].sup.+ 334.
[0651] Step 6: To a solution of
4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1--
carboxylic acid tert-butyl ester TFA salt (0.19 g, 0.44 mmol) in
DCM (20 mL) and TEA (1 mL) was added (S)-Mosher's acid chloride
(0.14 g, 0.55 mmol). The mixture was stirred at room temperature
for 20 minutes. The solvent was removed and the residue was subject
to column chromatography, eluted by hexane/ethyl acetate
(4:1-3:1-2:1). The first spot gave
(7S)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-
-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylic
acid tert-butyl ester (84 mg, 34%). .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.27 (s, 1H), 7.28 (m, 2H), 7.12 (m, 3H), 4.74 (m,
1H), 3.90 (s, 3H), 3.40 (m, 4H), 3.08 (m, 4H), 2.32 (m, 1H), 2.20
(m, 1H), 1.94 (m, 1H), 1.49 (s, 9H), 1.27 (m, 1H), 1.11 (d, J=6.4
Hz, 3H). MS (ESI+) [M+H].sup.+ 550 The second spot gave
(7R)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-
-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylic
acid tert-butyl ester (85 mg, 35%). .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.20 (s, 1H), 7.57 (m, 2H), 7.40 (m, 3H), 4.54 (m,
1H), 3.52 (s, 3H), 3.50 (m, 4H), 3.31 (m, 4H), 2.55 (m, 1H), 2.42
(m, 1H), 1.91 (m, 1H), 1.48 (s, 9H), 1.27 (m, 1H), 1.19 (d, J=6.4
Hz, 3H). MS (APCI+) [M+H].sup.+ 550.
[0652] Step 7: To a solution of
(7S)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-
-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylic
acid tert-butyl ester (84 mg, 0.153 mmol) in MeOH (5 mL) was added
LiOH (3M, 4 mL). The mixture was stirred at room temperature for 4
days and then neutralized with 2N HCl. The solvent was removed and
the residue was subject to column chromatography, eluted by ethyl
acetate-DCM/MeOH (20:1) to afford
(7S)-4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl-
)-piperazine-1-carboxylic acid tert-butyl ester (32 mg, 63%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.11 (s, 1H), 5.20 (s,
1H), 3.54 (m, 5H), 3.30 (m, 4H), 2.57 (m, 2H), 1.95 (m, 1H), 1.48
(s, 9H), 1.40 (m, 1H), 1.27 (d, J=6.4 Hz, 3H). MS (APCI+)
[M+H].sup.+ 334.
[0653] Step 8: To a solution of
(7S)-4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazi-
ne-1-carboxylic acid tert-butyl ester (32 mg, 0.096 mmol) in DCM (2
mL) was added HCl in dioxane (4M, 2 mL). The mixture was stirred at
room temperature for 3 hours. The solvent was removed to give
(7S)-7-Methyl-4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine
as HCl salt (22 mg, 99%). MS (APCI+) [M+H].sup.+ 234.
[0654] Step 9: To a solution of
(7S)-7-Methyl-4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine
(7.5 mg, 0.032 mmol) in DCM (4 mL) and DIEA (0.5 mL) was added
D-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (10
mg, 0.033 mmol) and HBTU (13 mg, 0.034 mmol). The mixture was
stirred at room temperature for 1 hour. The solvent was removed and
the residue was subject to column chromatography, eluted by
hexane/ethyl acetate (2:1) to give
(7S,2R)-{1-(4-Chlorobenzyl)-2-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido-
[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid
tert-butyl ester (16 mg, 96%). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.10 (s, 1H), 7.26 (d, J=6.8 Hz, 2H), 7.12 (d, J=8.0 Hz,
2H), 5.45 (d, J=8.4 Hz, 1H), 5.10 (s, 1H), 4.84 (m, 1H), 3.60 (m,
4H), 3.30 (m, 2H), 3.00 (m, 4H), 2.50 (m, 2H), 1.96 (m, 1H), 1.80
(m, 1H), 1.44 (m, 1H), 1.41 (s, 9H), 1.27 (d, J=6.4 Hz, 3H). MS
(APCI+) [M+H].sup.+ 516.
[0655] Step 10: To a solution of
(7S,2R)-{1-(4-Chlorobenzyl)-2-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3--
d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid
tert-butyl ester (16 mg, 0.031 mmol) in DCM (5 mL) was added HCl in
dioxane (4M, 2 mL). The mixture was stirred at room temperature for
4 hours. The solvent was removed to give
(7S,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyri-
do[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one
dihydrochloride (13 mg, 99%). MS (APCI+) [M+H].sup.+ 416.
(7R,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyri-
do[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one
dihydrochloride was prepared in a similar manner from
(7R)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-
-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylic
acid tert-butyl ester.
Example 92
##STR00108##
[0656] Preparation of
3-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one dihydrochloride
[0657] Step 1: To a solution of
7-benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine
dihydrochloride (35 mg, 0.081 mmol),
3-Boc-amino-2-(4-chlorophenyl)-propionic acid (27 mg, 0.089 mmol),
and TEA (0.11 mL, 0.81 mmol) 1.2 mL DCM was added HBTU (34 mg,
0.089 mmol). The reaction mixture was stirred at room temperature 2
hours, after which 0.15 mL 3M LiOH and 1.0 mL MeOH were added. The
reaction mixture was stirred at 35.degree. C. for 3.5 hours, after
which saturated NaHCO.sub.3 was added. The mixture was extracted
with DCM, and the combined extracts were dried (Na.sub.2SO.sub.4),
filtered, and concentrated. The crude was purified on silica gel
(flushed with 1:1 DCM:EtOAc, then gradient to 1:4 DCM:EtOAc) to
give
3-Boc-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin--
4-yl)-piperazin-1-yl]-propan-1-one, which was used in the next
step.
[0658] Step 2: To a solution of
3-Boc-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin--
4-yl)-piperazin-1-yl]-propan-1-one in 1 mL dioxane was added 1.5 mL
4M HCl/dioxane. The resulting suspension was stirred at room
temperature 7 hours, after which it was concentrated to dryness.
The solids were then dissolved in minimal MeOH, and the product was
triturated by the addition of ether. The solids were isolated by
filtration through a fritted funnel with nitrogen pressure, rinsed
with ether, and dried in vacuo to afford
3-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl-
)-piperazin-1-yl]-propan-1-one dihydrochloride (21 mg, 55%) as a
beige powder. .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 8.08 (s, 1H),
7.29 (d, J=8.4 Hz, 2H), 7.17 (d, J=8.4 Hz, 2H), 7.07 (s, 1H), 4.30
(t, J=6.5 Hz, 1H), 4.03-3.94 (m, 1H), 3.82-3.73 (m, 1H), 3.68-3.46
(m, 5H), 3.28 (dd, J=12.9, 7.4 Hz, 1H), 3.23 (dd, J=12.9, 5.7 Hz,
1H), 3.04-2.94 (m, 1H), 2.13 (s, 3H). LCMS (APCI+) m/z 399
[M+H].sup.+; Rt: 2.11 min.
Example 93
##STR00109##
[0659] The Preparation of
2-(4-Chlorophenyl)-3-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one bis-hydrochloride salt
[0660] Step 1: The
7-benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine
bis-hydrochloride (200 mg, 0.465 mmol) and
3-tert-butoxycarbonylamino-2-(4-chlorophenyl)-propionic acid (146
mg, 0.488 mmol) were dissolved/suspended in 2.0 mL of DMF at room
temperature and treated with TEA (259 .mu.L, 1.86 mmol). The HBTU
(194 mg, 0.511 mmol) was added in one sum, and the reaction was
allowed to stir overnight at room temperature to completion. The
reaction was partitioned between ethyl acetate and diluted
NaHCO.sub.3 solution. The aqueous was extracted with ethyl acetate,
and the organics were combined. The organic was washed with water,
then brine, separated, dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by chromatography
(silica gel eluted with 70:30 ethyl acetate:hexanes) to afford the
pure
[3-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pipera-
zin-1-yl]-2-(4-chlorophenyl)-3-oxo-propyl]-carbamic acid tert-butyl
ester as colorless gel (295 mg, 99%). .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.44 (s, 1H), 8.16 (d, J=8.4 Hz, 2H), 7.59 (t, J=7.2
Hz, 1H), 7.49 (appt, J=7.6 Hz, 2H), 7.31 (m, 3H), 7.20 (d, J=8.0
Hz, 2H), 5.12 (m, 1H), 4.09 (m, 1H), 3.92 (m, 1H), 3.60 (m, 1H),
3.56-3.46 (m, 4H), 3.38 (d, J=10.4 Hz, 2H), 3.32 (m, 1H), 2.90 (m,
1H), 2.29 (s, 3H), 1.41 (s, 9H). LCMS (APCI+) m/z 639 [M+H].sup.+;
Rt=3.70 min.
[0661] Step 2: The
[3-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pipera-
zin-1-yl]-2-(4-chlorophenyl)-3-oxo-propyl]-carbamic acid tert-butyl
ester (295 mg, 0.464 mmol) was dissolved in 2.3 mL of 1,4-dioxane
and treated with 4M HCl in 1,4-dioxane (2.3 mL, 9.29 mmol). The
mixture was allowed to stir for 4 hours to completion affording a
yellow precipitate. The suspension was diluted with diethyl ether
and poured into water. More diethyl ether was added, and the layers
were shaken. The ether wash was discarded, and the aqueous was
treated with saturated NaHCO.sub.3 solution until basic to pH paper
(about 10) to afford a white precipitate. The aqueous was extracted
with ethyl acetate, and the organics were combined. The organic was
washed with water, then brine, separated, dried over MgSO4,
filtered, and concentrated in vacuo to afford the near-pure
3-amino-1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-
-piperazin-1-yl]-2-(4-chlorophenyl)-propan-1-one as a colorless oil
(194 mg, 77%). .sup.1H NMR (free-base, CDCl.sub.3, 400 MHz) .delta.
8.44 (s, 1H), 8.16 (d, J=8.4 Hz, 2H), 7.59 (t, J=7.2 Hz, 1H), 7.49
(appt, J=7.6 Hz, 2H), 7.31 (m, 3H), 7.19 (d, J=8.0 Hz, 2H), 3.92
(m, 1H), 3.85 (dd, J=8.4, 5.2 Hz, 1H), 3.61 (m, 1H), 3.56-3.50 (m,
4H), 3.43 (m, 2H), 3.32 (m, 2H), 2.90 (m, 2H), 2.29 (s, 3H). LCMS
(APCI+) m/z 539 [M+H].sup.+; Rt=2.40 min.
[0662] Step 3: The
3-amino-1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-
-piperazin-1-yl]-2-(4-chlorophenyl)-propan-1-one (75 mg, 0.139
mmol) was dissolved in 1.0 mL of 1,2-dichloroethane and treated
with 37% aqueous formaldehyde (31 .mu.L, 0.417 mmol). The solution
was allowed to stir at room temperature for 15 minutes before the
sodium triacetoxyborohydride (118 mg, 0.557 mmol) was added in one
sum. The reaction was complete in one hour, and the contents were
poured into diluted NaHCO.sub.3 solution. The aqueous was extracted
with ethyl acetate, and the organics were combined. The organic was
washed with water, then brine, separated, dried over MgSO.sub.4,
filtered, and concentrated in vacuo. The residue was purified by
chromatography (silica gel eluted with 1% TEA in 9:1 ethyl
acetate:methanol) to afford the pure
1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperaz-
in-1-yl]-2-(4-chlorophenyl)-3-dimethylamino-propan-1-one
intermediate as a colorless oil. The oil was dissolved in 0.5 mL of
each: THF, methanol, and water. The solution was treated with
lithium hydroxide-monohydrate (29 mg, 0.696 mmol) to afford an
opaque solution, which stirred overnight to completion. The
solution was partitioned between ethyl acetate and water, and the
aqueous was extracted with more ethyl acetate. The combined organic
was washed with water, then brine, separated, dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo to afford the
crude product as a yellow oil. The residue was purified by
chromatography (silica gel eluted with 1% TEA in 4:1 ethyl
acetate:methanol) to afford the titled free base as a colorless
oil. The material was dissolved in a minimal amount of THF (>1
mL) and treated with 2.0M HCl in ether. The resulting white
precipitate was filtered, washed with diethyl ether, and dried
under reduced pressure to afford the
2-(4-chlorophenyl)-3-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one bis-hydrochloride salt as a
white solid (24 mg, 35%). .sup.1H NMR (Free-Base, CDCl.sub.3, 400
MHz) .delta. 10.72 (brs, 1H), 8.34 (s, 1H), 7.30 (d, J=8.4 Hz, 2H),
7.24 (d, J=8.4 Hz, 2H), 6.93 (s, 1H), 4.11 (m, 1H), 3.95 (m, 1H),
3.65 (m, 2H), 3.55 (m, 3H), 3.33 (m, 2H), 3.05 (m, 1H), 2.48 (dd,
J=12.4, 4.8 Hz, 1H), 2.36 (s, 3H), 2.32 (s, 6H). LCMS (APCI+) m/z
427 [M+H].sup.+; Rt=1.97 min.
Example 94
##STR00110##
[0663] Preparation of
2-(4-Chlorophenyl)-4-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-butan-1-one bis-hydrochloride salt
[0664] The
2-(4-chlorophenyl)-4-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,-
3-d]pyrimidin-4-yl)-piperazin-1-yl]-butan-1-one bis-hydrochloride
salt (21 mg, 30%) was prepared by procedures described in Example
93, Steps 1-3 [substituting
3-tert-butoxycarbonylamino-2-(4-chlorophenyl)-propionic acid with
4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid in Step
1]. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.29 (s, 1H), 7.41
(d, J=8.4 Hz, 2H), 7.36 (d, J=8.4 Hz, 2H), 7.25 (s, 1H), 4.25 (t,
J=6.8 Hz, 1H), 3.97 (appd, J=8.8 Hz, 2H), 3.85-3.72 (m, 4H), 3.63
(m, 1H), 3.32 (m, 1H), 3.17 (m, 1H), 3.07 (m, 1H), 2.89 (s, 3H),
2.88 (s, 3H), 2.41 (m, 1H), 2.39 (s, 3H), 2.10 (m, 1H). LCMS
(APCI+) m/z 441 [M+H].sup.+; Rt=1.87 min.
Example 95
##STR00111##
[0665] Preparation of
N-(2-Amino-ethyl)-N-(4-chlorobenzyl)-4-pyridin-4-yl-benzamide
bis-hydrochloride salt
[0666] Step 1: The (2-amino-ethyl)-carbamic acid tert-butyl ester
(5.00 g, 31.2 mmol) and 4-chloro-benzaldehyde (4.61 g, 32.77 mmol)
were dissolved in 60 mL of 1,2-dichloroethane at room temperature.
The reaction mixture was allowed to stir for 40 minutes prior to
treatment with sodium triacetoxyborohydride (9.90 g, 46.8 mmol).
The mixture was allowed to stir overnight to completion and
quenched with a saturated NaHCO.sub.3 solution. The aqueous was
extracted with DCM, separated, dried over MgSO.sub.4, and
concentrated in vacuo. The residue was purified by chromatography
(silica gel eluted with hexanes/EtOAc plus 2% triethyl amine) to
afford the pure [2-(4-chlorobenzylamino)-ethyl]-carbamic acid
tert-butyl ester as a viscous yellow oil (5.38 g, 61%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.29 (d, J=8.0 Hz, 2H), 7.25 (d,
J=8.0 Hz, 2H), 4.92 (brs, 1H), 3.75 (s, 2H), 3.23 (appd, J=5.6 Hz,
2H), 2.73 (appt, J=6.0 Hz, 2H), 1.44 (s, 9H), 1.36 (brs, 1H). LCMS
(APCI+) m/z 285 [M+H].sup.+; Rt=2.30 min.
[0667] Step 2: The 4-bromo-benzoic acid ethyl ester (1.00 g, 4.37
mmol), pyridine-4-boronic acid (537 mg, 4.37 mmol), and
tetrakis(triphenylphosphine)palladium(0) (757 mg, 0.655 mmol) were
degassed under nitrogen. The solids were dissolved in 15 mL of
1,4-dioxane and 2.6 mL of 2M sodium carbonate solution. The mixture
was heated to 80.degree. C. overnight completion and allowed to
cool to room temperature. The contents were partitioned between
ethyl acetate and water, and the aqueous was extracted with ethyl
acetate. The combined organic was washed with water, brine, dried
over MgSO.sub.4, filtered, and concentrated in vacuo. The residue
was purified by chromatography (silica gel eluted with 1:1
hexanes:EtOAc, Rf=0.2) to afford the pure 4-pyridin-4-yl-benzoic
acid ethyl ester as a pale yellow solid (350 mg, 35%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.71 (d, J=4.4 Hz, 2H), 8.16 (d,
J=8.0 Hz, 2H), 7.71 (d, J=8.0 Hz, 2H), 7.53 (d, J=4.4 Hz, 2H), 4.42
(q, J=7.2 Hz, 2H), 1.43 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 228
[M+H].sup.+; Rt=2.81 min.
[0668] Step 3: The 4-pyridin-4-yl-benzoic acid ethyl ester (430 mg,
1.89 mmol) was heated to 100.degree. C. in 6.5 mL of 3M HCl
solution overnight to completion. The mixture was cooled to room
temperature and diluted with water (dissolves precipitate). The
solution was filtered and concentrated in vacuo to afford the
4-pyridin-4-yl-benzoic acid hydrochloride salt as a pale yellow
solid (380 mg, 85%). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
8.93 (d, J=5.6 Hz, 2H), 8.27 (d, J=5.6 Hz, 2H), 8.13 (d, J=8.0 Hz,
2H), 8.09 (d, J=8.0 Hz, 2H), --CO.sub.2H or --NH.sup.+ not
observed. LCMS (APCI+) m/z 200 [M+H].sup.+; Rt=1.05 min.
[0669] Step 4: The 4-pyridin-4-yl-benzoic acid hydrochloride salt
(100 mg, 0.502 mmol), [2-(4-chlorobenzylamino)-ethyl]-carbamic acid
tert-butyl ester (143 mg, 0.502 mmol), HOBt (77 mg, 0.502 mmol),
and EDCI (56 mg, 0.552 mmol) were dissolved in 1.7 mL DMF, then
treated with triethyl amine (77 .mu.L, 0.552 mmol) to room
temperature. The reaction was allowed to stir overnight to
completion and partitioned between ethyl acetate and diluted
NaHCO.sub.3 solution. The aqueous was extracted with ethyl acetate,
and the organics were combined. The organic was washed with water,
brine, dried over MgSO.sub.4, filtered, and concentrated in vacuo.
The residue was purified by chromatography (silica gel eluted with
hexanes/ethyl acetate gradients plus 2% triethylamine) to afford
the
{2-[(4-chlorobenzyl)-(4-pyridin-4-yl-benzoyl)-amino]-ethyl}-carbamic
acid tert-butyl ester as a colorless oil (184 mg, 93%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.68 (d, J=4.8 Hz, 2H), 7.65 (m, 2H),
7.54 (d, J=8.4 Hz, 2H), 7.48 (d, J=4.4 Hz, 2H), 7.34 (m, 2H), 7.13
(m, 2H), 4.91 (m, 1H), 4.59 (brs, 2H), 3.62 (m, 2H), 3.44 (m, 2H),
1.46 (s, 9H). LCMS (APCI+) m/z 466 [M+H].sup.+; Rt=3.19 min.
[0670] Step 5: The
{2-[(4-chlorobenzyl)-(4-pyridin-4-yl-benzoyl)-amino]-ethyl}-carbamic
acid tert-butyl ester (184 mg, 0.395 mmol) was dissolved in 2.0 mL
of 1,4-dioxane and treated with 2.0 mL of 4M HCl in 1,4-dioxane.
The reaction was allowed to stir for one hour to completion and
diluted with diethyl ether. The product was isolated by vacuum
filtration to afford the
N-(2-amino-ethyl)-N-(4-chlorobenzyl)-4-pyridin-4-yl-benzamide
bis-hydrochloride salt as a fine pale-yellow powder (166 mg, 96%).
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 8.91 (d, J=6.0 Hz, 2H),
8.26 (d, J=6.0 Hz, 2H), 8.13 (brs, 3H), 8.05 (d, J=8.0 Hz, 2H),
7.76 (d, J=7.6 Hz, 2H), 7.45 (brs, 1H), 7.44 (d, J=8.0 Hz, 2H),
7.26 (d, J=7.6 Hz, 2H), 4.57 (brs, 2H), 3.61 (m, 2H), 3.08 (m, 2H).
LCMS (APCI+) m/z 366 [M+H].sup.+; Rt=2.13 min.
Example 96
##STR00112##
[0671] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyr-
imidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0672] Step 1: A mixture of 2-aminonicotinic acid (7.00 g, 50.7
mmol) and formamide (22.8 g, 506 mmol) was heated to 167.degree. C.
for 2.5 hours. After cooling, the solid was recrystallized from hot
water (100 mL) to afford the pure product (4.80 g, 64%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.94 (m, 1H), 8.64 (d, 1H, J=7.6
Hz), 8.30 (s, 1H), 7.59 (m, 1H). MS (APCI+) [M+H].sup.+ 148.
[0673] Step 2: The solution of 4-hydroxypyrido[2,3-d]pyrimidine
(2.00 g, 13.6 mmol) in POCl.sub.3 (40 mL) was refluxed for 2 hours.
After cooling, the excess POCl.sub.3 was removed under vacuum. The
residue was quenched with saturated NaHCO.sub.3 and extracted with
ethyl acetate (3.times.100 mL). The organic phase was dried and
concentrated. The residue was subject to column chromatography,
eluted by hexane/ethyl acetate (1:1) to give
4-chloropyrido[2,3-d]pyrimidine (0.72 g, 32%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 9.35 (m, 1H), 9.30 (s, 1H), 8.66 (m,
1H), 7.73 (m, 1H). MS (APCI+) [M+H].sup.+ 166.
[0674] Step 3: To a solution of 4-chloropyrido[2,3-d]pyrimidine
(0.72 g, 4.4 mmol) and 1-Boc piperazine (0.84 g, 4.5 mmol) in DCE
(10 mL) and IPA (10 mL) was added triethylamine (4 mL). The mixture
was refluxed for 4 hours. After cooling, the solvent was removed.
The residue was subject to column chromatography by ethyl acetate
to give 4-pyrido[2,3-d]pyrimidin-4-yl-piperazine-1-carboxylic acid
tert-butyl ester (1.26 g, 92%). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 9.07 (dd, J=4.30 Hz, J=1.98 Hz, 1H), 8.88 (s, 1H), 8.25
(dd, J=7.99 Hz, J=1.98 Hz, 1H), 7.41 (dd, J=8.18 Hz, J=3.74 Hz,
1H), 3.82 (m, 4H), 3.65 (m, 4H), 1.50 (s, 9H). MS (APCI+)
[M+H].sup.+ 316.
[0675] Step 4: To a solution of
4-pyrido[2,3-d]pyrimidin-4-yl-piperazine-1-carboxylic acid
tert-butyl ester (0.24 g, 0.76 mmol) in MeOH (10 mL) and TFA (1 mL)
was added PtO.sub.2 (10 mg). The mixture was stirred under H.sub.2
(1 atm) at room temperature overnight. The catalyst was filtered
off and the solvent was removed. The residue was subject to column
chromatography, eluted by DCM/MeOH (20:1) to afford
4-(5,6,7,8-Tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxyli-
c acid tert-butyl ester (0.15 g, 62%). .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 8.10 (s, 1H), 5.92 (s, 1H), 3.50 (m, 4H), 3.40 (m,
2H), 3.25 (m, 4H), 2.55 (m, 2H), 1.85 (m, 2H), 1.48 (s, 9H). MS
(APCI+) [M+H].sup.+ 320.
[0676] Step 5: To a solution of
4-(5,6,7,8-Tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxyli-
c acid tert-butyl ester (0.88 g, 2.75 mmol) in DCM (10 mL) was
added HCl in dioxane (4M, 5 mL). The mixture was stirred at RT for
4 hours. The solvent was removed to give
4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine as HCl
salt (0.80 g, 99%). MS (APCI+) [M+H].sup.+ 220.
[0677] Step 6: To a solution of
4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine
dihydrochloride (20 mg, 0.068 mmol) and
(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(21 mg, 0.070 mmol) in DCM (5 mL) and TEA (1 mL) was added HBTU (30
mg, 0.079 mmol). The mixture was stirred at room temperature for 4
hours. The solvent was removed and the residue was subject to
column chromatography, eluted with ethyl acetate/DCM/MeOH (20:1) to
give
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyri-
midin-4-yl)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester
(18 mg, 53%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.10 (s,
1H), 7.25 (m, 3H), 7.12 (m, 2H), 5.42 (d, J=8.8 Hz, 1H), 5.14 (s,
1H), 4.84 (m, 1H), 3.67 (m, 3H), 3.50 (m, 1H), 3.40 (m, 2H), 3.23
(m, 5H), 2.95 (m, 4H), 2.50 (m, 2H), 1.86 (m, 2H), 1.67 (s, 1H),
1.42 (s, 9H). MS (APCI+) [M+H].sup.+ 502.
[0678] Step 7: To a solution of
(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyri-
midin-4-yl)-piperazin-1-yl]-ethyl}-carbamic acid tert-butyl ester
(18 mg, 0.036 mmol) in DCM (4 mL) was added HCl in dioxane (4M, 2
mL). The mixture was stirred at room temperature for 4 hours. The
solvent was removed to give
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyr-
imidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride (14 mg,
99%). MS (APCI+) [M+H].sup.+ 401.
Example 97
##STR00113##
[0679] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0680] Step 1: To a solution of formamide HCl salt (21.6 g, 268
mmol) in MeOH (300 mL) was added NaOMe (25%, in MeOH, 120 mL, 555
mmol). The mixture was stirred at room temperature for 1 hour. Then
triethyl 1,1,2-ethanetricarboxylate (64.4 g, 60 mL, 262 mmol) in
MeOH (90 mL) was added slowly. After addition, the mixture was
stirred at room temperature for 20 hours. The solvent was removed
and the residue was dissolved in ice water (200 mL) and neutralized
with 2N HCl (140 mL) until pH=1-2. The solid formed was filtered,
washed with water (50 mL) and dried under vacuum to afford
(4,6-Dihydroxy-pyrimidin-5-yl)-acetic acid methyl ester (45 g,
93%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.03 (s, 1H), 3.67
(s, 3H), 3.42 (s, 2H). MS (APCI+) [M+H].sup.+ 185.
[0681] Step 2: To a solution of
(4,6-Dihydroxy-pyrimidin-5-yl)-acetic acid methyl ester (45 g, 244
mmol) in DCE (800 mL) was added DIEA (72 mL, 413 mmol), followed by
POCl.sub.3 (80 mL, 874 mmol) slowly. After addition, the mixture
was stirred at room temperature for 2 hours and then refluxed
overnight. After cooling, the solvent was removed and the residue
was dissolved in ice water (400 mL), neutralized with 10N NaOH
until pH 6. Extracted with ethyl acetate (3.times.500 mL). The
organic phase was dried and concentrated. The residue was subject
to column chromatography, eluted by hexane/ethyl acetate (4:1) to
afford (4,6-Dichloro-pyrimidin-5-yl)-acetic acid methyl ester (38
g, 70%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.73 (s, 1H),
4.00 (s, 2H), 3.77 (s, 3H). MS (APCI+) [M+H].sup.+ 222.
[0682] Step 3: To a solution of
(4,6-Dichloro-pyrimidin-5-yl)-acetic acid methyl ester (0.52 g,
2.35 mmol) in ether (40 mL) at -78.degree. C. was added DIBAL-H
(1.5M, 4 mL, 6.05 mmol) dropwise. After addition, the mixture was
allowed to warm up to room temperature and stirred for 3 hours.
Then quenched with 2N HCl (10 mL) at -78.degree. C. Extracted with
ethyl acetate (3.times.50 mL). The organic phase was dried and
concentrated to give fairly pure
2-(4,6-Dichloro-pyrimidin-5-yl)-ethanol (0.44 g, 97%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.66 (s, 1H), 3.96 (t, J=6.8 Hz, 2H),
3.23 (t, J=6.8 Hz, 2H). 1.65 (s, 1H). MS (APCI+) [M+H].sup.+
194.
[0683] Step 4: To a solution of
2-(4,6-Dichloro-pyrimidin-5-yl)-ethanol (0.46 g, 2.4 mmol) in DCM
(40 mL) and was added MsCl (0.50 g, 4.36 mmol), triethylamine (1
mL) and cat. amount of DMAP. After stirring at room temperature
overnight, the solvent was removed and the residue was subject to
column chromatography, eluted by hexane/ethyl acetate (4:1) to
afford methanesulfonic acid 2-(4,6-dichloro-pyrimidin-5-yl)-ethyl
ester (0.52 g, 80%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.71
(s, 1H), 4.50 (t, J=6.4 Hz, 2H), 3.42 (t, J=6.8 Hz, 2H), 3.02 (s,
3H).
[0684] Step 5: A solution of methanesulfonic acid
2-(4,6-dichloro-pyrimidin-5-yl)-ethyl ester (0.50 g, 1.84 mmol) and
4-methoxybenzylamine (0.60 g, 4.37 mmol) in DCE (30 mL) and TEA (4
mL) was refluxed overnight. After cooling, the solvent was removed
and the residue was subject to column chromatography, eluted by
hexane/ethyl acetate (4:1) to give
4-Chloro-7-(4-methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
(0.36 g, 71%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.35 (s,
1H), 7.26 (dd, J=2.8 Hz, J=1.6 Hz, 2H), 6.88 (d, J=4.4 Hz, 2H),
5.33 (s, 1H), 4.62 (d, J=5.6 Hz, 2H), 3.81 (s, 3H), 3.73 (t, J=7.2
Hz, 2H), 3.04 (t, J=6.8 Hz, 2H). MS (APCI+) [M+H].sup.+ 276.
[0685] Step 6: A solution of
4-Chloro-7-(4-methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
(0.36 g, 1.31 mmol), 1-Boc-piperazine (1.0 g, 5.37 mmol) and tBuOK
(0.18 g, 1.60 mmol) in NMP (20 mL) was heated to 128.degree. C. for
20 hours. After cooling, the mixture was diluted with ethyl acetate
(500 mL) and washed with water (5.times.150 mL). The organic phase
was dried and concentrated. The residue was subject to column
chromatography, eluted by hexane/ethyl acetate (1:1) to give
4-[7-(4-Methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl]-pipe-
razine-1-carboxylic acid tert-butyl ester (0.32 g, 57%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.17 (s, 1H), 7.20 (d, J=8.4 Hz,
2H), 6.84 (d, J=8.4 Hz, 2H), 4.50 (s, 2H), 3.79 (s, 3H), 3.59 (m,
4H), 3.48 (m, 4H), 3.35 (m, 2H), 3.30 (m, 2H), 1.49 (s, 9H). MS
(APCI+) [M+H].sup.+ 426.
[0686] Step 7: A solution of
4-[7-(4-Methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl]-pipe-
razine-1-carboxylic acid tert-butyl ester (0.32 g, 0.74 mmol) in
TFA (20 mL) was stirred at 65.degree. C. for 20 hours. After
cooling, the TFA was evaporated under vacuum to afford
4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
bis-trifluoroacetate (0.15 g, 99%). MS (APCI+) [M+H].sup.+ 206.
[0687] Step 8: To a solution of
4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
bis-trifluoroacetate (20 mg, 0.097 mmol) in DCM (10 mL) and TEA (2
mL) were added
(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(30 mg, 0.10 mmol) and HBTU (30 mg, 0.079 mmol). The mixture was
stirred at room temperature for 2 hours. The solvent was removed
and the residue was subject to column chromatography to afford
(2R)-{1-(4-Chlorobenzyl)-2-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-y-
l)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (19
mg, 40%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.03 (s, 1H),
7.25 (m, 2H), 7.12 (m, 2H), 5.40 (m, 1H), 4.83 (m, 2H), 3.50 (m,
10H), 3.20 (m, 5H), 2.96 (m, 2H), 2.28 (m, 3H), 1.42 (s, 9H). MS
(APCI+) [M+H].sup.+ 488.
[0688] Step 9: To a solution of
(2R)-{1-(4-Chlorobenzyl)-2-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-y-
l)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl ester (19
mg, 0.039 mmol) in DCM (4 mL) was added HCl in dioxane (4M, 2 mL).
The mixture was stirred at room temperature for 6 hours. The
solvent was removed to afford
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride (15 mg,
99%). MS (APCI+) [M+H].sup.+ 388.
Example 98
##STR00114##
[0689] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-
-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0690] Step 1: To a solution of KH (30%, 13.3 g, 125 mmol) in THF
(200 mL) at 0.degree. C. was added
(4,6-Dichloro-pyrimidin-5-yl)-acetic acid methyl ester (20.0 g,
90.0 mmol) and MeI (290.0 g, 8.8 mL, 141.0 mmol) in THF (200 mL)
slowly. After addition, the mixture was stirred at room temperature
for 30 min, and then heated to reflux for 1 hour. After cooling to
0.degree. C., the mixture was quenched with saturated aqueous
NH.sub.4Cl. The organic phase was separated and the aqueous phase
was extracted with ethyl acetate (3.times.200 mL). The organic
phase was combined and dried. After removal of the solvent, the
residue was subject to column chromatography, eluted by
hexane/ethyl acetate (5:1) to give
2-(4,6-Dichloro-pyrimidin-5-yl)-propionic acid methyl ester (17.6
g, 83%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.69 (s, 1H),
4.39 (dd, J=14.4 Hz, J=7.2 Hz, 1H), 3.73 (s, 3H), 1.57 (d, J=7.2
Hz, 3H).
[0691] Step 2: To a solution of
2-(4,6-Dichloro-pyrimidin-5-yl)-propionic acid methyl ester (0.5 g,
2.13 mmol) in ether (40 mL) at -78.degree. C. was added DIBAL-H
(1.5M, 4 mL, 6.0 mmol) dropwise. The mixture was allowed to warm up
to room temperature and stirred for 3 hours. Then quenched with 2N
HCl (10 mL) at -78.degree. C. The aqueous phase was extracted with
ethyl acetate (3.times.50 mL). The organic phase was dried and
concentrated. The residue was subject to column chromatography,
eluted by hexane/ethyl acetate (4:1) to give
2-(4,6-Dichloro-pyrimidin-5-yl)-propan-1-ol (0.40 g, 91%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.62 (s, 1H), 4.16 (m, 1H), 3.90
(m, 2H), 1.93 (s, 1H), 1.40 (d, J=7.2 Hz, 3H).
[0692] Step 3: To a solution of
2-(4,6-Dichloro-pyrimidin-5-yl)-propan-1-ol (0.40 g, 1.93 mmol) in
DCM (40 mL) were added MsCl (0.50 g, 4.36 mmol), TEA (1 mL) and
catalytic amount of DMAP. The mixture was stirred at room
temperature for 2 hours. The solvent was removed and the residue
was subject to column chromatography, eluted by hexane/ethyl
acetate (4:1) to give methanesulfonic acid
2-(4,6-dichloro-pyrimidin-5-yl)-propyl ester (0.54 g, 98%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.68 (s, 1H), 4.74 (m, 1H), 4.55
(m, 1H), 4.14 (m, 1H), 2.99 (s, 3H), 1.49 (dd, J=7.2 Hz, J=1.2 Hz,
3H).
[0693] Step 4: To a solution of methanesulfonic acid
2-(4,6-dichloro-pyrimidin-5-yl)-propyl ester (0.54 g, 1.89 mmol) in
DCM (30 mL) and TEA (4 mL) was added 4-methoxybenzyl amine (0.80 g,
5.83 mmol). The mixture was refluxed overnight. After cooling, the
solvent was removed and the residue was subject to column
chromatography, eluted by hexane/ethyl acetate (4:1) to give
4-Chloro-7-(4-methoxy-benzyl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrim-
idine (0.55 g, 99%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.24
(s, 1H), 7.17 (d, J=7.6 Hz, 2H), 6.86 (d, J=7.6 Hz, 2H), 4.55 (s,
2H), 3.80 (s, 3H), 3.66 (m, 1H), 3.37 (m, 1H), 3.07 (m, 1H), 1.31
(dd, J=7.2 Hz, J=1.2 Hz, 3H). MS (APCI+) [M+H].sup.+ 291.
[0694] Step 5: To a solution of
4-Chloro-7-(4-methoxy-benzyl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrim-
idine (0.55 g, 1.88 mmol) in NMP (20 mL) were added
1-Boc-piperazine (1.0 g, 5.40 mmol) and .sup.tBuOK (0.21 g, 1.88
mmol). The mixture was heated to 128.degree. C. for 30 hours. After
cooling, the mixture was diluted by ethyl acetate (500 mL) and
washed with water (5.times.150 mL). The organic phase was dried and
concentrated. The residue was subject to column chromatography,
eluted by hexane/ethyl acetate (1:1) to give
4-(5-Methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-car-
boxylic acid tert-butyl ester (0.30 g, 36%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.19 (s, 1H), 7.18 (d, J=8.4 Hz, 2H),
6.84 (d, J=8.4 Hz, 2H), 4.52 (dd, J=27.6 Hz, J=14.8 Hz, 2H), 3.79
(s, 3H), 3.58 (m, 12H), 3.34 (m 1H), 2.95 (m, 1H), 1.48 (s, 9H),
1.15 (d, J=6.8 Hz, 3H). MS (APCI+) [M+H].sup.+ 440.
[0695] Step 6: A solution of
4-(5-Methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-car-
boxylic acid tert-butyl ester (0.30 g, 0.68 mmol) in TFA (20 mL)
was heated to 65.degree. C. overnight. After cooling, the excess
TFA was evaporated under vacuum to give
5-Methyl-4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
as TFA salt (0.15 g, 99%). MS (APCI+) [M+H].sup.+ 220.
[0696] Step 7: To a solution of
5-Methyl-4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine
(18 mg, 0.082 mmol) in DCM (10 mL) and TEA (2 mL) were added
(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid
(25 mg, 0.082 mmol) and HBTU (31 mg, 0.082 mmol). The mixture was
stirred at room temperature for 2 hours. The solvent was removed
and the residue was subject to column chromatography, eluted by
ethyl acetate-DCM/MeOH (30:1) to give
(2R)-{1-(4-Chlorobenzyl)-2-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,-
3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid
tert-butyl ester (32 mg, 78%). .sup.1H NMR (CDCl.sub.3, 400 MHz)
.delta. 8.06 (s, 1H), 7.26 (d, J=6.0 Hz, 2 hours), 7.14 (d, J=8.0
Hz, 2H), 5.39 (d, J=8.0 Hz, 1H), 4.82 (m, 2H), 3.45 (m, 10H), 3.18
(m, 4H), 3.00 (m, 2H), 1.83 (m, 4H), 1.42 (s, 9H), 1.19 (m, 3H). MS
(APCI+) [M+H].sup.+ 502.
[0697] Step 8: To a solution of
(2R)-{1-(4-Chlorobenzyl)-2-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyri-
midin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamic acid tert-butyl
ester (32 mg, 0.064 mmol) in DCM/MeOH (5:1, 6 mL) was added HCl in
dioxane (4M, 2 mL). The mixture was stirred at room temperature for
6 hours. The solvent was removed to afford
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-
-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride (26
mg, 99%). MS (APCI+) [M+H].sup.+ 402.
Example 99
##STR00115##
[0698] Preparation of
(2R)--N-{4-[4-(2-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-y-
l}-methanesulfonamide dihydrochloride
[0699] Step 1: To a solution of 4-chloro-6-nitroquinazoline
(prepared according to the literature: Alexander J. Bridges et al.
J. Med. Chem. 1996, 39, 267-276, and references therein; 12 g, 57.5
mmol) and DIEA (10 mL, 57.5 mmol) in 230 mL IPA was added
Boc-piperazine (10.7 g, 57.5 mmol). The reaction mixture was heated
to 60.degree. C. and stirred for 13 hours, after which it was
cooled to room temperature and concentrated by rotary evaporation.
The residue was dissolved in dichloromethane (DCM) and washed with
1N NaOH. The organic layer was dried (Na.sub.2SO.sub.4), filtered,
and concentrated by rotary evaporation. The resulting oil was
purified on silica gel (1:1 to 1:4 DCM:EtOAc gradient) to furnish
4-(6-nitro-quinazolin-4-yl)-1-Boc-piperazine as a pale yellow oil
(18.3 g, 89%). LCMS (APCI+) m/z 360 [M+H].sup.+. HPLC Rt 3.06
min.
[0700] Step 2: To a suspension of Pd/C (5% w/w, 800 mg, 0.38 mmol)
in 100 mL 2-methoxyethanol (degassed with nitrogen prior to use)
was added a solution of
4-(6-nitro-quinazolin-4-yl)-1-Boc-piperazine (4.0 g, 11.1 mmol) in
10 mL 2-methoxyethanol. A balloon of H.sub.2 was bubbled through
the reaction mixture, and the reaction mixture was stirred at room
temperature under an atmosphere of H.sub.2 for 13 hours. Celite was
then added, and the reaction mixture was filtered through a pad of
celite and rinsed with MeOH. The filtrate was concentrated, and the
resulting oil was filtered through a short plug of silica gel with
EtOAc. The resulting filtrate was concentrated to give
4-(6-amino-quinazolin-4-yl)-1-Boc-piperazine (3.47 g, 95%). LCMS
(APCI+) m/z 330 [M+H].sup.+. HPLC Rt 2.31 min.
[0701] Step 3: To a 0.degree. C. solution of
(6-aminoquinazolin-4-yl)1-Boc-piperazine (1.0 g, 3.04 mmol),
triethylamine (1.7 mL, 12.2 mmol) and DMAP (93 mg, 0.76 mmol) in 20
mL DCM was added dropwise by addition funnel a solution of
methanesulfonyl chloride (0.59 mL, 7.6 mmol) in 6 mL DCM. The
reaction mixture was stirred 5 minutes, warmed to room temperature,
and stirred an additional 1.5 hours, after which the reaction
mixture was cooled to 0.degree. C., and NaOMe (5.4M in MeOH, 5.6
mL, 30.4 mmol) was added slowly by syringe. The reaction mixture
was stirred 10 minutes, warmed to room temperature, and stirred
another 2 hours, after which saturated NH.sub.4Cl was added. The
reaction mixture was extracted with DCM, and the combined extracts
were washed with brine, dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The resulting residue was purified on silica gel
(20:1 DCM:MeOH) to give
N-(4-Boc-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide (1.05
g, 85%) as a beige powder. LCMS (APCI+) m/z 408 [M+H].sup.+. HPLC
Rt 2.64 min.
[0702] Step 4: To a solution of
N-(4-Boc-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide (1.05
g, 2.58 mmol) in 15 mL dioxane was added 10 mL 4M HCl/dioxane. The
resulting suspension was stirred at room temperature 17 hours,
after which it was diluted with ether, and the solids were isolated
by filtration through a fritted funnel with nitrogen pressure,
rinsed with ether, and dried in vacuo to furnish
N-(4-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide
dihydrochloride (969 mg, 99%) as a white powder. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.80 (s, 1H), 8.12 (s, 1H), 7.87-7.84
(m, 2H), 4.50 (dd, J=5.2, 5.2 Hz, 4H), 3.57 (dd, J=5.2, 5.2 Hz,
4H), 3.13 (s, 3H). LCMS (APCI+) m/z 308 [M+H].sup.+. HPLC Rt 1.55
min.
[0703] Step 5: To a Jones tube containing PS-CDI (Argonaut, 1.04
mmol/g, 56 mg, 2.0 equiv) suspended in a solution of
N-(4-piperazin-1-yl-qyinazolin-6-yl)-mathanesulfonamide
dihydrochloride (11 mg, 0.029 mmol, 1.0 equiv) and DIEA (25 .mu.L,
0.15 mmol, 5.0 equiv) in 1.6 mL 9:1 CHCl.sub.3:THF were added
successively HOBt.H.sub.2O (6 mg, 0.038 mmol, 1.3 equiv) and
(D)-Boc-phenylalanine (8 mg, 0.032 mmol, 1.1 equiv.). The reaction
mixture was shaken for 15 hours at room temperature, after which
Si-trisamine (Silicycle, 1.21 mmol/g, 48 mg, 2.0 equiv) was added.
The reaction mixture was shaken an additional 1 hour, after which
it was vacuum filtered, the resins rinsed with CHCl.sub.3, and the
filtrate concentrated by rotary evaporation. The crude was purified
on silica gel (19:1 DCM:MeOH) to afford
(2R)--N-{4-[4-(2-Boc-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-
-6-yl}-methanesulfonamide as a clear, colorless residue.
[0704] Step 6: To a solution of
(2R)--N-{4-[4-(2-Boc-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-
-6-yl}-methanesulfonamide in 1.0 mL dioxane was added 1.2 mL 4M
HCl/dioxane. The resulting suspension was stirred at room
temperature another 13 hours, after which it was concentrated to
dryness. The resulting solids were dissolved in minimal MeOH, and
the product was triturated by the addition of ether. The resulting
suspension was diluted with ether, and the solids were isolated by
filtration through a fritted funnel with nitrogen pressure, rinsed
with ether, and dried further in vacuo to afford
(2R)--N-{4-[4-(2-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-y-
l}-methanesulfonamide dihydrochloride (13 mg, 84%) as a yellow
powder. LCMS (APCI+) m/z 455 [M+H].sup.+. HPLC Rt 2.18 min.
Example 100
##STR00116##
[0705] Preparation of
4-Amino-2-(2-chlorophenyl)-1-[4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-pipera-
zin-1-yl]-butan-1-one bis-hydrochloride salt
[0706] Step 1: The 1H-Pyrazolo[3,4-d]pyrimidin-4-ol (5.00 g, 36.73
mmol) was dissolved in 68.5 mL of phosphorous oxychloride and 9.31
mL of N,N-dimethyl aniline (73.47 mmol). This mixture was heated to
reflux (120 C) for 90 minutes to completion affording a dark red
solution. The mixture was concentrated in vacuo and cooled to
0.degree. C. in an ice bath. The residue was poured into ice water
and stirred for three minutes. The acidic melt was extracted with
ether, and the organics were combined. The organic was washed with
cold water, cold half saturated NaHCO.sub.3 solution, brine,
separated, dried over MgSO.sub.4, filtered, and concentrated in
vacuo to afford the 4-chloro-1H-pyrazolo[3,4-d]pyrimidine as a
light yellow powder (2.30 g, 41%). .sup.1H NMR (DMSO-d.sub.6, 400
MHz) .delta. 8.84 (s, 1H), 8.46 (s, 1H), NH not observed.
[0707] Step 2: The 4-chloro-1H-pyrazolo[3,4-d]pyrimidine (500 mg,
3.24 mmol) and piperazine-1-carboxylic acid tert-butyl ester (603
mg, 3.24 mmol) were dissolved in 11.0 mL of NMP then treated with
diisopropylethyl amine (845 .mu.L, 4.85 mmol). The yellow solution
was heated to 80 C. overnight to completion and was allowed to cool
to room temperature. The solution was diluted with ethyl acetate,
poured into diluted NaHCO.sub.3 solution, and extracted with ethyl
acetate. The combined organic was washed with water, brine,
separated, dried over MgSO.sub.4, filtered, and concentrated in
vacuo to afford the crude material as a tan solid. The material was
triturated with DCM/hexanes to afford the
4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester as a cream-colored solid (824 mg, 84%). .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.48 (s, 1H), 8.06 (s, 1H), 4.05
(m, 4H), 3.67 (m, 4H). LCMS (APCI+) m/z 305 [M+H].sup.+; Rt=2.14
min.
[0708] Step 3: The
4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylic acid
tert-butyl ester (600 mg, 1.97 mmol) was dissolved in 4 mL of
1,4-dioxane and treated with 10 mL of 4M HCl in 1,4-dioxane at room
temperature. The solution was allowed to stir for two hours to
afford a light-yellow suspension of product. The solvent was
diluted with diethyl ether, stirred for ten minutes, and filtered.
The pad of product was washed with diethyl ether and allowed to dry
under a stream of nitrogen to give the
4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine bis-hydrochloride
salt as a light-yellow solid (539 mg, 99%). .sup.1H NMR (D.sub.6O,
400 MHz) .delta. 8.59 (s, 1H), 8.42 (s, 1H), 4.29 (appt, J=5.6 Hz,
4H), 3.42 (appt, J=5.6 Hz, 4H). LCMS (APCI+) m/z 205 [M+H].sup.+;
Rt=0.34 min.
[0709] Step 4: The 4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
bis-hydrochloride salt (60 mg, 0.22 mmol), HOBt (29 mg, 0.22 mmol),
EDCI (46 mg, 0.24 mmol), and
4-tert-butoxycarbonylamino-2-(2-chlorophenyl)-butyric acid
[prepared by procedures described in Example 61 (68 mg. 0.22 mmol)]
were suspended/dissolved in 1.5 mL of DMF then treated with
triethylamine (121 L, 0.87 mmol). The mixture was allowed to stir
for four hours to completion then partitioned between ethyl acetate
and diluted NaHCO.sub.3 solution. The aqueous was extracted with
ethyl acetate, and the organics were combined. The organic was
washed with water, brine, separated, dried over MgSO.sub.4,
filtered, and concentrated in vacuo. The residue was dissolved in 1
mL of 1,4-dioxane and treated with 1 mL of 4M HCl in 1,4-dioxane.
The solution stirred overnight to completion at room temperature
and diluted with diethyl ether to afford a precipitate. This
material was broken up to afford a suspended granular solid which
was stirred for 30 minutes. The suspension was filtered, washed
with diethyl ether, and dried over a stream of nitrogen to afford
the
4-amino-2-(2-chlorophenyl)-1-[4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-pipera-
zin-1-yl]-butan-1-one bis-hydrochloride salt as a tan solid (69.2
mg, 67%). .sup.1H NMR (D.sub.6O, 400 MHz) .delta. 8.52 (brs, 1H),
8.31 (s, 1H), 7.40 (d, J=7.2 Hz, 1H), 7.12 (m, 3H), 4.42 (t, J=6.8
Hz, 1H), 4.11 (m, 1H), 3.98 (m, 3H), 3.70 (m, 2H), 3.61 (m, 1H),
3.41 (m, 1H), 2.96 (m, 1H), 2.80 (m, 1H), 2.21 (m, 1H), 1.94 (m,
1H). LCMS (APCI+) m/z 400 [M+H].sup.+; Rt=1.52 min.
Example 101
##STR00117##
[0710] Preparation of
4-Amino-2-(4-methylbenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-o-
ne, dihydrochloride
[0711] Step 1: To a solution containing LiHMDS (1.0M, 11.3 mL, 11
mmol) in 40 mL of THF under a nitrogen atmosphere at -78 C was
added a solution of 2-Oxo-pyrrolidine-1-carboxylic acid tert-butyl
ester (2 .gg, 11 mmol) in 15 mL of THF dropwise over 5 minutes.
After complete addition, the reaction was allowed to stir at
-78.degree. C. for 45 minutes, followed by the addition of a
solution containing 4-methyl benzyl bromide (2.1 g, 11 mmol) in 15
mL of THF dropwise over 5 minutes. The reaction was allowed to stir
at -78 C for 1 hour then warmed to 0 C and stirred for 1 hour. The
mixture was quenched with 36 mL of 3M LiOH and allowed to stir at
room temperature overnight. The reaction was diluted with water and
washed with ether. The aqueous phase was acidified with 1N HCl and
extracted with DCM. The organic phase was dried over magnesium
sulfate. Filtration and removal of solvent gave
4-tert-Butoxycarbonylamino-2-(4-methylbenzyl)-butyric acid (1.61 g,
49%.) LCMS (APCI--) m/z 306 [M-Boc-H].sup.-; Rt: 2.14 min.
[0712] Step 2: To a solution containing
4-tert-Butoxycarbonylamino-2-(4-methylbenzyl)-butyric acid (0.24 g,
0.77 mmol) in 25 mL of DMF under a nitrogen atmosphere was added
EDCI (0.16 g, 0.84 mmol), HOBT (130 mg, 0.84 mmol) and NMM (0.28 g,
2.8 mmol.) After stirring at room temperature for 15 minutes,
4-piperazin-1-yl-quinazoline (200 mg, 0.93 mmol) was added and the
reaction allowed to stir at room temperature. The reaction was
diluted with ethyl acetate and washed with water, saturated sodium
bicarbonate and water. The organic phase was dried over magnesium
sulfate. Filtration, removal of solvent and purification of the
residue via biotage eluting with 10% MeOH/DCM gave
[3-(4-Methyl-benzyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-ca-
rbamic acid tert-butyl ester (0.225 g, 64%) as a white solid. LCMS
(APCI+) m/z 504 [M+H].sup.+; Rt: 3.03 min. .sup.1H NMR (CDCl.sub.3,
400 MHz). 8.69 (1H, s), 7.92 (1H, d, J 8.3 Hz), 7.76-7.71 (2H, m),
7.45 (1H, t, J 7.8 Hz), 7.06 (4H, m), 4.53 (1H, br. s), 3.96-3.76
(2H, m), 3.63-3.54 (2H, m), 3.47-3.40 (2H, m), 3.24-2.84 (6H, m),
2.76-2.70 (1H, m), 2.27 (3H, s), 2.09-2.00 (1H, m), 1.73-1.65 (1H,
m), 1.40 (9H, s.)
[0713] Step 3: A mixture containing
[3-(4-Methyl-benzyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-ca-
rbamic acid tert-butyl ester (0.22 g, 0.44 mmol) in 10 mL of DCM
and mL of 4NHC in dioxane was allowed to stir at room temperature
under a nitrogen atmosphere overnight. The reaction was
concentrated under reduced pressure. The residue was dissolved in
methanol and ether added to precipitate the product. The solids
were filtered and dried to afford
4-Amino-2-(4-methylbenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-o-
ne dihydrochloride (167 mg.) LCMS (APCI+) m/z 404 [M+H].sup.+; Rt:
1.87 min. .sup.1H NMR (D2, 400 MHz). 8.45 (1H, s), 7.87 (2H, t, J
8.1 Hz), 7.65-7.57 (2H, m), 7.01 (4H, s), 4.18-4.12 (1H, m),
3.98-3.92 (1H, m), 3.82-3.71 (2H, m), 3.52-3.37 (3H, m), 3.25-2.76
(6H, m), 2.64-2.54 (1H, m), 2.02 (3H, s), 1.98-1.80 (1H, m.)
Example 102
##STR00118##
[0714] Preparation of
(2R,3'R)-2-(3'-Amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazo-
lin-4-yl-piperazin-1-yl)-ethanone trihydrochloride and
(2S,3'R)-2-(3'-Amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazo-
lin-4-yl-piperazin-1-yl)-ethanone trihydrochloride
[0715] Step 1: To a solution of glyoxylic acid monohydrate (1.0 g,
10.9 mmol) and (3R)-3-Boc-amino-pyrrolidine (2.06 g, 11.1 mmol) in
65 mL DCE was added 3,4-dichlorophenyl boronic acid (2.11 g, 11.1
mmol). The reaction mixture was heated to reflux and stirred 16
hours, after which it was cooled to room temperature, diluted with
DCM, and extracted with 1M Na.sub.2CO.sub.3. The basic aqueous
layer was extracted with EtOAc, and the combined extracts were
washed with 1N NaOH, dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The resulting residue was dissolved in minimal DCM,
and the product was triturated by the addition of ether. The
resulting solids were isolated by vacuum filtration, washed with
ether, and dried in vacuo to give one diastereomer of
(3'R)-(3'-Boc-amino-pyrrolidin-1-yl)-(3,4-difluorophenyl)-acetic
acid (1.53 g, 36%) as a tan powder, which will be referred to as
Diastereomer 1. .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 7.63 (s,
1H), 7.45 (d, J=8.2 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.09 (d, J=6.4
Hz, 0.2H), 6.95 (d, J=6.4 Hz, 0.8H), 3.92-3.78 (m, 1H), 3.51 (s,
0.25H), 3.49 (s, 0.75H), 3.40-3.30 (m, 1H), 2.77-2.69 (m, 0.2H),
2.64-2.53 (m, 1.8H), 2.42-2.33 (m, 1H), 2.31-2.17 (m, 1H),
2.01-1.88 (m, 1H), 1.58-1.46 (m, 1H), 1.36 (s, 9H). LCMS (APCI)+m/z
389 [M+H].sub.+; HPLC Rt 2.17 min.
[0716] The basic aqueous layer from above was carefully acidified
to about pH 6.5 with solid KHSO.sub.4 until CO.sub.2 evolution
ceased. The oily mixture was then extracted with EtOAc, and the
combined extracts were dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The resulting residue was dissolved in minimal DCM,
and the product was triturated by the addition of ether. The
resulting solids were isolated by vacuum filtration, washed with
ether, and dried in vacuo to give the other diastereomer of
(3'R)-(3'-Boc-amino-pyrrolidin-1-yl)-(3,4-difluorophenyl)-acetic
acid (0.67 g, 16%) as a tan powder, which will be referred to as
Diastereomer 2. .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 7.68 (s,
1H), 7.67-7.58 (m, 1.25H), 7.46-7.37 (m, 1H), 7.13-7.03 (m, 0.75H),
4.20 (d, J=6.6 Hz, 1H), 3.97 (br s, 1H), 3.11-3.00 (m, 0.5H),
2.90-2.76 (m, 1.5H), 2.74-2.63 (m, 0.5H), 2.63-2.52 (m, 1H),
2.47-2.37 (m, 0.5H), 2.13-1.97 (m, 1H), 1.74-1.60 (m, 1H), 1.36 (s,
9H). LCMS (APCI)+m/z 389 [M+H].sub.+; HPLC Rt 2.17 min.
[0717] Step 2, (Diastereomer 1): To a solution of
4-piperazin-1-ylquinazoline (30 mg, 0.14 mmol) and Diastereomer 1
(65 mg, 0.17 mmol) in 1.2 mL 3:1 DCM:THF were added successively
HOBt.H.sub.2O (21 mg, 0.14 mmol) and DCC (34 mg, 0.17 mmol). The
reaction mixture was stirred at room temperature 3.5 hours, after
which it was diluted with DCM, vacuum filtered through compressed
celite, and rinsed with DCM. The filtrate was then stirred with 2N
NaOH and extracted with DCM. The combined extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude was
purified on silica (1:4 DCM:ethyl acetate to 30:1 DCM:MeOH
gradient) to give
(3'R)-2-(3'-Boc-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinaz-
olin-4-yl-piperazin-1-yl)-ethanone. This was then dissolved in 1.2
mL dioxane, and 1.5 mL 4M HCl/dioxane was added. The resulting
suspension was stirred at room temperature 16 hours, after which it
was concentrated to dryness. The solids were dissolved in minimal
MeOH, and the product was triturated with ether. The resulting
solids were isolated by filtration through a fritted funnel with
nitrogen pressure, rinsed with ether, and dried in vacuo to give
one diasteomer of
(3'R)-2-(3'-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-
-4-yl-piperazin-1-yl)-ethanone trihydrochloride (67 mg, 81%) as a
pale yellow powder. .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 8.45
(s, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.89 (d, J=7.8 Hz, 1H), 7.67 (d,
J=8.4 Hz, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.56 (s, 1H), 7.51 (d, J=8.4
Hz, 1H), 7.33 (d, J=8.4 Hz, 1H), 5.54 (s, 0.75H), 5.52 (s, 0.25H),
4.25-3.95 (m, 5H), 3.77-3.35 (m, 7H), 3.26-3.06 (m, 1H), 2.60-2.40
(m, 1H), 2.19-1.94 (m, 1H). LCMS (APCI)+m/z 485 [M+H].sup.+; HPLC
Rt 1.71 min.
[0718] Step 2, Diastereomer 2: 4-piperazin-1-ylquinazoline was
acylated with Diastereomer 2 following Step 2 for Diastereomer 1
above, to furnish the other diastereomer of
(3'R)-2-(3-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin--
4-yl-piperazin-1-yl)-ethanone trihydrochloride (60 mg, 72%) as a
pale yellow powder. .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 8.45
(s, 1H), 7.91 (d, J=7.8 Hz, 1H), 7.89 (d, J=7.8 Hz, 1H), 7.67 (d,
J=8.4 Hz, 1H), 7.59 (d, J=8.0 Hz, 1H), 7.55 (s, 1H), 7.49 (d, J=8.2
Hz, 1H), 7.32 (d, J=7.6 Hz, 1H), 5.46 (d, J=12.7 Hz, 1H), 4.24-3.95
(m, 5H), 3.82-3.44 (m, 6H), 3.40-3.28 (m, 0.5H), 3.20-3.10 (m, 1H),
3.10-2.98 (m, 0.5H), 2.56-2.36 (m, 1H), 2.15-1.92 (m, 1H). LCMS
(APCI)+m/z 485 [M+H].sub.+; HPLC Rt 1.69 min.
Example 103
##STR00119##
[0719] The Preparation of
(2R)-2-amino-3-phenyl-1-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-1-yl)-pr-
opan-1-one trihydrochloride
[0720] Step 1: 2-aminonicotinic acid (7 g) and formamide (22.8 g)
were heated at 167 C (internal temperature) for 2.5 hours. After
cooling, the solid was recrystallized from 100 mL of hot water to
give Pyrido[2,3-d]pyrimidin-4-ol as pale yellow powder (5.2 g,
69.7%). .sup.1H NMR (DMSO, 400 MHz) .delta. 12.60 (br, 1H), 8.90
(br, 1H), 8.50 (m, 1H), 8.37 (s, 1H), 7.50 (m, 1H). R.sub.t 0.87
min. MS (ESI+) [M+H].sup.+ 148.
[0721] Step 2: The Pyrido[2,3-d]pyrimidin-4-ol (3 g) in POCl.sub.3
(45 mL) was stirred at reflux for 3 hours. The excess POCl.sub.3
was removed. The residue was added 10 mL of cold water and
extracted with EtOAc (2.times.30 mL). The combined organic layer
was washed with brine and dried over sodium sulfate. After removal
of solvent, it gave 4-chloropyrido[2,3-d]pyrimidine as yellow solid
(0.3 g, 7.6%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.38 (br,
1H), 9.31 (s, 1H), 8.62 (d, 1H), 7.75 (m, 1H).
[0722] Step 3: The 4-chloro-pyrido[2,3-d]pyrimidine (0.3 g) and
piperazine (1.6 g) in Ethanol (10 mL) was refluxed for 1 hour. The
solvent was removed and 50 mL of toluene was added. The toluene was
removed in vacuo. The resulting solid was used directly for the
next step without purification. R.sub.t 1.93 min. MS (ESI+)
[M+H].sup.+ 216.
[0723] Step 4: DIEA (0.74 mL) and HBTU (1.3 g) was added to the
solution of (2R)-2-tert-butoxycarbonylamino-3-phenyl-propionic acid
(0.092 g) in THF (5 mL) at 0.degree. C. The mixture was stirred at
room temperature for 20 minutes, and then
4-piperazin-1-yl-pyrido[2,3-d]pyrimidinene (0.31 g) was added. The
reaction was stirred at room temperature for 1 hour. 20 mL of EtOAc
was added and the organic layer was separated. The aqueous layer
extracted with EtOAc (20 mL). The combined organic layer was washed
with saturated sodium bicarbonate (20 mL) and dried over sodium
sulfate. After removal of solvent, the residue was purified by
flash chromatography (10:1=DCM:MeOH) to give
[1-benzyl-2-oxo-2-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-yl)-ethyl-carb-
amic acid tert-butyl ester as white foam solid (0.392 g, 58.9%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.11 (br, 1H), 8.92 (s,
1H), 8.15 (d, 1H), 7.40 (m, 1H), 7.20-7.33 (m, 5H), 5.42 (m, 1H),
4.85 (m, 1H), 3.78-3.82 (m, 2H), 3.61-3.70 (m, 2H), 3.50-3.58 (m,
2H), 3.15-3.20 (m, 1H), 3.08-3.12 (m, 2H), 1.42 (s, 9H). R.sub.t
2.23 min. MS (ESI+) [M+H].sup.+ 463.
[0724] Step 5:
[1-benzyl-2-oxo-2-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-yl)-ethyl-carb-
amic acid tert-butyl ester (0.046 mg) was dissolved in DCM (5 mL)
and HCl/dioxane (0.5 mL) was added. The suspension was stirred at
room temperature for 3 hours, after which it was concentrated to
give
(2R)-2-amino-3-phenyl-1-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-1-yl)-pr-
opan-1-one trihydrochloride (0.031 g, 86%). R.sub.t 1.55 min. MS
(ESI+) [M+H].sup.+ 363.
Example 104
##STR00120##
[0725] Preparation of
5-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1--
one dihydrochloride
[0726] Step 1: To a -78.degree. C. solution of LHMDS (1.0M, 5.3 mL,
5.3 mmol) in 10 mL THF was added by syringe a solution of
Boc-2-piperidone (1.0 g, 5.0 mmol) in 8 mL THF. The reaction
mixture was stirred at -78.degree. C. for 1 hour, after which a
0.degree. C. solution of 4-chlorobenzyl bromide (1.1 g, 5.3 mmol)
in 5 mL THF was added quickly by syringe. The reaction mixture was
stirred 1 hour at -78.degree. C., warmed to 0.degree. C., stirred
another 1 hour, then quenched with 17 mL 3M LiOH solution. The
reaction mixture was then stirred 15 hours at room temperature,
after which it was diluted with H.sub.2O and washed with ether. The
aqueous layer was acidified with solid KHSO.sub.4, extracted with
DCM, and the extracts were dried (Na.sub.2SO.sub.4), filtered, and
concentrated to give 5-Boc-amino-2-(4-chlorobenzyl)-pentanoic acid
(1.0 g, 60%) as a clear, colorless syrup. LCMS (APCI--) m/z 340
[M-H].sup.-; Rt: 2.37 min.
[0727] Step 2: A solution of EDCI (100 mg, 0.54 mmol),
HOBt.H.sub.2O (82 mg, 0.54 mmol),
5-Boc-amino-2-(4-chlorobenzyl)-pentanoic acid (170 mg, 0.50 mmol),
and TEA (190 .mu.L, 1.4 mmol) in 3 mL DMF was stirred 10 minutes,
and solid 4-piperazin-1-yl-quinazoline dihydrochloride (130 mg,
0.45 mmol) was added. The reaction mixture was stirred at room
temperature 15 hours, after which water was added. The reaction
mixture was extracted with DCM, and combined extracts washed with
sat NaHCO.sub.3, dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The crude was purified on silica gel (1:1 to 1:9
DCM:EtOAc) to give
5-Boc-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-penta-
n-1-one. LCMS (APCI+) m/z 538 [M+H].sup.+; Rt: 3.12 min.
[0728] Step 3: To a solution of
5-Boc-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-penta-
n-1-one in 1.5 mL dioxane was added 2.5 mL 4M HCl/dioxane. The
resulting suspension was stirred at room temperature 17 hours,
after which it was concentrated to dryness. The solids were
dissolved in minimal MeOH, and the product was triturated by the
addition of ether. The solids were isolated by filtration through a
fritted funnel with nitrogen pressure, rinsed with ether, and dried
in vacuo to give
5-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1--
one dihydrochloride (110 mg, 46%) as a white powder. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.71 (1H, d, J 2.0 Hz), 8.20 (1H, d,
J 8.7 Hz), 8.05 (1H, t, J 7.8 Hz), 7.84-7.75 (2H, m), 7.31-7.22
(4H, m), 4.40-4.30 (1H, m), 4.29-4.20 (1H, m), 4.12-4.02 (1H, m),
3.92-3.66 (4H, m), 3.53-3.42 (1H, m), 3.29-3.21 (1H, m), 2.97-2.81
(4H, m), 1.87-1.62 (4H, m). LCMS (APCI+) m/z 438 [M+H].sup.+; Rt:
1.98 min.
Example 105
##STR00121##
[0729] Preparation of
2-(2-Amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one dihydrochloride
[0730] The PS-CDI (175 mg, 0.181 mmol), HOBt monohydrate (214 mg,
0.140 mmol), 4-piperazin-1-yl-quinazoline dihydrochloride (52 mg,
0.181 mmol), and
2-(2-tert-butoxycarbonylamino-ethoxy)-3-(4-chlorophenyl)-propionic
acid (48 mg, 0.140 mmol; prepared by alkylation of
(2-Boc-amino-ethoxy)-acetic acid ethyl ester with
4-chlorobenzylbromide according to procedures described in the
literature: Nizal S. Chandrakumar et al. J. Med. Chem. 1992, 35,
2928-2938) were suspended/dissolved in 2.5 mL of chloroform (plus
3-5 drops THF). The mixture was shaken at room temperature
overnight, then treated with MP-CO.sub.3 (330 mg, 0.838 mmol) for
two hours. The mixture was vacuum filtered, rinsed with chloroform,
and concentrated in vacuo. The residue was purified on silica gel
(1:19 DCM:EtOAc) to afford
2-(2-Boc-amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin--
1-yl)-propan-1-one. This material was dissolved in 1.0 mL of
1,4-dioxane and treated with 1.5 mL of 4M HCl in 1,4-dioxane
affording slow precipitation. The mixture was sonicated briefly and
stirred at room temperature overnight to completion. The resulting
suspension was concentrated in vacuo, and the solids were suspended
in diethyl ether, filtered under nitrogen pressure, and dried in
vacuo to afford
2-(2-amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl-
)-propan-1-one dihydrochloride as a yellow powder (13 mg, 18%).
LCMS (APCI+) m/z 440 [M+H].sup.+. HPLC R.sub.t=1.75 min.
Example 106
##STR00122##
[0731] Preparation of
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyr-
imidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0732] Step 1: To a stirred solution of LDA (9.85 mL, 1.5 M, 15.8
mmol) in THF (20 mL) was added dropwise a solution of
4,6-dichloropyrimidine (2.00 g, 13.4 mmol) in THF (12 mL) at
-78.degree. C. After stirring for 1.5 hours, a solution of
cyclopropanecarbaldehyde (1.05 g, 15.0 mmol) in THF (10 mL) was
added dropwise. The solution was stirred at -78.degree. C. for 1
hour and then quenched by addition of water (10 mL). The reaction
mixture was allowed to warm to room temperature and partitioned
between EtOAc and water. The organic layer was washed with brine,
dried and concentrated. The residue was purified by column
chromatography (hexanes/EtOAc, 3:1) to give
Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanol (2.36 g, 80%) as
a yellow oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.70 (s,
1H), 4.56 (m, 1H), 2.63 (d, J=8.0 Hz, 1H), 1.70 (m, 1H), 0.76 (m,
1H), 0.55 (m, 2H).
[0733] Step 2: To a vigorously stirred solution of
Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanol (0.84 g, 3.8
mmol) in anhydrous acetone (12 mL) was added portionwise chromium
(VI) oxide (1.2 g, 12 mmol) at 0.degree. C. The mixture was stirred
at 0.degree. C. for 30 minutes. The excess of the oxidizing agent
was destroyed by the addition of isopropanol (2 mL). After stirring
for 15 minutes, the reaction mixture was poured into saturated
NaHCO.sub.3 solution and filtered through Celite. The filtrate was
extracted with EtOAc. The combined organic layers were washed with
brine, dried and concentrated. The residue was purified by column
chromatography (hexanes:EtOAc, 6:1) to give
Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanone (0.80 g, 96%)
as a colorless oil. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.83
(s, 1H), 2.26 (m, 1H), 1.46 (m, 2H), 1.26 (m, 2H).
[0734] Step 3: A mixture of
Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanone (0.75 g, 3.5
mmol), anhydrous hydrazine (0.13 mL, 4.1 mmol) and THF (35 mL) was
stirred at room temperature for 4 hours. The reaction was
partitioned between water and EtOAc. The organic layer was washed
with brine, dried and passed through a short silica gel pad to give
4-Chloro-3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine (0.50 g, 74%)
as a white solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 11.87
(s, 1H), 8.80 (s, 1H), 2.55 (m, 1H), 1.15 (m, 2H), 1.14 (m,
2H).
[0735] Step 4:
4-(3-Cyclopropyl-11H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxyl-
ic acid tert-butyl ester was prepared by the procedures described
in Example 40, Step 1, substituting 4-chloro-5-iodopyrimidine with
4-Chloro-3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine. LCMS (APCI+)
m/z 345 [M+H].sup.+; Rt=2.52 min.
[0736] Step 5:
3-Cyclopropyl-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
dihydrochloride was prepared by the procedures described in Example
34, Step 3, substituting
(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-eth-
yl}-carbamic acid tert-butyl ester with
4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxyli-
c acid tert-butyl ester. .sup.1H NMR (DMSO-d.sub.6, 400 MHz)
.delta. 8.58 (s, 1H), 4.60 (m, 4H), 3.53 (m, 4H), 2.36 (m, 1H),
1.31 (m, 2H), 1.14 (m, 2H). LCMS (APCI+) m/z 245 [M+H].sup.+;
Rt=1.02 min.
[0737] Step 6:
(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyr-
imidin-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride was
prepared by substituting
4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorene
dihydrochloride with
3-cyclopropyl-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine
dihydrochloride in Example 81, Step 3. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.48 (s, 1H), 7.40 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.4
Hz, 2H), 4.77 (m, 1H), 4.24 (m, 3H), 3.87 (m, 2H), 3.74 (m, 2H),
3.25 (m, 1H), 3.16 (m, 2H), 2.27 (m, 1H), 1.28 (m, 2H), 1.10 (m,
2H). LCMS (APCI+) m/z 426, 428 [M+H].sup.+; Rt=1.88 min.
Example 107
##STR00123##
[0738] Preparation of
2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one dihydrochloride
[0739] Step 1:
4-[2-(3,4-Dichlorophenyl)-2-methoxycarbonyl-ethyl]-imidazole-1-carboxylic
acid tert-butyl ester was prepared by the procedures described in
Example 78, Step 1, substituting
3-tert-butoxycarbonylamino-propionic acid tert-butyl ester with
(3,4-Dichlorophenyl)-acetic acid methyl ester and substituting
4-Bromo-1-bromomethyl-2-fluoro-benzene with
4-Bromomethyl-imidazole-1-carboxylic acid tert-butyl ester
(prepared from 4(5)-hydroxymethylimidazole hydrochloride according
to the literature: J. Med. Chem. 1997, 40, 2208). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.96 (s, 1H), 7.42 (s, 1H), 7.38 (d,
J=8.0 Hz, 1H), 7.16 (d, J=8.0 Hz, 1H), 7.02 (s, 1H), 4.08 (t, J=7.6
Hz, 1H), 3.66 (s, 3H), 3.32 (dd, J=14.4 Hz, J=8.4 Hz, 1H), 2.92
(dd, J=14.4 Hz, J=6.8 Hz, 1H). LCMS (APCI+) m/z 299, 301, 303
[M-Boc+H].sup.+; Rt=3.72 min.
[0740] Step 2:
2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-propionic acid was
prepared by the procedures described in Example 78, Step 2,
substituting
3-tert-Butoxycarbonylamino-2-(4-trifluoromethylbenzyl)-propionic
acid ethyl ester with
4-[2-(3,4-Dichlorophenyl)-2-methoxycarbonyl-ethyl]-imidazole-1-carboxylic
acid tert-butyl ester. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.25 (s, 1H), 7.51 (s, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.26 (d, J=8.0
Hz, 1H), 7.00 (s, 1H), 3.84 (m, 1H), 3.38 (m, 1H), 3.00 (dd, J=14.4
Hz, J=6.8 Hz, 1H). LCMS (APCI+) m/z 285, 287, 289 [M+H].sup.+;
Rt=1.54 min.
[0741] Step 3:
2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimi-
din-4-yl)-piperazin-1-yl]-propan-1-one was prepared by substituting
5-piperazin-1-yl-1H-indazole with
4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride and
substituting (D)-Boc-4-chlorophenylalanine with
2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-propionic acid in
Example B-1, Step 2. The free amine was converted to HCl salt by
treatment with HCl in Ether. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.76 (s, 1H), 8.32 (s, 1H), 7.52 (m, 2H), 7.38 (s, 1H),
7.29 (m, 2H), 6.92 (s, 1H), 4.58 (m, 1H), 3.70-4.20 (m, 7H), 3.63
(m, 1H), 3.43 (m, 1H), 3.16 (m, 1H). LCMS (APCI+) m/z 470, 472, 474
[M+H].sup.+; Rt=2.15 min.
[0742] The words "comprise," "comprising," "include," "including,"
and "includes" when used in this specification and in the following
claims are intended to specify the presence of stated features,
integers, components, or steps, but they do not preclude the
presence or addition of one or more other features, integers,
components, steps, or groups.
* * * * *