U.S. patent application number 15/162453 was filed with the patent office on 2016-11-17 for substituted benzene compounds.
The applicant listed for this patent is Epizyme, Inc.. Invention is credited to Richard Chesworth, Kenneth W. Duncan, Heike Keilhack, Christine Klaus, Sarah K. Knutson, Kevin W. Kuntz, Masashi Seki, Natalie Warholic, Timothy J. N. Wigle.
Application Number | 20160332969 15/162453 |
Document ID | / |
Family ID | 46000399 |
Filed Date | 2016-11-17 |
United States Patent
Application |
20160332969 |
Kind Code |
A1 |
Kuntz; Kevin W. ; et
al. |
November 17, 2016 |
SUBSTITUTED BENZENE COMPOUNDS
Abstract
The present invention relates to substituted benzene compounds.
The present invention also relates to pharmaceutical compositions
containing these compounds and methods of treating cancer by
administering these compounds and pharmaceutical compositions to
subjects in need thereof. The present invention also relates to the
use of such compounds for research or other non-therapeutic
purposes.
Inventors: |
Kuntz; Kevin W.; (Woburn,
MA) ; Chesworth; Richard; (Concord, MA) ;
Duncan; Kenneth W.; (Norwood, MA) ; Keilhack;
Heike; (Belmont, MA) ; Warholic; Natalie;
(Brighton, MA) ; Klaus; Christine; (Waban, MA)
; Knutson; Sarah K.; (Cambridge, MA) ; Wigle;
Timothy J. N.; (Waltham, MA) ; Seki; Masashi;
(Tsukuba-shi, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Epizyme, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
46000399 |
Appl. No.: |
15/162453 |
Filed: |
May 23, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14110873 |
Jan 30, 2014 |
9376422 |
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PCT/US2012/033662 |
Apr 13, 2012 |
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15162453 |
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61474825 |
Apr 13, 2011 |
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61505676 |
Jul 8, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 401/12 20130101;
C07D 491/10 20130101; C07D 213/64 20130101; A61P 35/00 20180101;
C07D 401/14 20130101; C07D 409/12 20130101; C07D 407/12 20130101;
C07D 413/14 20130101; C07D 491/048 20130101; C07D 491/08 20130101;
C07D 405/14 20130101; C07D 413/12 20130101; C07D 405/12 20130101;
C07D 491/113 20130101 |
International
Class: |
C07D 213/64 20060101
C07D213/64; C07D 405/12 20060101 C07D405/12; C07D 491/113 20060101
C07D491/113; C07D 491/048 20060101 C07D491/048; C07D 405/14
20060101 C07D405/14; C07D 409/12 20060101 C07D409/12; C07D 491/08
20060101 C07D491/08; C07D 401/12 20060101 C07D401/12; C07D 401/14
20060101 C07D401/14 |
Claims
1. A compound of Formula (I) or a pharmaceutically acceptable salt
thereof: ##STR01099## wherein X.sub.1 is N or CR.sub.11; X.sub.2 is
N or CR.sub.13; X.sub.3 is N or C, and when X.sub.3 is N, R.sub.6
is absent; Z is NR.sub.7R.sub.8, OR.sub.7, S(O).sub.aR.sub.7, or
CR.sub.7R.sub.8R.sub.14, in which a is 0, 1, or 2; each of R.sub.1,
R.sub.5, R.sub.9, and R.sub.10, independently, is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl; each
of R.sub.2, R.sub.3, and R.sub.4, independently, is
-Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or C.sub.1-C.sub.3
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo, hydroxyl, COOH,
cyano, azido, or R.sub.S1, in which R.sub.S1 is C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
R.sub.6 is H, halo, cyano, azido, OR.sub.a, --NR.sub.aR.sub.b,
--C(O)R.sub.a, --C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b,
--NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl, b is 0, 1, or 2, each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; R.sub.7 is
-Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4 alkyl
linker, or C.sub.2-C.sub.4 alkenyl linker, each linker optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.4 is H, halo, cyano, NR.sub.gR.sub.h, --OR.sub.g,
--C(O)R.sub.g, --C(O)OR.sub.g, --C(O)NR.sub.gR.sub.h,
--C(O)NR.sub.gOR.sub.h, --NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g,
or R.sub.S6, in which each of R.sub.g and R.sub.h, independently is
H or R.sub.S7, each of R.sub.S6 and R.sub.S7, independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
14-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
each of R.sub.S6 and R.sub.S7 is optionally substituted with one or
more -Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O),
C(O)NR.sub.k, NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl
linker, R.sub.k being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H,
halo, C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
each of R.sub.8, R.sub.11, R.sub.12, and R.sub.13, independently,
is H, halo, hydroxyl, COOH, cyano, R.sub.S8, OR.sub.S8, or
COOR.sub.S8, in which R.sub.S8 is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, 4 to 12-membered heterocycloalkyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, or di-C.sub.1-C.sub.6 alkylamino,
and R.sub.S8 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, COOH, C(O)O--C.sub.1-C.sub.6 alkyl,
cyano, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl; provided that
the compound is not
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide,
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide,
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimet-
hyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimeth-
oxybenzamide,
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide,
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide,
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, or
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
2. The compound of claim 1, wherein X.sub.1 is N or CR.sub.11;
X.sub.2 is N or CR.sub.13; X.sub.3 is N or C, and when X.sub.3 is
N, R.sub.6 is absent; Z is NR.sub.7R.sub.8, OR.sub.7,
S(O).sub.aR.sub.7, or CR.sub.7R.sub.8R.sub.14, in which a is 0, 1,
or 2; each of R.sub.1, R.sub.5, R.sub.9, and R.sub.10,
independently, is H or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 12-membered heterocycloalkyl, and 5- or
6-membered heteroaryl; each of R.sub.2, R.sub.3, and R.sub.4,
independently, is -Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or
C.sub.1-C.sub.3 alkyl linker optionally substituted with halo,
cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo,
hydroxyl, COOH, cyano, azido, or R.sub.S1, in which R.sub.S1 is
C.sub.1-C.sub.3 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
R.sub.6 is H, halo, cyano, azido, --NR.sub.aR.sub.b, --C(O)R.sub.a,
--C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a,
--S(O).sub.bR.sub.a, --S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in
which R.sub.S2 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
12-membered heterocycloalkyl, b is 0, 1, or 2, each of R.sub.a and
R.sub.b, independently is H or R.sub.S3, and R.sub.S3 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl; or
R.sub.a and R.sub.b, together with the N atom to which they are
attached, form a 4 to 12-membered heterocycloalkyl ring having 0 or
1 additional heteroatom; and each of R.sub.S2, R.sub.S3, and the 4
to 12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b,
is optionally substituted with one or more -Q.sub.2-T.sub.2,
wherein Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.2 is H, halo, cyano, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--C(O)NR.sub.cR.sub.d, --NR.sub.dC(O)R.sub.c,
--NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; R.sub.7 is
-Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4 alkyl
linker, or C.sub.2-C.sub.4 alkenyl linker, each linker optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.4 is H, halo, cyano, NR.sub.gR.sub.h, --OR.sub.g,
--C(O)R.sub.g, --C(O)OR.sub.g, --C(O)NR.sub.gR.sub.h,
--C(O)NR.sub.gOR.sub.h, --NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g,
or R.sub.S6, in which each of R.sub.g and R.sub.h, independently is
H or R.sub.S7, each of R.sub.S6 and R.sub.S7, independently is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
14-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
each of R.sub.S6 and R.sub.S7 is optionally substituted with one or
more -Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O),
C(O)NR.sub.k, NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl
linker, R.sub.k being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H,
halo, C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
provided that (i) R.sub.7 is not C(O)R.sub.g or --S(O).sub.2R.sub.g
when Z is NR.sub.7R.sub.8; (ii) R.sub.7 is not C.sub.1-C.sub.6
alkyl when Z is OR.sub.7, and (iii) R.sub.7 is not H; each of
R.sub.8, R.sub.11, R.sub.12, and R.sub.13, independently, is H,
halo, hydroxyl, COOH, cyano, R.sub.S8, OR.sub.S8, or COOR.sub.S8,
in which R.sub.S8 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, 4 to
12-membered heterocycloalkyl, amino, mono-C.sub.1-C.sub.6
alkylamino, or di-C.sub.1-C.sub.6 alkylamino, and R.sub.S8 is
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, COOH, C(O)O--C.sub.1-C.sub.6 alkyl,
cyano, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl.
3. The compound of claim 1, wherein the compound is of Formula
(Ia): ##STR01100##
4-7. (canceled)
8. The compound of claim 1, wherein Z is NR.sub.7R.sub.8.
9-11. (canceled)
12. The compound claim 1, wherein R.sub.6 is halo, C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.6 cycloalkyl, C(O)H, or --C(O)R.sub.a, in which
R.sub.a is C.sub.1-C.sub.6 alkyl or 4 to 12-membered
heterocycloalkyl, and each of the C.sub.1-C.sub.3 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, and
C.sub.3-C.sub.6 cycloalkyl is optionally substituted with one or
more -Q.sub.2-T.sub.2.
13-16. (canceled)
17. The compound of claim 1, wherein R.sub.7 is C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.8 cycloalkyl or 4 to 14-membered
heterocycloalkyl, each optionally substituted with one or more
-Q.sub.5-T.sub.5.
18. The compound of claim 1, wherein R.sub.7 is piperidinyl,
tetrahydropyran, tetrahydrothiopyran, oxetanyl, azetidinyl,
pyrrolidinyl, 1,4-dioxaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-3'-one-4-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-7'-one-4-yl,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-3'-one-4-yl,
1-azaspiro[4.5]decan-2-one-8-yl, cyclobutyl, cyclopentyl, or
cyclohexyl, each optionally substituted with one
-Q.sub.5-T.sub.5.
19-28. (canceled)
29. The compound of claim 1, wherein each of R.sub.2 and R.sub.4,
independently is C.sub.1-C.sub.3 alkyl optionally substituted with
C.sub.1-C.sub.6 alkoxyl.
30. (canceled)
31. The compound of claim 1, wherein R.sub.1 is H and R.sub.3 is
H.
32. The compound of claim 1, wherein R.sub.12 is H, methyl, ethyl,
ethenyl, or halo.
33-35. (canceled)
36. The compound of claim 1, wherein R.sub.8 is H, methyl, or
ethyl.
37-39. (canceled)
40. The compound of claim 1, wherein R.sub.13 is H or methyl and
R.sub.11 is H.
41-52. (canceled)
53. The compound of claim 1, wherein the compound is of Formula
(II): ##STR01101##
54. The compound of claim 1, wherein the compound is of Formula
(IIA): ##STR01102## wherein n is 0, 1, or 2; U is O, S,
N-Q.sub.5-T.sub.5, or CH-Q.sub.5-T.sub.5; and R.sub.12 is Cl, Br,
or methyl.
55-65. (canceled)
66. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1, or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable
carrier.
67. A method of treating cancer comprising administering to a
subject in need thereof a therapeutically effective amount of a
compound of claim 1,
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide,
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide,
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimet-
hyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimeth-
oxybenzamide,
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide,
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide,
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, or
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide, or a pharmaceutically acceptable salt
thereof.
68. (canceled)
69. The compound of claim 1, wherein Q.sub.2 is a bond or an
unsubstituted C.sub.1-C.sub.3 alkyl linker.
70. The compound of claim 1, wherein T.sub.2 is 4 to 7-membered
heterocycloalkyl optionally substituted with one or more
-Q.sub.3-T.sub.3.
71. The compound of claim 1, wherein R.sub.6 is ethynyl substituted
with one or more -Q.sub.2-T.sub.2, in which Q.sub.2 is a bond or
C.sub.1-C.sub.3 alkyl linker and T.sub.2 is C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.6 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
72. The compound of claim 1, wherein Q.sub.5 is a bond and T.sub.5
is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, or di-C.sub.1-C.sub.6 alkylamino,
T.sub.5 being optionally substituted with one or more substituents
selected from the group consisting of halo, hydroxyl,
C.sub.1-C.sub.6 alkoxyl, or C.sub.3-C.sub.8 cycloalkyl.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 14/110,873, filed Jan. 30, 2014 (now allowed), which is a
national stage application, filed under 35 U.S.C. .sctn.371, of
International Application No. PCT/US2012/033662, filed Apr. 13,
2012, which claims priority to, and the benefit of, U.S.
provisional application No. 61/474,825, filed Apr. 13, 2011, and
61/505,676 filed Jul. 8, 2011, the entire contents of each of which
are incorporated herein by reference in their entireties.
INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING
[0002] The contents of the text file named
"EPIZ-008C01US-ST25.txt," which was created on May 23, 2016 and is
2 KB in size, are hereby incorporated by reference in their
entireties.
BACKGROUND OF THE INVENTION
[0003] In eukaryotic cells DNA is packaged with histones to form
chromatin. Changes in the ordered structure of chromatin can lead
to alterations in transcription of associated genes. Control of
changes in chromatin structure (and hence of transcription) is
mediated by covalent modifications to histones, most notably of
their N-terminal tails. These modifications are often referred to
as epigenetic because they can lead to heritable changes in gene
expression, but do not affect the sequence of the DNA itself.
Covalent modifications (for example, methylation, acetylation,
phosphorylation, and ubiquitination) of the side chains of amino
acids are enzymatically mediated. The selective addition of methyl
groups to specific amino acid sites on histones is controlled by
the action of a unique family of enzymes known as histone
methyltransferases (HMTs).
[0004] The orchestrated collection of biochemical systems behind
transcriptional regulation must be tightly controlled in order for
cell growth and differentiation to proceed optimally. Disease
states result when these controls are disrupted by aberrant
expression and/or activity of the enzymes responsible for DNA and
histone modification. In human cancers, for example, there is a
growing body of evidence to suggest that dysregulated epigenetic
enzyme activity contributes to the uncontrolled cell proliferation
associated with cancer as well as other cancer-relevant phenotypes
such as enhanced cell migration and invasion. Beyond cancer, there
is growing evidence for a role of epigenetic enzymes in a number of
other human diseases, including metabolic diseases (such as
diabetes), inflammatory diseases (such as Crohn's disease),
neurodegenerative diseases (such as Alzheimer's disease) and
cardiovascular diseases. Therefore, selectively modulating the
aberrant action of epigenetic enzymes may hold promise for the
treatment of a range of diseases.
[0005] Polycomb group (PcG) and trithorax group (trxG) proteins are
known to be part of the cellular memory system. See, e.g., Francis
et al. (2001) Nat Rev Mol Cell Biol 2:409-21 and Simon et al.
(2002) Curr Opin Genet Dev 12:210-8. In general, PcG proteins are
transcriptional repressors that maintain the "off state," and trxG
proteins are transcriptional activators that maintain the "on
state." Because members of PcG and trxG proteins contain intrinsic
histone methyltransferase (HMTase) activity, PcG and trxG proteins
may participate in cellular memory through methylation of core
histones. See, e.g., Beisel et al. (2002) Nature 419:857-62; Cao et
al. (2002) Science 298:1039-43; Czermin et al. (2002) Cell
111:185-96; Kuzmichev et al. (2002) Genes Dev 16:2893-905; Milne et
al. (2002) Mol Cell 10:1107-17; Muller et al. (2002) Cell
111:197-208; and Nakamura et al. (2002) Mol Cell 10:1119-28.
[0006] Biochemical and genetic studies have provided evidence that
Drosophila PcG proteins function in at least two distinct protein
complexes, the Polycomb repressive complex 1 (PRC1) and the
ESC-E(Z) complex (also known as Polycomb repressive complex 2
(PRC2)). Otte et al. (2003) Curr Opin Genet Dev 13:448-54. Studies
in Drosophila have demonstrated that the ESC-E(Z)/EED-EZH2 (i.e.,
PRC2) complexes have intrinsic histone methyltransferase activity.
Although the compositions of the complexes isolated by different
groups are slightly different, they generally contain EED, EZH2,
SUZ12, and RbAp48 or Drosophila homologs thereof. However, a
reconstituted complex comprising only EED, EZH2, and SUZ12 retains
histone methyltransferase activity for lysine 27 of histone H3.
U.S. Pat. No. 7,563,589.
[0007] Of the various proteins making up PRC2 complexes, EZH2
(Enhancer of Zeste Homolog 2) is the catalytic subunit. The
catalytic site of EZH2 in turn is present within a SET domain, a
highly conserved sequence motif (named after Su(var)3-9, Enhancer
of Zeste, Trithorax) that is found in several chromatin-associated
proteins, including members of both the Trithorax group and
Polycomb group. SET domain is characteristic of all known histone
lysine methyltransferases except the H3-K79 methyltransferase
DOT1.
[0008] In addition to Hox gene silencing, PRC2-mediated histone
H3-K27 methylation has been shown to participate in X-inactivation.
Plath et al. (2003) Science 300:131-5; Silva et al. (2003) Dev Cell
4:481-95. Recruitment of the PRC2 complex to Xi and subsequent
trimethylation on histone H3-K27 occurs during the initiation stage
of X-inactivation and is dependent on Xist RNA. Furthermore, EZH2
and its associated histone H3-K27 methyltransferase activity were
found to mark differentially the pluripotent epiblast cells and the
differentiated trophectoderm, and consistent with a role of EZH2 in
maintaining the epigenetic modification patterns of pluripotent
epiblast cells, Cre-mediated deletion of EZH2 results in loss of
histone H3-K27 methylation in the cells. Erhardt et al. (2003)
Development 130:4235-48). Further, studies in prostate and breast
cancer cell lines and tissues have revealed a strong correlation
between the levels of EZH2 and SUZ12 and the invasiveness of these
cancers, indicating that dysfunction of the PRC2 complex may
contribute to cancer. Bracken et al. (2003) EMBO J 22:5323-35;
Kirmizis et al. (2003) Mol Cancer Ther 2:113-21; Kleer et al.
(2003) Proc Natl Acad Sci USA 100:11606-11; Varambally et al.
(2002) Nature 419:624-9.
[0009] Recently, somatic mutations of tyrosine 641 (Y641C, Y641F,
Y641N, Y641S and Y641H; sometimes also referred to as Y646C, Y646F,
Y646N, Y646S and Y646H, respectively) of EZH2 were reported to be
associated with follicular lymphoma (FL) and the germinal center B
cell-like (GCB) subtype of diffuse large B-cell lymphoma (DLBCL).
Morin et al. (2010) Nat Genet 42:181-5. In all cases, occurrence of
the mutant EZH2 gene was found to be heterozygous, and expression
of both wild-type and mutant alleles was detected in the mutant
samples profiled by transcriptome sequencing. It was also
demonstrated that all of the mutant forms of EZH2 could be
incorporated into the multi-protein PRC2 complex, but that the
resulting complexes lacked the ability to catalyze methylation of
the H3-K27 equivalent residue of a peptidic substrate. Hence, it
was concluded that the disease-associated changes at Tyr641 of EZH2
resulted in loss of function with respect to EZH2-catalyzed H3-K27
methylation.
SUMMARY OF THE INVENTION
[0010] In one aspect, the present invention features a substituted
benzene compound of Formula (I) below or a pharmaceutically
acceptable salt or ester thereof.
##STR00001##
In this formula: [0011] X.sub.1 is N or CR.sub.11; [0012] X.sub.2
is N or CR.sub.13; [0013] X.sub.3 is N or C, and when X.sub.3 is N,
R.sub.6 is absent; [0014] Z is NR.sub.7R.sub.8, OR.sub.7,
S(O).sub.aR.sub.7, or CR.sub.7R.sub.8R.sub.14, in which a is 0, 1,
or 2; [0015] each of R.sub.1, R.sub.5, R.sub.9, and R.sub.10,
independently, is H or C.sub.1-C.sub.6 alkyl optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano,
C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, 4 to 12-membered heterocycloalkyl, and 5- or
6-membered heteroaryl; [0016] each of R.sub.2, R.sub.3, and
R.sub.4, independently, is -Q.sub.1-T.sub.1, in which Q.sub.1 is a
bond or C.sub.1-C.sub.3 alkyl linker optionally substituted with
halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H,
halo, hydroxyl, COOH, cyano, azido, or R.sub.S1, in which R.sub.S1
is C.sub.1-C.sub.3 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0017] R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)R.sub.a, --C(O)OR.sub.a,
--C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl; b is 0, 1, or 2; each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; [0018] R.sub.7
is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4
alkyl linker, or C.sub.2-C.sub.4 alkenyl linker, each linker
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.h,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 14-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
[0019] each of R.sub.8, R.sub.11, R.sub.12, and R.sub.13,
independently, is H, halo, hydroxyl, COOH, cyano, R.sub.S8,
OR.sub.S8, or COOR.sub.S8, in which R.sub.S8 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered heterocycloalkyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, or di-C.sub.1-C.sub.6
alkylamino, and R.sub.S8 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl ring or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and [0020] R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0021] provided that the compound is not [0022]
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide, [0023]
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide, [0024]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide, [0025]
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimethyl--
2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide, [0026]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide, [0027]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimethoxybe-
nzamide, [0028]
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide, [0029]
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide, [0030]
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, or [0031]
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
[0032] In another aspect, the present invention features a
substituted benzene compound of Formula (I) above or a
pharmaceutically acceptable salt or ester thereof, in which [0033]
X.sub.1 is N or CR.sub.11; [0034] X.sub.2 is N or CR.sub.13; [0035]
X.sub.3 is N or C, and when X.sub.3 is N, R.sub.6 is absent; [0036]
Z is NR.sub.7R.sub.8, OR.sub.7, S(O).sub.aR.sub.7, or
CR.sub.7R.sub.8R.sub.14, in which a is 0, 1, or 2; [0037] each of
R.sub.1, R.sub.5, R.sub.9, and R.sub.10, independently, is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0038] each of R.sub.2, R.sub.3, and R.sub.4, independently, is
-Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or C.sub.1-C.sub.3
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo, hydroxyl, COOH,
cyano, azido, or R.sub.S1, in which R.sub.S1 is C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0039] R.sub.6 is H, halo, cyano, azido, --NR.sub.aR.sub.b,
--C(O)R.sub.a, --C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b,
--NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl, b is 0, 1, or 2, each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; [0040] R.sub.7
is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4
alkyl linker, or C.sub.2-C.sub.4 alkenyl linker, each linker
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.h,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 14-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
provided that (i) R.sub.7 is not C(O)R.sub.g or --S(O).sub.2R.sub.g
when Z is NR.sub.7R.sub.8; (ii) R.sub.7 is not C.sub.1-C.sub.6
alkyl when Z is OR.sub.7, and (iii) R.sub.7 is not H; [0041] each
of R.sub.8, R.sub.11, R.sub.12, and R.sub.13, independently, is H,
halo, hydroxyl, COOH, cyano, R.sub.S8, OR.sub.S8, or COOR.sub.S8,
in which R.sub.S8 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, 4 to
12-membered heterocycloalkyl, amino, mono-C.sub.1-C.sub.6
alkylamino, or di-C.sub.1-C.sub.6 alkylamino, and R.sub.S8 is
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and [0042] R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl.
[0043] One subset of the compounds of Formula (I) includes those of
Formula (Ia):
##STR00002##
[0044] Another subset of the compounds of Formula (I) includes
those of Formula (Ib), (Ic), or (Id):
##STR00003##
[0045] Still another subset of the compounds of Formula (I)
includes those of Formula (Ie), (II) or (IIA):
##STR00004##
[0046] The compounds of Formulae (I), (Ia), (Ib), (Ic), (Id), (Ie),
(II), and (IIA) can include one or more of the following
features:
[0047] X.sub.1 is CR.sub.11 and X.sub.2 is CR.sub.13.
[0048] X.sub.1 is CR.sub.11 and X.sub.2 is N.
[0049] X.sub.1 is N and X.sub.2 is CR.sub.13.
[0050] X.sub.1 is N and X.sub.2 is N.
[0051] Z is NR.sub.7R.sub.8.
[0052] Z is CR.sub.7R.sub.8R.sub.14.
[0053] Z is OR.sub.7.
[0054] Z is S(O).sub.aR.sub.7, in which a is 0, 1, or 2.
[0055] Z is SR.sub.7.
[0056] R.sub.6 is halo, C.sub.1-C.sub.3 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.3-C.sub.6 cycloalkyl, C(O)H, or --C(O)R.sub.a, in
which R.sub.a is C.sub.1-C.sub.6 alkyl or 4 to 12-membered (e.g., 4
to 7-membered) heterocycloalkyl.
[0057] R.sub.6 is 4 to 12-membered (e.g., 4 to 7-membered)
heterocycloalkyl optionally substituted with one or more
-Q.sub.2-T.sub.2, in which -Q.sub.2-T.sub.2 is oxo or Q.sub.2 is a
bond and T.sub.2 is C.sub.1-C.sub.6 alkyl, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, or 4 to 12-membered (e.g., 4 to 7-membered)
heterocycloalkyl optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0058] -Q.sub.2-T.sub.2 is not H.
[0059] Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker and
T.sub.3 is selected from the group consisting of C.sub.1-C.sub.3
alkyl, OR.sub.e, --S(O).sub.2R.sub.e, or --NR.sub.eR.sub.f.
[0060] R.sub.6 is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,
1,2,3,6-tetrahydropyridinyl,
2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,
morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, or
pyrrolidinyl.
[0061] R.sub.6 is F, Br, or Cl.
[0062] R.sub.7 is not H.
[0063] R.sub.7 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl
or 4 to 14-membered (e.g., 4 to 7-membered) heterocycloalkyl, each
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0064] R.sub.7 is piperidinyl, tetrahydropyran,
tetrahydrothiopyran, oxetanyl, azetidinyl, pyrrolidinyl,
1,4-dioxaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-3'-one-4-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-7'-one-4-yl,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-3'-one-4-yl,
1-azaspiro[4.5]decan-2-one-8-yl, cyclobutyl, cyclopentyl, or
cyclohexyl, each optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0065] T.sub.5 is H, halo, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkoxyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, or 4 to
12-membered (e.g., 4 to 7-membered) heterocycloalkyl.
[0066] Q.sub.5 is a bond and T.sub.5 is C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered (e.g., 4 to
7-membered) heterocycloalkyl.
[0067] Q.sub.5 is a bond and T.sub.5 is H, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered heterocycloalkyl, 5-
or 6-membered heteroaryl, amino, mono-C.sub.1-C.sub.6 alkylamino,
or di-C.sub.1-C.sub.6 alkylamino, T.sub.5 being optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, C.sub.1-C.sub.6 alkoxyl, or
C.sub.3-C.sub.8 cycloalkyl.
[0068] Q.sub.5 is CO, S(O).sub.2, or NHC(O); and T.sub.5 is
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8
cycloalkyl, or 4 to 12-membered (e.g., 4 to 7-membered)
heterocycloalkyl.
[0069] Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and T.sub.5 is H or
C.sub.6-C.sub.10 aryl.
[0070] Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and T.sub.5 is
C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered (e.g., 4 to
7-membered) heterocycloalkyl, or S(O).sub.qR.sub.q.
[0071] R.sub.11 is H.
[0072] R.sub.7 is cyclopentyl or cyclohexyl, each optionally
substituted with one -Q.sub.5-T.sub.5.
[0073] Q.sub.5 is NHC(O) and T.sub.5 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxy.
[0074] R.sub.7 is isopropyl.
[0075] Each of R.sub.2 and R.sub.4, independently is
C.sub.1-C.sub.3 alkyl optionally substituted with C.sub.1-C.sub.6
alkoxyl.
[0076] Each of R.sub.2 and R.sub.4 is methyl.
[0077] R.sub.1 is H.
[0078] R.sub.12 is H, methyl, ethyl, ethenyl, or halo.
[0079] R.sub.12 is methyl.
[0080] R.sub.12 is ethyl.
[0081] R.sub.12 is ethenyl.
[0082] R.sub.8 is H, methyl, or ethyl.
[0083] R.sub.8 is methyl.
[0084] R.sub.8 is ethyl.
[0085] R.sub.8 is ethenyl or propenyl.
[0086] R.sub.8 is 4 to 12-membered (e.g., 4 to 7-membered)
heterocycloalkyl, such as piperidinyl or tetrahydropyranyl.
[0087] Z is NR.sub.7R.sub.8 or CR.sub.7R.sub.8R.sub.14 wherein
R.sub.7 and R.sub.8, together with the atom to which they are
attached, form a ring selected from the group consisting of
piperidinyl, morpholinyl, piperazinyl, azetidinyl, pyrrolidinyl,
2-oxa-5-azabicyclo[2.2.1]heptan-5-yl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, each optionally
substituted with one -Q.sub.6-T.sub.6.
[0088] R.sub.13 is H or methyl.
[0089] R.sub.13 is H.
[0090] R.sub.3 is H.
[0091] Z is NR.sub.7R.sub.8, OR.sub.7, or S(O).sub.aR.sub.7;
R.sub.1 is H or C.sub.1-C.sub.6 alkyl optionally substituted with
one or more substituents selected from hydroxyl, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, and C.sub.6-C.sub.10 aryl; each of R.sub.2 and R.sub.4
independently, is -Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or
C.sub.1-C.sub.3 alkyl linker optionally substituted with one or
more substituents selected from halo and hydroxyl, and T.sub.1 is
H, halo, or azido; R.sub.3 is H or halo; R.sub.5 is H or
C.sub.1-C.sub.6 alkyl; R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)NR.sub.aR.sub.b, --S(O).sub.bR.sub.a, or
R.sub.S2; wherein R.sub.S2 is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, or 4 to
12-membered heterocycloalkyl, and wherein each of R.sub.a and
R.sub.b, independently is H, C.sub.1-C.sub.6 alkyl, or 4 to
12-membered heterocycloalkyl; or R.sub.a and R.sub.b, together with
the N atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 or 1 additional heteroatom; and each
of R.sub.a, R.sub.S2 and the 4 to 12-membered heterocycloalkyl ring
formed by R.sub.a and R.sub.b, is optionally substituted with one
or more -Q.sub.2-T.sub.2; wherein Q.sub.2 is a bond or
C.sub.1-C.sub.3 alkyl linker, and T.sub.2 is H, halo, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)OR.sub.c, or C.sub.6 alkyl, in which each
of R.sub.c and R.sub.d, independently is H or C.sub.1-C.sub.6
alkyl, or R.sub.c and R.sub.d, together with the N atom to which
they are attached, form a 4 to 12-membered heterocycloalkyl ring
having 0 or 1 additional heteroatom and optionally substituted with
C.sub.1-C.sub.6 alkyl; R.sub.7 is -Q.sub.4-T.sub.4, in which
Q.sub.4 is a bond or C.sub.1-C.sub.4 alkyl linker and T.sub.4 is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, C(O)--C.sub.1-6 alkyl, C(O)--C.sub.3-6 cycloalkyl, or 4 to
14-membered heterocycloalkyl, each optionally substituted with one
or more -Q.sub.5-T.sub.5; R.sub.8 is H, C.sub.1-C.sub.6 alkyl
optionally substituted with halo, hydroxyl or C.sub.1-C.sub.6
alkoxyl, C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.8 cycloalkyl, or 4
to 7-membered heterocycloalkyl; or R.sub.7 and R.sub.8, together
with the N atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 to 2 additional heteroatoms, and the
4 to 12-membered heterocycloalkyl ring formed by R.sub.7 and
R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6; and R.sub.12 is halo, C.sub.1-C.sub.6 alkoxyl, or
C.sub.1-C.sub.6 alkyl optionally substituted with halo or
C.sub.2-C.sub.6 alkenyl.
[0092] R.sub.1 is H or C.sub.1-C.sub.6 alkyl optionally substituted
one or more times with a substituent selected from hydroxyl,
C.sub.1-C.sub.6 alkoxyl and C.sub.6-C.sub.10 aryl; R.sub.7 is
-Q.sub.4-T.sub.4, in which Q.sub.4 is a bond or a C.sub.1-C.sub.4
alkyl linker, and T.sub.4 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, C(O)-- C.sub.1-C.sub.6 alkyl,
C(O)-- C.sub.3-C.sub.6 cycloalkyl, dihydropyranyl,
tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl,
pyrrolidinyl, azetidinyl and oxetanyl, each optionally substituted
with one or more substituents independently selected from oxo and
-Q.sub.5-T.sub.5; R.sub.8 is H or C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.2-C.sub.6 alkenyl or
C.sub.3-C.sub.8 cycloalkyl; and R.sub.12 is halo, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl or C.sub.1-C.sub.6 alkoxyl.
[0093] Z is NR.sub.7R.sub.8 or SR.sub.7; R.sub.6 is H, halo, cyano,
OR.sub.a, --C(O)NR.sub.aR.sub.b, --S(O).sub.2R.sub.a, or R.sub.S2;
wherein R.sub.S2 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, or 4 to 12-membered heterocycloalkyl, and
wherein each of R.sub.a and R.sub.b, independently is H or
C.sub.1-C.sub.6 alkyl; or R.sub.a and R.sub.b, together with the N
atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 or 1 additional heteroatom; and each
of R.sub.S2 and the 4 to 12-membered heterocycloalkyl ring formed
by R.sub.a and R.sub.b, is optionally substituted with one or more
-Q.sub.2-T.sub.2; wherein Q.sub.2 is a bond or C.sub.1-C.sub.3
alkyl linker, and T.sub.2 is H, halo, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)OC.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6
alkyl, in which each of R.sub.c and R.sub.d, independently is H or
C.sub.1-C.sub.6 alkyl, or R.sub.c and R.sub.d, together with the N
atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 or 1 additional heteroatom and 0 or
1 C.sub.1-C.sub.6 alkyl substituents; R.sub.7 is C.sub.1-C.sub.6
alkyl, C.sub.3-C.sub.8 cycloalkyl, dihydropyranyl,
tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl,
pyrrolidinyl, azetidinyl and oxetanyl, each optionally substituted
with one or more substituents independently selected from oxo and
-Q.sub.5-T.sub.5; and R.sub.12 is halo or C.sub.1-C.sub.6
alkyl.
[0094] R.sub.2, R.sub.4 and R.sub.12 are each independently
C.sub.1-C.sub.6 alkyl and R.sub.5 is H.
[0095] R.sub.7 is cyclohexyl, dihydropyranyl, tetrahydropyranyl,
tetrahydrothiopyranyl, piperidinyl, pyrrolidinyl, azetidinyl
oxetanyl, 1,4-dioxaspiro[4.5]decan-8-yl, 1-oxaspiro[4.5]decan-8-yl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-4-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-4-yl,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-4-yl, or
1-azaspiro[4.5]decan-8-yl, each substituted with one or more
-Q.sub.5-T.sub.5.
[0096] Z is selected from the group consisting of piperidinyl,
morpholinyl, piperazinyl, azetidinyl, pyrrolidinyl,
2-oxa-5-azabicyclo[2.2.1]heptan-5-yl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, and cyclohexenyl, each
optionally substituted with one -Q.sub.6-T.sub.6.
[0097] R.sub.6 is halo and Z is S(O).sub.aR.sub.7, in which a is 0,
1, or 2 and R.sub.7 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl or 4 to 7-membered heterocycloalkyl and R.sub.7 is
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0098] R.sub.6 is halo and Z is OR.sub.7, in which R.sub.7 is 4 to
7-membered heterocycloalkyl optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0099] R.sub.6 is --S(O).sub.bR.sub.a or azido, in which b is 0, 1,
or 2 and R.sub.a is C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.8
cycloalkyl; and Z is NR.sub.7R.sub.8, in which R.sub.7 is
C.sub.3-C.sub.8 cycloalkyl or 4 to 14-membered heterocycloalkyl,
each optionally substituted with one or more -Q.sub.5-T.sub.5; and
R.sub.8 is H or C.sub.1-C.sub.6 alkyl.
[0100] R.sub.6 is halo and Z is NR.sub.7R.sub.8 or
CR.sub.7R.sub.8R.sub.14 wherein R.sub.7 and R.sub.8, together with
the atom to which they are attached, form a 4 to 11-membered
heterocycloalkyl ring having 1 to 3 heteroatoms or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0101] U is O, S, N-Q.sub.5-T.sub.5, or CH-Q.sub.5-T.sub.5.
[0102] n is 0, 1, or 2.
[0103] R.sub.12 is Cl, Br, or methyl.
[0104] One or more -Q.sub.5-T.sub.5 are oxo.
[0105] U is CH-Q.sub.5-T.sub.5 and n is 0
[0106] One or more -Q.sub.6-T.sub.6 are oxo.
[0107] Q.sub.6 is a bond or C(O) and T.sub.6 is C.sub.1-C.sub.6
alkyl or C alkoxy.
[0108] The present invention also provides pharmaceutical
compositions comprising one or more pharmaceutically acceptable
carriers and one or more compounds selected from those of any of
the Formulae described herein, and
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide,
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide,
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimet-
hyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimeth-
oxybenzamide,
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide,
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide,
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, and
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
[0109] Another aspect of this invention is a method of treating or
preventing cancer. The method includes administering to a subject
in need thereof a therapeutically effective amount of one or more
compounds selected from those of any of the Formulae described
herein, and
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide,
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide,
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimet-
hyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide,
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimeth-
oxybenzamide,
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide,
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide,
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, and
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
[0110] Unless otherwise stated, any description of a method of
treatment includes uses of the compounds to provide such treatment
or prophylaxis as is described in the specification, as well as
uses of the compounds to prepare a medicament to treat or prevent
such condition. The treatment includes treatment of human or
non-human animals including rodents and other disease models.
[0111] For example, the method comprises the step of administering
to a subject having a cancer with aberrant H3-K27 methylation an
effective amount of one or more compounds of Formulae described
herein, wherein the compound(s) inhibits histone methyltransferase
activity of EZH2, thereby treating the cancer. Examples of aberrant
H3-K27 methylation may include a global increase in and/or altered
distribution of H3-K27 di or tri-methylation within the cancer cell
chromatin.
[0112] For example, the cancer is selected from the group
consisting of cancers that overexpress EZH2 or other PRC2 subunits,
contain loss-of-function mutations in H3-K27 demethylases such as
UTX, or overexpress accessory proteins such as PHF19/PCL3 capable
of increasing and or mislocalizing EZH2 activity (see references in
Sneeringer et al. Proc Natl Acad Sci USA 107(49):20980-5,
2010).
[0113] For example, the method comprises the step of administering
to a subject having a cancer overexpressing EZH2 a therapeutically
effective amount of one or more compounds of Formulae described
herein, wherein the compound(s) inhibits histone methyltransferase
activity of EZH2, thereby treating the cancer.
[0114] For example, the method comprises the step of administering
to a subject having a cancer with a loss-of-function mutation in
the H3-K27 demethylase UTX a therapeutically effective amount of
one or more compounds of Formulae described herein, wherein the
compound(s) inhibits histone methyltransferase activity of EZH2,
thereby treating the cancer
[0115] For example, the method comprises the step of administering
to a subject having a cancer overexpressing an accessory
component(s) of the PRC2, such as PHF19/PCL3, a therapeutically
effective amount of one or more compounds of Formulae described
herein, wherein the compound(s) inhibits histone methyltransferase
activity of EZH2, thereby treating the cancer
[0116] In still another aspect, this invention relates to a method
of modulating the activity of the wild-type EZH2, the catalytic
subunit of the PRC2 complex which catalyzes the mono-through
tri-methylation of lysine 27 on histone H3 (H3-K27). For example,
the present invention relates to a method of inhibiting the
activity of EZH2 in a cell. This method can be conducted either in
vitro or in vivo.
[0117] In yet another aspect, this invention features to a method
of inhibiting in a subject conversion of H3-K27 to trimethylated
H3-K27. The method comprises administering to a subject a
therapeutically effective amount of one or more of the compound of
Formulae described herein to inhibit histone methyltransferase
activity of EZH2, thereby inhibiting conversion of H3-K27 to
trimethylated H3-K27 in the subject.
[0118] For example, the method comprises the step of administering
to a subject having a cancer expressing a Y641 mutant of EZH2 a
therapeutically effective amount of one or more compounds of
Formulae described herein, wherein the compound(s) inhibits histone
methyltransferase activity of EZH2, thereby treating the
cancer.
[0119] For example, the cancer is selected from the group
consisting of follicular lymphoma and diffuse large B-cell lymphoma
(DLBCL) of germinal center B cell-like (GCB) subtype. For example,
the cancer is lymphoma, leukemia or melanoma. Preferably, the
lymphoma is non-Hodgkin lymphoma, follicular lymphoma or diffuse
large B-cell lymphoma. Alternatively, the leukemia is chronic
myelogenous leukemia (CML), acute myeloid leukemia, acute
lymphocytic leukemia or mixed lineage leukemia.
[0120] The precancerous condition is myelodysplastic syndromes
(MDS, formerly known as preleukemia).
[0121] For example, the cancer is a hematological cancer.
[0122] For example, the method comprises the step of administering
to a subject having a cancer expressing a Y641 mutant of EZH2 a
therapeutically effective amount of one or more compounds of
Formulae described herein, wherein the compound(s) selectively
inhibits histone methyltransferase activity of the Y641 mutant of
EZH2, thereby treating the cancer.
[0123] For example, the method further comprises the steps of
performing an assay to detect a Y641 mutant of EZH2 in a sample
comprising cancer cells from a subject having a cancer.
[0124] In still another aspect, this invention relates to a method
of modulating the activity of the wild-type and mutant histone
methyltransferase EZH2, the catalytic subunit of the PRC2 complex
which catalyzes the mono- through tri-methylation of lysine 27 on
histone H3 (H3-K27). For example, the present invention relates to
a method of inhibiting the activity of certain mutant forms of EZH2
in a cell. The mutant forms of EZH2 include a substitution of
another amino acid residue for tyrosine 641 (Y641, also Tyr641) of
wild-type EZH2. The method includes contacting the cell with an
effective amount of one or more of the compound of any Formula
described herein. This method can be conducted either in vitro or
in vivo.
[0125] In yet another aspect, this invention features to a method
of inhibiting in a subject conversion of H3-K27 to trimethylated
H3-K27. The method comprises administering to a subject expressing
a Y641 mutant of EZH2 a therapeutically effective amount of one or
more of the compound of any Formula described herein to inhibit
histone methyltransferase activity of EZH2, thereby inhibiting
conversion of H3-K27 to trimethylated H3-K27 in the subject. For
example, the histone methyltransferase activity inhibited is that
of the Y641 mutant of EZH2. For example, the compound of this
invention selectively inhibits histone methyltransferase activity
of the Y641 mutant of EZH2. For example, the Y641 mutant of EZH2 is
selected from the group consisting of Y641C, Y641F, Y641H, Y641N,
and Y641S.
[0126] The method of inhibiting in a subject conversion of H3-K27
to trimethylated H3-K27 may also comprise performing an assay to
detect a Y641 mutant of EZH2 in a sample from a subject before
administering to the subject expressing a Y641 mutant of EZH2 a
therapeutically effective amount of one or more of the compound of
any Formula described herein. For example, performing the assay to
detect the Y641 mutant of EZH2 includes whole-genome resequencing
or target region resequencing that detects a nucleic acid encoding
the Y641 mutant of EZH2. For example, performing the assay to
detect the Y641 mutant of EZH2 includes contacting the sample with
an antibody that binds specifically to a polypeptide or fragment
thereof characteristic of the Y641 mutant of EZH2. For example,
performing the assay to detect the Y641 mutant of EZH2 includes
contacting the sample under highly stringent conditions with a
nucleic acid probe that hybridizes to a nucleic acid encoding a
polypeptide or fragment thereof characteristic of the Y641 mutant
of EZH2.
[0127] Further, the invention also relates to a method of
identifying an inhibitor of a Y641 mutant of EZH2. The method
comprises the steps of combining an isolated Y641 mutant of EZH2
with a histone substrate, a methyl group donor, and a test
compound, wherein the histone substrate comprises a form of H3-K27
selected from the group consisting of unmethylated H3-K27,
monomethylated H3-K27, dimethylated H3-K27, and any combination
thereof; and performing an assay to detect methylation of H3-K27
(e.g., formation of trimethylated H3-K27) in the histone substrate,
thereby identifying the test compound as an inhibitor of the Y641
mutant of EZH2 when methylation of H3-K27 (e.g., formation of
trimethylated H3-K27) in the presence of the test compound is less
than methylation of H3-K27 (e.g., formation of trimethylated
H3-K27) in the absence of the test compound.
[0128] In one embodiment, performing the assay to detect
methylation of H3-K27 in the histone substrate comprises measuring
incorporation of labeled methyl groups.
[0129] In one embodiment, the labeled methyl groups are
isotopically labeled methyl groups.
[0130] In one embodiment, performing the assay to detect
methylation of H3-K27 in the histone substrate comprises contacting
the histone substrate with an antibody that binds specifically to
trimethylated H3-K27.
[0131] Also within the scope of the invention is a method of
identifying a selective inhibitor of a Y641 mutant of EZH2. The
method comprises the steps of combining an isolated Y641 mutant of
EZH2 with a histone substrate, a methyl group donor, and a test
compound, wherein the histone substrate comprises a form of H3-K27
selected from the group consisting of monomethylated H3-K27,
dimethylated H3-K27, and a combination of monomethylated H3-K27 and
dimethylated H3-K27, thereby forming a test mixture; combining an
isolated wild-type EZH2 with a histone substrate, a methyl group
donor, and a test compound, wherein the histone substrate comprises
a form of H3-K27 selected from the group consisting of
monomethylated H3-K27, dimethylated H3-K27, and a combination of
monomethylated H3-K27 and dimethylated H3-K27, thereby forming a
control mixture; performing an assay to detect trimethylation of
the histone substrate in each of the test mixture and the control
mixture; calculating the ratio of (a) trimethylation with the Y641
mutant of EZH2 and the test compound (M+) to (b) trimethylation
with the Y641 mutant of EZH2 without the test compound (M-);
calculating the ratio of (c) trimethylation with wild-type EZH2 and
the test compound (WT+) to (d) trimethylation with wild-type EZH2
without the test compound (WT-); comparing the ratio (a)/(b) with
the ratio (c)/(d); and identifying the test compound as a selective
inhibitor of the Y641 mutant of EZH2 when the ratio (a)/(b) is less
than the ratio (c)/(d).
[0132] The present invention further provides a method of
identifying a subject as a candidate for treatment with one or more
compounds of the invention. The method comprises the steps of
performing an assay to detect a Y641 mutant of EZH2 in a sample
from a subject; and identifying a subject expressing a Y641 mutant
of EZH2 as a candidate for treatment with one or more compounds of
the invention, wherein the compound(s) inhibits histone
methyltransferase activity of EZH2.
[0133] Still another aspect of the invention is a method of
inhibiting conversion of H3-K27 to trimethylated H3-K27. The method
comprises the step of contacting a Y641 mutant of EZH2 with a
histone substrate comprising H3-K27 and an effective amount of a
compound of the present invention, wherein the compound inhibits
histone methyltransferase activity of EZH2, thereby inhibiting
conversion of H3-K27 to trimethylated H3-K27.
[0134] Further, the compounds or methods described herein can be
used for research (e.g., studying epigenetic enzymes) and other
non-therapeutic purposes.
[0135] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. In the
specification, the singular forms also include the plural unless
the context clearly dictates otherwise. Although methods and
materials similar or equivalent to those described herein can be
used in the practice or testing of the present invention, suitable
methods and materials are described below. All publications, patent
applications, patents and other references mentioned herein are
incorporated by reference. The references cited herein are not
admitted to be prior art to the claimed invention. In the case of
conflict, the present specification, including definitions, will
control. In addition, the materials, methods and examples are
illustrative only and are not intended to be limiting.
[0136] Other features and advantages of the invention will be
apparent from the following detailed description and claims.
BRIEF DESCRIPTIONS OF FIGURES
[0137] FIG. 1 (A) is an idealized plot of cell count (i.e., cell
number) as a function of time showing exponential proliferation
during log-phase cell growth.
[0138] FIG. 1(B) is an idealized plot of ln(cell count) as a
function of time for the data from panel (A).
[0139] FIG. 2 is a graph showing biphasic cell growth curves in the
presence of an antiproliferative compound for which there is a
delay before the impact of the compound on cell growth is realized.
The compound begins to affect cell growth at the time point labeled
"start of impact." The solid circles represent idealized data for
the vehicle (or solvent) control sample that is not treated with
compound. The other symbols represent biphasic growth curves for
cells treated with different concentrations of compound (i.e.,
drug).
[0140] FIG. 3 is a replot of k.sub.p as a function of compound
concentration for (A) a cytostatic and (B) a cytotoxic compound,
illustrating the graphic determination of the LCC for a cytotoxic
agent. Note that for a cytostatic compound (panel A), the value of
k.sub.p can never drop below zero.
[0141] FIG. 4 is a diagram showing tumor growth of WSU-DLCL2
xenograft bearing mice treated with Compound 222 over 27 days.
[0142] FIG. 5 is a diagram showing lobal H3K27me3 methylation in
WSU-DLCL2 tumors from mice treated with Compound 222 or vehicle for
27 days.
DETAILED DESCRIPTION OF THE INVENTION
[0143] The present invention provides novel substituted benzene
compounds, synthetic methods for making the compounds,
pharmaceutical compositions containing them and various uses of the
compounds.
1. SUBSTITUTED BENZENE COMPOUNDS
[0144] The present invention provides the compounds of Formula
(I):
##STR00005##
[0145] In this formula: [0146] X.sub.1 is N or CR.sub.11; [0147]
X.sub.2 is N or CR.sub.13; [0148] X.sub.3 is N or C, and when
X.sub.3 is N, R.sub.6 is absent; [0149] Z is NR.sub.7R.sub.8,
OR.sub.7, S(O).sub.aR.sub.7, or CR.sub.7R.sub.8R.sub.14, in which a
is 0, 1, or 2; [0150] each of R.sub.1, R.sub.5, R.sub.9, and
R.sub.10, independently, is H or C.sub.1-C.sub.6 alkyl optionally
substituted with one or more substituents selected from the group
consisting of halo, hydroxyl, COOH, C(O)O--C.sub.1-C.sub.6 alkyl,
cyano, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl; [0151] each of
R.sub.2, R.sub.3, and R.sub.4, independently, is -Q.sub.1-T.sub.1,
in which Q.sub.1 is a bond or C.sub.1-C.sub.3 alkyl linker
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo, hydroxyl, COOH,
cyano, azido, or R.sub.S1, in which R.sub.S1 is C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0152] R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)R.sub.a, --C(O)OR.sub.a,
--C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl; b is 0, 1, or 2, each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.6 alkoxy, and
T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; [0153] R.sub.7
is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4
alkyl linker, or C.sub.2-C.sub.4 alkenyl linker, each linker
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.h,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
[0154] each of R.sub.8, R.sub.11, R.sub.12, and R.sub.13,
independently, is H, halo, hydroxyl, COOH, cyano, R.sub.S8,
OR.sub.S8, or COOR.sub.S8, in which R.sub.S8 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered heterocycloalkyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, or di-C.sub.1-C.sub.6
alkylamino, and R.sub.S8 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and [0155] R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0156] provided that the compound is not [0157]
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide, [0158]
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide, [0159]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide, [0160]
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimethyl--
2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide, [0161]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide, [0162]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimethoxybe-
nzamide, [0163]
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide, [0164]
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide, [0165]
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, or [0166]
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
[0167] For example, X.sub.1 is CR.sub.11 and X.sub.2 is
CR.sub.13.
[0168] For example, X.sub.1 is CR.sub.11 and X.sub.2 is N.
[0169] For example, X.sub.1 is N and X.sub.2 is CR.sub.13.
[0170] For example, X.sub.1 is N and X.sub.2 is N.
[0171] For example, X.sub.3 is C.
[0172] For example, X.sub.3 is N and R.sub.6 is absent.
[0173] For example, Z is NR.sub.7R.sub.8.
[0174] For example, Z is CR.sub.7R.sub.8R.sub.14.
[0175] For example, Z is OR.sub.7.
[0176] For example, Z is S(O).sub.aR.sub.7, in which a is 0, 1, or
2
[0177] For example, Z is SR.sub.7.
[0178] For example, R.sub.6 is H.
[0179] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0180] For example, R.sub.6 is C.sub.1-C.sub.3 alkyl optionally
substituted with one or more -Q.sub.2-T.sub.2.
[0181] For example, R.sub.6 is CF.sub.3.
[0182] For example, R.sub.6 is C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, or C.sub.3-C.sub.6 cycloalkyl each
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0183] For example, R.sub.6 is ethenyl.
[0184] For example, R.sub.6 is ethynyl.
[0185] For example, R.sub.6 is ethynyl substituted with one or more
-Q.sub.2-T.sub.2, in which Q.sub.2 is a bond or C.sub.1-C.sub.3
alkyl linker and T.sub.2 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6
cycloalkyl, or 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
and morpholinyl, and the like) optionally substituted with one or
more -Q.sub.3-T.sub.3.
[0186] For example, R.sub.6 is azido.
[0187] For example, R.sub.6 is cyano.
[0188] For example, R.sub.6 is C(O)H.
[0189] For example, R.sub.6 is OR.sub.a or --C(O)R.sub.a.
[0190] For example, R.sub.a is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.2-T.sub.2.
[0191] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0192] For example, R.sub.6 is piperidinyl,
2,2,6,6-tetramethyl-piperidinyl, 1,2,3,6-tetrahydropyridinyl,
2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,
morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, or
pyrrolidinyl, each of which is optionally substituted with one or
more -Q.sub.2-T.sub.2.
[0193] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.2-T.sub.2, and
-Q.sub.2-T.sub.2 is oxo or Q.sub.2 is a bond and T.sub.2 is
--OR.sub.c, --NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.1-C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0194] For example, R.sub.6 is --NR.sub.aR.sub.b, --C(O)R.sub.a,
--C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a,
--SR.sub.a, --S(O).sub.2R.sub.a, or
--S(O).sub.2NR.sub.aR.sub.b.
[0195] For example, each of R.sub.a and R.sub.b, independently is
H, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.8 cycloalkyl optionally
substituted with one or more -Q.sub.2-T.sub.2.
[0196] For example, one of R.sub.a and R.sub.b is H.
[0197] For example, R.sub.a and R.sub.b, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.2-T.sub.2.
[0198] For example, -Q.sub.2-T.sub.2 is not H.
[0199] For example, -Q.sub.2-T.sub.2 is oxo.
[0200] For example, Q.sub.2 is a bond.
[0201] For example, Q.sub.2 is an unsubstituted C.sub.1-C.sub.3
alkyl linker.
[0202] For example, T.sub.2 is C.sub.1-C.sub.6 alkyl or
C.sub.6-C.sub.10 aryl, each optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0203] For example, T.sub.2 is an unsubstituted substituted
straight chain C.sub.1-C.sub.6 or branched C.sub.3-C.sub.6 alkyl,
including but not limited to, methyl, ethyl, n-propyl, i-propyl,
n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl and n-hexyl.
[0204] For example, T.sub.2 is phenyl.
[0205] For example, T.sub.2 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0206] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.3-T.sub.3.
[0207] For example, T.sub.2 is --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, or --S(O).sub.2R.sub.c.
[0208] For example, R.sub.c is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.3-T.sub.3.
[0209] For example, each of R.sub.c and R.sub.d, independently is H
or C.sub.1-C.sub.6 alkyl optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0210] For example, R.sub.c is H.
[0211] For example, R.sub.d is H.
[0212] For example, R.sub.c and R.sub.d, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0213] For example, Q.sub.2 is a bond and T.sub.2 is --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0214] For example, -Q.sub.3-T.sub.3 is oxo.
[0215] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl and one or more -Q.sub.3-T.sub.3 are
oxo.
[0216] For example, Q.sub.3 is a bond or unsubstituted or
substituted C.sub.1-C.sub.3 alkyl linker.
[0217] For example, T.sub.3 is H, halo, 4 to 7-membered
heterocycloalkyl, C.sub.1-C.sub.3 alkyl, OR.sub.e, COOR.sub.e,
--S(O).sub.2R.sub.e, --NR.sub.eR.sub.f, or
--C(O)NR.sub.eR.sub.f.
[0218] For example, one of R.sub.d and R.sub.e is H.
[0219] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, halo, OR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f.
[0220] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, OR.sub.e, --S(O).sub.2R.sub.e, or
--NR.sub.eR.sub.f.
[0221] For example, R.sub.e is H.
[0222] For example, R.sub.f is H.
[0223] For example, R.sub.7 is not H.
[0224] For example, R.sub.7 is --C(O)R.sub.g.
[0225] For example, R.sub.7 is --C(O)R.sub.g, in which R.sub.g is
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered heterocycloalkyl,
C.sub.3-C.sub.8 cycloalkyl.
[0226] For example, R.sub.7 is C.sub.6-C.sub.10 aryl substituted
with one or more -Q.sub.5-T.sub.5.
[0227] For example, R.sub.7 is phenyl optionally substituted with
one or more -Q.sub.5-T.sub.5.
[0228] For example, R.sub.7 is C.sub.1-C.sub.6 alkyl optionally
substituted with one or more -Q.sub.5-T.sub.5.
[0229] For example, R.sub.7 is C.sub.3-C.sub.8 cycloalkyl
optionally substituted with one or more Q.sub.5-T.sub.5.
[0230] For example, R.sub.7 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0231] For example, R.sub.7 is 8 to 14-membered heterocycloalkyl
such as 1,4-dioxaspiro[4.5]decanyl (e.g.,
1,4-dioxaspiro[4.5]decan-8-yl), 1,4-dioxa-8-azaspiro[4.5]decanyl
(e.g., 1,4-dioxa-8-azaspiro[4.5]decan-8-yl), 1-oxaspiro[4.5]decanyl
(e.g., 1-oxaspiro[4.5]decan-8-yl or
1-oxaspiro[4.5]decan-2-one-8-yl), 1-azaspiro[4.5]decanyl (e.g.,
1-azaspiro[4.5]decan-8-yl or 1-azaspiro[4.5]decan-2-one-8-yl),
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-4-yl or
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-3'-one-4-yl),
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl (e.g.,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-4-yl or
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-7'-one-4-yl), or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-4-yl or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-3'-one-4-yl), each
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0232] For example, R.sub.7 is 5 to 6-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0233] For example, R.sub.7 is isopropyl.
[0234] For example, R.sub.7 is pyrrolidinyl, piperidinyl,
tetrahydropyran, cyclopentyl, or cyclohexyl, cycloheptyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0235] For example, R.sub.7 is cyclopentyl or cyclohexyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0236] For example, Q.sub.5 is NHC(O) and T.sub.5 is
C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy.
[0237] For example, -Q.sub.5-T.sub.5 is oxo.
[0238] For example, T.sub.4 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl or C.sub.6-C.sub.10 aryl, and one or
more -Q.sub.5-T.sub.5 are oxo.
[0239] For example, R.sub.7 is 1-oxide-tetrahydro-2H-thiopyranyl or
1,1-dioxide-tetrahydro-2H-thiopyranyl.
[0240] For example, R.sub.7 is cyclohexanonyl, e.g.,
cyclohexanon-4-yl.
[0241] For example, T.sub.5 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0242] For example, Q.sub.5 is a bond and T.sub.5 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0243] For example, Q.sub.5 is a bond and T.sub.5 is 5- or
6-membered heteroaryl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, T.sub.5 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, C.sub.1-C.sub.6 alkoxyl, or C.sub.3-C.sub.8
cycloalkyl.
[0244] For example, Q.sub.5 is CO, S(O).sub.2, or NHC(O); and
T.sub.5 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0245] For example, T.sub.5 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0246] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is H or C.sub.6-C.sub.10 aryl.
[0247] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.qR.sub.q.
[0248] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is S(O).sub.aR.sub.7, in which a is 0,
1, or 2 and R.sub.7 is C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl), C.sub.3-C.sub.8 cycloalkyl
(e.g., cyclopentyl, cyclohexyl, or cycloheptyl) or 4 to 14-membered
heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0249] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is OR.sub.7 in which R.sub.7 is 4 to
14-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 1,4-dioxaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0250] For example, R.sub.11 is H.
[0251] For example, each of R.sub.2 and R.sub.4, independently, is
H or C.sub.1-C.sub.6 alkyl optionally substituted with amino,
azido, halo, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, or C.sub.6-C.sub.10 aryl.
[0252] For example, each of R.sub.2 and R.sub.4, independently is
C.sub.1-C.sub.3 alkyl optionally substituted with C.sub.1-C.sub.6
alkoxyl.
[0253] For example, each of R.sub.2 and R.sub.4 is methyl.
[0254] For example, R.sub.1 is H.
[0255] For example, R.sub.1 is C.sub.1-C.sub.6 alkyl optionally
substituted with azido, halo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, or C.sub.6-C.sub.10
aryl.
[0256] For example, R.sub.12 is H, methyl, ethyl, ethenyl, or
halo.
[0257] For example, R.sub.12 is methyl.
[0258] For example, R.sub.12 is ethyl or propenyl.
[0259] For example, R.sub.12 is methoxyl.
[0260] For example, R.sub.12 is ethenyl.
[0261] For example, R.sub.8 is H, methyl, ethyl, or ethenyl.
[0262] For example, R.sub.8 is methyl.
[0263] For example, R.sub.8 is ethyl.
[0264] For example, R.sub.8 is propyl.
[0265] For example, R.sub.8 is ethenyl or propenyl.
[0266] For example, R.sub.8 is C.sub.1-C.sub.6 alkyl substituted
with one or more substituents selected from the group consisting of
halo (e.g., F, Cl, or Br), hydroxyl, or C.sub.1-C.sub.6
alkoxyl.
[0267] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and
morpholinyl, and the like).
[0268] For example, R.sub.8 is piperidinyl.
[0269] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl and R.sub.7 is -Q.sub.4-T.sub.4, in
which Q.sub.4 is a bond or C.sub.1-C.sub.4 alkyl linker and T.sub.4
is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl or 4 to
7-membered heterocycloalkyl.
[0270] For example, Z is NR.sub.7R.sub.8 or CR.sub.7R.sub.8R.sub.14
wherein R.sub.7 and R.sub.8, together with the atom to which they
are attached, form a 4 to 11-membered heterocycloalkyl ring having
1 to 3 heteroatoms (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, morpholinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0271] For example, the ring formed by R.sub.7 and R.sub.8 is
selected from the group consisting of azetidinyl, pyrrolidinyl,
piperidinyl, morpholinyl, piperazinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, each optionally
substituted with one -Q.sub.6-T.sub.6.
[0272] For example, Z is 1,4-dioxa-8-azaspiro[4.5]decan-8-yl,
pyrrolidine-2,5-dione-1-yl, or piperidine-2,6-dione-1-yl.
[0273] For example, one or more -Q.sub.6-T.sub.6 is oxo.
[0274] For example, T.sub.6 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0275] For example, Q.sub.6 is a bond and T.sub.6 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0276] For example, Q.sub.6 is CO, S(O).sub.2, or NHC(O); and
T.sub.6 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0277] For example, T.sub.6 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0278] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is H or C.sub.6-C.sub.10 aryl.
[0279] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.pR.sub.p.
[0280] For example, each of R.sub.p and R.sub.q, independently, is
C.sub.1-C.sub.6 alkyl.
[0281] For example, R.sub.6 is --S(O).sub.bR.sub.a or azido, in
which b is 0, 1, or 2 and R.sub.a is C.sub.1-C.sub.6 alkyl or
C.sub.3-C.sub.8 cycloalkyl; and Z is NR.sub.7R.sub.8, in which
R.sub.7 is C.sub.3-C.sub.8 cycloalkyl (e.g., cyclopentyl,
cyclohexyl, or cycloheptyl) or 4 to 14-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like),
each optionally substituted with one or more -Q.sub.5-T.sub.5; and
R.sub.8 is H or C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl).
[0282] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is NR.sub.7R.sub.8 or
CR.sub.7R.sub.8R.sub.14 wherein R.sub.7 and R.sub.8, together with
the atom to which they are attached, form a 4 to 11-membered
heterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, pyrrolidine-2,5-dione-1-yl,
piperidine-2,6-dione-1-yl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0283] For example, R.sub.13 is H or methyl.
[0284] For example, R.sub.13 is H.
[0285] For example, R.sub.3 is H.
[0286] For example, each of R.sub.5, R.sub.9, and R.sub.10 is
H.
[0287] In another aspect, the present invention features a
substituted benzene compound of Formula (I) above or a
pharmaceutically acceptable salt or ester thereof, in which [0288]
X.sub.1 is N or CR.sub.11; [0289] X.sub.2 is N or CR.sub.13; [0290]
X.sub.3 is N or C, and when X.sub.3 is N, R.sub.6 is absent; [0291]
Z is NR.sub.7R.sub.8, OR.sub.7, S(O).sub.aR.sub.7, or
CR.sub.7R.sub.8R.sub.14, in which a is 0, 1, or 2; [0292] each of
R.sub.1, R.sub.5, R.sub.9, and R.sub.10, independently, is H or
C.sub.1-C.sub.6 alkyl optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0293] each of R.sub.2, R.sub.3, and R.sub.4, independently, is
-Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or C.sub.1-C.sub.3
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo, hydroxyl, COOH,
cyano, azido, or R.sub.S1, in which R.sub.S1 is C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0294] R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)R.sub.a, --C(O)OR.sub.a,
--C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl; b is 0, 1, or 2, each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo;
[0295] R.sub.7 is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond,
C.sub.1-C.sub.4 alkyl linker, or C.sub.2-C.sub.4 alkenyl linker,
each linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.h,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
provided that (i) R.sub.7 is not C(O)R.sub.g or --S(O).sub.2R.sub.g
when Z is NR.sub.7R.sub.8; (ii) R.sub.7 is not C.sub.1-C.sub.6
alkyl when Z is OR.sub.7, and (iii) R.sub.7 is not H; [0296] each
of R.sub.8, R.sub.11, R.sub.12, and R.sub.13, independently, is H,
halo, hydroxyl, COOH, cyano, R.sub.S8, OR.sub.S8, or COOR.sub.S8,
in which R.sub.S8 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, 4 to
12-membered heterocycloalkyl, amino, mono-C.sub.1-C.sub.6
alkylamino, or di-C.sub.1-C.sub.6 alkylamino, and R.sub.S8 is
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and [0297] R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl.
[0298] For example, X.sub.1 is CR.sub.11 and X.sub.2 is
CR.sub.13.
[0299] For example, X.sub.1 is CR.sub.11 and X.sub.2 is N.
[0300] For example, X.sub.1 is N and X.sub.2 is CR.sub.13.
[0301] For example, X.sub.1 is N and X.sub.2 is N.
[0302] For example, X.sub.3 is C.
[0303] For example, X.sub.3 is N and R.sub.6 is absent.
[0304] For example, Z is NR.sub.7R.sub.8.
[0305] For example, Z is CR.sub.7R.sub.8R.sub.14.
[0306] For example, Z is OR.sub.7.
[0307] For example, Z is S(O).sub.aR.sub.7, in which a is 0, 1, or
2.
[0308] For example, Z is SR.sub.7
[0309] For example, R.sub.6 is H.
[0310] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0311] For example, R.sub.6 is C.sub.1-C.sub.3 alkyl optionally
substituted with one or more -Q.sub.2-T.sub.2.
[0312] For example, R.sub.6 is CF.sub.3.
[0313] For example, R.sub.6 is C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, or C.sub.3-C.sub.6 cycloalkyl each
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0314] For example, R.sub.6 is ethenyl.
[0315] For example, R.sub.6 is ethynyl.
[0316] For example, R.sub.6 is ethynyl substituted with one or more
-Q.sub.2-T.sub.2, in which Q.sub.2 is a bond or C.sub.1-C.sub.3
alkyl linker and T.sub.2 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6
cycloalkyl, or 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
and morpholinyl, and the like) optionally substituted with one or
more -Q.sub.3-T.sub.3.
[0317] For example, R.sub.6 is azido.
[0318] For example, R.sub.6 is cyano.
[0319] For example, R.sub.6 is C(O)H.
[0320] For example, R.sub.6 is --C(O)R.sub.a.
[0321] For example, R.sub.a is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.2-T.sub.2.
[0322] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0323] For example, R.sub.6 is piperidinyl,
2,2,6,6-tetramethyl-piperidinyl, 1,2,3,6-tetrahydropyridinyl,
2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,
morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, or
pyrrolidinyl, each of which is optionally substituted with one or
more -Q.sub.2-T.sub.2.
[0324] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.2-T.sub.2, and
-Q.sub.2-T.sub.2 is oxo or Q.sub.2 is a bond and T.sub.2 is
--OR.sub.c, --NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.1-C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0325] For example, R.sub.6 is --NR.sub.aR.sub.b, --C(O)R.sub.a,
--C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a,
--SR.sub.a, --S(O).sub.2R.sub.a, or
--S(O).sub.2NR.sub.aR.sub.b.
[0326] For example, each of R.sub.a and R.sub.b, independently is
H, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.8 cycloalkyl optionally
substituted with one or more -Q.sub.2-T.sub.2.
[0327] For example, one of R.sub.a and R.sub.b is H.
[0328] For example, R.sub.a and R.sub.b, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.2-T.sub.2.
[0329] For example, -Q.sub.2-T.sub.2 is not H.
[0330] For example, -Q.sub.2-T.sub.2 is oxo.
[0331] For example, Q.sub.2 is a bond.
[0332] For example, Q.sub.2 is an unsubstituted C.sub.1-C.sub.3
alkyl linker.
[0333] For example, T.sub.2 is C.sub.1-C.sub.6 alkyl or
C.sub.6-C.sub.10 aryl, each optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0334] For example, T.sub.2 is an unsubstituted substituted
straight chain C.sub.1-C.sub.6 or branched C.sub.3-C.sub.6 alkyl,
including but not limited to, methyl, ethyl, n-propyl, i-propyl,
n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl and n-hexyl.
[0335] For example, T.sub.2 is phenyl.
[0336] For example, T.sub.2 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0337] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.3-T.sub.3.
[0338] For example, T.sub.2 is --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, or --S(O).sub.2R.sub.c.
[0339] For example, R.sub.c is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.3-T.sub.3.
[0340] For example, each of R.sub.c and R.sub.d, independently is H
or C.sub.1-C.sub.6 alkyl optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0341] For example, R.sub.c is H.
[0342] For example, R.sub.d is H.
[0343] For example, R.sub.c and R.sub.d, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0344] For example, Q.sub.2 is a bond and T.sub.2 is --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.1-C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0345] For example, -Q.sub.3-T.sub.3 is oxo.
[0346] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl and one or more -Q.sub.3-T.sub.3 are
oxo.
[0347] For example, Q.sub.3 is a bond or unsubstituted or
substituted C.sub.1-C.sub.3 alkyl linker.
[0348] For example, T.sub.3 is H, halo, 4 to 7-membered
heterocycloalkyl, C.sub.1-C.sub.3 alkyl, OR.sub.e, COOR.sub.e,
--S(O).sub.2R.sub.e, --NR.sub.eR.sub.f, or
--C(O)NR.sub.eR.sub.f.
[0349] For example, one of R.sub.d and R.sub.e is H.
[0350] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, halo, OR.sub.e, --S(O).sub.2R.sub.e, and
--C(O)NR.sub.eR.sub.f.
[0351] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, OR.sub.e, --S(O).sub.2R.sub.e, or
--NR.sub.eR.sub.f.
[0352] For example, R.sub.e is H.
[0353] For example, R.sub.f is H.
[0354] For example, R.sub.7 is not H.
[0355] For example, R.sub.7 is --C(O)R.sub.g.
[0356] For example, R.sub.7 is --C(O)R.sub.g, in which R.sub.g is
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered heterocycloalkyl,
C.sub.3-C.sub.8 cycloalkyl.
[0357] For example, R.sub.7 is C.sub.6-C.sub.10 aryl substituted
with one or more -Q.sub.5-T.sub.5.
[0358] For example, R.sub.7 is phenyl optionally substituted with
one or more -Q.sub.5-T.sub.5.
[0359] For example, R.sub.7 is C.sub.1-C.sub.6 alkyl optionally
substituted with one or more -Q.sub.5-T.sub.5.
[0360] For example, R.sub.7 is C.sub.3-C.sub.8 cycloalkyl
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0361] For example, R.sub.7 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0362] For example, R.sub.7 is 8 to 14-membered heterocycloalkyl
such as 1,4-dioxaspiro[4.5]decanyl (e.g.,
1,4-dioxaspiro[4.5]decan-8-yl), 1,4-dioxa-8-azaspiro[4.5]decanyl
(e.g., 1,4-dioxa-8-azaspiro[4.5]decan-8-yl), 1-oxaspiro[4.5]decanyl
(e.g., 1-oxaspiro[4.5]decan-8-yl or
1-oxaspiro[4.5]decan-2-one-8-yl), 1-azaspiro[4.5]decanyl (e.g.,
1-azaspiro[4.5]decan-8-yl or 1-azaspiro[4.5]decan-2-one-8-yl),
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-4-yl or
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-3'-one-4-yl),
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl (e.g.,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-4-yl or
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-7'-one-4-yl), or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-4-yl or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-3'-one-4-yl), each
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0363] For example, R.sub.7 is 5 to 6-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0364] For example, R.sub.7 is isopropyl.
[0365] For example, R.sub.7 is pyrrolidinyl, piperidinyl,
tetrahydropyran, cyclopentyl, or cyclohexyl, cycloheptyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0366] For example, R.sub.7 is cyclopentyl or cyclohexyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0367] For example, Q.sub.5 is NHC(O) and T.sub.5 is
C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy.
[0368] For example, -Q.sub.5-T.sub.5 is oxo.
[0369] For example, T.sub.4 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl or C.sub.6-C.sub.10 aryl, and one or
more -Q.sub.5-T.sub.5 are oxo.
[0370] For example, R.sub.7 is 1-oxide-tetrahydro-2H-thiopyranyl or
1,1-dioxide-tetrahydro-2H-thiopyranyl.
[0371] For example, R.sub.7 is cyclohexanonyl, e.g.,
cyclohexanon-4-yl.
[0372] For example, T.sub.5 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0373] For example, Q.sub.5 is a bond and T.sub.5 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0374] For example, Q.sub.5 is a bond and T.sub.5 is 5- or
6-membered heteroaryl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, T.sub.5 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, C.sub.1-C.sub.6 alkoxyl, or C.sub.3-C.sub.8
cycloalkyl.
[0375] For example, Q.sub.5 is CO, S(O).sub.2, or NHC(O); and
T.sub.5 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0376] For example, T.sub.5 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0377] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is H or C.sub.6-C.sub.10 aryl.
[0378] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.qR.sub.q.
[0379] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is S(O).sub.aR.sub.7, in which a is 0,
1, or 2 and R.sub.7 is C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl), C.sub.3-C.sub.8 cycloalkyl
(e.g., cyclopentyl, cyclohexyl, or cycloheptyl) or 4 to 14-membered
heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0380] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is OR.sub.7 in which R.sub.7 is 4 to
14-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 1,4-dioxaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0381] For example, R.sub.11 is H.
[0382] For example, each of R.sub.2 and R.sub.4, independently, is
H or C.sub.1-C.sub.6 alkyl optionally substituted with amino,
azido, halo, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, or C.sub.6-C.sub.10 aryl.
[0383] For example, each of R.sub.2 and R.sub.4, independently is
C.sub.1-C.sub.3 alkyl optionally substituted with C.sub.1-C.sub.6
alkoxyl.
[0384] For example, each of R.sub.2 and R.sub.4 is methyl.
[0385] For example, R.sub.1 is H.
[0386] For example, R.sub.1 is C.sub.1-C.sub.6 alkyl optionally
substituted with azido, halo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, or C.sub.6-C.sub.10
aryl.
[0387] For example, R.sub.12 is H, methyl, ethyl, ethenyl, or
halo.
[0388] For example, R.sub.12 is methyl.
[0389] For example, R.sub.12 is ethyl or propenyl.
[0390] For example, R.sub.12 is methoxyl.
[0391] For example, R.sub.12 is ethenyl.
[0392] For example, R.sub.8 is H, methyl, ethyl, or ethenyl.
[0393] For example, R.sub.8 is methyl.
[0394] For example, R.sub.8 is ethyl.
[0395] For example, R.sub.8 is propyl.
[0396] For example, R.sub.8 is ethenyl or propenyl.
[0397] For example, R.sub.8 is C.sub.1-C.sub.6 alkyl substituted
with one or more substituents selected from the group consisting of
halo (e.g., F, Cl, or Br), hydroxyl, or C.sub.1-C.sub.6
alkoxyl.
[0398] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and
morpholinyl, and the like).
[0399] For example, R.sub.8 is piperidinyl.
[0400] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl and R.sub.7 is -Q.sub.4-T.sub.4, in
which Q.sub.4 is a bond or C.sub.1-C.sub.4 alkyl linker and T.sub.4
is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl or 4 to
7-membered heterocycloalkyl.
[0401] For example, Z is NR.sub.7R.sub.8 or CR.sub.7R.sub.8R.sub.14
wherein R.sub.7 and R.sub.8, together with the atom to which they
are attached, form a 4 to 11-membered heterocycloalkyl ring having
1 to 3 heteroatoms (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, morpholinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0402] For example, the ring formed by R.sub.7 and R.sub.8 is
selected from the group consisting of azetidinyl, pyrrolidinyl,
piperidinyl, morpholinyl, piperazinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, each optionally
substituted with one -Q.sub.6-T.sub.6.
[0403] For example, Z is 1,4-dioxa-8-azaspiro[4.5]decan-8-yl,
pyrrolidine-2,5-dione-1-yl, or piperidine-2,6-dione-1-yl.
[0404] For example, one or more -Q.sub.6-T.sub.6 is oxo.
[0405] For example, T.sub.6 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0406] For example, Q.sub.6 is a bond and T.sub.6 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0407] For example, Q.sub.6 is CO, S(O).sub.2, or NHC(O); and
T.sub.6 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0408] For example, T.sub.6 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0409] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is H or C.sub.6-C.sub.10 aryl.
[0410] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.pR.sub.p.
[0411] For example, each of R.sub.p and R.sub.q, independently, is
C.sub.1-C.sub.6 alkyl.
[0412] For example, R.sub.6 is --S(O).sub.bR.sub.a or azido, in
which b is 0, 1, or 2 and R.sub.a is C.sub.1-C.sub.6 alkyl or
C.sub.3-C.sub.8 cycloalkyl; and Z is NR.sub.7R.sub.8, in which
R.sub.7 is C.sub.3-C.sub.8 cycloalkyl (e.g., cyclopentyl,
cyclohexyl, or cycloheptyl) or 4 to 14-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like),
each optionally substituted with one or more -Q.sub.5-T.sub.5; and
R.sub.8 is H or C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl).
[0413] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is NR.sub.7R.sub.8 or
CR.sub.7R.sub.8R.sub.14 wherein R.sub.7 and R.sub.8, together with
the atom to which they are attached, form a 4 to 11-membered
heterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, pyrrolidine-2,5-dione-1-yl
piperidine-2,6-dione-1-yl and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0414] For example, R.sub.13 is H or methyl.
[0415] For example, R.sub.13 is H.
[0416] For example, R.sub.3 is H.
[0417] For example, each of R.sub.5, R.sub.9, and R.sub.10 is
H.
[0418] The present invention provides the compounds of Formula
(Ia)
##STR00006##
or a pharmaceutically acceptable salt or ester thereof, wherein:
[0419] X.sub.1 is N or CR.sub.11; [0420] X.sub.2 is N or CR.sub.13;
[0421] X.sub.3 is N or C, and when X.sub.3 is N, R.sub.6 is absent;
[0422] Z is NR.sub.7R.sub.8, OR.sub.7, S(O).sub.aR.sub.7, or
CR.sub.7R.sub.8R.sub.14, in which a is 0, 1, or 2; [0423] each of
R.sub.1 and R.sub.5, independently, is H or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0424] each of R.sub.2, R.sub.3, and R.sub.4, independently, is
-Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or C.sub.1-C.sub.3
alkyl linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.1 is H, halo, hydroxyl, COOH,
cyano, azido, or R.sub.S1, in which R.sub.S1 is C.sub.1-C.sub.3
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.1-C.sub.6 alkoxyl, C(O)O--C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, 4
to 12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl,
and R.sub.S1 is optionally substituted with one or more
substituents selected from the group consisting of halo, hydroxyl,
oxo, COOH, C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6
alkoxyl, amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0425] R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)R.sub.a, --C(O)OR.sub.a,
--C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a, --S(O).sub.bR.sub.a,
--S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in which R.sub.S2 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl, b is 0, 1, or 2, each of R.sub.a and R.sub.b,
independently is H or R.sub.S3, and R.sub.S3 is C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl; or R.sub.a and
R.sub.b, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom; and each of R.sub.S2, R.sub.S3, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2, wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.2 is H, halo, cyano, --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, --C(O)NR.sub.cR.sub.d,
--NR.sub.dC(O)R.sub.c, --NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.6 alkoxy, and
T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; or [0426]
R.sub.6 is H, halo, cyano, azido, --NR.sub.aR.sub.b, --C(O)R.sub.a,
--C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a,
--S(O).sub.bR.sub.a, --S(O).sub.bNR.sub.aR.sub.b, or R.sub.S2, in
which R.sub.S2 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
12-membered heterocycloalkyl, b is 0, 1, or 2, each of R.sub.a and
R.sub.b, independently is H or R.sub.S3, and R.sub.S3 is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl; or
R.sub.a and R.sub.b, together with the N atom to which they are
attached, form a 4 to 12-membered heterocycloalkyl ring having 0 or
1 additional heteroatom; and each of R.sub.S2, R.sub.S3, and the 4
to 12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b,
is optionally substituted with one or more -Q.sub.2-T.sub.2,
wherein Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker each
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.2 is H, halo, cyano, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--C(O)NR.sub.cR.sub.d, --NR.sub.dC(O)R.sub.c,
--NR.sub.dC(O)OR.sub.c, --S(O).sub.2R.sub.c,
--S(O).sub.2NR.sub.cR.sub.d, or R.sub.S4, in which each of R.sub.c
and R.sub.d, independently is H or R.sub.S5, each of R.sub.S4 and
R.sub.S5, independently, is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, or R.sub.c and
R.sub.d, together with the N atom to which they are attached, form
a 4 to 12-membered heterocycloalkyl ring having 0 or 1 additional
heteroatom, and each of R.sub.S4, R.sub.S5, and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.c and R.sub.d, is
optionally substituted with one or more -Q.sub.3-T.sub.3, wherein
Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl linker each optionally
substituted with halo, cyano, hydroxyl or C.sub.1-C.sub.6 alkoxy,
and T.sub.3 is selected from the group consisting of halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, 4 to 12-membered heterocycloalkyl, 5- or 6-membered
heteroaryl, OR.sub.e, COOR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f, each of R.sub.e and
R.sub.f independently being H or C.sub.1-C.sub.6 alkyl, or
-Q.sub.3-T.sub.3 is oxo; or -Q.sub.2-T.sub.2 is oxo; [0427] R.sub.7
is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond, C.sub.1-C.sub.4
alkyl linker, or C.sub.2-C.sub.4 alkenyl linker, each linker
optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.h,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 14-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
or [0428] R.sub.7 is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond,
C.sub.1-C.sub.4 alkyl linker, or C.sub.2-C.sub.4 alkenyl linker,
each linker optionally substituted with halo, cyano, hydroxyl or
C.sub.1-C.sub.6 alkoxy, and T.sub.4 is H, halo, cyano,
NR.sub.gR.sub.h, --OR.sub.g, --C(O)R.sub.g, --C(O)OR.sub.g,
--C(O)NR.sub.gR.sub.h, --C(O)NR.sub.gOR.sub.n,
--NR.sub.gC(O)R.sub.h, --S(O).sub.2R.sub.g, or R.sub.S6, in which
each of R.sub.g and R.sub.h, independently is H or R.sub.S7, each
of R.sub.S6 and R.sub.S7, independently is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8
cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, or 5- or 6-membered heteroaryl, and each of
R.sub.S6 and R.sub.S7 is optionally substituted with one or more
-Q.sub.5-T.sub.5, wherein Q.sub.5 is a bond, C(O), C(O)NR.sub.k,
NR.sub.kC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.k
being H or C.sub.1-C.sub.6 alkyl, and T.sub.5 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.qR.sub.q in which q is 0, 1, or 2 and R.sub.q is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.5 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.5 is H, halo, hydroxyl, or cyano; or -Q.sub.5-T.sub.5 is oxo;
provided that (i) R.sub.7 is not C(O)R.sub.g or --S(O).sub.2R.sub.g
when Z is NR.sub.7R.sub.8; (ii) R.sub.7 is not C.sub.1-C.sub.6
alkyl when Z is OR.sub.7, and (iii) R.sub.7 is not H; [0429] each
of R.sub.8, R.sub.11, R.sub.12, and R.sub.13, independently, is H,
halo, hydroxyl, COOH, cyano, R.sub.S8, OR.sub.S8, or COOR.sub.S8,
in which R.sub.S8 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, 4 to
12-membered heterocycloalkyl, amino, mono-C.sub.1-C.sub.6
alkylamino, or di-C.sub.1-C.sub.6 alkylamino, and R.sub.S8 is
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, and di-C.sub.1-C.sub.6
alkylamino; or R.sub.7 and R.sub.8, together with the N atom to
which they are attached, form a 4 to 12-membered heterocycloalkyl
ring having 0 to 2 additional heteroatoms, or R.sub.7 and R.sub.8,
together with the C atom to which they are attached, form
C.sub.3-C.sub.8 cycloalkyl or a 4 to 12-membered heterocycloalkyl
ring having 1 to 3 heteroatoms, and each of the 4 to 12-membered
heterocycloalkyl rings or C.sub.3-C.sub.8 cycloalkyl formed by
R.sub.7 and R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6, wherein Q.sub.6 is a bond, C(O), C(O)NR.sub.m,
NR.sub.mC(O), S(O).sub.2, or C.sub.1-C.sub.3 alkyl linker, R.sub.m
being H or C.sub.1-C.sub.6 alkyl, and T.sub.6 is H, halo,
C.sub.1-C.sub.6 alkyl, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, 5- or 6-membered heteroaryl, or
S(O).sub.pR.sub.p in which p is 0, 1, or 2 and R.sub.p is
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, or 5- or 6-membered heteroaryl, and
T.sub.6 is optionally substituted with one or more substituents
selected from the group consisting of halo, C.sub.1-C.sub.6 alkyl,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to 12-membered
heterocycloalkyl, and 5- or 6-membered heteroaryl except when
T.sub.6 is H, halo, hydroxyl, or cyano; or -Q.sub.6-T.sub.6 is oxo;
and [0430] R.sub.14 is absent, H, or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
the group consisting of halo, hydroxyl, COOH,
C(O)O--C.sub.1-C.sub.6 alkyl, cyano, C.sub.1-C.sub.6 alkoxyl,
amino, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, 4 to
12-membered heterocycloalkyl, and 5- or 6-membered heteroaryl;
[0431] provided that the compound is not [0432]
N-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-2-me-
thylphenyl)furan-2-carboxamide, [0433]
N,N'-(5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-
,3-phenylene)diacetamide, [0434]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-pivalamidobenzam-
ide, [0435]
3-(3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-sulfonamido)-N-((4,6-dimethyl--
2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide, [0436]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,5-dimethoxybenza-
mide, [0437]
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4,5-trimethoxybe-
nzamide, [0438]
3-allyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4,5-dimeth-
oxybenzamide, [0439]
4-(2-amino-2-oxoethoxy)-3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)-5-methoxybenzamide, [0440]
3-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-hydroxy-
-5-methoxybenzamide, or [0441]
3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-methoxy--
4-propoxybenzamide.
[0442] For example, X.sub.1 is CR.sub.11 and X.sub.2 is
CR.sub.13.
[0443] For example, X.sub.1 is CR.sub.11 and X.sub.2 is N.
[0444] For example, X.sub.1 is N and X.sub.2 is CR.sub.13.
[0445] For example, X.sub.1 is N and X.sub.2 is N.
[0446] For example, X.sub.3 is C.
[0447] For example, X.sub.3 is N and R.sub.6 is absent.
[0448] For example, Z is NR.sub.7R.sub.8.
[0449] For example, Z is CR.sub.7R.sub.8R.sub.14.
[0450] For example, Z is OR.sub.7.
[0451] For example, Z is S(O).sub.aR.sub.7, in which a is 0, 1, or
2
[0452] For example, Z is SR.sub.7.
[0453] For example, R.sub.6 is H.
[0454] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0455] For example, R.sub.6 is C.sub.1-C.sub.3 alkyl optionally
substituted with one or more -Q.sub.2-T.sub.2.
[0456] For example, R.sub.6 is CF.sub.3.
[0457] For example, R.sub.6 is C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, or C.sub.3-C.sub.6 cycloalkyl each
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0458] For example, R.sub.6 is ethenyl.
[0459] For example, R.sub.6 is ethynyl.
[0460] For example, R.sub.6 is ethynyl substituted with one or more
-Q.sub.2-T.sub.2, in which Q.sub.2 is a bond or C.sub.1-C.sub.3
alkyl linker and T.sub.2 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6
cycloalkyl, or 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
and morpholinyl, and the like) optionally substituted with one or
more -Q.sub.3-T.sub.3.
[0461] For example, R.sub.6 is azido.
[0462] For example, R.sub.6 is cyano.
[0463] For example, R.sub.6 is C(O)H.
[0464] For example, R.sub.6 is OR.sub.a or --C(O)R.sub.a.
[0465] For example, R.sub.a is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.2-T.sub.2.
[0466] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.2-T.sub.2.
[0467] For example, R.sub.6 is piperidinyl,
2,2,6,6-tetramethyl-piperidinyl, 1,2,3,6-tetrahydropyridinyl,
2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,
morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, or
pyrrolidinyl, each of which is optionally substituted with one or
more -Q.sub.2-T.sub.2.
[0468] For example, R.sub.6 is 4 to 7-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.2-T.sub.2, and
-Q.sub.2-T.sub.2 is oxo or Q.sub.2 is a bond and T.sub.2 is
--OR.sub.c, --NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.1-C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0469] For example, R.sub.6 is --NR.sub.aR.sub.b, --C(O)R.sub.a,
--C(O)OR.sub.a, --C(O)NR.sub.aR.sub.b, --NR.sub.bC(O)R.sub.a,
--SR.sub.a, --S(O).sub.2R.sub.a, or
--S(O).sub.2NR.sub.aR.sub.b.
[0470] For example, each of R.sub.a and R.sub.b, independently is
H, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.8 cycloalkyl optionally
substituted with one or more -Q.sub.2-T.sub.2
[0471] For example, one of R.sub.a and R.sub.b is H.
[0472] For example, R.sub.a and R.sub.b, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.2-T.sub.2
[0473] For example, -Q.sub.2-T.sub.2 is not H.
[0474] For example, -Q.sub.2-T.sub.2 is oxo.
[0475] For example, Q.sub.2 is a bond.
[0476] For example, Q.sub.2 is an unsubstituted C.sub.1-C.sub.3
alkyl linker.
[0477] For example, T.sub.2 is C.sub.1-C.sub.6 alkyl or
C.sub.6-C.sub.10 aryl, each optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0478] For example, T.sub.2 is an unsubstituted substituted
straight chain C.sub.1-C.sub.6 or branched C.sub.3-C.sub.6 alkyl,
including but not limited to, methyl, ethyl, n-propyl, i-propyl,
n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl and n-hexyl.
[0479] For example, T.sub.2 is phenyl.
[0480] For example, T.sub.2 is halo (e.g., fluorine, chlorine,
bromine, and iodine).
[0481] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.3-T.sub.3.
[0482] For example, T.sub.2 is --OR.sub.c, --NR.sub.cR.sub.d,
--C(O)R.sub.c, --C(O)OR.sub.c, or --S(O).sub.2R.sub.c.
[0483] For example, R.sub.c is C.sub.1-C.sub.6 alkyl or 4 to
7-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like), which
is optionally substituted with one or more -Q.sub.3-T.sub.3.
[0484] For example, each of R.sub.c and R.sub.d, independently is H
or C.sub.1-C.sub.6 alkyl optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0485] For example, R.sub.c is H.
[0486] For example, R.sub.d is H.
[0487] For example, R.sub.c and R.sub.d, together with the N atom
to which they are attached, form a 4 to 7-membered heterocycloalkyl
ring having 0 or 1 additional heteroatoms to the N atom (e.g.,
azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like) and the
ring is optionally substituted with one or more
-Q.sub.3-T.sub.3.
[0488] For example, Q.sub.2 is a bond and T.sub.2 is --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)R.sub.c, --C(O)OR.sub.c,
--S(O).sub.2R.sub.c, C.sub.6 alkyl, or 4 to 7-membered
heterocycloalkyl, each of which is optionally substituted with one
or more -Q.sub.3-T.sub.3 when R.sub.c or R.sub.d is not H.
[0489] For example, -Q.sub.3-T.sub.3 is oxo.
[0490] For example, T.sub.2 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl and one or more -Q.sub.3-T.sub.3 are
oxo.
[0491] For example, Q.sub.3 is a bond or unsubstituted or
substituted C.sub.1-C.sub.3 alkyl linker.
[0492] For example, T.sub.3 is H, halo, 4 to 7-membered
heterocycloalkyl, C.sub.1-C.sub.3 alkyl, OR.sub.e, COOR.sub.e,
--S(O).sub.2R.sub.e, --NR.sub.eR.sub.f, or
--C(O)NR.sub.eR.sub.f.
[0493] For example, one of R.sub.d and R.sub.e is H.
[0494] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, halo, OR.sub.e, --S(O).sub.2R.sub.e,
--NR.sub.eR.sub.f, and --C(O)NR.sub.eR.sub.f.
[0495] For example, Q.sub.3 is a bond or C.sub.1-C.sub.3 alkyl
linker and T.sub.3 is selected from the group consisting of
C.sub.1-C.sub.3 alkyl, OR.sub.e, --S(O).sub.2R.sub.e, or
--NR.sub.eR.sub.f.
[0496] For example, R.sub.e is H.
[0497] For example, R.sub.f is H.
[0498] For example, R.sub.7 is not H.
[0499] For example, R.sub.7 is --C(O)R.sub.g.
[0500] For example, R.sub.7 is --C(O)R.sub.g, in which R.sub.g is
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered heterocycloalkyl,
C.sub.3-C.sub.8 cycloalkyl.
[0501] For example, R.sub.7 is C.sub.6-C.sub.10 aryl substituted
with one or more -Q.sub.5-T.sub.5.
[0502] For example, R.sub.7 is phenyl optionally substituted with
one or more -Q.sub.5-T.sub.5.
[0503] For example, R.sub.7 is C.sub.1-C.sub.6 alkyl optionally
substituted with one or more -Q.sub.5-T.sub.5.
[0504] For example, R.sub.7 is C.sub.3-C.sub.8 cycloalkyl
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0505] For example, R.sub.7 is 4 to 7-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, and morpholinyl, and the like)
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0506] For example, R.sub.7 is 8 to 14-membered heterocycloalkyl
such as 1,4-dioxaspiro[4.5]decanyl (e.g.,
1,4-dioxaspiro[4.5]decan-8-yl), 1,4-dioxa-8-azaspiro[4.5]decanyl
(e.g., 1,4-dioxa-8-azaspiro[4.5]decan-8-yl), 1-oxaspiro[4.5]decanyl
(e.g., 1-oxaspiro[4.5]decan-8-yl or
1-oxaspiro[4.5]decan-2-one-8-yl), 1-azaspiro[4.5]decanyl (e.g.,
1-azaspiro[4.5]decan-8-yl or 1-azaspiro[4.5]decan-2-one-8-yl),
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-4-yl or
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-3'-one-4-yl),
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl (e.g.,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-4-yl or
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-7'-one-4-yl), or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl (e.g.,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-4-yl or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-3'-one-4-yl), each
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0507] For example, R.sub.7 is 5 to 6-membered heterocycloalkyl
optionally substituted with one or more -Q.sub.5-T.sub.5.
[0508] For example, R.sub.7 is isopropyl.
[0509] For example, R.sub.7 is pyrrolidinyl, piperidinyl,
tetrahydropyran, cyclopentyl, or cyclohexyl, cycloheptyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0510] For example, R.sub.7 is cyclopentyl or cyclohexyl, each
optionally substituted with one -Q.sub.5-T.sub.5.
[0511] For example, Q.sub.5 is NHC(O) and T.sub.5 is
C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy.
[0512] For example, -Q.sub.5-T.sub.5 is oxo.
[0513] For example, T.sub.4 is 4 to 7-membered heterocycloalkyl or
C.sub.3-C.sub.8 cycloalkyl or C.sub.6-C.sub.10 aryl, and one or
more -Q.sub.5-T.sub.5 are oxo.
[0514] For example, R.sub.7 is 1-oxide-tetrahydro-2H-thiopyranyl or
1,1-dioxide-tetrahydro-2H-thiopyranyl.
[0515] For example, R.sub.7 is cyclohexanonyl, e.g.,
cyclohexanon-4-yl.
[0516] For example, T.sub.5 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0517] For example, Q.sub.5 is a bond and T.sub.5 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0518] For example, Q.sub.5 is a bond and T.sub.5 is 5- or
6-membered heteroaryl, amino, mono-C.sub.1-C.sub.6 alkylamino,
di-C.sub.1-C.sub.6 alkylamino, T.sub.5 being optionally substituted
with one or more substituents selected from the group consisting of
halo, hydroxyl, C.sub.1-C.sub.6 alkoxyl, or C.sub.3-C.sub.8
cycloalkyl.
[0519] For example, Q.sub.5 is CO, S(O).sub.2, or NHC(O); and
T.sub.5 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0520] For example, T.sub.5 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0521] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is H or C.sub.6-C.sub.10 aryl.
[0522] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.qR.sub.q.
[0523] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is S(O).sub.aR.sub.7, in which a is 0,
1, or 2 and R.sub.7 is C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl), C.sub.3-C.sub.8 cycloalkyl
(e.g., cyclopentyl, cyclohexyl, or cycloheptyl) or 4 to 14-membered
heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0524] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is OR.sub.7 in which R.sub.7 is 4 to
14-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 1,4-dioxaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0525] For example, R.sub.11 is H.
[0526] For example, each of R.sub.2 and R.sub.4, independently, is
H or C.sub.1-C.sub.6 alkyl optionally substituted with amino,
azido, halo, mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6
alkylamino, or C.sub.6-C.sub.10 aryl.
[0527] For example, each of R.sub.2 and R.sub.4, independently is
C.sub.1-C.sub.3 alkyl optionally substituted with C.sub.1-C.sub.6
alkoxyl.
[0528] For example, each of R.sub.2 and R.sub.4 is methyl.
[0529] For example, R.sub.1 is H.
[0530] For example, R.sub.1 is C.sub.1-C.sub.6 alkyl optionally
substituted with azido, halo, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, or C.sub.6-C.sub.10
aryl.
[0531] For example, R.sub.12 is H, methyl, ethyl, ethenyl, or
halo.
[0532] For example, R.sub.12 is methyl.
[0533] For example, R.sub.12 is ethyl or propenyl.
[0534] For example, R.sub.12 is methoxyl.
[0535] For example, R.sub.12 is ethenyl.
[0536] For example, R.sub.8 is H, methyl, ethyl, or ethenyl.
[0537] For example, R.sub.8 is methyl.
[0538] For example, R.sub.8 is ethyl.
[0539] For example, R.sub.8 is propyl.
[0540] For example, R.sub.8 is ethenyl or propenyl.
[0541] For example, R.sub.8 is C.sub.1-C.sub.6 alkyl substituted
with one or more substituents selected from the group consisting of
halo (e.g., F, Cl, or Br), hydroxyl, or C.sub.1-C.sub.6
alkoxyl.
[0542] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, diazabicyclo[2.2.1]heptanyl,
2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, and
morpholinyl, and the like).
[0543] For example, R.sub.8 is piperidinyl.
[0544] For example, R.sub.8 is 4 to 7-membered optionally
substituted heterocycloalkyl and R.sub.7 is -Q.sub.4-T.sub.4, in
which Q.sub.4 is a bond or C.sub.1-C.sub.4 alkyl linker and T.sub.4
is H, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl or 4 to
7-membered heterocycloalkyl.
[0545] For example, Z is NR.sub.7R.sub.8 or CR.sub.7R.sub.8R.sub.14
wherein R.sub.7 and R.sub.8, together with the atom to which they
are attached, form a 4 to 11-membered heterocycloalkyl ring having
1 to 3 heteroatoms (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, morpholinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0546] For example, the ring formed by R.sub.7 and R.sub.8 is
selected from the group consisting of azetidinyl, pyrrolidinyl,
piperidinyl, morpholinyl, piperazinyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, and cyclohexenyl, each optionally
substituted with one -Q.sub.6-T.sub.6.
[0547] For example, Z is 1,4-dioxa-8-azaspiro[4.5]decan-8-yl,
pyrrolidine-2,5-dione-1-yl, or piperidine-2,6-dione-1-yl.
[0548] For example, one or more -Q.sub.6-T.sub.6 is oxo.
[0549] For example, T.sub.6 is H, halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.6-C.sub.10 aryl, or 4 to 7-membered heterocycloalkyl.
[0550] For example, Q.sub.6 is a bond and T.sub.6 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, or 4 to
7-membered heterocycloalkyl.
[0551] For example, Q.sub.6 is CO, S(O).sub.2, or NHC(O); and
T.sub.6 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 7-membered
heterocycloalkyl.
[0552] For example, T.sub.6 is C.sub.1-C.sub.6 alkyl or
C.sub.1-C.sub.6 alkoxyl, each optionally substituted with halo,
hydroxyl, cyano, C.sub.1-C.sub.6 alkoxyl, amino,
mono-C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino, or
C.sub.3-C.sub.8 cycloalkyl.
[0553] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is H or C.sub.6-C.sub.10 aryl.
[0554] For example, Q.sub.6 is C.sub.1-C.sub.3 alkyl linker and
T.sub.6 is C.sub.3-C.sub.8 cycloalkyl, 4 to 7-membered
heterocycloalkyl, or S(O).sub.pR.sub.p.
[0555] For example, each of R.sub.p and R.sub.q, independently, is
C.sub.1-C.sub.6 alkyl.
[0556] For example, R.sub.6 is --S(O).sub.bR.sub.a or azido, in
which b is 0, 1, or 2 and R.sub.a is C.sub.1-C.sub.6 alkyl or
C.sub.3-C.sub.8 cycloalkyl; and Z is NR.sub.7R.sub.8, in which
R.sub.7 is C.sub.3-C.sub.8 cycloalkyl (e.g., cyclopentyl,
cyclohexyl, or cycloheptyl) or 4 to 14-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like),
each optionally substituted with one or more -Q.sub.5-T.sub.5; and
R.sub.8 is H or C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl).
[0557] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is NR.sub.7R.sub.8 or
CR.sub.7R.sub.8R.sub.14 wherein R.sub.7 and R.sub.8, together with
the atom to which they are attached, form a 4 to 11-membered
heterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, pyrrolidine-2,5-dione-1-yl,
piperidine-2,6-dione-1-yl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0558] For example, R.sub.13 is H or methyl.
[0559] For example, R.sub.13 is H.
[0560] For example, R.sub.3 is H.
[0561] For example, each of R.sub.5, R.sub.9, and R.sub.10 is
H.
[0562] Another subset of the compounds of Formula (I) includes
those of Formula (Ib), (Ic), or (Id):
##STR00007##
or a pharmaceutically acceptable salt or ester thereof, wherein Z,
X.sub.2, X.sub.3, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.11, and R.sub.12 are as defined herein for Formula
(I).
[0563] Yet another subset of the compounds of Formula (I) is those
of Formula (Ie):
##STR00008##
[0564] wherein [0565] Z is NR.sub.7R.sub.8, OR.sub.7, or
S(O).sub.aR.sub.7; [0566] R.sub.1 is H or C.sub.1-C.sub.6 alkyl
optionally substituted with one or more substituents selected from
hydroxyl, C.sub.1-C.sub.6 alkoxyl, amino, mono-C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, and C.sub.6-C.sub.10
aryl; [0567] each of R.sub.2 and R.sub.4 independently, is
-Q.sub.1-T.sub.1, in which Q.sub.1 is a bond or C.sub.1-C.sub.3
alkyl linker optionally substituted with one or more substituents
selected from halo and hydroxyl, and T.sub.1 is H, halo, or azido;
[0568] R.sub.3 is H or halo; [0569] R.sub.5 is H or C.sub.1-C.sub.6
alkyl; [0570] R.sub.6 is H, halo, cyano, azido, OR.sub.a,
--NR.sub.aR.sub.b, --C(O)NR.sub.aR.sub.b, --S(O).sub.bR.sub.a, or
R.sub.S2; wherein R.sub.S2 is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, or 4 to
12-membered heterocycloalkyl, and wherein each of R.sub.a and
R.sub.b, independently is H, C.sub.1-C.sub.6 alkyl, or 4 to
12-membered heterocycloalkyl; or R.sub.a and R.sub.b, together with
the N atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 or 1 additional heteroatom; and each
of R.sub.a, R.sub.S2 and the 4 to 12-membered heterocycloalkyl ring
formed by R.sub.a and R.sub.b, is optionally substituted with one
or more -Q.sub.2-T.sub.2; wherein Q.sub.2 is a bond or
C.sub.1-C.sub.3 alkyl linker, and T.sub.2 is H, halo, --OR.sub.c,
--NR.sub.cR.sub.d, --C(O)OR.sub.c, or C.sub.1-C.sub.6 alkyl, in
which each of R.sub.c and R.sub.d, independently is H or
C.sub.1-C.sub.6 alkyl, or R.sub.c and R.sub.d, together with the N
atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 or 1 additional heteroatom and
optionally substituted with C.sub.1-C.sub.6 alkyl; [0571] R.sub.7
is -Q.sub.4-T.sub.4, in which Q.sub.4 is a bond or C.sub.1-C.sub.4
alkyl linker and T.sub.4 is H, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10 aryl, C(O)--C.sub.1-6
alkyl, C(O)--C.sub.3-6 cycloalkyl, or 4 to 14-membered
heterocycloalkyl, each optionally substituted with one or more
-Q.sub.5-T.sub.5; [0572] R.sub.8 is H, C.sub.1-C.sub.6 alkyl
optionally substituted with halo, hydroxyl or C.sub.1-C.sub.6
alkoxyl, C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.8 cycloalkyl, or 4
to 7-membered heterocycloalkyl; or R.sub.7 and R.sub.8, together
with the N atom to which they are attached, form a 4 to 12-membered
heterocycloalkyl ring having 0 to 2 additional heteroatoms, and the
4 to 12-membered heterocycloalkyl ring formed by R.sub.7 and
R.sub.8 is optionally substituted with one or more
-Q.sub.6-T.sub.6; and [0573] R.sub.12 is halo, C.sub.1-C.sub.6
alkoxyl, or C.sub.1-C.sub.6 alkyl optionally substituted with halo
or C.sub.2-C.sub.6 alkenyl.
[0574] In addition to the above-described features of the compounds
of this invention where applicable, the compounds of Formula (Ie)
can include one or more of the following features:
[0575] For example, R.sub.1 is H or C.sub.1-C.sub.6 alkyl
optionally substituted one or more times with a substituent
selected from hydroxyl, C.sub.1-C.sub.6 alkoxyl and
C.sub.6-C.sub.10 aryl; R.sub.7 is -Q.sub.4-T.sub.4, in which
Q.sub.4 is a bond or a C.sub.1-C.sub.4 alkyl linker, and T.sub.4 is
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.10
aryl, C(O)-- C.sub.1-C.sub.6 alkyl, C(O)-- C.sub.3-C.sub.6
cycloalkyl, dihydropyranyl, tetrahydropyranyl,
tetrahydrothiopyranyl, piperidinyl, pyrrolidinyl, azetidinyl and
oxetanyl, each optionally substituted with one or more substituents
independently selected from oxo and -Q.sub.5-T.sub.5; R.sub.8 is H
or C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.2-C.sub.6 alkenyl or C.sub.3-C.sub.8 cycloalkyl; and R.sub.12
is halo, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl or
C.sub.1-C.sub.6 alkoxyl.
[0576] For example, Z is NR.sub.7R.sub.8 or SR.sub.7; R.sub.6 is H,
halo, cyano, OR.sub.a, --C(O)NR.sub.aR.sub.b, --S(O).sub.2R.sub.a,
or R.sub.S2; wherein R.sub.S2 is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, or 4 to
12-membered heterocycloalkyl, and wherein each of R.sub.a and
R.sub.b, independently is H or C.sub.1-C.sub.6 alkyl; or R.sub.a
and R.sub.b, together with the N atom to which they are attached,
form a 4 to 12-membered heterocycloalkyl ring having 0 or 1
additional heteroatom; and each of R.sub.S2 and the 4 to
12-membered heterocycloalkyl ring formed by R.sub.a and R.sub.b, is
optionally substituted with one or more -Q.sub.2-T.sub.2; wherein
Q.sub.2 is a bond or C.sub.1-C.sub.3 alkyl linker, and T.sub.2 is
H, halo, --OR.sub.c, --NR.sub.cR.sub.d, --C(O)OC.sub.1-C.sub.6
alkyl, or C.sub.1-C.sub.6 alkyl, in which each of R.sub.c and
R.sub.d, independently is H or C.sub.1-C.sub.6 alkyl, or R.sub.c
and R.sub.d, together with the N atom to which they are attached,
form a 4 to 12-membered heterocycloalkyl ring having 0 or 1
additional heteroatom and 0 or 1 C.sub.1-C.sub.6 alkyl
substituents; R.sub.7 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkyl, dihydropyranyl, tetrahydropyranyl,
tetrahydrothiopyranyl, piperidinyl, pyrrolidinyl, azetidinyl and
oxetanyl, each optionally substituted with one or more substituents
independently selected from oxo and -Q.sub.5-T.sub.5; and R.sub.12
is halo or C.sub.1-C.sub.6 alkyl.
[0577] For example, R.sub.2, R.sub.4 and R.sub.12 are each
independently C.sub.1-C.sub.6 alkyl and R.sub.5 is H.
[0578] For example, R.sub.7 is cyclohexyl, dihydropyranyl,
tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl,
pyrrolidinyl, azetidinyl oxetanyl, 1,4-dioxaspiro[4.5]decan-8-yl,
1-oxaspiro[4.5]decan-8-yl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-4-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-4-yl,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-4-yl, or
1-azaspiro[4.5]decan-8-yl, each substituted with one or more
-Q.sub.5-T.sub.5.
[0579] For example, Z is selected from the group consisting of
piperidinyl, morpholinyl, piperazinyl, azetidinyl, pyrrolidinyl,
2-oxa-5-azabicyclo[2.2.1]heptan-5-yl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, and cyclohexenyl, each
optionally substituted with one -Q.sub.6-T.sub.6.
[0580] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is S(O).sub.aR.sub.7, in which a is 0,
1, or 2 and R.sub.7 is C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl), C.sub.3-C.sub.8 cycloalkyl
(e.g., cyclopentyl, cyclohexyl, or cycloheptyl) or 4 to 14-membered
heterocycloalkyl (e.g., azetidinyl, oxetanyl, thietanyl,
pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,
isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0581] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is OR.sub.7 in which R.sub.7 is 4 to
14-membered heterocycloalkyl (e.g., azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 1,4-dioxaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like)
and R.sub.7 is optionally substituted with one or more
-Q.sub.5-T.sub.5.
[0582] For example, R.sub.6 is --S(O).sub.bR.sub.a or azido, in
which b is 0, 1, or 2 and R.sub.a is C.sub.1-C.sub.6 alkyl or
C.sub.3-C.sub.8 cycloalkyl; and Z is NR.sub.7R.sub.8, in which
R.sub.7 is C.sub.3-C.sub.8 cycloalkyl (e.g., cyclopentyl,
cyclohexyl, or cycloheptyl) or 4 to 14-membered heterocycloalkyl
(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, piperidinyl,
1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,
3,6-dihydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, 1,4-diazepanyl,
1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, morpholinyl,
1,4-dioxaspiro[4.5]decanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1-oxaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl, or
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the like),
each optionally substituted with one or more -Q.sub.5-T.sub.5; and
R.sub.8 is H or C.sub.1-C.sub.6 alkyl (e.g., methyl, ethyl,
n-propyl, i-propyl, butyl, or t-butyl).
[0583] For example, R.sub.6 is halo (e.g., fluorine, chlorine,
bromine, and iodine) and Z is NR.sub.7R.sub.8 or
CR.sub.7R.sub.8R.sub.14 wherein R.sub.7 and R.sub.8, together with
the atom to which they are attached, form a 4 to 11-membered
heterocycloalkyl ring having 1 to 3 heteroatoms (e.g., azetidinyl,
oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,
oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,
piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,
tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl,
tetrahydro-2H-thiopyranyl, 1,4-diazepanyl, 1,4-oxazepanyl,
2-oxa-5-azabicyclo[2.2.1]heptanyl, 2,5-diazabicyclo[2.2.1]heptanyl,
morpholinyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1,4-dioxa-8-azaspiro[4.5]decan-8-yl, pyrrolidine-2,5-dione-1-yl,
piperidine-2,6-dione-1-yl, and the like) or C.sub.3-C.sub.8
cycloalkyl, each optionally substituted with one or more
-Q.sub.6-T.sub.6.
[0584] Another subset of the compounds of Formula (I) includes
those of Formula (II):
##STR00009##
or a pharmaceutically acceptable salts or esters thereof, wherein
R.sub.6, R.sub.7, R.sub.8, and R.sub.12 are defined herein.
[0585] Yet another subset of the compounds of Formula (I) includes
those of Formula (IIA):
##STR00010##
or a pharmaceutically acceptable salts or esters thereof, wherein n
is 0, 1, or 2; U is O, S, N-Q.sub.5-T.sub.5, or CH-Q.sub.5-T.sub.5;
R.sub.12 is Cl, Br, or methyl; and R.sub.6, R.sub.7, R.sub.8,
Q.sub.5, and T.sub.5 are defined herein.
[0586] In addition to the above-described features of the compounds
of this invention where applicable, the compounds of Formula (II)
or (IIA) can include one or more of the following features:
[0587] For example, Q.sub.5 is a bond and T.sub.5 is H,
C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered
heterocycloalkyl, 5- or 6-membered heteroaryl, amino,
mono-C.sub.1-C.sub.6 alkylamino, or di-C.sub.1-C.sub.6 alkylamino,
T.sub.5 being optionally substituted with one or more substituents
selected from the group consisting of halo, hydroxyl,
C.sub.1-C.sub.6 alkoxyl, or C.sub.3-C.sub.8 cycloalkyl.
[0588] For example, Q.sub.5 is CO, S(O).sub.2, or NHC(O); and
T.sub.5 is C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxyl,
C.sub.3-C.sub.8 cycloalkyl, or 4 to 12-membered
heterocycloalkyl.
[0589] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is H or C.sub.6-C.sub.10 aryl.
[0590] For example, Q.sub.5 is C.sub.1-C.sub.3 alkyl linker and
T.sub.5 is C.sub.3-C.sub.8 cycloalkyl, 4 to 12-membered
heterocycloalkyl, or S(O).sub.qR.sub.q.
[0591] For example, Q.sub.5 is NHC(O) and T.sub.5 is
C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy.
[0592] For example, one or more -Q.sub.5-T.sub.5 are oxo.
[0593] For example, U is CH-Q.sub.5-T.sub.5 and n is 0
[0594] For example, one or more -Q.sub.6-T.sub.6 are oxo.
[0595] For example, Q.sub.6 is a bond or C(O) and T.sub.6 is
C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 alkoxy.
[0596] Still another subset of the compounds of Formula (I)
includes those of Formula (III):
##STR00011##
or a pharmaceutically acceptable salts or esters thereof, wherein
R.sub.7 is a 4 or 6-membered heterocycloalkyl having one nitrogen
atom in the ring and is substituted with one or two methyl groups
or one i-propyl group; R.sup.3 is H or F; R.sub.4 is methyl, ethyl,
n-propyl, isopropyl, or CF.sub.3, and R.sub.6 is CF.sub.3, Cl, or
F, provided that when R.sub.4 is methyl, (1) R.sub.6 is CF.sub.3,
or (2) R.sub.3 is F, or (3) R.sub.6 is CF.sub.3 and R.sub.3 is F,
or (4) R.sub.6 is F or Cl and R.sub.7 is a 6-membered
heterocycloalkyl having only one nitrogen and is substituted with
two methyl groups.
[0597] In addition to the above-described features of the compounds
of this invention where applicable, the compounds of Formula (III)
can include one or more of the following features:
[0598] For example, R.sub.7 is a 4 or 6-membered heterocycloalkyl
having one and only one heteroatom in the ring and the heteroatom
is nitrogen.
[0599] For example, R.sub.7 is not further substituted besides the
one or two methyl groups or the one i-propyl group.
[0600] For example, R.sub.7 is
##STR00012##
[0601] For example, R.sub.7 is
##STR00013##
[0602] For example, R.sub.7 is
##STR00014##
[0603] Representative compounds of the present invention include
compounds listed in Table 1. In the table below, each occurrence
of
##STR00015##
should be construed as
##STR00016##
TABLE-US-00001 TABLE 1 Compound Number Structure Name MS.dagger. 1
##STR00017## 5-bromo-3- (cyclopentyl(methyl)amino)-
N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 446.2 2 ##STR00018## 5-bromo-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(piperidin-4- yl)amino)benzamide 462.2 3 ##STR00019##
5-chloro-3- (cyclohexyl(methyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 416.4 4
##STR00020## 5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl(1- pivaloylpiperidin-4-
yl)amino)benzamide 545.2 5 ##STR00021## 5-bromo-3-
(cyclopentyl(ethyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 460.1 6
##STR00022## 3-((4- acetamidocyclohexyl) (methyl)amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 516.9 7 ##STR00023## 5-chloro-3-
(cyclopentyl(methyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 402.2 8
##STR00024## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(piperidin-4-
yl)amino)benzamide 417.3 9 ##STR00025## 3-((4-
aminocyclohexyl)(methyl) amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 475.1 10 ##STR00026## 5-bromo-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(tetrahydro-2H- pyran-4- yl)amino)benzamide 462.2 11
##STR00027## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (N-methylisobutyramido)
benzamide 404.2 12 ##STR00028## 5-bromo-3-
(cyclohexyl(methyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 460.1 13
##STR00029## 2,5-dichloro-3- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)benzamide 422.1 14 ##STR00030## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(tetrahydro-2H- pyran-4- yl)amino)benzamide 418.3 15
##STR00031## tert-butyl (4-((5-bromo-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(methyl)
amino)cyclohexyl)carbamate 575.8 16 ##STR00032##
3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2-methyl-5- (1-methyl-1,2,3,6-
tetrahydropyridin-4- yl)benzamide 463.4 17 ##STR00033##
3-((1-acetylpiperidin-4- yl)(methyl)amino)-5-
bromo-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 503.2 18 ##STR00034## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (1-methylpiperidin-4- yl)benzamide 465.4
19 ##STR00035## 5-bromo-2-chloro-3- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)benzamide 466.1 20 ##STR00036## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (trifluoromethyl)benzamide 436.2 21
##STR00037## 3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2-methyl-5-
(trifluoromethyl)benzamide 368.2 22 ##STR00038## 3-((4-
acetamidocyclohexyl) amino)-5-bromo-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 503.1 23
##STR00039## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(1-
methylpiperidin-4- yl)amino)benzamide 431.3 24 ##STR00040##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- ((1-pivaloylpiperidin-4- yl)amino)benzamide
531.1 25 ##STR00041## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (2,2,6,6-tetramethyl-
1,2,3,6-tetrahydropyridin- 4-yl)benzamide 505.4 26 ##STR00042##
tert-butyl (4-((5-bromo-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)amino)
cyclohexyl)carbamate 562.2 27 ##STR00043## 3-((4-
aminocyclohexyl)amino)- 5-bromo-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 461.1 28
##STR00044## 5-bromo-3- (cyclohexylamino)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 446.2 29 ##STR00045## 3-((1-acetylpiperidin-4-
yl)amino)-5-bromo-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 489.2 30 ##STR00046##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl(1- methylpiperidin-4-
yl)amino)benzamide 475.02 31 ##STR00047## 5-bromo-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(isopropyl(methyl)amino)- 2-methylbenzamide 420.2 32 ##STR00048##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- ((tetrahydro-2H-pyran-4- yl)amino)benzamide
447.9 33 ##STR00049## 5-bromo-3- (cyclopentylamino)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 432.1 34 ##STR00050## tert-butyl 4-(2-chloro-3-
(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)carbamoyl)
phenoxy)piperidine-1- carboxylate 490.3 35 ##STR00051##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (piperidin-4- ylamino)benzamide 447.1 36
##STR00052## 5-bromo-3- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2,4- dimethylbenzamide 460.1 37 ##STR00053##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- ((1-methylpiperidin-4- yl)amino)benzamide
460.96 38 ##STR00054## 2-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(piperidin- 4-yloxy)benzamide 390.1
39 ##STR00055## 2-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-((1- (methylsulfonyl)piperidin-
4-yl)oxy)benzamide 468.1 40 ##STR00056## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(N-methylacetamido) benzamide 376.2 41 ##STR00057##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (isopropylamino)-2- methylbenzamide 406.1 42
##STR00058## 3-(cyclopentylamino)-N- ((4,6-dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-5- (2,2,6,6-tetramethyl-
1,2,3,6-tetrahydropyridin- 4-yl)benzamide 491.2 43 ##STR00059##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (dimethylamino)-2- methylbenzamide 348.2 44
##STR00060## 3-bromo-5- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)benzamide 432 45 ##STR00061## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2-(4- methylpiperazin-1-yl)-6-
(piperidin-1- yl)isonicotinamide 438.3 46 ##STR00062##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-(4-
methylpiperazin-1-yl)-6- morpholinoisonicotinamide 441.2 47
##STR00063## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-2,6-bis(4- methylpiperazin-1- yl)isonicotinamide 454.2
48 ##STR00064## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-2,6-bis(4- methylpiperazin-1- yl)isonicotinamide 430.2
49 ##STR00065## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3-(N- methylcyclopentane
carboxamido)benzamide 377.2 50 ##STR00066## tert-butyl
(2-((5-chloro-3- (((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(methyl) amino)ethyl)carbamate
477.3 51 ##STR00067## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-((2- (dimethylamino)ethyl)
(methyl)amino)-2- methylbenzamide 405.3 52 ##STR00068##
3-(allyl(cyclopentyl)amino)- 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 428.3 53
##STR00069## 3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2-methyl-5- (piperazin-1-
yl)benzamide 452.4 54 ##STR00070## 3-(cyclopentyl(methyl)
amino-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (4-methylpiperazin-1- yl)benzamide 466.4
55 ##STR00071## 5-chloro-3- (cyclohexyl(ethyl)amino)-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-
methylbenzamide 430.2 56 ##STR00072## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 432.3 57
##STR00073## 5-chloro-3- (cycloheptyl(ethyl)amino)-
N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 444.3 58 ##STR00074## 3-(((1s,4s)-4-
aminocyclohexyl)(methyl) amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 475.2 59 ##STR00075## 3-(((1r,4r)-4-
aminocyclohexyl)(methyl) amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 475.2 60 ##STR00076## 3-(((1s,4s)-4-
acetamidocyclohexyl) (methyl)amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 517.3 61 ##STR00077## 3-(((1r,4r)-4-
acetamidocyclohexyl) (methyl)amino)-5-bromo-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 517.3 62 ##STR00078## 2-bromo-3-
(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)benzamide 432.1 63 ##STR00079##
3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- vinylbenzamide 380.3 64
##STR00080## 5-chloro-3- (cyclopentyl(ethyl)amino)-
N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 416.3 65 ##STR00081## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(m-tolylamino)benzamide 410.3 66 ##STR00082## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(piperidin-1- yl)benzamide 388.2
67 ##STR00083## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- morpholinobenzamide 390.2
68 ##STR00084## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (pyrrolidin-1-
yl)benzamide 374.2 69 ##STR00085## 3-(azetidin-1-yl)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 360.2 70 ##STR00086## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(3-methylpiperidin-1- yl)benzamide 402.3 71 ##STR00087##
3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- isopropylbenzamide 396.25
72 ##STR00088## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (4-methylpiperidin-1-
yl)benzamide 402.2 73 ##STR00089## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(4-methylpiperazin-1- yl)benzamide 403.2 74 ##STR00090##
3-(cyclopentyl (methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- ethylbenzamide 382.3 75
##STR00091## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(m-
tolyl)amino)benzamide 424.2 76 ##STR00092## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(1,4-dioxa-8- azaspiro[4.5]decan-8- yl)benzamide 446.2 77
##STR00093## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(1-(2-(methylsulfonyl)ethyl) piperidin-4- yl)amino)benzamide
523.2 78 ##STR00094## 3-((1-acetylpiperidin-4- yl)(methyl)amino)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 459.3 79 ##STR00095## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(1- pivaloylpiperidin-4- yl)amino)benzamide 501.2 80
##STR00096## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (pyrrolidin-3-
ylamino)benzamide 389.2 81 [[Duplicate with #19]] 82 ##STR00097##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (piperazin-1- yl)benzamide 389.1 83
##STR00098## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-((1- isopropylpiperidin-4-
yl)(methyl)amino)-2- methylbenzamide 459.3 84 ##STR00099##
3-((1-benzylpiperidin-4- yl)(methyl)amino)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 507.2 85 ##STR00100## 3-([1,4'-bipiperidin]-4-
yl(methyl)amino)-5- chloro-N-((4,6-dimethyl- 2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 500.2 86
##STR00101## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- ((1-methylpyrrolidin-3-
yl)amino)benzamide 403.1 87 ##STR00102## tert-butyl
(2-((5-chloro-3- (((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(methyl) amino)ethyl)(methyl)
carbamate 391.2 88 ##STR00103## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(1- (methylsulfonyl)piperidin- 4-yl)amino)benzamide 494.9 89
##STR00104## 5-chloro-3-((1- (cyclopropylmethyl) piperidin-4-
yl)(methyl)amino)-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 471.2 90 ##STR00105##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-((1-(2- hydroxyethyl)piperidin- 4-yl)(methyl)amino)-2-
methylbenzamide 461.2 91 ##STR00106## 3-(allyl(piperidin-4-
yl)amino)-5-chloro-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 443.3 92 ##STR00107##
3-allyl(tetrahydro-2H- pyran-4-yl)amino)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 444.3 93 ##STR00108## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(2- (methylamino)ethyl) amino)benzamide 391.2 94
##STR00109## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (piperidin-3-
ylamino)benzamide 403.3 95 ##STR00110## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(piperidin-3- yl)amino)benzamide 417.3 96 ##STR00111##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl(1- methylpiperidin-3-
yl)amino)benzamide 431.2 97 ##STR00112## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(piperidin-4- yl(propyl)amino) benzamide 445.3 98 ##STR00113##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (propyl(tetrahydro-2H- pyran-4-
yl)amino)benzamide 445.3 99 ##STR00114## tert-butyl 3-((5-chloro-3-
(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(ethyl) amino)pyrrolidine-1-
carboxylate 517.4 100 ##STR00115## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(pyrrolidin-3- yl)amino)-2- methylbenzamide 417.3 101
##STR00116## tert-butyl 4-((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)
amino)piperidine-1- carboxylate 531.4 102 ##STR00117##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(piperidin-4- yl)amino)-2- methylbenzamide
431.2 103 ##STR00118## 3-(azetidin-3- yl(ethyl)amino)-5-chloro-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-
methylbenzamide 403.2 104 ##STR00119## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(isobutyl(methyl)amino)- 2-methylbenzamide 390.3 105 ##STR00120##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(isobutyl)amino)-2- methylbenzamide 404.3 106
##STR00121## 3-bromo-6-chloro-4- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)picolinamide 469.2 107 ##STR00122## tert-butyl
((1s,4s)-4-((5- chloro-3-(((4,6-dimethyl- 2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)
amino)cyclohexyl)carbamate 545.4 108 ##STR00123##
tert-butyl((1r,4r-4-((5- chloro-3-(((4,6-dimethyl- 2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)
amino)cyclohexyl)carbamate 545.3 109 ##STR00124## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(1-
pivaloylpiperidin-4- yl)amino)-2- methylbenzamide 515.4 110
##STR00125## 4-((5-chloro-3-(((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(methyl)
amino)-N,N- dimethylpiperidine-1- carboxamide 488.2 111
##STR00126## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(1-(oxetan-3-
yl)piperidin-4- yl)amino)benzamide 473.3 112 ##STR00127##
tert-butyl 3-((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(methyl)
amino)azetidine-1- carboxylate 489.3 113 ##STR00128##
3-(azetidin-3- yl(methyl)amino)-5- chloro-N-((4,6-dimethyl-
2-oxo-1,2- dihydropyridin-3- yl)methyl)-2- methylbenzamide 389.3
114 ##STR00129## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(1-
pivaloylazetidin-3- yl)amino)benzamide 473.2 115 ##STR00130##
tert-butyl 3-((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)
amino)azetidine-1- carboxylate 503.3 116 ##STR00131##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(1- pivaloylazetidin-3- yl)amino)-2-
methylbenzamide 487.2 117 ##STR00132## 5-chloro-3-
((cyclopentylmethyl) (methyl)amino)-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 416.2 118
##STR00133## 5-chloro-3- ((cyclopentylmethyl)
(ethyl)amino)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 430.2 119 ##STR00134## 3-(((1s,4s)-4-
aminocyclohexyl)(ethyl) amino)-5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2- methylbenzamide
445.3 120 ##STR00135## 3-(((1r,4r)-4- aminocyclohexyl)(ethyl)
amino)-5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 445.2 121 ##STR00136## 3-(((1s,4s)-4-
acetamidocyclohexyl) (ethyl)amino)-5-chloro-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 487.3 122 ##STR00137## 3-(((1r,4r)-4-
acetamidocyclohexyl) (ethyl)amino)-5-chloro-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 487.3 123 ##STR00138## 5-chloro-3-
(cyclobutyl(methyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 388.1 124
##STR00139## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl(oxetan-3-
yl)amino)benzamide 390.2 125 ##STR00140## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(oxetan-3- yl)amino)-2- methylbenzamide 404.2 126
##STR00141## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (methyl((tetrahydro-2H-
pyran-4- yl)methyl)amino)benzamide 432.3 127 ##STR00142##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((tetrahydro-2H- pyran-4- yl)methyl)amino)-2-
methylbenzamide 446.2 128 ##STR00143## 5-chloro-3-
((cyclohexylmethyl) (ethyl)amino)-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 444.3 129
##STR00144## 3-(((1-acetylpiperidin-4- yl)methyl)(methyl)amino)-
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 473.3 130 ##STR00145##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl((1- pivaloylpiperidin-4-
yl)methyl)amino) benzamide 515.3 131 ##STR00146## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(piperidin-4- ylmethyl)amino)-2- methylbenzamide 445.2 132
##STR00147## 3-(((1-acetylpiperidin-4- yl)methyl)(ethyl)amino)-
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 487.3 133 ##STR00148## 5-chloro-3-
((cyclopropylmethyl) (piperidin-4-yl)amino)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 457.2 134 ##STR00149## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methylbenzamide 135 ##STR00150##
2-bromo-5-chloro-3- (cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)benzamide
466.1 136 ##STR00151## 5-chloro-3- ((cyclohexylmethyl)
(methyl)amino)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 430.2 137 ##STR00152## 5-chloro-3-
(cyclobutyl(ethyl)amino)- N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2- methylbenzamide 402.2 138
##STR00153## tert-butyl 4-(((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(methyl)
amino)methyl)piperidine- 1-carboxylate 531.3 139 ##STR00154##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl(piperidin-4- ylmethyl)amino)
benzamide 431.2 140 ##STR00155## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl((1- methylpiperidin-4- yl)methyl)amino) benzamide 445.2 141
##STR00156## tert-butyl 4-(((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)
amino)methyl)piperidine-1- carboxylate 545.3 142 ##STR00157##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl((1- pivaloylpiperidin-4- yl)methyl)amino)-2-
methylbenzamide 530.4 143 ##STR00158## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl((1-
methylpiperidin-4- yl)methyl)amino)-2- methylbenzamide 459.3 144
##STR00159## 3,6-dichloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-4- (methyl(piperidin-4-
yl)amino)picolinamide 439.8 145 ##STR00160## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-5-methoxy- 2-methylbenzamide 398.1 146 ##STR00161##
3-((3-aminopropyl)(methyl) amino)-5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2- methylbenzamide
391.1 147 ##STR00162## tert-butyl (3-((5-chloro-3-
(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(methyl)
amino)propyl)carbamate 491.1 148 ##STR00163## tert-butyl 4-(3-
(cyclopentyl(methyl) amino)-5-(((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)carbamoyl)-4-
methylphenoxy)piperidine- 1-carboxylate 567.3 149 ##STR00164##
3-(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2-methyl-5- (piperidin-4-
yloxy)benzamide 467.2 150 ##STR00165## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-5- fluoro-2- methylbenzamide 416.1 151
##STR00166## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-5-ethyl-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 446.2 152 ##STR00167## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-5- isopropyl-2- methylbenzamide 440.2 153
##STR00168## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-5-(tetrahydro-2H- pyran-4-yl)benzamide 482.2 154
##STR00169## tert-butyl 4-(3-(((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)carbamoyl)-5- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-4- methylphenyl)-5,6- dihydropyridine-1(2H)-
carboxylate 579.3 155 ##STR00170## tert-butyl 4-(3-(((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)carbamoyl)-5-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-4- methylphenyl)piperidine-
1-carboxylate 581.2 156 ##STR00171## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- ((1-methylpiperidin-4- yl)oxy)benzamide
481.3 157 ##STR00172## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2,5- dimethylbenzamide 412.2 158 ##STR00173##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-5-vinylbenzamide
424.1 159 ##STR00174## 5-(3,6-dihydro-2H-pyran-
4-yl)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 480.3
160 ##STR00175## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-5-(piperidin-4- yl)benzamide 481.3 161 ##STR00176##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-5-(1-
methylpiperidin-4- yl)benzamide 495.3 162 ##STR00177##
5-cyano-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 423.2 163 ##STR00178## 6-chloro-4-
(cyclopentyl(methyl) amino)-N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-3- methylpicolinamide 403.2
164 ##STR00179## 5-acetyl-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methylbenzamide 440.2 165 ##STR00180##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-5-
(trifluoromethyl)benzamide 466.2 166 ##STR00181##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3 yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-5-(2- hydroxyethoxy)-2-
methylbenzamide 458.2 167 ##STR00182## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-5- ethynyl-2- methylbenzamide 422.3 168
##STR00183## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
thiopyran-4-yl)amino)-2- methylbenzamide 448.1 169 ##STR00184##
5-chloro-3- (diethylamino)-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 376 170
##STR00185## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl(propyl)amino)-2-
methylbenzamide 390.2 171 ##STR00186## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(isopropyl)amino)- 2-methylbenzamide 390.1 172 ##STR00187##
5-bromo-3- (cyclopentylamino)-N- ((4,6-dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 432.1 173
##STR00188## 5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (isopropylamino)-2- methylbenzamide 406.1 174
##STR00189## 5-bromo-3- (cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 446.2 175 ##STR00190## 3-acetamido-5-chloro-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 362.1 176 ##STR00191## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
isobutyramido-2- methylbenzamide 390.1 177 ##STR00192##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methylamino)benzamide 334.2 178
##STR00193## 5-chloro-N-((4,6- dmethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (methylamino)benzamide 334.2 179
##STR00194## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (N-methylpivalamido)
benzamide 418.2 180 ##STR00195## 5-chloro-3-(N-
cyclopentylacetamido)- N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2- methylbenzamide 430.3 181
##STR00196## 5-chloro-3- (cyclopentyl(2,3- dihydroxypropyl)amino)-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-
methylbenzamide 462.2 182 ##STR00197## 3-(cyclopentyl(methyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-(prop-1- en-2-yl)benzamide 394.3 183 ##STR00198##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-2-methyl-3- (pyrrolidin-1- ylmethyl)benzamide 354.3 184
##STR00199## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (m- tolylamino)benzamide
410.3 185 [[Duplicate with #184]] 186 ##STR00200##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- (pyrrolidin-3- ylamino)benzamide 389.2 187
##STR00201## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- ((1-methylpyrrolidin-3-
yl)amino)benzamide 403.1 188 ##STR00202## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-((1-
ethylpiperidin-4- yl)(methyl)amino)-2- methylbenzamide 445.4 189
##STR00203## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (piperidin-3-
ylamino)benzamide 403.3 190 ##STR00204## 5-chloro-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-N-((1,4,6-
trimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)benzamide 446.3
191 ##STR00205## 2-bromo-5-chloro-N- ((4,6-dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-
yl)amino)benzamide 498 192 ##STR00206## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methoxybenzamide 449.5
193 ##STR00207## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-5- methoxy-2-
methylbenzamide 428.2 194 ##STR00208## tert-butyl 3-(3-(((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)carbamoyl)-5-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-4- methylphenoxy)azetidine-
1-carboxylate 569.3 195 ##STR00209## 5-(azetidin-3-yloxy)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 469.3
196 ##STR00210## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-5-(2-
methoxyethoxy)-2- methylbenzamide 472.2 197 ##STR00211##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-((1,1- dioxidotetrahydro-2H- thiopyran-4-
yl)(ethyl)amino)-2- methylbenzamide 480.1 198 ##STR00212##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(2- hydroxyethyl)amino)-2- methylbenzamide 392.1
199 ##STR00213## 3-(bis(2- hydroxyethyl)amino)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 408.1 200 ##STR00214## 5-bromo-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 476.1
201 ##STR00215## 5-chloro-N-((4- ((dimethylamino)methyl)-
6-methyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 475.2
202 ##STR00216## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-5-(2- morpholinoethoxy) benzamide 527.4
203 ##STR00217## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-5-(3- morpholinopropoxy) benzamide 541.4 204 ##STR00218##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-N,2-
dimethylbenzamide 446.1 205 ##STR00219## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-ethyl-3-
(methyl(tetrahydro-2H- pyran-4- yl)amino)benzamide 432.1 206
##STR00220## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-ethyl-3- (ethyl(tetrahydro-2H-
pyran-4- yl)amino)benzamide 446.1 207 ##STR00221## 5-chloro-3-
(cyclohexylthio)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 419.2 208 ##STR00222## 5-chloro-3-
(cyclohexylsulfinyl)-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 435.1 209 ##STR00223## 5-chloro-3-
(cyclohexylsulfonyl)-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 451.1 210 ##STR00224##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(1- (methylsulfonyl)piperidin- 4-yl)amino)-2-
methylbenzamide 509.2 211 ##STR00225## 3-((1-acetylpiperidin-4-
yl)(ethyl)amino)-5- chloro-N-((4,6-dimethyl- 2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 495.1 212
##STR00226## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methoxy-3- (methyl(tetrahydro-2H-
pyran-4- yl)amino)benzamide 434.1 213 ##STR00227##
5-(azetidin-3-yl)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 453.2 214 ##STR00228## N-((4,6-dimethyl-2-oxo-
I,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-5- (ethylsulfonyl)-2- methylbenzamide 490.3 215
##STR00229## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-5-
(ethylthio)-2- methylbenzamide 458.2 216 ##STR00230##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-((2,2- dimethyltetrahydro-2H-
pyran-4-yl)(ethyl)amino)- 2-methylbenzamide 460.3 217 ##STR00231##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(2,2,6,6- tetramethyltetrahydro-
2H-pyran-4-yl)amino)-2- methylbenzamide 488.5 218 ##STR00232##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(1- oxidotetrahydro-2H- thiopyran-4-yl)amino)-2-
methylbenzamide 464.1 219 ##STR00233## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(N-
ethylpropionamido)-2- methylbenzamide 404.2 220 ##STR00234##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(N- ethylisobutyramido)-2- methylbenzamide 418.2 221
##STR00235## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(((1s,4s)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methylbenzamide 473.3
222 ##STR00236## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(((1r,4r)-4- (dimethylamino)
cyclohexyl)(ethyl)amino)-2- methylbenzamide 473.4 223 ##STR00237##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((2S,4S)-2- methyltetrahydro-2H-
pyran-4-yl)amino)-2- methylbenzamide 446.2 224 ##STR00238##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-5- formyl-2-
methylbenzamide 426.2 225 ##STR00239## N-((4,6-dimethyl-2-oxo
1,2-dihydropyridin-3- yl)methyl)-5-(3- (dimethylamino)prop-1-
yn-1-yl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 479.2 226 ##STR00240## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methyl-5-(3- morpholinoprop-1-yn-1-
yl)benzamide 521.3 227 ##STR00241## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methyl-5-(3-(4- methylpiperazin-1-
yl)prop-1-yn-1- yl)benzamide 534.4 228 ##STR00242##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1r,4r)-1- oxidotetrahydro-2H-
thiopyran-4-yl)amino)-2- methylbenzamide 464.1 229 ##STR00243##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl(methyl)-3- (ethyl((1s,4s)-1- oxidotetrahydro-2H-
thiopyran-4-yl)amino)-2- methylbenzamide 464.2 230 ##STR00244##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(N- ethylacetamido)-2- methylbenzamide 390.2 231
##STR00245## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl(methyl)-3- (isopropylthio)-2- methylbenzamide
379.1 232 ##STR00246## 5-chloro-3- (cyclopentylthio)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 405.1 233 ##STR00247## terl-butyl 4-(5-chloro-3-
(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)-5,6- dihydropyridine-1(2H)-
carboxylate 486.2 234 ##STR00248## 5-chloro-3-(3,6-dihydro-
2H-pyran-4-yl)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 387.1 235 ##STR00249##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-5-((4-
methylpiperazin-1- yl)methyl)benzamide 510.4 236 ##STR00250##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methyl-5-(pyrrolidin-1-
ylmethyl(benzamide 481.3 237 ##STR00251## N-(5-chloro-3-(((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)carbamoyl)-2-
methylphenyl)-N- methyltetrahydro-2H- pyran-4-carboxamide 446.1 238
##STR00252## 3-((1S,4S)-2-oxa-5- azabicyclo[2.2.1]heptan-
5-yl)-5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 402.1 239 ##STR00253## tert-butyl
4-(3-(((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)piperidine- 1-carboxylate
454.2 240 ##STR00254## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2-methyl-3- (piperidin-4-
yl)benzamide 354.2 241 ##STR00255## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2-methyl-3-
(tetrahydro-2H-pyran-4- yl)benzamide 355.2 242 ##STR00256##
5-chloro-3-(ethyl(1- (methylsulfonyl)piperidin-
4-yl)amino)-2-methyl- N-((1,4,6-trimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)benzamide 522.2 243
##STR00257## 5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2-methyl-3- ((tetrahydro-2H-pyran-4- yl)(2,2,2-
trifluoroethyl)amino)benzamide 531 244 ##STR00258##
5-bromo-3-((2,2- difluoroethyl)(tetrahydro- 2H-pyran-4-yl)amino)-N-
((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 513 245 ##STR00259## 5-(azetidin-3-yl)-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
(methyl(tetrahydro-2H- pyran-4- yl)amino)benzamide 439.3 246
##STR00260## 3-(bis(2- methoxyethyl)amino)-5-
chloro-N-((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 436.2 247 ##STR00261## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((2R,4S)-2- methyltetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 446.2 248 ##STR00262## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(((2R,4r,6S)-2,6- dimethyltetrahydro-2H- pyran-4-yl)(ethyl)amino)-
2-methylbenzamide 460.3 249 ##STR00263## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(((2R,4s,6S)-2,6- dimethyltetrahydro-2H- pyran-4-yl)(ethyl)amino)-
2-methylbenzamide 460.3 250 ##STR00264## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methyl-5- (morpholinomethyl) benzamide 497.3
251 ##STR00265## 3-((1R,4R)-2-oxa-5- azabicyclo[2.2.1]heptan-
5-yl)-5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 402.2 252 ##STR00266##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethylthio)- 2-methylbenzamide 365.2 253 ##STR00267##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl(methyl)-2-methyl-3- ((2- methyltetrahydrofuran-3-
yl)thio)benzamide 421.1 254 ##STR00268## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2-methyl-3- (1-methylpiperidin-4-
yl)benzamide 368.3 255 ##STR00269## N-((4-(azidomethyl)-6-
methyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-5-chloro-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-2- methylbenzamide 473.2
256 ##STR00270## 5-chloro-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methyl-N-((1,4,6- trimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)benzamide 487.4 257 ##STR00271##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(2- methoxyethyl)amino)-2- methylbenzamide 406.2
258 ##STR00272## 3-cyclohexyl-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 353.2 259
##STR00273## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(4,4- dimethylcyclohexyl)-2- methylbenzamide 381.3 260
##STR00274## 5-azido-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methylbenzamide 439.3 261 ##STR00275## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-N,2- dimethylbenzamide
487.2 262 ##STR00276## 5-chloro-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-N-((4- ethyl-6-methyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2- methylbenzamide 446.1 263
##STR00277## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- ((tetrahydro-2H-pyran-4-
yl)thio)benzamide 421 264 ##STR00278## tert-butyl 4-((5-chloro-3-
(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)thio) piperidine-1-carboxylate
542.2 265 ##STR00279## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (piperidin-4-
ylthio)benzamide 420.1 266 ##STR00280## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl(methyl)-2-methyl-3-
((1-methylpiperidin-4- yl)thio)benzamide 434.1 267 ##STR00281##
5-chloro-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-N-((6-methyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)benzamide 418.1 268 ##STR00282##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methylbenzamide 439.3
269 ##STR00283## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-2-methyl-3- (methyl(telrahydro-2H-
pyran-4-yl)amino)-5-(1- methylazetidin-3- yl)benzamide 453.2 270
##STR00284## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-3- (2-oxopiperidin-1-
yl)benzamide 402.1 271 ##STR00285## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3- ((1-
(methylsulfonyl)piperidin- 4-yl)thio)benzamide 498 272 ##STR00286##
5-chloro-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-N-((4-methyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)benzamide 418.2 273 ##STR00287##
5-chloro-3-(((1r,4r)-4- (diethylamino)cyclohexyl)
(ethyl)amino)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 501.5 274 ##STR00288##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(2,5- dioxopyrrolidin-1-yl)-2- methylbenzamide 402.3
275 ##STR00289## 5-chloro-N-(1-(4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)ethyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methylbenzamide 446.2 276 ##STR00290##
5-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methylbenzamide 519.3 277 ##STR00291##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5- ethynyl-2- methylbenzamide 463.5 278 ##STR00292##
5-cyano-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methylbenzamide 464.3 279 ##STR00293##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1r,4r)-4- (methyl(tetrahydro-2H- pyran-4-
yl)amino)cyclohexyl) amino)-2-methylbenzamide 543.7 280
##STR00294## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(1,4- dioxaspiro[4.5]decan-8-
yl)amino)-2- methylbenzamide 488.3 281 ##STR00295##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(2,6- dioxopiperidin-1-yl)-2- methylbenzamide 416.1
282 ##STR00296## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl(methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5-ethyl- 2-methylbenzamide 467.4 283 ##STR00297##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methyl-5-(3-(4- methylpiperazin-1- yl)prop-1-yn-1-
yl)benzamide 575.5 284 ##STR00298## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((1r,4r)-4- (methylamino)cyclohexyl) amino)-2-
methylbenzamide 459.4 285 ##STR00299## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((1r,4r)-4- (ethyl(methyl)amino) cyclohexyl)amino)-2-
methylbenzamide 487.4 286 ##STR00300## 5-chloro-3-(((1r,4r)-4-
((cyclopropylmethyl) (methyl)amino)cyclohexyl)
(ethyl)amino)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 513.4 287 ##STR00301##
5-chloro-3-(((1r,4r)-4- (cyclobutyl(methyl)amino)
cyclohexyl)(ethyl)amino)- N-((4,6-dimethyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 513.3 288
##STR00302## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(4- oxocyclohexyl)amino)-2-
methylbenzamide 444.3 289 ##STR00303## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-3-
((tetrahydro-2H-pyran-4- yl)oxy)benzamide 405.2 290 ##STR00304##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5-(3- (dimethylamino)prop-1- yn-1-yl)-2-
methylbenzamide 520.6 291 ##STR00305## 5-chloro-N-((4,6-
dimethyl-2-oxo-l,2- dihydropyridin-3- yl(methyl)-3-
(ethyl((1r,4r)-4-((2- hydroxyethyl)(methyl)
amino)cyclohexyl)amino)- 2-methylbenzamide 503.3 292 ##STR00306##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1r,4r)-4- (pyrrolidin-1-
yl)cyclohexyl)amino)-2- methylbenzamide 499.3 293 ##STR00307##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1s,4s)-4- (pyrrolidin-1-
yl)cyclohexyl)amino)-2- methylbenzamide 499.3 294 ##STR00308##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1r,4r)-4- morpholinocyclohexyl) amino)-2-
methylbenzamide 515.3 295 ##STR00309## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((1s,4s)-4- morpholinocyclohexyl) amino)-2- methylbenzamide
515.3 296 ##STR00310## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1r,4r)-4-(4-
methylpiperazin-1- yl)cyclohexyl)amino)-2- methylbenzamide 528.4
297 ##STR00311## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1s,4s)-4-(4-
methylpiperazin-1- yl)cyclohexyl)amino)-2- methylbenzamide 528.3
298 ##STR00312## 5-chloro-N-((4,6- dimethyl-2-oxo-1-propyl-
1,2-dihydropyridin-3- yl)methyl)-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methylbenzamide 515.3
299 ##STR00313## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl(methyl)-3- (ethyl((1r,4r)-4-((2-
methoxyethyl)(methyl) amino(cyclohexyl)amino)- 2-methylbenzamide
517.4 300 ##STR00314## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1s,4s)-4- (methyl(2,2,2-
trifluoroethyl)amino) cyclohexyl)amino)-2- methylbenzamide 541.4
301 ##STR00315## N-((1-benzyl-4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-5-chloro-3- (((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methylbenzamide 563.4
302 ##STR00316## 5-chloro-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-N-((1- (2-hydroxyethyl)-4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 517.4 303
##STR00317## 5-chloro-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-N-((1- (2-methoxyethyl)-4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-2- methylbenzamide 531.4 304
##STR00318## 3-(((1s,4s)-4-(azetidin-1- yl)cyclohexyl)(ethyl)
amino)-5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 485.2 305 ##STR00319##
3-(((1r,4r)-4-(azetidin-1- yl)cyclohexyl)(ethyl
(amino)-5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 485.2 306 ##STR00320##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methyl-5- (trifluoromethyl)benzamide 507.5 307
##STR00321## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1r,4r)-4-
hydroxycyclohexyl)amino)- 2-methylbenzamide 446.3 308 ##STR00322##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5- methoxy-2- methylbenzamide 469.8 309 ##STR00323##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl(methyl)-3-((4-
(dimethylamino)cyclohexyl) (ethyl)amino)-5-(2- hydroxyethoxy)-2-
methylbenzamide 499.6 310 ##STR00324## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-((4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5-(2- methoxyethoxy)-2- methylbenzamide 513.6 311
##STR00325## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5- (ethylthio)-2- methylbenzamide 499.6 312
##STR00326## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1r,4r)-4- (methyl(2,2,2-
trifluoroethyl)amino) cyclohexyl)amino)-2- methylbenzamide 541.5
313 ##STR00327## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(4- hydroxycyclohexyl)amino)-
2-methylbenzamide 446.3 314 ##STR00328## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2,5',5'-
trimethyl-2'-oxo- 2',3',4',5'-tetrahydro-[1,1'-
biphenyl]-3-carboxamide 427.4 315 ##STR00329##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methyl-5-(tetrahydro-2H- pyran-4-yl)benzamide
523.6 316 ##STR00330## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methyl-N-((6-methyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)benzamide 459.5 317
##STR00331## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(5,5- dimethyl-2- oxocyclohexyl)-2- methylbenzamide
395.4 318 ##STR00332## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(5,5- dimethyl-2- oxocyclohexyl)-2-
methylbenzamide 429.4 319 ##STR00333## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-5- (ethylsulfonyl)-2-
methylbenzamide 531.6 320 ##STR00334## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-5- isopropyl-2-
methylbenzamide 481.4 321 ##STR00335## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-N-((4-
ethyl-6-methyl-2-oxo- 1,2-dihydropyridin-3- yl(methyl)-2-
methylbenzamide 487.5 322 ##STR00336## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methyl-5-(1- methylazetidin-3- yl)benzamide
467.3 323 ##STR00337## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-N-((1-
ethyl-4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 501.5 324 ##STR00338## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-N-((1-(2-
(dimethylamino)ethyl)- 4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 544.6 325 ##STR00339## 5-chloro-3-
(ethyl(tetrahydro-2H- pyran-4-yl)amino)-N-((4- (hydroxymethyl)-6-
methyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2- methylbenzamide
448.4 326 ##STR00340## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-N-((4- (hydroxymethyl)-6-
methyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2- methylbenzamide
489.5 327 ##STR00341## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methyl-N-((4-methyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)benzamide 459.4 328
##STR00342## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-2- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-3-
methylisonicotinamide 399.1 329 ##STR00343## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(4-
methoxycyclohexyl)amino)- 2-methylbenzamide 460.4 330 ##STR00344##
5-chloro-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-2-
methyl-N-((6-methyl-2- oxo-4-(trifluoromethyl)-
1,2-dihydropyridin-3- yl(methyl)benzamide 486.4 331 ##STR00345##
5-chloro-3-(((1r,4r)-4- (dimethylamino)cyclohexyl) (ethyl)amino)-2-
methyl-N-((6-methyl-2- oxo-4-(trifluoromethyl)-
1,2-dihydropyridin-3- yl(methyl)benzamide 527.5 332 ##STR00346##
3-(benzyl(methyl)amino)- N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-2- (trifluoromethyl)benzamide
444.2 333 ##STR00347## 3-(benzyl(ethyl)amino)-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-
(trifluoromethyl)benzamide 458.3 334 ##STR00348##
3-(benzyl(ethyl)amino)- N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl(methyl)-2- methylbenzamide 404.3
335 ##STR00349## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-((4- (dimethylamino)
cyclohexyl)thio)-2- methylbenzamide 462.4 336 ##STR00350##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-((4- hydroxycyclohexyl)thio)- 2-methylbenzamide 435.3
337 ##STR00351## 5-chloro-3- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-2- methyl-N-((2-oxo-1,2- dihydropyridin-3-
yl)methyl)benzamide 404.3 338 ##STR00352## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methyl-N-((2-oxo-1,2-
dihydropyridin-3- yl)methyl)benzamide 445.5 339 ##STR00353##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(ethyl(4- hydroxy-4- methylcyclohexyl)amino)-
2-methylbenzamide 426.5 340 ##STR00354## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(ethyl(4- methoxy-4-
methylcyclohexyl(amino)- 2-methylbenzamide 440.45 341 ##STR00355##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(ethyl(4- hydroxy-4-(6- methoxypyridin-3-
yl)cyclohexyl)amino)-2- methylbenzamide 519.3 342 ##STR00356##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-((4- (dimethylamino) cyclohexyl)oxy)-2-
methylbenzamide 446.3 343 ##STR00357## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((1r,4r)-4- methoxycyclohexyl)amino)- 2-methylbenzamide 460.4
344 ##STR00358## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1s,4s)-4-
methoxycyclohexyl)amino)- 2-methylbenzamide 460.3 345 ##STR00359##
5-chloro-3- (ethyl(tetrahydro-2H- pyran-4-yl)amino)-N-((6-
ethyl-4-methyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-2-
methylbenzamide 446.3 346 ##STR00360## 5-(cyclopropylethynyl)-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methylbenzamide 503.5 347 ##STR00361##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- ethylbenzamide 487.2 348 ##STR00362##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((1s,4s)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- ethylbenzamide 487.2 349 ##STR00363##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-((4-
(dmethylamino)cyclohexyl) (ethyl)amino)-2- methyl-5-((1-
methylazetidin-3- yl)oxy)benzamide 524.6 350 ##STR00364##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5-(3- hydroxy-3-methylbut-1- yn-1-yl)-2-
methylbenzamide 521.5 351 ##STR00365## 5-(azetidine-1-carbonyl)-
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2- methylbenzamide 522.8 352 ##STR00366##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(ethyl(3- methoxypropyl)amino)- 2-methylbenzamide
386.4 353 ##STR00367## 5-chloro-3-(((1r,4r)-4-
(dimethylamino)cyclohexyl) (ethyl)amino)-2- methyl-N-((6-methyl-2-
oxo-4-propyl-1,2- dihydropyridin-3- yl)methyl)benzamide 501.5 354
##STR00368## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-5-(3,3- dimethylbut-1-yn-1-yl)- 2-methylbenzamide
519.6 355 ##STR00369## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(ethyl(3-
hydroxypropyl)amino)-2- methylbenzamide 372.3 356 ##STR00370##
5-chloro-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-N-((5- fluoro-4,6-dimethyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2- methylbenzamide 491.1 357 ##STR00371##
5-bromo-2-chloro-N- ((4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)benzamide 539.25 358 ##STR00372##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-((3-
(dimethylamino)propyl) (ethyl)amino)-2- methylbenzamide 399.4 359
##STR00373## 3-bromo-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-5-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-2,6- dimethylbenzamide 531.4 360 ##STR00374##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(4- methylcyclohexyl)amino)- 2-methylbenzamide
444.4 361 ##STR00375## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(4- ethylcyclohexyl)amino)-
2-methylbenzamide 458.4 362 ##STR00376## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(1-
methylpiperidin-4- yl)amino)-2- methylbenzamide 445.4 363
##STR00377## 5-chloro-3-((4,4- difluorocyclohexyl)(ethyl)
amino)-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-
methylbenzamide 466.2 364 ##STR00378## 5-chloro-3-(((1r,4r)-4-
((2,2- difluoroethyl)(methyl) amino)cyclohexyl)(ethyl)
amino)-N-((4,6-dimethyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 523.5 365 ##STR00379## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((1r,4r)-4- (methyl(oxetan-3- yl)amino)cyclohexyl)
amino)-2-methylbenzamide 515.4 366 ##STR00380## 5-chloro-N-((4,6-
dimethyl-2-oxo-l,2- dihydropyridin-3- yl)methyl)-3-((4,4-
dimethylcyclohexyl) (ethyl)amino)-2- methylbenzamide 458.4 367
##STR00381## 3-((4-(tert-butyl) cyclohexyl)(ethyl)amino)-
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 486.5 368 ##STR00382##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(3'- oxo-3'H- spiro[cyclohexane-1,1'-
isobenzofuran]-4- yl)amino)-2- methylbenzamide 548.5 369
##STR00383## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(1- isopropylpiperidin-4-
yl)amino)-2- methylbenzamide 473.5 370 ##STR00384##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(4- isopropylcyclohexyl) amino)-2-
methylbenzamide 472.3 371 ##STR00385## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(1,4-
dioxaspiro[4.5]decan-7- yl)amino)-2- methylbenzamide 488.4 372
##STR00386## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(1- oxaspiro[4.5]decan-8-
yl)amino)-2- methylbenzamide 486.35 373 ##STR00387##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1s,4s)-3'-oxo-3'H- spiro[cyclohexane-1,1'-
isobenzofuran]-4- yl)amino)-2- methylbenzamide 548.4 374
##STR00388## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1r,4r)-3'-oxo-3'H-
spiro[cyclohexane-1,1'- isobenzofuran]-4- yl)amino)-2-
methylbenzamide 548.4 375 ##STR00389## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(7'-
oxo-7'H- spiro[cyclohexane-1,5- furo[3,4-b]pyridin]-4- yl)amino)-2-
methylbenzamide 549.3 376 ##STR00390## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-(ethyl(3'-
oxo-3'H- spiro[cyclohexane-1,1'- furo[3,4-c]pyridin]-4-
yl)amino)-2- methylbenzamide 549.5 377 ##STR00391##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(ethyl(1- methylazetidin-3- yl)amino)-2-
methylbenzamide 417.3 378 ##STR00392## (1r,4r)-ethyl 4-((5-chloro-
3-(((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)amino)
cyclohexanecarboxylate 502.5 379 ##STR00393## (1s,4s)-ethyl 4-((5-
chloro-3-(((4,6-dimethyl- 2-oxo-1,2- dihydropyridin-3-
yl)methyl)carbamoyl)-2- methylphenyl)(ethyl)amino)
cyclohexanecarboxylate 502.4 380 ##STR00394##
(lr,4r)-4-((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2-
methylphenyl)(ethyl)amino) cyclohexanecarboxylic acid 474.3 381
##STR00395## (1s,4s)-4-((5-chloro-3- (((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)carbamoyl)-2-
methylphenyl)(ethyl)amino) cyclohexanecarboxylic acid 474.4 382
##STR00396## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1r,4r)-4- (pyrrolidine-1-
carbonyl)cyclohexyl)amino)- 2-methylbenzamide 527.5 383
##STR00397## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1s,4s)-4- (pyrrolidine-1-
carbonyl)cyclohexyl)amino)- 2-methylbenzamide 527.4 384
##STR00398## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3-(ethyl(4- (hydroxymethyl)cyclohexyl)
amino)-2-methylbenzamide 460.2 385 ##STR00399##
5-chloro-3-(ethyl(1- methylpiperidin-4- yl)amino)-2-methyl-N-
((6-methyl-2-oxo-4- propyl-1,2- dihydropyridin-3-
yl)methyl)benzamide 473.4 386 ##STR00400## 3,6-dichloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-4-
(methyl(tetrahydro-2H- pyran-4- yl)amino)picolinamide 440.8 387
##STR00401## 3-bromo-6-chloro-N- ((4,6-dimethyl-2-oxo-1,2-
dihydropyridin-3- yl(methyl)-4- (methyl(tetrahydro-2H- pyran-4-
yl)amino)picolinamide 485 388 ##STR00402## 5-chloro-N-((4,6-
dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-3-
(ethyl((5s,8s)-2-oxo-1- azaspiro[4.5]decan-8- yl)amino)-2-
methylbenzamide 499.35 389 ##STR00403## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-5- (ethyl(tetrahydro-2H-
pyran-4-yl)amino)-4- methylnicotinamide 399.5 390 ##STR00404##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-5-(((1r,4r)-4- (dimethylamino)cyclohexyl)
(ethyl)amino)-4- methylnicotinamide 440.5 391 ##STR00405##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((5r,8r)-2-oxo-1- azaspiro[4.5]decan-8-
yl)amino)-2- methylbenzamide 499.4 392 ##STR00406##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-(ethyl(1- methylpiperidin-4- yl)amino)-2-methyl-5-
(trifluoromethyl) benzamide 501.3 393 ##STR00407## 3-(ethyl(1-
methylpiperidin-4- yl)amino)-2-methyl-N- ((6-methyl-2-oxo-4-
propyl-1,2- dihydropyridin-3- yl)methyl)-5- (trifluoromethyl)
benzamide 505.2 394 ##STR00408## 3-(ethyl(1- methylpiperidin-4-
yl)amino)-N-((4- isopropyl-6-methyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (trifluoromethyl) benzamide 529.2 395
##STR00409## 3-(ethyl(1- methylpiperidin-4- yl)amino)-2-methyl-N-
((6-methyl-2-oxo-4- (trifluoromethyl)-1,2- dihydropyridin-3-
yl)methyl)-5- (trifluoromethyl) benzamide 555.2 396 ##STR00410##
3-(ethyl(1- methylpiperidin-4- yl)amino)-N-((5-fluoro-
6-methyl-2-oxo-4-propyl- 1,2-dihydropyridin-3-
yl)methyl)-2-methyl-5- (trifluoromethyl) benzamide 525.6 397
##STR00411## 3-(ethyl(1- methylpiperidin-4- yl)amino)-N-((5-fluoro-
4-isopropyl-6-methyl-2- oxo-1,2-dihydropyridin-
3-yl)methyl)-2-methyl-5- (trifluoromethyl) benzamide 525.6 398
##STR00412## 3-(ethyl(1- methylpiperidin-4- yl)amino)-N-((5-fluoro-
4,6-dimethyl-2-oxo-1,2- dihydropyridin-3- yl)methyl)-2-methyl-5-
(trifluoromethyl) benzamide 497.5
399 ##STR00413## 3-(ethyl(1- methylpiperidin-4-
yl)amino)-N-((5-fluoro- 6-methyl-2-oxo-4- (trifluoromethyl)-1,2-
dihydropyridin-3- yl)methyl)-2-methyl-5- (trifluoromethyl)
benzamide 551.5 400 ##STR00414## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(ethyl(1- isopropylpiperidin-4-
yl)amino)-2-methyl-5- (trifluoromethyl) benzamide 507.2 401
##STR00415## N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3-
yl)methyl)-3-((2R,6R)- 2,6-dimethylpiperidin-4- yl)(ethyl)amino)-2-
methyl-5- (trifluoromethyl)benzamide 493.1 402 ##STR00416##
N-((4,6-dimethyl-2-oxo- 1,2-dihydropyridin-3- yl)methyl)-3-
(((2S,4s,6R)-2,6- dimethylpiperidin-4- yl)(ethyl)amino)-2-
methyl-5- (trifluoromethyl) benzamide 493.2 403 ##STR00417##
3-(ethyl(1- methylazetidin-3- yl)amino)-2-methyl-N-
((6-methyl-2-oxo-4- propyl-1,2- dihydropyridin-3- yl)methyl)-5-
(trifluoromethyl) benzamide 480.5 404 ##STR00418## 3-(ethyl(1-
methylazetidin-3- yl)amino)-N-((4- isopropyl-6-methyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2-methyl-5- (trifluoroniethyl)
benzamide 479.5 405 ##STR00419## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(ethyl(1- methylazetidin-3-
yl)amino)-2-methyl-5- (trifluoromethyl) benzamide 451.5 406
##STR00420## 5-chloro-3-(ethyl(1- methylpiperidin-4-
yl)amino)-2-methyl-N- ((6-methyl-2-oxo-4- propyl-1,2-
dihydropyridin-3- yl)methyl)benzamide 473.1 407 ##STR00421##
5-chloro-3-(ethyl(1- methylpiperidin-4- yl)amino)-N-((4-
isopropyl-6-methyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 495.2 408 ##STR00422## 5-chloro-3-(ethyl(1-
methylpiperidin-4- yl)amino)-2-methyl-N- ((6-methyl-2-oxo-4-
(trifluoromethyl)-1,2- dihydropyridin-3- yl)methylbenzamide 499.1
409 ##STR00423## 5-chloro-3-(ethyl(1- methylpiperidin-4-
yl)amino)-N-((5-fluoro- 4,6-dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-2- methylbenzamide 463.5 410 ##STR00424##
5-chloro-3-(ethyl(1- methylpiperidin-4- yl)amino)-N-((5-fluoro-
6-methyl-2-oxo-4-propyl- 1,2-dihydropyridin-3- yl(methyl)-2-
methylbenzamide 491.5 411 ##STR00425## 5-chloro-3-(ethyl(1-
methylpiperidin-4- yl)amino)-N-((5-fluoro- 4-isopropyl-6-methyl-2-
oxo-1,2-dihydropyridin- 3-yl)methyl)-2- methylbenzamide 491.5 412
##STR00426## 5-chloro-3-(ethyl(1- methylazetidin-3-
yl)amino)-2-methyl-N- ((6-methyl-2-oxo-4- propyl-1,2-
dihydropyridin-3- yl)methyl)benzamide 445.5 413 ##STR00427##
5-chloro-3-(ethyl(1- methylazetidin-3- yl)amino)-N-((4-
isopropyl-6-methyl-2- oxo-1,2-dihydropyridin- 3-yl)methyl)-2-
methylbenzamide 445.6 414 ##STR00428## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3-(((2R,6R)-
2,6-dimethylpiperidin-4- yl)(ethyl)amino)-5- fluoro-2-
methylbenzamide 443.1 415 ##STR00429## N-((4,6-dimethyl-2-oxo-
1,2-dihydropyridin-3- yl)methyl)-3- (((2S,4s,6R)-2,6-
dimethylpiperidin-4- yl)(ethyl)amino)-5- fluoro-2- methylbenzamide
443.2 416 ##STR00430## 5-chloro-N-((4,6- dimethyl-2-oxo-1,2-
dihydropyridin-3- yl)methyl)-3- (ethyl((1s,4s)-4- (methylsulfonyl)
cyclohexyl)amino)-2- methylbenzamide 508.1 417 ##STR00431##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3- (ethyl((1r,4r)-4- (methylsulfonyl)
cyclohexyl)amino)-2- methylbenzamide 508 418 ##STR00432##
5-chloro-N-((4,6- dimethyl-2-oxo-1,2- dihydropyridin-3-
yl)methyl)-3-(((2S,6R)- 2,6-dimethylpiperidin-4-
yl)(ethyl)amino)-2- methylbenzamide *The cis/trans stereochemical
assignments of Compounds 58/59, 60/61, 107/108, and 416/417 are
arbitratily assigned. .dagger.MS-Mass spectrometry of sample. This
can be recorded in positive or negative ion modes and potential
ions include M, M + 1, M + 23 (M + Na) and M - 1.
[0604] As used herein, "alkyl", "C.sub.1, C.sub.2, C.sub.3,
C.sub.4, C.sub.5 or C.sub.6 alkyl" or "C.sub.1-C.sub.6 alkyl" is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 or
C.sub.6 straight chain (linear) saturated aliphatic hydrocarbon
groups and C.sub.3, C.sub.4, C.sub.5 or C.sub.6 branched saturated
aliphatic hydrocarbon groups. For example, C.sub.1-C.sub.6 alkyl is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 and
C.sub.6 alkyl groups. Examples of alkyl include, moieties having
from one to six carbon atoms, such as, but not limited to, methyl,
ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl,
s-pentyl or n-hexyl.
[0605] In certain embodiments, a straight chain or branched alkyl
has six or fewer carbon atoms (e.g., C.sub.1-C.sub.6 for straight
chain, C.sub.3-C.sub.6 for branched chain), and in another
embodiment, a straight chain or branched alkyl has four or fewer
carbon atoms.
[0606] As used herein, the term "cycloalkyl" refers to a saturated
or unsaturated nonaromatic hydrocarbon mono- or multi-ring (e.g.,
fused, bridged, or spiro rings) system having 3 to 30 carbon atoms
(e.g., C.sub.3-C.sub.10). Examples of cycloalkyl include, but are
not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl,
cycloheptenyl, and adamantyl. The term "heterocycloalkyl" refers to
a saturated or unsaturated nonaromatic 3-8 membered monocyclic,
7-12 membered bicyclic (fused, bridged, or spiro rings), or 11-14
membered tricyclic ring system (fused, bridged, or spiro rings)
having one or more heteroatoms (such as O, N, S, or Se), unless
specified otherwise. Examples of heterocycloalkyl groups include,
but are not limited to, piperidinyl, piperazinyl, pyrrolidinyl,
dioxanyl, tetrahydrofuranyl, isoindolinyl, indolinyl,
imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,
triazolidinyl, tetrahyrofuranyl, oxiranyl, azetidinyl, oxetanyl,
thietanyl, 1,2,3,6-tetrahydropyridinyl, tetrahydropyranyl,
dihydropyranyl, pyranyl, morpholinyl, tetrahydrothiopyranyl,
1,4-diazepanyl, 1,4-oxazepanyl, 2-oxa-5-azabicyclo[2.2.1]heptanyl,
2,5-diazabicyclo[2.2.1]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,
2,6-diazaspiro[3.3]heptanyl, 1,4-dioxa-8-azaspiro[4.5]decanyl,
1,4-dioxaspiro[4.5]decanyl, 1-oxaspiro[4.5]decanyl,
1-azaspiro[4.5]decanyl,
3'H-spiro[cyclohexane-1,1'-isobenzofuran]-yl,
7'H-spiro[cyclohexane-1,5'-furo[3,4-b]pyridin]-yl,
3'H-spiro[cyclohexane-1,1'-furo[3,4-c]pyridin]-yl, and the
like.
[0607] The term "optionally substituted alkyl" refers to
unsubstituted alkyl or alkyl having designated substituents
replacing one or more hydrogen atoms on one or more carbons of the
hydrocarbon backbone. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0608] An "arylalkyl" or an "aralkyl" moiety is an alkyl
substituted with an aryl (e.g., phenylmethyl (benzyl)). An
"alkylaryl" moiety is an aryl substituted with an alkyl (e.g.,
methylphenyl).
[0609] As used herein, "alkyl linker" is intended to include
C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 or C.sub.6 straight
chain (linear) saturated divalent aliphatic hydrocarbon groups and
C.sub.3, C.sub.4, C.sub.5 or C.sub.6 branched saturated aliphatic
hydrocarbon groups. For example, C.sub.1-C.sub.6 alkyl linker is
intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5 and
C.sub.6 alkyl linker groups. Examples of alkyl linker include,
moieties having from one to six carbon atoms, such as, but not
limited to, methyl (--CH.sub.2--), ethyl (--CH.sub.2CH.sub.2--),
n-propyl (--CH.sub.2CH.sub.2CH.sub.2--), i-propyl
(--CHCH.sub.3CH.sub.2--), n-butyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), s-butyl
(--CHCH.sub.3CH.sub.2CH.sub.2--), i-butyl
(--C(CH.sub.3).sub.2CH.sub.2--), n-pentyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--), s-pentyl
(--CHCH.sub.3CH.sub.2CH.sub.2CH.sub.2--) or n-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2--).
[0610] "Alkenyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
that contain at least one double bond. For example, the term
"alkenyl" includes straight chain alkenyl groups (e.g., ethenyl,
propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl,
decenyl), and branched alkenyl groups. In certain embodiments, a
straight chain or branched alkenyl group has six or fewer carbon
atoms in its backbone (e.g., C.sub.2-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain). The term "C.sub.2-C.sub.6"
includes alkenyl groups containing two to six carbon atoms. The
term "C.sub.3-C.sub.6" includes alkenyl groups containing three to
six carbon atoms.
[0611] The term "optionally substituted alkenyl" refers to
unsubstituted alkenyl or alkenyl having designated substituents
replacing one or more hydrogen atoms on one or more hydrocarbon
backbone carbon atoms. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, heterocyclyl, alkylaryl, or an aromatic or
heteroaromatic moiety.
[0612] "Alkynyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
which contain at least one triple bond. For example, "alkynyl"
includes straight chain alkynyl groups (e.g., ethynyl, propynyl,
butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl),
and branched alkynyl groups. In certain embodiments, a straight
chain or branched alkynyl group has six or fewer carbon atoms in
its backbone (e.g., C.sub.2-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain). The term "C.sub.2-C.sub.6"
includes alkynyl groups containing two to six carbon atoms. The
term "C.sub.3-C.sub.6" includes alkynyl groups containing three to
six carbon atoms.
[0613] The term "optionally substituted alkynyl" refers to
unsubstituted alkynyl or alkynyl having designated substituents
replacing one or more hydrogen atoms on one or more hydrocarbon
backbone carbon atoms. Such substituents can include, for example,
alkyl, alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
amino (including alkylamino, dialkylamino, arylamino, diarylamino
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0614] Other optionally substituted moieties (such as optionally
substituted cycloalkyl, heterocycloalkyl, aryl, or heteroaryl)
include both the unsubstituted moieties and the moieties having one
or more of the designated substituents. For example, substituted
heterocycloalkyl includes those substituted with one or more alkyl
groups, such as 2,2,6,6-tetramethyl-piperidinyl and
2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl.
[0615] "Aryl" includes groups with aromaticity, including
"conjugated," or multicyclic systems with at least one aromatic
ring and do not contain any heteroatom in the ring structure.
Examples include phenyl, benzyl, 1,2,3,4-tetrahydronaphthalenyl,
etc.
[0616] "Heteroaryl" groups are aryl groups, as defined above,
except having from one to four heteroatoms in the ring structure,
and may also be referred to as "aryl heterocycles" or
"heteroaromatics." As used herein, the term "heteroaryl" is
intended to include a stable 5-, 6-, or 7-membered monocyclic or
7-, 8-, 9-, 10-, 11- or 12-membered bicyclic aromatic heterocyclic
ring which consists of carbon atoms and one or more heteroatoms,
e.g., 1 or 1-2 or 1-3 or 1-4 or 1-5 or 1-6 heteroatoms, or e.g. 2,
3, 4, 5, or 6 heteroatoms, independently selected from the group
consisting of nitrogen, oxygen and sulfur. The nitrogen atom may be
substituted or unsubstituted (i.e., N or NR wherein R is H or other
substituents, as defined). The nitrogen and sulfur heteroatoms may
optionally be oxidized (i.e., N.fwdarw.O and S(O).sub.p, where p=1
or 2). It is to be noted that total number of S and O atoms in the
aromatic heterocycle is not more than 1.
[0617] Examples of heteroaryl groups include pyrrole, furan,
thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole,
pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine,
pyrimidine, and the like.
[0618] Furthermore, the terms "aryl" and "heteroaryl" include
multicyclic aryl and heteroaryl groups, e.g., tricyclic, bicyclic,
e.g., naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,
isoquinoline, naphthrydine, indole, benzofuran, purine, benzofuran,
deazapurine, indolizine.
[0619] In the case of multicyclic aromatic rings, only one of the
rings needs to be aromatic (e.g., 2,3-dihydroindole), although all
of the rings may be aromatic (e.g., quinoline). The second ring can
also be fused or bridged.
[0620] The cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring
can be substituted at one or more ring positions (e.g., the
ring-forming carbon or heteroatom such as N) with such substituents
as described above, for example, alkyl, alkenyl, alkynyl, halogen,
hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, amino (including alkylamino,
dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino
(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and
ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,
sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkylaryl, or an aromatic or heteroaromatic moiety. Aryl and
heteroaryl groups can also be fused or bridged with alicyclic or
heterocyclic rings, which are not aromatic so as to form a
multicyclic system (e.g., tetralin, methylenedioxyphenyl).
[0621] As used herein, "carbocycle" or "carbocyclic ring" is
intended to include any stable monocyclic, bicyclic or tricyclic
ring having the specified number of carbons, any of which may be
saturated, unsaturated, or aromatic. Carbocycle includes cycloalkyl
and aryl. For example, a C.sub.3-C.sub.14 carbocycle is intended to
include a monocyclic, bicyclic or tricyclic ring having 3, 4, 5, 6,
7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms. Examples of carbocycles
include, but are not limited to, cyclopropyl, cyclobutyl,
cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl,
cycloheptenyl, cycloheptyl, cycloheptenyl, adamantyl, cyclooctyl,
cyclooctenyl, cyclooctadienyl, fluorenyl, phenyl, naphthyl,
indanyl, adamantyl and tetrahydronaphthyl. Bridged rings are also
included in the definition of carbocycle, including, for example,
[3.3.0]bicyclooctane, [4.3.0]bicyclononane, and [4.4.0]
bicyclodecane and [2.2.2] bicyclooctane. A bridged ring occurs when
one or more carbon atoms link two non-adjacent carbon atoms. In one
embodiment, bridge rings are one or two carbon atoms. It is noted
that a bridge always converts a monocyclic ring into a tricyclic
ring. When a ring is bridged, the substituents recited for the ring
may also be present on the bridge. Fused (e.g., naphthyl,
tetrahydronaphthyl) and spiro rings are also included.
[0622] As used herein, "heterocycle" or "heterocyclic group"
includes any ring structure (saturated, unsaturated, or aromatic)
which contains at least one ring heteroatom (e.g., N, O or S).
Heterocycle includes heterocycloalkyl and heteroaryl. Examples of
heterocycles include, but are not limited to, morpholine,
pyrrolidine, tetrahydrothiophene, piperidine, piperazine, oxetane,
pyran, tetrahydropyran, azetidine, and tetrahydrofuran.
[0623] Examples of heterocyclic groups include, but are not limited
to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl,
carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,
2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran,
furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl,
1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl,
3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl,
isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl,
methylenedioxyphenyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,
1,2,4-oxadiazol5(4H)-one, oxazolidinyl, oxazolyl, oxindolyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl,
pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,
pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,
1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and
xanthenyl.
[0624] The term "substituted," as used herein, means that any one
or more hydrogen atoms on the designated atom is replaced with a
selection from the indicated groups, provided that the designated
atom's normal valency is not exceeded, and that the substitution
results in a stable compound. When a substituent is oxo or keto
(i.e., .dbd.O), then 2 hydrogen atoms on the atom are replaced.
Keto substituents are not present on aromatic moieties. Ring double
bonds, as used herein, are double bonds that are formed between two
adjacent ring atoms (e.g., C.dbd.C, C.dbd.N or N.dbd.N). "Stable
compound" and "stable structure" are meant to indicate a compound
that is sufficiently robust to survive isolation to a useful degree
of purity from a reaction mixture, and formulation into an
efficacious therapeutic agent.
[0625] When a bond to a substituent is shown to cross a bond
connecting two atoms in a ring, then such substituent may be bonded
to any atom in the ring. When a substituent is listed without
indicating the atom via which such substituent is bonded to the
rest of the compound of a given formula, then such substituent may
be bonded via any atom in such formula. Combinations of
substituents and/or variables are permissible, but only if such
combinations result in stable compounds.
[0626] When any variable (e.g., R.sub.1) occurs more than one time
in any constituent or formula for a compound, its definition at
each occurrence is independent of its definition at every other
occurrence. Thus, for example, if a group is shown to be
substituted with 0-2 R.sub.1 moieties, then the group may
optionally be substituted with up to two R.sub.1 moieties and
R.sub.1 at each occurrence is selected independently from the
definition of R.sub.1. Also, combinations of substituents and/or
variables are permissible, but only if such combinations result in
stable compounds.
[0627] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or
[0628] As used herein, "halo" or "halogen" refers to fluoro,
chloro, bromo and iodo. The term "perhalogenated" generally refers
to a moiety wherein all hydrogen atoms are replaced by halogen
atoms. The term "haloalkyl" or "haloalkoxyl" refers to an alkyl or
alkoxyl substituted with one or more halogen atoms.
[0629] The term "carbonyl" includes compounds and moieties which
contain a carbon connected with a double bond to an oxygen atom.
Examples of moieties containing a carbonyl include, but are not
limited to, aldehydes, ketones, carboxylic acids, amides, esters,
anhydrides, etc.
[0630] The term "carboxyl" refers to --COOH or its C.sub.1-C.sub.6
alkyl ester.
[0631] "Acyl" includes moieties that contain the acyl radical
(R--C(O)--) or a carbonyl group. "Substituted acyl" includes acyl
groups where one or more of the hydrogen atoms are replaced by, for
example, alkyl groups, alkynyl groups, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, amino (including alkylamino,
dialkylamino, arylamino, diarylamino and alkylarylamino), acylamino
(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and
ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,
sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkylaryl, or an aromatic or heteroaromatic moiety.
[0632] "Aroyl" includes moieties with an aryl or heteroaromatic
moiety bound to a carbonyl group. Examples of aroyl groups include
phenylcarboxy, naphthyl carboxy, etc.
[0633] "Alkoxyalkyl," "alkylaminoalkyl," and "thioalkoxyalkyl"
include alkyl groups, as described above, wherein oxygen, nitrogen,
or sulfur atoms replace one or more hydrocarbon backbone carbon
atoms.
[0634] The term "alkoxy" or "alkoxyl" includes substituted and
unsubstituted alkyl, alkenyl and alkynyl groups covalently linked
to an oxygen atom. Examples of alkoxy groups or alkoxyl radicals
include, but are not limited to, methoxy, ethoxy, isopropyloxy,
propoxy, butoxy and pentoxy groups. Examples of substituted alkoxy
groups include halogenated alkoxy groups. The alkoxy groups can be
substituted with groups such as alkenyl, alkynyl, halogen,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy and trichloromethoxy.
[0635] The term "ether" or "alkoxy" includes compounds or moieties
which contain an oxygen bonded to two carbon atoms or heteroatoms.
For example, the term includes "alkoxyalkyl," which refers to an
alkyl, alkenyl, or alkynyl group covalently bonded to an oxygen
atom which is covalently bonded to an alkyl group.
[0636] The term "ester" includes compounds or moieties which
contain a carbon or a heteroatom bound to an oxygen atom which is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc.
[0637] The term "thioalkyl" includes compounds or moieties which
contain an alkyl group connected with a sulfur atom. The thioalkyl
groups can be substituted with groups such as alkyl, alkenyl,
alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid,
alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl,
alkoxyl, amino (including alkylamino, dialkylamino, arylamino,
diarylamino and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moieties.
[0638] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom.
[0639] The term "thioether" includes moieties which contain a
sulfur atom bonded to two carbon atoms or heteroatoms. Examples of
thioethers include, but are not limited to alkthioalkyls,
alkthioalkenyls, and alkthioalkynyls. The term "alkthioalkyls"
include moieties with an alkyl, alkenyl, or alkynyl group bonded to
a sulfur atom which is bonded to an alkyl group. Similarly, the
term "alkthioalkenyls" refers to moieties wherein an alkyl, alkenyl
or alkynyl group is bonded to a sulfur atom which is covalently
bonded to an alkenyl group; and alkthioalkynyls" refers to moieties
wherein an alkyl, alkenyl or alkynyl group is bonded to a sulfur
atom which is covalently bonded to an alkynyl group.
[0640] As used herein, "amine" or "amino" refers to unsubstituted
or substituted --NH.sub.2. "Alkylamino" includes groups of
compounds wherein nitrogen of --NH.sub.2 is bound to at least one
alkyl group. Examples of alkylamino groups include benzylamino,
methylamino, ethylamino, phenethylamino, etc. "Dialkylamino"
includes groups wherein the nitrogen of --NH.sub.2 is bound to at
least two additional alkyl groups. Examples of dialkylamino groups
include, but are not limited to, dimethylamino and diethylamino.
"Arylamino" and "diarylamino" include groups wherein the nitrogen
is bound to at least one or two aryl groups, respectively.
"Aminoaryl" and "aminoaryloxy" refer to aryl and aryloxy
substituted with amino. "Alkylarylamino," "alkylaminoaryl" or
"arylaminoalkyl" refers to an amino group which is bound to at
least one alkyl group and at least one aryl group. "Alkaminoalkyl"
refers to an alkyl, alkenyl, or alkynyl group bound to a nitrogen
atom which is also bound to an alkyl group. "Acylamino" includes
groups wherein nitrogen is bound to an acyl group. Examples of
acylamino include, but are not limited to, alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido groups.
[0641] The term "amide" or "aminocarboxy" includes compounds or
moieties that contain a nitrogen atom that is bound to the carbon
of a carbonyl or a thiocarbonyl group. The term includes
"alkaminocarboxy" groups that include alkyl, alkenyl or alkynyl
groups bound to an amino group which is bound to the carbon of a
carbonyl or thiocarbonyl group. It also includes "arylaminocarboxy"
groups that include aryl or heteroaryl moieties bound to an amino
group that is bound to the carbon of a carbonyl or thiocarbonyl
group. The terms "alkylaminocarboxy", "alkenylaminocarboxy",
"alkynylaminocarboxy" and "arylaminocarboxy" include moieties
wherein alkyl, alkenyl, alkynyl and aryl moieties, respectively,
are bound to a nitrogen atom which is in turn bound to the carbon
of a carbonyl group. Amides can be substituted with substituents
such as straight chain alkyl, branched alkyl, cycloalkyl, aryl,
heteroaryl or heterocycle. Substituents on amide groups may be
further substituted.
[0642] Compounds of the present invention that contain nitrogens
can be converted to N-oxides by treatment with an oxidizing agent
(e.g., 3-chloroperoxybenzoic acid (mCPBA) and/or hydrogen
peroxides) to afford other compounds of the present invention.
Thus, all shown and claimed nitrogen-containing compounds are
considered, when allowed by valency and structure, to include both
the compound as shown and its N-oxide derivative (which can be
designated as N.fwdarw.O or N.sup.+--O.sup.-). Furthermore, in
other instances, the nitrogens in the compounds of the present
invention can be converted to N-hydroxy or N-alkoxy compounds. For
example, N-hydroxy compounds can be prepared by oxidation of the
parent amine by an oxidizing agent such as m-CPBA. All shown and
claimed nitrogen-containing compounds are also considered, when
allowed by valency and structure, to cover both the compound as
shown and its N-hydroxy (i.e., N--OH) and N-alkoxy (i.e., N--OR,
wherein R is substituted or unsubstituted C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkynyl, 3-14-membered
carbocycle or 3-14-membered heterocycle) derivatives.
[0643] In the present specification, the structural formula of the
compound represents a certain isomer for convenience in some cases,
but the present invention includes all isomers, such as geometrical
isomers, optical isomers based on an asymmetrical carbon,
stereoisomers, tautomers, and the like, it being understood that
not all isomers may have the same level of activity. In addition, a
crystal polymorphism may be present for the compounds represented
by the formula. It is noted that any crystal form, crystal form
mixture, or anhydride or hydrate thereof is included in the scope
of the present invention. Furthermore, so-called metabolite which
is produced by degradation of the present compound in vivo is
included in the scope of the present invention.
[0644] "Isomerism" means compounds that have identical molecular
formulae but differ in the sequence of bonding of their atoms or in
the arrangement of their atoms in space. Isomers that differ in the
arrangement of their atoms in space are termed "stereoisomers."
Stereoisomers that are not mirror images of one another are termed
"diastereoisomers," and stereoisomers that are non-superimposable
mirror images of each other are termed "enantiomers" or sometimes
optical isomers. A mixture containing equal amounts of individual
enantiomeric forms of opposite chirality is termed a "racemic
mixture."
[0645] A carbon atom bonded to four nonidentical substituents is
termed a "chiral center."
[0646] "Chiral isomer" means a compound with at least one chiral
center. Compounds with more than one chiral center may exist either
as an individual diastereomer or as a mixture of diastereomers,
termed "diastereomeric mixture." When one chiral center is present,
a stereoisomer may be characterized by the absolute configuration
(R or S) of that chiral center. Absolute configuration refers to
the arrangement in space of the substituents attached to the chiral
center. The substituents attached to the chiral center under
consideration are ranked in accordance with the Sequence Rule of
Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit.
1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413;
Cahn and Ingold, J. Chem. Soc. 1951 (London), 612; Cahn et al.,
Experientia 1956, 12, 81; Cahn, J. Chem. Educ. 1964, 41, 116).
[0647] "Geometric isomer" means the diastereomers that owe their
existence to hindered rotation about double bonds or a cycloalkyl
linker (e.g., 1,3-cylcobutyl). These configurations are
differentiated in their names by the prefixes cis and trans, or Z
and E, which indicate that the groups are on the same or opposite
side of the double bond in the molecule according to the
Cahn-Ingold-Prelog rules.
[0648] It is to be understood that the compounds of the present
invention may be depicted as different chiral isomers or geometric
isomers. It should also be understood that when compounds have
chiral isomeric or geometric isomeric forms, all isomeric forms are
intended to be included in the scope of the present invention, and
the naming of the compounds does not exclude any isomeric forms, it
being understood that not all isomers may have the same level of
activity.
[0649] Furthermore, the structures and other compounds discussed in
this invention include all atropic isomers thereof, it being
understood that not all atropic isomers may have the same level of
activity. "Atropic isomers" are a type of stereoisomer in which the
atoms of two isomers are arranged differently in space. Atropic
isomers owe their existence to a restricted rotation caused by
hindrance of rotation of large groups about a central bond. Such
atropic isomers typically exist as a mixture, however as a result
of recent advances in chromatography techniques, it has been
possible to separate mixtures of two atropic isomers in select
cases.
[0650] "Tautomer" is one of two or more structural isomers that
exist in equilibrium and is readily converted from one isomeric
form to another. This conversion results in the formal migration of
a hydrogen atom accompanied by a switch of adjacent conjugated
double bonds. Tautomers exist as a mixture of a tautomeric set in
solution. In solutions where tautomerization is possible, a
chemical equilibrium of the tautomers will be reached. The exact
ratio of the tautomers depends on several factors, including
temperature, solvent and pH. The concept of tautomers that are
interconvertable by tautomerizations is called tautomerism.
[0651] Of the various types of tautomerism that are possible, two
are commonly observed. In keto-enol tautomerism a simultaneous
shift of electrons and a hydrogen atom occurs. Ring-chain
tautomerism arises as a result of the aldehyde group (--CHO) in a
sugar chain molecule reacting with one of the hydroxy groups (--OH)
in the same molecule to give it a cyclic (ring-shaped) form as
exhibited by glucose.
[0652] Common tautomeric pairs are: ketone-enol, amide-nitrile,
lactam-lactim, amide-imidic acid tautomerism in heterocyclic rings
(e.g., in nucleobases such as guanine, thymine and cytosine),
imine-enamine and enamine-enamine. An example of keto-enol
equilibria is between pyridin-2(1H)-ones and the corresponding
pyridin-2-ols, as shown below.
##STR00433##
[0653] It is to be understood that the compounds of the present
invention may be depicted as different tautomers. It should also be
understood that when compounds have tautomeric forms, all
tautomeric forms are intended to be included in the scope of the
present invention, and the naming of the compounds does not exclude
any tautomer form. It will be understood that certain tautomers may
have a higher level of activity than others.
[0654] The term "crystal polymorphs", "polymorphs" or "crystal
forms" means crystal structures in which a compound (or a salt or
solvate thereof) can crystallize in different crystal packing
arrangements, all of which have the same elemental composition.
Different crystal forms usually have different X-ray diffraction
patterns, infrared spectral, melting points, density hardness,
crystal shape, optical and electrical properties, stability and
solubility. Recrystallization solvent, rate of crystallization,
storage temperature, and other factors may cause one crystal form
to dominate. Crystal polymorphs of the compounds can be prepared by
crystallization under different conditions.
[0655] The compounds of any Formula described herein include the
compounds themselves, as well as their salts, their solvates, and
their prodrugs, if applicable. A salt, for example, can be formed
between an anion and a positively charged group (e.g., amino) on a
substituted benzene compound. Suitable anions include chloride,
bromide, iodide, sulfate, bisulfate, sulfamate, nitrate, phosphate,
citrate, methanesulfonate, trifluoroacetate, glutamate,
glucuronate, glutarate, malate, maleate, succinate, fumarate,
tartrate, tosylate, salicylate, lactate, naphthalenesulfonate, and
acetate (e.g., trifluoroacetate). The term "pharmaceutically
acceptable anion" refers to an anion suitable for forming a
pharmaceutically acceptable salt. Likewise, a salt can also be
formed between a cation and a negatively charged group (e.g.,
carboxylate) on a substituted benzene compound. Suitable cations
include sodium ion, potassium ion, magnesium ion, calcium ion, and
an ammonium cation such as tetramethylammonium ion. The substituted
benzene compounds also include those salts containing quaternary
nitrogen atoms.
[0656] Examples of prodrugs include esters and other
pharmaceutically acceptable derivatives, which, upon administration
to a subject, are capable of providing active substituted benzene
compounds.
[0657] Additionally, the compounds of the present invention, for
example, the salts of the compounds, can exist in either hydrated
or unhydrated (the anhydrous) form or as solvates with other
solvent molecules. Nonlimiting examples of hydrates include
monohydrates, dihydrates, etc. Nonlimiting examples of solvates
include ethanol solvates, acetone solvates, etc.
[0658] "Solvate" means solvent addition forms that contain either
stoichiometric or non stoichiometric amounts of solvent. Some
compounds have a tendency to trap a fixed molar ratio of solvent
molecules in the crystalline solid state, thus forming a solvate.
If the solvent is water the solvate formed is a hydrate; and if the
solvent is alcohol, the solvate formed is an alcoholate. Hydrates
are formed by the combination of one or more molecules of water
with one molecule of the substance in which the water retains its
molecular state as H.sub.2O.
[0659] As used herein, the term "analog" refers to a chemical
compound that is structurally similar to another but differs
slightly in composition (as in the replacement of one atom by an
atom of a different element or in the presence of a particular
functional group, or the replacement of one functional group by
another functional group). Thus, an analog is a compound that is
similar or comparable in function and appearance, but not in
structure or origin to the reference compound.
[0660] As defined herein, the term "derivative" refers to compounds
that have a common core structure, and are substituted with various
groups as described herein. For example, all of the compounds
represented by Formula (I) are substituted benzene compounds, and
have Formula (I) as a common core.
[0661] The term "bioisostere" refers to a compound resulting from
the exchange of an atom or of a group of atoms with another,
broadly similar, atom or group of atoms. The objective of a
bioisosteric replacement is to create a new compound with similar
biological properties to the parent compound. The bioisosteric
replacement may be physicochemically or topologically based.
Examples of carboxylic acid bioisosteres include, but are not
limited to, acyl sulfonimides, tetrazoles, sulfonates and
phosphonates. See, e.g., Patani and LaVoie, Chem. Rev. 96,
3147-3176, 1996.
[0662] The present invention is intended to include all isotopes of
atoms occurring in the present compounds. Isotopes include those
atoms having the same atomic number but different mass numbers. By
way of general example and without limitation, isotopes of hydrogen
include tritium and deuterium, and isotopes of carbon include C-13
and C-14.
2. SYNTHESIS OF SUBSTITUTED BENZENE COMPOUNDS
[0663] The present invention provides methods for the synthesis of
the compounds of any of the Formulae described herein. The present
invention also provides detailed methods for the synthesis of
various disclosed compounds of the present invention according to
the following schemes as shown in the Examples.
[0664] Throughout the description, where compositions are described
as having, including, or comprising specific components, it is
contemplated that compositions also consist essentially of, or
consist of, the recited components. Similarly, where methods or
processes are described as having, including, or comprising
specific process steps, the processes also consist essentially of,
or consist of, the recited processing steps. Further, it should be
understood that the order of steps or order for performing certain
actions is immaterial so long as the invention remains operable.
Moreover, two or more steps or actions can be conducted
simultaneously.
[0665] The synthetic processes of the invention can tolerate a wide
variety of functional groups, therefore various substituted
starting materials can be used. The processes generally provide the
desired final compound at or near the end of the overall process,
although it may be desirable in certain instances to further
convert the compound to a pharmaceutically acceptable salt, ester,
or prodrug thereof.
[0666] Compounds of the present invention can be prepared in a
variety of ways using commercially available starting materials,
compounds known in the literature, or from readily prepared
intermediates, by employing standard synthetic methods and
procedures either known to those skilled in the art, or which will
be apparent to the skilled artisan in light of the teachings
herein. Standard synthetic methods and procedures for the
preparation of organic molecules and functional group
transformations and manipulations can be obtained from the relevant
scientific literature or from standard textbooks in the field.
Although not limited to any one or several sources, classic texts
such as Smith, M. B., March, J., March's Advanced Organic
Chemistry: Reactions, Mechanisms, and Structure, 5.sup.th edition,
John Wiley & Sons: New York, 2001; Greene, T. W., Wuts, P. G.
M., Protective Groups in Organic Synthesis, 3.sup.rd edition, John
Wiley & Sons: New York, 1999; R. Larock, Comprehensive Organic
Transformations, VCH Publishers (1989); L. Fieser and M. Fieser,
Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and
Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for
Organic Synthesis, John Wiley and Sons (1995), incorporated by
reference herein, are useful and recognized reference textbooks of
organic synthesis known to those in the art. The following
descriptions of synthetic methods are designed to illustrate, but
not to limit, general procedures for the preparation of compounds
of the present invention.
[0667] Compounds of the present invention can be conveniently
prepared by a variety of methods familiar to those skilled in the
art. The compounds of this invention with any of the Formulae
described herein may be prepared according to the procedures
illustrated in Schemes 1-10 below, from commercially available
starting materials or starting materials which can be prepared
using literature procedures. The Z and R groups (such as R.sub.1,
R.sub.2, R.sub.3, R.sub.4, R.sub.6, R.sub.7, R.sub.8, and R.sub.12)
in Schemes 1-10 are as defined in any Formula described herein,
unless otherwise specified.
[0668] One of ordinary skill in the art will note that, during the
reaction sequences and synthetic schemes described herein, the
order of certain steps may be changed, such as the introduction and
removal of protecting groups.
[0669] One of ordinary skill in the art will recognize that certain
groups may require protection from the reaction conditions via the
use of protecting groups. Protecting groups may also be used to
differentiate similar functional groups in molecules. A list of
protecting groups and how to introduce and remove these groups can
be found in Greene, T. W., Wuts, P. G. M., Protective Groups in
Organic Synthesis, 3.sup.rd edition, John Wiley & Sons: New
York, 1999.
[0670] Preferred protecting groups include, but are not limited
to:
[0671] For a hydroxyl moiety: TBS, benzyl, THP, Ac
[0672] For carboxylic acids: benzyl ester, methyl ester, ethyl
ester, allyl ester
[0673] For amines: Cbz, BOC, DMB
[0674] For diols: Ac (.times.2) TBS (.times.2), or when taken
together acetonides
[0675] For thiols: Ac
[0676] For benzimidazoles: SEM, benzyl, PMB, DMB
[0677] For aldehydes: di-alkyl acetals such as dimethoxy acetal or
diethyl acetyl.
[0678] In the reaction schemes described herein, multiple
stereoisomers may be produced. When no particular stereoisomer is
indicated, it is understood to mean all possible stereoisomers that
could be produced from the reaction. A person of ordinary skill in
the art will recognize that the reactions can be optimized to give
one isomer preferentially, or new schemes may be devised to produce
a single isomer. If mixtures are produced, techniques such as
preparative thin layer chromatography, preparative HPLC,
preparative chiral HPLC, or preparative SFC may be used to separate
the isomers.
[0679] The following abbreviations are used throughout the
specification and are defined below: [0680] AA ammonium acetate
[0681] ACN acetonitrile [0682] Ac acetyl [0683] AcOH acetic acid
[0684] atm atmosphere [0685] aq. Aqueous [0686] BID or b.i.d. bis
in die (twice a day) [0687] tBuOK potassium t-butoxide [0688] Bn
benzyl [0689] BOC tert-butoxy carbonyl [0690] BOP
(benzotriazol-1-yloxy)tris(dimethylamino)-phosphoniumhexafluorophosphate
[0691] Cbz benzyloxy carbonyl [0692] CDCl.sub.3 deuterated
chloroform [0693] CH.sub.2Cl.sub.2 dichloromethane [0694] COMU
(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethyl-amino-morpholino-carben-
ium hexafluorophosphate [0695] d days [0696] DBU
1,8-diazabicyclo[5.4.0]undec-7-ene [0697] DCE 1,2 dichloroethane
[0698] DCM dichloromethane [0699] DEAD Diethyl azodicarboxylate
[0700] DIAD Diisopropyl azodicarboxylate [0701] DiBAL-H diisobutyl
aluminium hydride [0702] DIPEA N,N-diisopropylethylamine (Hunig's
base) [0703] DMA Dimethylacetamide [0704] DMAP N,N
dimethyl-4-aminopyridine [0705] DMB 2,4 dimethoxy benzyl [0706] DMF
N,N-Dimethylformamide [0707] DMSO Dimethyl sulfoxide [0708] DPPA
Diphenylphosphonic azide [0709] EA or EtOAc Ethyl acetate [0710]
EDC or EDCI N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide [0711]
Et.sub.2O diethyl ether [0712] ELS Evaporative Light Scattering
[0713] ESI- Electrospray negative mode [0714] ESI+ Electrospray
positive mode [0715] Et.sub.3N or TEA triethylamine [0716] EtOH
ethanol [0717] FA formic acid [0718] FC or FCC Flash chromatogrpahy
[0719] h hours [0720] H.sub.2O water [0721] HATU
O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate [0722] HOAT 1-Hydroxy-7-azabenzotriazole [0723]
HOBt 1-Hydroxybenzotriazole [0724] HO-Su N-Hydroxysuccinimide
[0725] HCl hydrogen chloride or hydrochloric acid [0726] HPLC High
performance liquid chromatography [0727] K.sub.2CO.sub.3 potassium
carbonate [0728] KHMDs Potassium hexamethyldisilazide [0729] LC/MS
or LC-MS Liquid chromatography mass spectrum [0730] LDA Lithium
diisopropylamide [0731] LiHMDs Lithium hexamethyldisilazide [0732]
LG leaving group [0733] M Molar [0734] m/z mass/charge ratio [0735]
m-CPBA meta-chloroperbenzoic acid [0736] MeCN Acetonitrile [0737]
MeOD d4-methanol [0738] MeI Methyl iodide [0739] MS3 .ANG. 3 .ANG.
molecular sieves [0740] MgSO.sub.4 Magnesium Sulfate [0741] min
minutes [0742] Ms Mesyl [0743] MsCl Mesyl chloride [0744] MsO
Mesylate [0745] MS Mass Spectrum [0746] MWI microwave irradiation
[0747] Na.sub.2CO.sub.3 sodium carbonate [0748] Na.sub.2SO.sub.4
sodium sulfate [0749] NaHCO.sub.3 sodium bicarbonate [0750] NaHMDs
Sodium hexamethyldisilazide [0751] NaOH sodium hydroxide [0752]
NaHCO.sub.3 sodium bicarbonate [0753] Na.sub.2SO.sub.4 sodium
sulfate [0754] NIS N-iodosuccinimide [0755] NMR Nuclear Magnetic
Resonance [0756] o/n or O/N overnight [0757] Pd/C Palladium on
carbon [0758] Pd(dppf)Cl.sub.2.DCM
[1,1'-Bis(diphenylphosphino)ferrocene] dichloropalladium(II),
complex with dichloromethane [0759] PPAA 1-Propanephosphonic acid
cyclic anhydride [0760] Pd(OH).sub.2 Palladium dihydroxide [0761]
PE Petroleum Ether [0762] PG protecting group [0763] PMB para
methoxybenzyl [0764] ppm parts per million [0765] p.o. per os (oral
administration) [0766] prep HPLC preparative High Performance
Liquid Chromatography [0767] prep TLC preparative thin layer
chromatography [0768] p-TsOH para-toluenesulfonic acid [0769] PyBOP
(Benzotriazol-1-yloxy)tripyrrolidinophosphonium [0770]
Hexafluorophosphate [0771] QD or q.d. quaque die (once a day)
[0772] RBF round bottom flask [0773] RP-HPLC Reverse phase High
Performance liquid chromatography [0774] Rt or RT Room temperature
[0775] SEM (Trimethylsilyl)ethoxymethyl [0776] SEMC1
(Trimethylsilyl)ethoxymethyl chloride [0777] SFC Super critical
chromatography [0778] SGC silica gel chromatography [0779] STAB
Sodium triacetoxy borohydride [0780] TBAF tetra-n-butylammonium
fluoride [0781] TBME tert-Butyl methyl ether [0782] TEA
Triethylamine [0783] TFA trifluoroacetic acid [0784] TfO triflate
[0785] THF tetrahydrofuran [0786] THP tetrahydropyran [0787] TID or
t.i.d ter in die (three times a day) [0788] TLC thin layer
chromatography [0789] TMSCl Trimethylsilyl chloride [0790] Ts tosyl
[0791] TsOH tosic acid [0792] UV ultraviolet
##STR00434##
[0793] Scheme 1 shows the synthesis of benzene analogs wherein
Z.dbd.--N(R.sub.7)(R.sub.8) following a general route that utilizes
well-established chemistry. Substituted nitrobenzoic acids, many of
which are commercially available or can be prepared by nitrations
of the appropriate substituted benzoic acids or other chemistry
known to one skilled in the art, can be converted to their methyl
esters by treatment with methyliodide in a polar solvent such as
DMF in the presence of an appropriate base such as sodium carbonate
at an appropriate temperature such as 60.degree. C. (Step 1). The
nitro group can be reduced to an amine using an appropriate
reducing agent such as iron in the presence of an acid such as
ammonium chloride in a protic solvent such as ethanol at an
appropriate temperature such as 80.degree. C. (Step 2).
Introduction of the R.sub.7 can be done using a reductive amination
with an appropriate R.sub.7-ketone or R.sub.7-aldehyde in the
presence of an appropriate reducing agent such as sodium
cyanoborohydride and catalytic acid such as acetic acid in an
appropriate solvent such as methanol. A variety of R.sub.8 groups
can be introduced by alkylation using R.sub.8-LG, where LG is a
leaving group such as iodine, in the presence of a mild base such
as cesium carbonate in an appropriate polar solvent such as
acetonitrile at an appropriate temperature such as 80.degree. C.
(Step 4). Alternatively, R.sub.8 groups can be introduced by
reductive amination with R.sub.8-ketone or R.sub.8-aldehyde in the
presence of an appropriate reducing agent such as sodium
cyanoborohydride and catalytic acid such as acetic acid in an
appropriate solvent such as methanol. The ester moiety can be
converted to an amide using a standard two step protocol. The ester
can be hydrolyzed to the corresponding acid using a suitable base
such as sodium hydroxide in a polar solvent such as ethanol (Step
5). The acid would then be subjecting to a standard amide coupling
reaction whereupon the appropriate amine would be added along with
a suitable amide coupling reagent such as PYBOP in a suitable
solvent such as DMSO to give the desired amide (Step 6).
##STR00435##
[0794] Depending upon the nature of the R.sub.6 substituent,
further chemical modification could be employed to convert the
R.sub.6 substituent into an alternative R.sub.6 substituent. A
representative sampling of such modifications could include
hydrogenation, protecting group removal followed by additional
amide coupling reactions, palladium catalyzed coupling reactions,
reductive amination reactions or alkylation reactions. For example,
if R.sub.6 is a bromide, alternative R.sub.6 substituents could
then be introduced using standard transition metal-based protocols
that rely upon a leaving group such as a bromide as a connection
point.
[0795] In one such protocol as depicted in Scheme 2 non-aromatic
R.sub.6 substituents attached via a carbon-carbon bond may be
introduced by Suzuki reaction of a compound where R.sub.6.dbd.Br
with an appropriate unsaturated non-aromatic boronic ester
derivative (e.g. an olefinic boronic ester derivative such as vinyl
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane) in the presence of
a mild base and a palladium catalyst in a polar solvent such as
dioxane/water, at elevated temperature to give the desired new
R.sub.6 substituent. Depending upon the nature of the R.sub.6
substituent, further chemical modification could be employed to
convert the unsaturated R.sub.6 substituent into an alternative
R.sub.6 substituent. A representative sampling of such
modifications could include hydrogenation, protecting group removal
followed by additional amide coupling reactions, palladium
catalyzed coupling reactions, reductive amination reactions or
alkylation reactions. For example, in cases where an unsaturated
non-aromatic R.sub.6 group is introduced, further modification by
hydrogenation can give the corresponding saturated R.sub.6 group
(e.g. conversion of a vinyl group to an ethyl group). In cases of
where R.sub.6 groups introduced have protected amine functionality
further modifications include deprotection to give amines which may
in subsequent steps be further modified for example by amide
formation or reductive amination reactions.
[0796] In another protocol as depicted in Scheme 3, non-aromatic
R.sub.6 substituents attached via a carbon-carbon bond may be
introduced by Sonogashira reaction of a compound where
R.sub.6.dbd.Br optionally followed by further modification of the
introduced alkynyl group. In the Sonogashira reaction, a compound
where R.sub.6.dbd.Br is coupled with a terminal alkyne derivative
in the presence of a mild base, a copper catalyst and a palladium
catalyst in an organic solvent such as toluene at elevated
temperature. This results in the replacement of the Br group with
an alkynyl group. The resulting compound wherein the R.sub.6
substituent is an alkynyl group may be subject to subsequent
suitable modifications to give an alternative R.sub.6 substituent.
A representative sampling of such modifications could include
hydrogenation, protecting group removal followed by additional
amide coupling reactions, reductive amination reactions or
alkylation reactions.
##STR00436##
[0797] In another protocol non-aromatic R.sub.6 substituents
attached via a carbon-carbon bond may be prepared by other
substitution reactions of the bromine atom compounds where
R.sub.6.dbd.Br, optionally followed by further modification of the
introduced R.sub.6 group. Examples of such substitution reactions
include coupling reactions with zinc reagents such as cyanation and
Negishi reactions. In the case of cyanation reaction, compounds
where R.sub.6.dbd.Br may be reacted with zinc cyanide under
standard palladium catalyst mediated reaction conditions to give
compounds where R.sub.6.dbd.CN. The cyano group in such compounds
may be subject to further modification to give other R.sub.6
groups. Such cyano modifications include i. reduction to an amine
which may be subsequently converted to an amide by acylation or
alkylation, reduction to an aldehyde which may be subjected to
reductive amination reaction to give corresponding derivatives. In
Negishi reactions alkylzinc reagents which may be prepared from
alkyl iodides (e.g. N-Boc-3-iodoazetidine) are coupled to compounds
where R.sub.6.dbd.Br using palladium or nickel catalysts. In the
resulting products the introduced R.sub.6 group may be converted to
an alternative group by further modifications of the R.sub.6 group
in subsequent steps such as deprotection, amide formation or
alkylation.
[0798] Compounds with R.sub.6 substituents which are amines
attached via a nitrogen-carbon bond may be introduced by Buchwald
coupling reaction of compounds where R.sub.6.dbd.Br followed by
optional modification of the R.sub.6 group as depicted in Scheme 4.
In the Buchwald reaction compounds where R.sub.6.dbd.Br are treated
with a primary or secondary amine (e.g. tert-butyl
piperazine-1-carboxylate) in the presence of a palladium catalyst
(e.g. Pd(dba)2/BINAP) and a base (e.g. cesium carbonate) in an
organic solvent (e.g. toluene) at elevated temperature. The
Buchwald coupling product may be subjected to subsequent suitable
modifications to give an alternative R.sub.6 substituent. Such
modifications are exemplified by protecting group removal, amide
coupling reactions, reductive amination reactions or alkylation
reactions.
##STR00437##
[0799] Compounds with R.sub.6 substituents which are alkylthio
groups attached via a sulfur-carbon bond may be prepared by
coupling reaction of compounds where R.sub.6.dbd.Br with thiols in
the presence of a palladium catalyst and a weak base (e.g. DIPEA)
in an organic solvent at elevated temperature. The coupling product
sulfides may be subject to subsequent suitable modifications to
give an alternative R.sub.6 substituent. Such modifications include
sulfur oxidation reactions to give sulfoxides and sulfones,
protecting group removal, amide coupling reactions, reductive
amination reactions or alkylation reactions.
[0800] In a modification of the general synthesis in Scheme 1,
depending upon the nature of the R.sub.7 substituent, further
chemical modification subsequent to Step 6 of Scheme 1 could be
employed to convert the R.sub.7 substituent into an alternative
R.sub.7 substituent. For example a protected amino group contained
within R.sub.7 may be subjected to deprotection reaction (e.g. Boc
group cleavage) to give free amino groups. Such free amino groups
may be subjected to reductive amination reactions or alkylation
reactions to give substituted amines.
[0801] Scheme 5 shows the general synthesis of picolinamide
compounds. Starting from methyl 3-bromo-6-chloropicolinate
oxidation to the N-oxide followed by chlorination with phosphorus
oxychloride gives methyl 3-bromo-4,6-dichloropicolinate. The
4-chloro group can be selectively substituted with diverse mono and
dialkyl amines which may also contain functional or protected
functional groups that may be unmasked at a later stage. The
3-bromo group may be retained or may be optionally converted into
an alternative R.sub.12 group by suitable substitution reaction and
further functional group modifications. Such reactions include
coupling reactions mediated with palladium catalysts. For example
the 3-bromo group may be converted to an R.sub.12=methyl group by
Stille reaction with tetramethyltin. Ester hydrolysis followed by
amide coupling with appropriate 3-(aminomethyl)-pyridin-2-ones
yields picolinamide compounds wherein R.sub.6 is a chloro group.
The chloro group may optionally be converted to alternative R.sub.6
groups by suitable substitution reactions either in a final step or
alternatively prior to ester hydrolysis Step 6. Examples of such
substitution reactions include cyanation and amination reactions
either directly or mediated with palladium catalysts. Analogous
compounds wherein R.sub.12 is chloro may be prepared in analogous
fashion from methyl 3,4, 6-trichloropyridine-2-carboxylate.
##STR00438##
[0802] General syntheses of 3-(aminomethyl)-pyridin-2(1H)-ones
intermediates for the amide coupling reaction from Scheme 1 are
depicted in Scheme 6. In one method, a diketone can be condensed
with 2-cyanoacetamide in the presence of an appropriate reagent
such as piperidine acetate in a polar solvent such as ethanol to
provide a cyanopyridone (Step 9). In another method, when R.sub.3
is H, an appropriately substituted alkynyl ketone can be condensed
with 2-cyanoacetamide in the presence of an appropriate reagent
such as piperidine acetate in a polar solvent such as ethanol to
provide a cyanopyridone (Step 11). The cyano group can be reduced
under appropriate conditions such as hydrogenation in the presence
of catalytic Raney nickel in a polar solvent such as ammonium in
methanol to provide the amine (Step 10).
##STR00439##
[0803] Additionally, depending upon the nature of the R.sub.2,
R.sub.3, or R.sub.4 group, further chemical modification can be
employed to convert each of them independently into an alternative
substituent. A representative sampling of such modifications can
include hydrogenation, protecting group removal followed by
additional amide coupling reactions, palladium catalyzed coupling
reactions, reductive amination reactions, and alkylation
reactions.
[0804] Scheme 7 depicts a variant of the general synthesis route of
Scheme 1 based on 2-substituted (substituent is an R.sub.12 group)
methyl 3-amino-5-bromo-benzoate starting materials. These starting
materials can in turn be prepared from 2-substituted
3-nitro-benzoic acids which are commercially available or can be
prepared by nitration of 2-substituted benzoic acids. Thus,
bromination of 2-substituted 3-nitro-benzoic acids with a suitable
reagent such as 1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione
yields the appropriate 2-substituted 3-nitro-5-bromo-benzoic acids.
A variety of esterification and then nitro group reduction methods
can then be sequentially implemented to prepare the 2-substituted
methyl 3-amino-5-bromo-benzoate starting materials from the
2-substituted 3-nitro-5-bromo-benzoic acids.
##STR00440##
[0805] As depicted in Scheme 7 the R.sub.7 group can be introduced
from 2-substituted methyl 3-amino-5-bromo-benzoates in Step 1 using
a reductive amination with an appropriate R.sub.7-ketone or
R.sub.7-aldehyde in the presence of an appropriate reducing agent
such as sodium cyanoborohydride and catalytic acid such as acetic
acid in an appropriate solvent such as methanol. Similarly, R.sub.8
groups can be introduced in Step 2 by reductive amination with
R.sub.8-ketone or R.sub.8-aldehyde in the presence of an
appropriate reducing agent such as sodium cyanoborohydride and
catalytic acid such as acetic acid in an appropriate solvent such
as methanol. Alternatively, a variety of R.sub.8 groups can be
introduced by alkylation using R.sub.8-LG, where LG is a leaving
group such as iodine, in the presence of a mild base such as cesium
carbonate in an appropriate polar solvent such as acetonitrile at
an appropriate temperature such as 80.degree. C. In Step 3, R.sub.6
groups other than bromine can be introduced via palladium catalyzed
coupling reactions. Examples of such R.sub.6 groups and methods
have been described above. For example amines may be introduced by
Buchwald reactions and unsaturated groups may be introduced by
Suzuki or Sonogashiri reactions. The R.sub.6 substituent may be
subject to subsequent suitable modifications to give an alternative
R.sub.6 substituent. A representative sampling of such
modifications could include hydrogenation (e.g. to saturate
unsaturated groups), protecting group removal followed by
additional amide coupling reactions, reductive amination reactions
or alkylation reactions. In Step 4 the ester moiety can be
hydrolyzed to the corresponding acid using a suitable base such as
sodium hydroxide in a polar solvent such as ethanol. In Step 5, the
acid can be subjected to a standard amide coupling reaction
whereupon the appropriate 3-(aminomethyl)-pyridin-2-one would be
added along with a suitable amide coupling reagent such as PYBOP in
a suitable solvent such as DMSO to give the desired amide.
Depending upon the nature of the R.sub.7 substituent, further
chemical modification subsequent to Step 5 of Scheme 4 could be
employed to convert the R.sub.7 substituent into an alternative
R.sub.7 substituent. For example a protected amino group contained
within R.sub.7 may be subjected to deprotection reaction (e.g. Boc
group cleavage) to give free amino groups. Such free amino groups
may be subjected to reductive amination reactions or alkylation
reactions to give substituted amines.
[0806] Scheme 8 below depicts the general synthesis of
2-monoalkylamino and
2-dialkylamino-3,6-disubstituted-isonicotinamides wherein the
3-substituent corresponds to R.sub.12 and the 6-substituent
corresponds to R.sub.6. In Step 1 the 3-substituent may be
introduced by the method described by Epsztain J. et al.
Tetrahedron, 1991, v. 47, 1697-16708, by metalation of
2-chloro-isonicotinanilide with n-butyllithium followed by trapping
with an alkyliodide such as methyliodide or aldehyde or other
electrophilic group.
##STR00441## ##STR00442##
[0807] In cases where the trapping reagent yields a substituent
with a functional group this group may be masked or converted into
another functional group compatible with the subsequent chemical
steps. In Step 2 anilide amide hydrolysis under standard acidic
conditions may be conducted followed by methyl ester synthesis
under standard conditions for example as shown with methyl iodide
and base gives corresponding methyl 2-chloro-3-substituted
isonicotinates. In Step 4 an alkylamino group can be introduced by
Buchwald coupling reaction of an R.sub.7NH.sub.2 monoalkylamine
with the methyl 2-chloro-3-substituted isonicotinates. This
reaction is well precedented for diverse 2-chloropyridine systems
in the chemical literature. In an optional Step 5 for dialkylamino
compounds R.sub.8 groups can be introduced by reductive amination
with R.sub.8-ketone or R.sub.8-aldehyde in the presence of an
appropriate reducing agent such as sodium cyanoborohydride and
catalytic acid such as acetic acid in an appropriate solvent such
as methanol. Alternatively, a variety of R.sub.8 groups can be
introduced by alkylation using R.sub.8-LG, where LG is a leaving
group such as iodine, in the presence of a mild base such as cesium
carbonate in an appropriate polar solvent such as acetonitrile at
an appropriate temperature such as 80.degree. C. In Step 6,
oxidation to the N-oxide followed by chlorination with phosphorus
oxychloride gives methyl 6-chloro-2-mono or
dialkylamino-3-substituted isonicotinates. In Step 7 the ester
moiety can be hydrolyzed to the corresponding acid using a suitable
base such as sodium hydroxide in a polar solvent such as ethanol.
In Step 8, the acid can be subjected to a standard amide coupling
reaction whereupon the appropriate substituted
3-(aminomethyl)-pyridin-2-one would be added along with a suitable
amide coupling reagent such as PYBOP in a suitable solvent such as
DMSO to give the desired amide. In Step 9, the chloro group may
optionally be converted to alternative R.sub.6 groups by suitable
substitution reactions either in a final step or alternatively
prior to ester hydrolysis Step 6. Examples of such substitution
reactions include cyanation and amination reactions either directly
or mediated with palladium catalysts. The R.sub.6 substituent may
be subject to subsequent suitable modifications to give an
alternative R.sub.6 substituent. A representative sampling of such
modifications could include hydrogenation (e.g. to saturate
unsaturated groups), protecting group removal followed by
additional amide coupling reactions, reductive amination reactions
or alkylation reactions. Depending upon the nature of the R.sub.7
substituent, further chemical modification steps may be employed to
convert the R.sub.7 substituent into an alternative R.sub.7
substituent. For example a protected amino group contained within
R.sub.7 may be subjected to deprotection reaction (e.g. Boc group
cleavage) to give free amino groups. Such free amino groups may be
subjected to reductive amination reactions or alkylation reactions
to give substituted amines.
##STR00443## ##STR00444##
[0808] Scheme 9 depicts a synthesis of benzene analogs wherein Z is
a sulfide, sulfoxide or sulfone group following a general route
that utilizes well-established chemistry. Starting with a
substituted benzoic acid such as 5-chloro-2-methylbenzoic acid,
nitration using standard conditions such as treatment with conc.
H.sub.2SO.sub.4 and conc. HNO.sub.3 can provide the nitro analog.
Esterification of the acid can be achieved using an alkylating
agent such as methyl iodide in the presence of a base such as
sodium carbonate in a polar solvent such as DMF. The nitro group
can be reduced using conditions such iron and ammonium chloride in
a protic solvent such as ethanol with heating to a temperature such
as 80.degree. C. The resulting aniline can be converted to a
bromide using a Sandmeyer reaction such treatment with CuBr.sub.2
and t-butyl nitrite in a solvent such as acetonitrile. A palladium
catalyzed coupling of a thiol with the bromide can be achieved
using a palladium source such as Pd(OAc).sub.2 with a ligand such
as Xanthphos in the presence of a base such as N,N-diisopropyl
ethylamine in a solvent such as 1,4-dioxane optionally heating to a
temperature such as 100.degree. C. The ester can be hydrolyzed with
an aqueous base such as NaOH in water. The resulting acid can be
coupled to the appropriate substituted
3-(aminomethyl)-pyridin-2-one (e.g. 3-(amino methyl)-4,
6-dimethylpyridin-2(1H)-one as depicted in Scheme 9) using standard
amino acid coupling conditions such as PyBOP in DMSO. The resulting
thioether may be oxidized to the corresponding sulfoxide or sulfone
by using the appropriate equivalents of an oxidant such as m-CPBA
in a solvent such as DCM. The R.sub.6 chloro group may be replaced
with an alternative R.sub.6 group in an additional Step 10 or after
Step 5 or prior to amide coupling. Examples of alternative R.sub.6
groups include substituents that can be incorporated by using
palladium couplings such as a Buchwald reaction to give amine
groups (e.g. morpholino). The R.sub.6 substituent may be subject to
subsequent suitable modifications to give an alternative R.sub.6
substituent. A representative sampling of such modifications
includes protecting group removal followed by additional amide
coupling reactions, reductive amination reactions or alkylation
reactions.
##STR00445## ##STR00446##
[0809] Scheme 10 depicts a synthesis of modified benzene analogs
wherein Z is an ether group following a general route that utilizes
well-established chemistry. Starting with a substituted aniline
such as methyl 3-amino-5-chloro-2-methylbenzoate, the aniline can
be converted to a phenol using a Sandmeyer reaction such as
treatment with aqueous NaNO.sub.2 solution in a aqueous acid such
as 50% H.sub.2SO.sub.4. The phenol can be alkylated using an
alkylating agent such as tetrahydro-2H-pyran-4-yl
4-methylbenzenesulfonate in the presence of an appropriate base
such as cesium carbonate in as polar solvent such as DMF optionally
heating to a temperature such as 80.degree. C. The ester can be
hydrolyzed with an aqueous base such as NaOH in water. The
resulting acid can be coupled to the appropriate substituted
3-(aminomethyl)-pyridin-2-one using standard amino acid coupling
conditions such as PyBOP in DMSO. The R.sub.6 chloro group may be
replaced with an alternative R.sub.6 group in an additional after
Step 5 or prior to amide coupling. Examples of alternative R.sub.6
groups include substituents that can be incorporated by using
palladium couplings such as a Buchwald reaction to give amine
groups (e.g. morpholino). The R.sub.6 substituent may be subject to
subsequent suitable modifications to give an alternative R.sub.6
substituent. A representative sampling of such modifications
includes protecting group removal followed by additional amide
coupling reactions, reductive amination reactions or
alkylation.
[0810] A person of ordinary skill in the art will recognize that in
the above schemes the order of many of the steps are
interchangeable.
3. METHODS OF TREATMENT
[0811] Compounds of the present invention inhibit the histone
methyltransferase activity of EZH2 or a mutant thereof and,
accordingly, the present invention also provides methods for
treating conditions and diseases the course of which can be
influenced by modulating the methylation status of histones or
other proteins, wherein said methylation status is mediated at
least in part by the activity of EZH2. In one aspect of the
invention, certain compounds disclosed herein are candidates for
treating, or preventing certain conditions and diseases. Modulation
of the methylation status of histones can in turn influence the
level of expression of target genes activated by methylation,
and/or target genes suppressed by methylation. The method includes
administering to a subject in need of such treatment, a
therapeutically effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph, solvate, or stereoisomeror thereof.
[0812] The disorder in which EZH2-mediated protein methylation
plays a part can be cancer or a precancerous condition. The present
invention further provides the use of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof in the treatment of cancer
or pre-cancer the course of which can be influenced by modulating
EZH2-mediated protein methylation, or, for the preparation of a
medicament useful for the treatment of such cancer or pre-cancer.
Exemplary cancers that may be treated include lymphomas, including
non-Hodgkin lymphoma, follicular lymphoma (FL) and diffuse large
B-cell lymphoma (DLBCL); melanoma; and leukemia, including CML.
Exemplary precancerous condition includes myelodysplastic syndrome
(MDS; formerly known as preleukemia).
[0813] The present invention also provides methods of protecting
against a disorder in which EZH2-mediated protein methylation plays
a part in a subject in need thereof by administering a
therapeutically effective amount of compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, to a subject in need of
such treatment. The disorder can be cancer, e.g., cancer in which
EZH2-mediated protein methylation plays a role. The present
invention also provides the use of compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph, solvate, or stereoisomeror thereof, for the
preparation of a medicament useful for the prevention of a cell
proliferative disorder associated, at least in part, with
EZH2-mediated protein methylation.
[0814] The compounds of this invention can or may be used to
modulate protein (e.g., histone) methylation, e.g., to modulate
histone methyltransferase or histone demethylase enzyme activity.
At least some of the compounds of the invention can be used in vivo
or in vitro for modulating protein methylation. Histone methylation
has been reported to be involved in aberrant expression of certain
genes in cancers, and in silencing of neuronal genes in
non-neuronal cells. At least some compounds described herein are
suitable candidates for treating these diseases, i.e., to decrease
methylation or restore methylation to roughly its level in
counterpart normal cells.
[0815] Compounds that are methylation modulators can or may be used
for modulating cell proliferation. For example, in some cases
excessive proliferation may be reduced with agents that decrease
methylation, whereas insufficient proliferation may be stimulated
with agents that increase methylation. Accordingly, diseases that
may be treated by the compounds of the invention could include
hyperproliferative diseases, such as benign cell growth and
malignant cell growth.
[0816] As used herein, a "subject in need thereof" is a subject
having a disorder in which EZH2-mediated protein methylation plays
a part, or a subject having an increased risk of developing such
disorder relative to the population at large. A subject in need
thereof can have a precancerous condition. Preferably, a subject in
need thereof has cancer. A "subject" includes a mammal. The mammal
can be e.g., a human or appropriate non-human mammal, such as
primate, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or a
pig. The subject can also be a bird or fowl. In one embodiment, the
mammal is a human.
[0817] As used herein, the term "cell proliferative disorder"
refers to conditions in which unregulated or abnormal growth, or
both, of cells can lead to the development of an unwanted condition
or disease, which may or may not be cancerous. Exemplary cell
proliferative disorders that may be treated with the compounds of
the invention encompass a variety of conditions wherein cell
division is deregulated. Exemplary cell proliferative disorder
include, but are not limited to, neoplasms, benign tumors,
malignant tumors, pre-cancerous conditions, in situ tumors,
encapsulated tumors, metastatic tumors, liquid tumors, solid
tumors, immunological tumors, hematological tumors, cancers,
carcinomas, leukemias, lymphomas, sarcomas, and rapidly dividing
cells. The term "rapidly dividing cell" as used herein is defined
as any cell that divides at a rate that exceeds or is greater than
what is expected or observed among neighboring or juxtaposed cells
within the same tissue. A cell proliferative disorder includes a
precancer or a precancerous condition. A cell proliferative
disorder includes cancer. The methods and uses provided herein can
be or may be used to treat or alleviate a symptom of cancer or to
identify suitable candidates for such purposes. The term "cancer"
includes solid tumors, as well as, hematologic tumors and/or
malignancies. A "precancer cell" or "precancerous cell" is a cell
manifesting a cell proliferative disorder that is a precancer or a
precancerous condition. A "cancer cell" or "cancerous cell" is a
cell manifesting a cell proliferative disorder that is a cancer.
Any reproducible means of measurement may be used to identify
cancer cells or precancerous cells. Cancer cells or precancerous
cells can be identified by histological typing or grading of a
tissue sample (e.g., a biopsy sample). Cancer cells or precancerous
cells can be identified through the use of appropriate molecular
markers.
[0818] Exemplary non-cancerous conditions or disorders that may be
treated using one or more compounds of the present invention
include, but are not limited to, rheumatoid arthritis;
inflammation; autoimmune disease; lymphoproliferative conditions;
acromegaly; rheumatoid spondylitis; osteoarthritis; gout, other
arthritic conditions; sepsis; septic shock; endotoxic shock;
gram-negative sepsis; toxic shock syndrome; asthma; adult
respiratory distress syndrome; chronic obstructive pulmonary
disease; chronic pulmonary inflammation; inflammatory bowel
disease; Crohn's disease; psoriasis; eczema; ulcerative colitis;
pancreatic fibrosis; hepatic fibrosis; acute and chronic renal
disease; irritable bowel syndrome; pyresis; restenosis; cerebral
malaria; stroke and ischemic injury; neural trauma; Alzheimer's
disease; Huntington's disease; Parkinson's disease; acute and
chronic pain; allergic rhinitis; allergic conjunctivitis; chronic
heart failure; acute coronary syndrome; cachexia; malaria; leprosy;
leishmaniasis; Lyme disease; Reiter's syndrome; acute synovitis;
muscle degeneration, bursitis; tendonitis; tenosynovitis;
herniated, ruptures, or prolapsed intervertebral disk syndrome;
osteopetrosis; thrombosis; restenosis; silicosis; pulmonary
sarcosis; bone resorption diseases, such as osteoporosis;
graft-versus-host reaction; Multiple Sclerosis; lupus;
fibromyalgia; AIDS and other viral diseases such as Herpes Zoster,
Herpes Simplex I or II, influenza virus and cytomegalovirus; and
diabetes mellitus.
[0819] Exemplary cancers that may be treated using one or more
compounds of the present invention include, but are not limited to,
adrenocortical carcinoma, AIDS-related cancers, AIDS-related
lymphoma, anal cancer, anorectal cancer, cancer of the anal canal,
appendix cancer, childhood cerebellar astrocytoma, childhood
cerebral astrocytoma, basal cell carcinoma, skin cancer
(non-melanoma), biliary cancer, extrahepatic bile duct cancer,
intrahepatic bile duct cancer, bladder cancer, urinary bladder
cancer, bone and joint cancer, osteosarcoma and malignant fibrous
histiocytoma, brain cancer, brain tumor, brain stem glioma,
cerebellar astrocytoma, cerebral astrocytoma/malignant glioma,
ependymoma, medulloblastoma, supratentorial primitive
neuroectodeimal tumors, visual pathway and hypothalamic glioma,
breast cancer, bronchial adenomas/carcinoids, carcinoid tumor,
gastrointestinal, nervous system cancer, nervous system lymphoma,
central nervous system cancer, central nervous system lymphoma,
cervical cancer, childhood cancers, chronic lymphocytic leukemia,
chronic myelogenous leukemia, chronic myeloproliferative disorders,
colon cancer, colorectal cancer, cutaneous T-cell lymphoma,
lymphoid neoplasm, mycosis fungoides, Seziary Syndrome, endometrial
cancer, esophageal cancer, extracranial germ cell tumor,
extragonadal germ cell tumor, extrahepatic bile duct cancer, eye
cancer, intraocular melanoma, retinoblastoma, gallbladder cancer,
gastric (stomach) cancer, gastrointestinal carcinoid tumor,
gastrointestinal stromal tumor (GIST), germ cell tumor, ovarian
germ cell tumor, gestational trophoblastic tumor glioma, head and
neck cancer, hepatocellular (liver) cancer, Hodgkin lymphoma,
hypopharyngeal cancer, intraocular melanoma, ocular cancer, islet
cell tumors (endocrine pancreas), Kaposi Sarcoma, kidney cancer,
renal cancer, kidney cancer, laryngeal cancer, acute lymphoblastic
leukemia, acute myeloid leukemia, chronic lymphocytic leukemia,
chronic myelogenous leukemia, hairy cell leukemia, lip and oral
cavity cancer, liver cancer, lung cancer, non-small cell lung
cancer, small cell lung cancer, AIDS-related lymphoma, non-Hodgkin
lymphoma, primary central nervous system lymphoma, Waldenstram
macroglobulinemia, medulloblastoma, melanoma, intraocular (eye)
melanoma, merkel cell carcinoma, mesothelioma malignant,
mesothelioma, metastatic squamous neck cancer, mouth cancer, cancer
of the tongue, multiple endocrine neoplasia syndrome, mycosis
fungoides, myelodysplastic syndromes,
myelodysplastic/myeloproliferative diseases, chronic myelogenous
leukemia, acute myeloid leukemia, multiple myeloma, chronic
myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma,
oral cancer, oral cavity cancer, oropharyngeal cancer, ovarian
cancer, ovarian epithelial cancer, ovarian low malignant potential
tumor, pancreatic cancer, islet cell pancreatic cancer, paranasal
sinus and nasal cavity cancer, parathyroid cancer, penile cancer,
pharyngeal cancer, pheochromocytoma, pineoblastoma and
supratentorial primitive neuroectodermal tumors, pituitary tumor,
plasma cell neoplasm/multiple myeloma, pleuropulmonary blastoma,
prostate cancer, rectal cancer, renal pelvis and ureter,
transitional cell cancer, retinoblastoma, rhabdomyosarcoma,
salivary gland cancer, ewing family of sarcoma tumors, Kaposi
Sarcoma, soft tissue sarcoma, uterine cancer, uterine sarcoma, skin
cancer (non-melanoma), skin cancer (melanoma), merkel cell skin
carcinoma, small intestine cancer, soft tissue sarcoma, squamous
cell carcinoma, stomach (gastric) cancer, supratentorial primitive
neuroectodermal tumors, testicular cancer, throat cancer, thymoma,
thymoma and thymic carcinoma, thyroid cancer, transitional cell
cancer of the renal pelvis and ureter and other urinary organs,
gestational trophoblastic tumor, urethral cancer, endometrial
uterine cancer, uterine sarcoma, uterine corpus cancer, vaginal
cancer, vulvar cancer, and Wilm's Tumor.
[0820] A "cell proliferative disorder of the hematologic system" is
a cell proliferative disorder involving cells of the hematologic
system. A cell proliferative disorder of the hematologic system can
include lymphoma, leukemia, myeloid neoplasms, mast cell neoplasms,
myelodysplasia, benign monoclonal gammopathy, lymphomatoid
granulomatosis, lymphomatoid papulosis, polycythemia vera, chronic
myelocytic leukemia, agnogenic myeloid metaplasia, and essential
thrombocythemia. A cell proliferative disorder of the hematologic
system can include hyperplasia, dysplasia, and metaplasia of cells
of the hematologic system. In one aspect, compositions of the
present invention may be used to treat a cancer selected from the
group consisting of a hematologic cancer of the present invention
or a hematologic cell proliferative disorder of the present
invention, or used to identify suitable candidates for such
purposes. A hematologic cancer of the present invention can include
multiple myeloma, lymphoma (including Hodgkin's lymphoma,
non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas of
lymphocytic and cutaneous origin), leukemia (including childhood
leukemia, hairy-cell leukemia, acute lymphocytic leukemia, acute
myelocytic leukemia, chronic lymphocytic leukemia, chronic
myelocytic leukemia, chronic myelogenous leukemia, and mast cell
leukemia), myeloid neoplasms and mast cell neoplasms.
[0821] A "cell proliferative disorder of the lung" is a cell
proliferative disorder involving cells of the lung. Cell
proliferative disorders of the lung can include all forms of cell
proliferative disorders affecting lung cells. Cell proliferative
disorders of the lung can include lung cancer, a precancer or
precancerous condition of the lung, benign growths or lesions of
the lung, and malignant growths or lesions of the lung, and
metastatic lesions in tissue and organs in the body other than the
lung. In one aspect, compositions of the present invention may be
used to treat lung cancer or cell proliferative disorders of the
lung, or used to identify suitable candidates for such purposes.
Lung cancer can include malignant lung neoplasms, carcinoma in
situ, typical carcinoid tumors, and atypical carcinoid tumors. Lung
cancer can include small cell lung cancer ("SCLC"), non-small cell
lung cancer ("NSCLC"), squamous cell carcinoma, adenocarcinoma,
small cell carcinoma, large cell carcinoma, adenosquamous cell
carcinoma, and mesothelioma. Lung cancer can include "scar
carcinoma," bronchioalveolar carcinoma, giant cell carcinoma,
spindle cell carcinoma, and large cell neuroendocrine carcinoma.
Lung cancer can include lung neoplasms having histologic and
ultrastructual heterogeneity (e.g., mixed cell types).
[0822] Cell proliferative disorders of the lung can include all
forms of cell proliferative disorders affecting lung cells. Cell
proliferative disorders of the lung can include lung cancer,
precancerous conditions of the lung. Cell proliferative disorders
of the lung can include hyperplasia, metaplasia, and dysplasia of
the lung. Cell proliferative disorders of the lung can include
asbestos-induced hyperplasia, squamous metaplasia, and benign
reactive mesothelial metaplasia. Cell proliferative disorders of
the lung can include replacement of columnar epithelium with
stratified squamous epithelium, and mucosal dysplasia. Individuals
exposed to inhaled injurious environmental agents such as cigarette
smoke and asbestos may be at increased risk for developing cell
proliferative disorders of the lung. Prior lung diseases that may
predispose individuals to development of cell proliferative
disorders of the lung can include chronic interstitial lung
disease, necrotizing pulmonary disease, scleroderma, rheumatoid
disease, sarcoidosis, interstitial pneumonitis, tuberculosis,
repeated pneumonias, idiopathic pulmonary fibrosis, granulomata,
asbestosis, fibrosing alveolitis, and Hodgkin's disease.
[0823] A "cell proliferative disorder of the colon" is a cell
proliferative disorder involving cells of the colon. In one aspect,
compositions of the present invention may be used to treat colon
cancer or cell proliferative disorders of the colon, or used to
identify suitable candidates for such purposes. Colon cancer can
include all forms of cancer of the colon. Colon cancer can include
sporadic and hereditary colon cancers. Colon cancer can include
malignant colon neoplasms, carcinoma in situ, typical carcinoid
tumors, and atypical carcinoid tumors. Colon cancer can include
adenocarcinoma, squamous cell carcinoma, and adenosquamous cell
carcinoma. Colon cancer can be associated with a hereditary
syndrome selected from the group consisting of hereditary
nonpolyposis colorectal cancer, familial adenomatous polyposis,
Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndrome and
juvenile polyposis. Colon cancer can be caused by a hereditary
syndrome selected from the group consisting of hereditary
nonpolyposis colorectal cancer, familial adenomatous polyposis,
Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndrome and
juvenile polyposis.
[0824] Cell proliferative disorders of the colon can include all
forms of cell proliferative disorders affecting colon cells. Cell
proliferative disorders of the colon can include colon cancer,
precancerous conditions of the colon, adenomatous polyps of the
colon and metachronous lesions of the colon. A cell proliferative
disorder of the colon can include adenoma. Cell proliferative
disorders of the colon can be characterized by hyperplasia,
metaplasia, and dysplasia of the colon. Prior colon diseases that
may predispose individuals to development of cell proliferative
disorders of the colon can include prior colon cancer. Current
disease that may predispose individuals to development of cell
proliferative disorders of the colon can include Crohn's disease
and ulcerative colitis. A cell proliferative disorder of the colon
can be associated with a mutation in a gene selected from the group
consisting of p53, ras, FAP and DCC. An individual can have an
elevated risk of developing a cell proliferative disorder of the
colon due to the presence of a mutation in a gene selected from the
group consisting of p53, ras, FAP and DCC.
[0825] A "cell proliferative disorder of the pancreas" is a cell
proliferative disorder involving cells of the pancreas. Cell
proliferative disorders of the pancreas can include all forms of
cell proliferative disorders affecting pancreatic cells. Cell
proliferative disorders of the pancreas can include pancreas
cancer, a precancer or precancerous condition of the pancreas,
hyperplasia of the pancreas, and dysaplasia of the pancreas, benign
growths or lesions of the pancreas, and malignant growths or
lesions of the pancreas, and metastatic lesions in tissue and
organs in the body other than the pancreas. Pancreatic cancer
includes all forms of cancer of the pancreas. Pancreatic cancer can
include ductal adenocarcinoma, adenosquamous carcinoma, pleomorphic
giant cell carcinoma, mucinous adenocarcinoma, osteoclast-like
giant cell carcinoma, mucinous cystadenocarcinoma, acinar
carcinoma, unclassified large cell carcinoma, small cell carcinoma,
pancreatoblastoma, papillary neoplasm, mucinous cystadenoma,
papillary cystic neoplasm, and serous cystadenoma. Pancreatic
cancer can also include pancreatic neoplasms having histologic and
ultrastructual heterogeneity (e.g., mixed cell types).
[0826] A "cell proliferative disorder of the prostate" is a cell
proliferative disorder involving cells of the prostate. Cell
proliferative disorders of the prostate can include all forms of
cell proliferative disorders affecting prostate cells. Cell
proliferative disorders of the prostate can include prostate
cancer, a precancer or precancerous condition of the prostate,
benign growths or lesions of the prostate, and malignant growths or
lesions of the prostate, and metastatic lesions in tissue and
organs in the body other than the prostate. Cell proliferative
disorders of the prostate can include hyperplasia, metaplasia, and
dysplasia of the prostate.
[0827] A "cell proliferative disorder of the skin" is a cell
proliferative disorder involving cells of the skin. Cell
proliferative disorders of the skin can include all forms of cell
proliferative disorders affecting skin cells. Cell proliferative
disorders of the skin can include a precancer or precancerous
condition of the skin, benign growths or lesions of the skin,
melanoma, malignant melanoma and other malignant growths or lesions
of the skin, and metastatic lesions in tissue and organs in the
body other than the skin. Cell proliferative disorders of the skin
can include hyperplasia, metaplasia, and dysplasia of the skin.
[0828] A "cell proliferative disorder of the ovary" is a cell
proliferative disorder involving cells of the ovary. Cell
proliferative disorders of the ovary can include all forms of cell
proliferative disorders affecting cells of the ovary. Cell
proliferative disorders of the ovary can include a precancer or
precancerous condition of the ovary, benign growths or lesions of
the ovary, ovarian cancer, malignant growths or lesions of the
ovary, and metastatic lesions in tissue and organs in the body
other than the ovary. Cell proliferative disorders of the skin can
include hyperplasia, metaplasia, and dysplasia of cells of the
ovary.
[0829] A "cell proliferative disorder of the breast" is a cell
proliferative disorder involving cells of the breast. Cell
proliferative disorders of the breast can include all forms of cell
proliferative disorders affecting breast cells. Cell proliferative
disorders of the breast can include breast cancer, a precancer or
precancerous condition of the breast, benign growths or lesions of
the breast, and malignant growths or lesions of the breast, and
metastatic lesions in tissue and organs in the body other than the
breast. Cell proliferative disorders of the breast can include
hyperplasia, metaplasia, and dysplasia of the breast.
[0830] A cell proliferative disorder of the breast can be a
precancerous condition of the breast. Compositions of the present
invention may be used to treat a precancerous condition of the
breast. A precancerous condition of the breast can include atypical
hyperplasia of the breast, ductal carcinoma in situ (DCIS),
intraductal carcinoma, lobular carcinoma in situ (LCIS), lobular
neoplasia, and stage 0 or grade 0 growth or lesion of the breast
(e.g., stage 0 or grade 0 breast cancer, or carcinoma in situ). A
precancerous condition of the breast can be staged according to the
TNM classification scheme as accepted by the American Joint
Committee on Cancer (AJCC), where the primary tumor (T) has been
assigned a stage of T0 or Tis; and where the regional lymph nodes
(N) have been assigned a stage of N0; and where distant metastasis
(M) has been assigned a stage of M0.
[0831] The cell proliferative disorder of the breast can be breast
cancer. In one aspect, compositions of the present invention may be
used to treat breast cancer, or used to identify suitable
candidates for such purposes. Breast cancer may include all forms
of cancer of the breast. Breast cancer includes all forms of cancer
of the breast. Breast cancer can include primary epithelial breast
cancers. Breast cancer can include cancers in which the breast is
involved by other tumors such as lymphoma, sarcoma or melanoma.
Breast cancer can include carcinoma of the breast, ductal carcinoma
of the breast, lobular carcinoma of the breast, undifferentiated
carcinoma of the breast, cystosarcoma phyllodes of the breast,
angiosarcoma of the breast, and primary lymphoma of the breast.
Breast cancer can include Stage I, II, IIIA, IIIB, IIIC and IV
breast cancer. Ductal carcinoma of the breast can include invasive
carcinoma, invasive carcinoma in situ with predominant intraductal
component, inflammatory breast cancer, and a ductal carcinoma of
the breast with a histologic type selected from the group
consisting of comedo, mucinous (colloid), medullary, medullary with
lymphcytic infiltrate, papillary, scirrhous, and tubular. Lobular
carcinoma of the breast can include invasive lobular carcinoma with
predominant in situ component, invasive lobular carcinoma, and
infiltrating lobular carcinoma. Breast cancer can include Paget's
disease, Paget's disease with intraductal carcinoma, and Paget's
disease with invasive ductal carcinoma. Breast cancer can include
breast neoplasms having histologic and ultrastructual heterogeneity
(e.g., mixed cell types).
[0832] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph, or solvate
thereof, may be used to treat breast cancer, or used to identify
suitable candidates for such purposes. A breast cancer that is to
be treated can include familial breast cancer. A breast cancer that
is to be treated can include sporadic breast cancer. A breast
cancer that is to be treated can arise in a male subject. A breast
cancer that is to be treated can arise in a female subject. A
breast cancer that is to be treated can arise in a premenopausal
female subject or a postmenopausal female subject. A breast cancer
that is to be treated can arise in a subject equal to or older than
30 years old, or a subject younger than 30 years old. A breast
cancer that is to be treated has arisen in a subject equal to or
older than 50 years old, or a subject younger than 50 years old. A
breast cancer that is to be treated can arise in a subject equal to
or older than 70 years old, or a subject younger than 70 years
old.
[0833] A breast cancer that is to be treated can be typed to
identify a familial or spontaneous mutation in BRCA1, BRCA2, or
p53. A breast cancer that is to be treated can be typed as having a
HER2/neu gene amplification, as overexpressing HER2/neu, or as
having a low, intermediate or high level of HER2/neu expression. A
breast cancer that is to be treated can be typed for a marker
selected from the group consisting of estrogen receptor (ER),
progesterone receptor (PR), human epidermal growth factor
receptor-2, Ki-67, CA15-3, CA 27-29, and c-Met. A breast cancer
that is to be treated can be typed as ER-unknown, ER-rich or
ER-poor. A breast cancer that is to be treated can be typed as
ER-negative or ER-positive. ER-typing of a breast cancer may be
performed by any reproducible means. ER-typing of a breast cancer
may be performed as set forth in Onkologie 27: 175-179 (2004). A
breast cancer that is to be treated can be typed as PR-unknown,
PR-rich, or PR-poor. A breast cancer that is to be treated can be
typed as PR-negative or PR-positive. A breast cancer that is to be
treated can be typed as receptor positive or receptor negative. A
breast cancer that is to be treated can be typed as being
associated with elevated blood levels of CA 15-3, or CA 27-29, or
both.
[0834] A breast cancer that is to be treated can include a
localized tumor of the breast. A breast cancer that is to be
treated can include a tumor of the breast that is associated with a
negative sentinel lymph node (SLN) biopsy. A breast cancer that is
to be treated can include a tumor of the breast that is associated
with a positive sentinel lymph node (SLN) biopsy. A breast cancer
that is to be treated can include a tumor of the breast that is
associated with one or more positive axillary lymph nodes, where
the axillary lymph nodes have been staged by any applicable method.
A breast cancer that is to be treated can include a tumor of the
breast that has been typed as having nodal negative status (e.g.,
node-negative) or nodal positive status (e.g., node-positive). A
breast cancer that is to be treated can include a tumor of the
breast that has metastasized to other locations in the body. A
breast cancer that is to be treated can be classified as having
metastasized to a location selected from the group consisting of
bone, lung, liver, or brain. A breast cancer that is to be treated
can be classified according to a characteristic selected from the
group consisting of metastatic, localized, regional,
local-regional, locally advanced, distant, multicentric, bilateral,
ipsilateral, contralateral, newly diagnosed, recurrent, and
inoperable.
[0835] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
may be used to treat or prevent a cell proliferative disorder of
the breast, or to treat or prevent breast cancer, in a subject
having an increased risk of developing breast cancer relative to
the population at large, or used to identify suitable candidates
for such purposes. A subject with an increased risk of developing
breast cancer relative to the population at large is a female
subject with a family history or personal history of breast cancer.
A subject with an increased risk of developing breast cancer
relative to the population at large is a female subject having a
germ-line or spontaneous mutation in BRCA1 or BRCA2, or both. A
subject with an increased risk of developing breast cancer relative
to the population at large is a female subject with a family
history of breast cancer and a germ-line or spontaneous mutation in
BRCA1 or BRCA2, or both. A subject with an increased risk of
developing breast cancer relative to the population at large is a
female who is greater than 30 years old, greater than 40 years old,
greater than 50 years old, greater than 60 years old, greater than
70 years old, greater than 80 years old, or greater than 90 years
old. A subject with an increased risk of developing breast cancer
relative to the population at large is a subject with atypical
hyperplasia of the breast, ductal carcinoma in situ (DCIS),
intraductal carcinoma, lobular carcinoma in situ (LCIS), lobular
neoplasia, or a stage 0 growth or lesion of the breast (e.g., stage
0 or grade 0 breast cancer, or carcinoma in situ).
[0836] A breast cancer that is to be treated can histologically
graded according to the Scarff-Bloom-Richardson system, wherein a
breast tumor has been assigned a mitosis count score of 1, 2, or 3;
a nuclear pleiomorphism score of 1, 2, or 3; a tubule formation
score of 1, 2, or 3; and a total Scarff-Bloom-Richardson score of
between 3 and 9. A breast cancer that is to be treated can be
assigned a tumor grade according to the International Consensus
Panel on the Treatment of Breast Cancer selected from the group
consisting of grade 1, grade 1-2, grade 2, grade 2-3, or grade
3.
[0837] A cancer that is to be treated can be staged according to
the American Joint Committee on Cancer (AJCC) TNM classification
system, where the tumor (T) has been assigned a stage of TX, T1,
T1mic, T1a, T1b, T1c, T2, T3, T4, T4a, T4b, T4c, or T4d; and where
the regional lymph nodes (N) have been assigned a stage of NX, N0,
N1, N2, N2a, N2b, N3, N3a, N3b, or N3c; and where distant
metastasis (M) can be assigned a stage of MX, M0, or M1. A cancer
that is to be treated can be staged according to an American Joint
Committee on Cancer (AJCC) classification as Stage I, Stage IIA,
Stage IIB, Stage IIIA, Stage IIIB, Stage IIIC, or Stage IV. A
cancer that is to be treated can be assigned a grade according to
an AJCC classification as Grade GX (e.g., grade cannot be
assessed), Grade 1, Grade 2, Grade 3 or Grade 4. A cancer that is
to be treated can be staged according to an AJCC pathologic
classification (pN) of pNX, pN0, PN0 (I-), PN0 (I+), PN0 (mol-),
PN0 (mol+), PN1, PN1(mi), PN1a, PN1b, PN1c, pN2, pN2a, pN2b, pN3,
pN3a, pN3b, or pN3c.
[0838] A cancer that is to be treated can include a tumor that has
been determined to be less than or equal to about 2 centimeters in
diameter. A cancer that is to be treated can include a tumor that
has been determined to be from about 2 to about 5 centimeters in
diameter. A cancer that is to be treated can include a tumor that
has been determined to be greater than or equal to about 3
centimeters in diameter. A cancer that is to be treated can include
a tumor that has been determined to be greater than 5 centimeters
in diameter. A cancer that is to be treated can be classified by
microscopic appearance as well differentiated, moderately
differentiated, poorly differentiated, or undifferentiated. A
cancer that is to be treated can be classified by microscopic
appearance with respect to mitosis count (e.g., amount of cell
division) or nuclear pleiomorphism (e.g., change in cells). A
cancer that is to be treated can be classified by microscopic
appearance as being associated with areas of necrosis (e.g., areas
of dying or degenerating cells). A cancer that is to be treated can
be classified as having an abnormal karyotype, having an abnormal
number of chromosomes, or having one or more chromosomes that are
abnormal in appearance. A cancer that is to be treated can be
classified as being aneuploid, triploid, tetraploid, or as having
an altered ploidy. A cancer that is to be treated can be classified
as having a chromosomal translocation, or a deletion or duplication
of an entire chromosome, or a region of deletion, duplication or
amplification of a portion of a chromosome.
[0839] A cancer that is to be treated can be evaluated by DNA
cytometry, flow cytometry, or image cytometry. A cancer that is to
be treated can be typed as having 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, or 90% of cells in the synthesis stage of cell division
(e.g., in S phase of cell division). A cancer that is to be treated
can be typed as having a low S-phase fraction or a high S-phase
fraction.
[0840] As used herein, a "normal cell" is a cell that cannot be
classified as part of a "cell proliferative disorder". A normal
cell lacks unregulated or abnormal growth, or both, that can lead
to the development of an unwanted condition or disease. Preferably,
a normal cell possesses normally functioning cell cycle checkpoint
control mechanisms.
[0841] As used herein, "contacting a cell" refers to a condition in
which a compound or other composition of matter is in direct
contact with a cell, or is close enough to induce a desired
biological effect in a cell.
[0842] As used herein, "candidate compound" refers to a compound of
the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph or solvate thereof, that has been or
will be tested in one or more in vitro or in vivo biological
assays, in order to determine if that compound is likely to elicit
a desired biological or medical response in a cell, tissue, system,
animal or human that is being sought by a researcher or clinician.
A candidate compound is a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof. The biological or medical response can be the
treatment of cancer. The biological or medical response can be
treatment or prevention of a cell proliferative disorder. The
biological response or effect can also include a change in cell
proliferation or growth that occurs in vitro or in an animal model,
as well as other biological changes that are observable in vitro.
In vitro or in vivo biological assays can include, but are not
limited to, enzymatic activity assays, electrophoretic mobility
shift assays, reporter gene assays, in vitro cell viability assays,
and the assays described herein.
[0843] As used herein, "monotherapy" refers to the administration
of a single active or therapeutic compound to a subject in need
thereof. Preferably, monotherapy will involve administration of a
therapeutically effective amount of an active compound. For
example, cancer monotherapy with one of the compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, to a subject in need of
treatment of cancer. Monotherapy may be contrasted with combination
therapy, in which a combination of multiple active compounds is
administered, preferably with each component of the combination
present in a therapeutically effective amount. Monotherapy with a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, may be
more effective than combination therapy in inducing a desired
biological effect.
[0844] As used herein, "treating" or "treat" describes the
management and care of a patient for the purpose of combating a
disease, condition, or disorder and includes the administration of
a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
to alleviate the symptoms or complications of a disease, condition
or disorder, or to eliminate the disease, condition or disorder.
The term "treat" can also include treatment of a cell in vitro or
an animal model.
[0845] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
can or may also be used to prevent a relevant disease, condition or
disorder, or used to identify suitable candidates for such
purposes. As used herein, "preventing" or "prevent" describes
reducing or eliminating the onset of the symptoms or complications
of such disease, condition or disorder.
[0846] As used herein, the term "alleviate" is meant to describe a
process by which the severity of a sign or symptom of a disorder is
decreased. Importantly, a sign or symptom can be alleviated without
being eliminated. The administration of pharmaceutical compositions
of the invention can or may lead to the elimination of a sign or
symptom, however, elimination is not required. Effective dosages
should be expected to decrease the severity of a sign or symptom.
For instance, a sign or symptom of a disorder such as cancer, which
can occur in multiple locations, is alleviated if the severity of
the cancer is decreased within at least one of multiple
locations.
[0847] As used herein, the term "severity" is meant to describe the
potential of cancer to transform from a precancerous, or benign,
state into a malignant state. Alternatively, or in addition,
severity is meant to describe a cancer stage, for example,
according to the TNM system (accepted by the International Union
Against Cancer (UICC) and the American Joint Committee on Cancer
(AJCC)) or by other art-recognized methods. Cancer stage refers to
the extent or severity of the cancer, based on factors such as the
location of the primary tumor, tumor size, number of tumors, and
lymph node involvement (spread of cancer into lymph nodes).
Alternatively, or in addition, severity is meant to describe the
tumor grade by art-recognized methods (see, National Cancer
Institute, www.cancer.gov). Tumor grade is a system used to
classify cancer cells in terms of how abnormal they look under a
microscope and how quickly the tumor is likely to grow and spread.
Many factors are considered when determining tumor grade, including
the structure and growth pattern of the cells. The specific factors
used to determine tumor grade vary with each type of cancer.
Severity also describes a histologic grade, also called
differentiation, which refers to how much the tumor cells resemble
normal cells of the same tissue type (see, National Cancer
Institute, www.cancer.gov). Furthermore, severity describes a
nuclear grade, which refers to the size and shape of the nucleus in
tumor cells and the percentage of tumor cells that are dividing
(see, National Cancer Institute, www.cancer.gov).
[0848] Severity can also describe the degree to which a tumor has
secreted growth factors, degraded the extracellular matrix, become
vascularized, lost adhesion to juxtaposed tissues, or metastasized.
Moreover, severity can describe the number of locations to which a
primary tumor has metastasized. Finally, severity can include the
difficulty of treating tumors of varying types and locations. For
example, inoperable tumors, those cancers which have greater access
to multiple body systems (hematological and immunological tumors),
and those which are the most resistant to traditional treatments
are considered most severe. In these situations, prolonging the
life expectancy of the subject and/or reducing pain, decreasing the
proportion of cancerous cells or restricting cells to one system,
and improving cancer stage/tumor grade/histological grade/nuclear
grade are considered alleviating a sign or symptom of the
cancer.
[0849] As used herein the term "symptom" is defined as an
indication of disease, illness, injury, or that something is not
right in the body. Symptoms are felt or noticed by the individual
experiencing the symptom, but may not easily be noticed by others.
Others are defined as non-health-care professionals.
[0850] As used herein the term "sign" is also defined as an
indication that something is not right in the body. But signs are
defined as things that can be seen by a doctor, nurse, or other
health care professional.
[0851] Cancer is a group of diseases that may cause almost any sign
or symptom. The signs and symptoms will depend on where the cancer
is, the size of the cancer, and how much it affects the nearby
organs or structures. If a cancer spreads (metastasizes), then
symptoms may appear in different parts of the body.
[0852] As a cancer grows, it begins to push on nearby organs, blood
vessels, and nerves. This pressure creates some of the signs and
symptoms of cancer. If the cancer is in a critical area, such as
certain parts of the brain, even the smallest tumor can cause early
symptoms.
[0853] But sometimes cancers start in places where it does not
cause any symptoms until the cancer has grown quite large. Pancreas
cancers, for example, do not usually grow large enough to be felt
from the outside of the body. Some pancreatic cancers do not cause
symptoms until they begin to grow around nearby nerves (this causes
a backache). Others grow around the bile duct, which blocks the
flow of bile and leads to a yellowing of the skin known as
jaundice. By the time a pancreatic cancer causes these signs or
symptoms, it has usually reached an advanced stage.
[0854] A cancer may also cause symptoms such as fever, fatigue, or
weight loss. This may be because cancer cells use up much of the
body's energy supply or release substances that change the body's
metabolism. Or the cancer may cause the immune system to react in
ways that produce these symptoms.
[0855] Sometimes, cancer cells release substances into the
bloodstream that cause symptoms not usually thought to result from
cancers. For example, some cancers of the pancreas can release
substances which cause blood clots to develop in veins of the legs.
Some lung cancers make hormone-like substances that affect blood
calcium levels, affecting nerves and muscles and causing weakness
and dizziness.
[0856] Cancer presents several general signs or symptoms that occur
when a variety of subtypes of cancer cells are present. Most people
with cancer will lose weight at some time with their disease. An
unexplained (unintentional) weight loss of 10 pounds or more may be
the first sign of cancer, particularly cancers of the pancreas,
stomach, esophagus, or lung.
[0857] Fever is very common with cancer, but is more often seen in
advanced disease. Almost all patients with cancer will have fever
at some time, especially if the cancer or its treatment affects the
immune system and makes it harder for the body to fight infection.
Less often, fever may be an early sign of cancer, such as with
leukemia or lymphoma.
[0858] Fatigue may be an important symptom as cancer progresses. It
may happen early, though, in cancers such as with leukemia, or if
the cancer is causing an ongoing loss of blood, as in some colon or
stomach cancers.
[0859] Pain may be an early symptom with some cancers such as bone
cancers or testicular cancer. But most often pain is a symptom of
advanced disease.
[0860] Along with cancers of the skin (see next section), some
internal cancers can cause skin signs that can be seen. These
changes include the skin looking darker (hyperpigmentation), yellow
(jaundice), or red (erythema); itching; or excessive hair
growth.
[0861] Alternatively, or in addition, cancer subtypes present
specific signs or symptoms. Changes in bowel habits or bladder
function could indicate cancer. Long-term constipation, diarrhea,
or a change in the size of the stool may be a sign of colon cancer.
Pain with urination, blood in the urine, or a change in bladder
function (such as more frequent or less frequent urination) could
be related to bladder or prostate cancer.
[0862] Changes in skin condition or appearance of a new skin
condition could indicate cancer. Skin cancers may bleed and look
like sores that do not heal. A long-lasting sore in the mouth could
be an oral cancer, especially in patients who smoke, chew tobacco,
or frequently drink alcohol. Sores on the penis or vagina may
either be signs of infection or an early cancer.
[0863] Unusual bleeding or discharge could indicate cancer. Unusual
bleeding can happen in either early or advanced cancer. Blood in
the sputum (phlegm) may be a sign of lung cancer. Blood in the
stool (or a dark or black stool) could be a sign of colon or rectal
cancer. Cancer of the cervix or the endometrium (lining of the
uterus) can cause vaginal bleeding. Blood in the urine may be a
sign of bladder or kidney cancer. A bloody discharge from the
nipple may be a sign of breast cancer.
[0864] A thickening or lump in the breast or in other parts of the
body could indicate the presence of a cancer. Many cancers can be
felt through the skin, mostly in the breast, testicle, lymph nodes
(glands), and the soft tissues of the body. A lump or thickening
may be an early or late sign of cancer. Any lump or thickening
could be indicative of cancer, especially if the formation is new
or has grown in size.
[0865] Indigestion or trouble swallowing could indicate cancer.
While these symptoms commonly have other causes, indigestion or
swallowing problems may be a sign of cancer of the esophagus,
stomach, or pharynx (throat).
[0866] Recent changes in a wart or mole could be indicative of
cancer. Any wart, mole, or freckle that changes in color, size, or
shape, or loses its definite borders indicates the potential
development of cancer. For example, the skin lesion may be a
melanoma.
[0867] A persistent cough or hoarseness could be indicative of
cancer. A cough that does not go away may be a sign of lung cancer.
Hoarseness can be a sign of cancer of the larynx (voice box) or
thyroid.
[0868] While the signs and symptoms listed above are the more
common ones seen with cancer, there are many others that are less
common and are not listed here.
[0869] Treating cancer may result in or can result in a reduction
in size of a tumor. A reduction in size of a tumor may also be
referred to as "tumor regression". Preferably, after treatment,
tumor size would be reduced by 5% or greater relative to its size
prior to treatment; more preferably, tumor size is reduced by 10%
or greater; more preferably, reduced by 20% or greater; more
preferably, reduced by 30% or greater; more preferably, reduced by
40% or greater; even more preferably, reduced by 50% or greater;
and most preferably, reduced by greater than 75% or greater. Size
of a tumor may be measured by any reproducible means of
measurement. The size of a tumor may be measured as a diameter of
the tumor.
[0870] Treating cancer may result in or can result in a reduction
in tumor volume. Preferably, after treatment, tumor volume would be
reduced by 5% or greater relative to its size prior to treatment;
more preferably, tumor volume is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75% or greater. Tumor volume may be
measured by any reproducible means of measurement.
[0871] Treating cancer may result in or can result in a decrease in
number of tumors. Preferably, after treatment, tumor number would
be reduced by 5% or greater relative to number prior to treatment;
more preferably, tumor number is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75%. Number of tumors may be measured by
any reproducible means of measurement. The number of tumors may be
measured by counting tumors visible to the naked eye or at a
specified magnification. Preferably, the specified magnification is
2.times., 3.times., 4.times., 5.times., 10.times., or
50.times..
[0872] Treating cancer may result in or can result in a decrease in
number of metastatic lesions in other tissues or organs distant
from the primary tumor site. Preferably, after treatment, the
number of metastatic lesions would be reduced by 5% or greater
relative to number prior to treatment; more preferably, the number
of metastatic lesions is reduced by 10% or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by
30% or greater; more preferably, reduced by 40% or greater; even
more preferably, reduced by 50% or greater; and most preferably,
reduced by greater than 75%. The number of metastatic lesions may
be measured by any reproducible means of measurement. The number of
metastatic lesions may be measured by counting metastatic lesions
visible to the naked eye or at a specified magnification.
Preferably, the specified magnification is 2.times., 3.times.,
4.times., 5.times., 10.times., or 50.times..
[0873] Treating cancer may result in or can result in an increase
in average survival time of a population of treated subjects in
comparison to a population receiving carrier alone. Preferably, the
average survival time would be increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. An increase in average survival time of a
population may be measured, for example, by calculating for a
population the average length of survival following initiation of
treatment with an active compound. An increase in average survival
time of a population may also be measured, for example, by
calculating for a population the average length of survival
following completion of a first round of treatment with an active
compound.
[0874] Treating cancer may result in or can result in an increase
in average survival time of a population of treated subjects in
comparison to a population of untreated subjects. Preferably, the
average survival time would be increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. An increase in average survival time of a
population may be measured, for example, by calculating for a
population the average length of survival following initiation of
treatment with an active compound. An increase in average survival
time of a population may also be measured, for example, by
calculating for a population the average length of survival
following completion of a first round of treatment with an active
compound.
[0875] Treating cancer may result in or can result in increase in
average survival time of a population of treated subjects in
comparison to a population receiving monotherapy with a drug that
is not a compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof.
Preferably, the average survival time would be increased by more
than 30 days; more preferably, by more than 60 days; more
preferably, by more than 90 days; and most preferably, by more than
120 days. An increase in average survival time of a population may
be measured by any reproducible means. An increase in average
survival time of a population may be measured, for example, by
calculating for a population the average length of survival
following initiation of treatment with an active compound. An
increase in average survival time of a population may also be
measured, for example, by calculating for a population the average
length of survival following completion of a first round of
treatment with an active compound.
[0876] Treating cancer may result in or can result in a decrease in
the mortality rate of a population of treated subjects in
comparison to a population receiving carrier alone. Treating cancer
may result in or can result in a decrease in the mortality rate of
a population of treated subjects in comparison to an untreated
population. Treating cancer may result in or can result in a
decrease in the mortality rate of a population of treated subjects
in comparison to a population receiving monotherapy with a drug
that is not a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof. Preferably, the mortality rate would be
decreased by more than 2%; more preferably, by more than 5%; more
preferably, by more than 10%; and most preferably, by more than
25%. A decrease in the mortality rate of a population of treated
subjects may be measured by any reproducible means. A decrease in
the mortality rate of a population may be measured, for example, by
calculating for a population the average number of disease-related
deaths per unit time following initiation of treatment with an
active compound. A decrease in the mortality rate of a population
may also be measured, for example, by calculating for a population
the average number of disease-related deaths per unit time
following completion of a first round of treatment with an active
compound.
[0877] Treating cancer may result in or can result in a decrease in
tumor growth rate. Preferably, after treatment, tumor growth rate
would be reduced by at least 5% relative to number prior to
treatment; more preferably, tumor growth rate would be reduced by
at least 10%; more preferably, reduced by at least 20%; more
preferably, reduced by at least 30%; more preferably, reduced by at
least 40%; more preferably, reduced by at least 50%; even more
preferably, reduced by at least 50%; and most preferably, reduced
by at least 75%. Tumor growth rate may be measured by any
reproducible means of measurement. Tumor growth rate can be
measured according to a change in tumor diameter per unit time.
[0878] Treating cancer may result in or can result in a decrease in
tumor regrowth. Preferably, after treatment, tumor regrowth would
be less than 5%; more preferably, tumor regrowth would be less than
10%; more preferably, less than 20%; more preferably, less than
30%; more preferably, less than 40%; more preferably, less than
50%; even more preferably, less than 50%; and most preferably, less
than 75%. Tumor regrowth may be measured by any reproducible means
of measurement. Tumor regrowth is measured, for example, by
measuring an increase in the diameter of a tumor after a prior
tumor shrinkage that followed treatment. A decrease in tumor
regrowth is indicated by failure of tumors to reoccur after
treatment has stopped.
[0879] Treating or preventing a cell proliferative disorder may
result in or can result in a reduction in the rate of cellular
proliferation. Preferably, after treatment, the rate of cellular
proliferation would be reduced by at least 5%; more preferably, by
at least 10%; more preferably, by at least 20%; more preferably, by
at least 30%; more preferably, by at least 40%; more preferably, by
at least 50%; even more preferably, by at least 50%; and most
preferably, by at least 75%. The rate of cellular proliferation may
be measured by any reproducible means of measurement. The rate of
cellular proliferation is measured, for example, by measuring the
number of dividing cells in a tissue sample per unit time.
[0880] Treating or preventing a cell proliferative disorder may
result in or can result in a reduction in the proportion of
proliferating cells. Preferably, after treatment, the proportion of
proliferating cells would be reduced by at least 5%; more
preferably, by at least 10%; more preferably, by at least 20%; more
preferably, by at least 30%; more preferably, by at least 40%; more
preferably, by at least 50%; even more preferably, by at least 50%;
and most preferably, by at least 75%. The proportion of
proliferating cells may be measured by any reproducible means of
measurement. Preferably, the proportion of proliferating cells is
measured, for example, by quantifying the number of dividing cells
relative to the number of nondividing cells in a tissue sample. The
proportion of proliferating cells can be equivalent to the mitotic
index.
[0881] Treating or preventing a cell proliferative disorder may
result in or can result in a decrease in size of an area or zone of
cellular proliferation. Preferably, after treatment, size of an
area or zone of cellular proliferation would be reduced by at least
5% relative to its size prior to treatment; more preferably,
reduced by at least 10%; more preferably, reduced by at least 20%;
more preferably, reduced by at least 30%; more preferably, reduced
by at least 40%; more preferably, reduced by at least 50%; even
more preferably, reduced by at least 50%; and most preferably,
reduced by at least 75%. Size of an area or zone of cellular
proliferation may be measured by any reproducible means of
measurement. The size of an area or zone of cellular proliferation
may be measured as a diameter or width of an area or zone of
cellular proliferation.
[0882] Treating or preventing a cell proliferative disorder may
result in or can result in a decrease in the number or proportion
of cells having an abnormal appearance or morphology. Preferably,
after treatment, the number of cells having an abnormal morphology
would be reduced by at least 5% relative to its size prior to
treatment; more preferably, reduced by at least 10%; more
preferably, reduced by at least 20%; more preferably, reduced by at
least 30%; more preferably, reduced by at least 40%; more
preferably, reduced by at least 50%; even more preferably, reduced
by at least 50%; and most preferably, reduced by at least 75%. An
abnormal cellular appearance or morphology may be measured by any
reproducible means of measurement. An abnormal cellular morphology
can be measured by microscopy, e.g., using an inverted tissue
culture microscope. An abnormal cellular morphology can take the
form of nuclear pleiomorphism.
[0883] As used herein, the term "selectively" means tending to
occur at a higher frequency in one population than in another
population. The compared populations can be cell populations. A
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, may or can
act selectively on a cancer or precancerous cell but not on a
normal cell. A compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, may or can act selectively to modulate one
molecular target (e.g., a target protein methyltransferase) but
does not significantly modulate another molecular target (e.g., a
non-target protein methyltransferase). The invention also provides
a method for selectively inhibiting the activity of an enzyme, such
as a protein methyltransferase. Preferably, an event occurs
selectively in population A relative to population B if it occurs
greater than two times more frequently in population A as compared
to population B. An event occurs selectively if it occurs greater
than five times more frequently in population A. An event occurs
selectively if it occurs greater than ten times more frequently in
population A; more preferably, greater than fifty times; even more
preferably, greater than 100 times; and most preferably, greater
than 1000 times more frequently in population A as compared to
population B. For example, cell death would be said to occur
selectively in cancer cells if it occurred greater than twice as
frequently in cancer cells as compared to normal cells.
[0884] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
may or can modulate the activity of a molecular target (e.g., a
target protein methyltransferase). Modulating refers to stimulating
or inhibiting an activity of a molecular target. Preferably, a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, modulates
the activity of a molecular target if it stimulates or inhibits the
activity of the molecular target by at least 2-fold relative to the
activity of the molecular target under the same conditions but
lacking only the presence of said compound. More preferably, a
compound of the present invention, or a pharmaceutically acceptable
salt, prodrug, metabolite, polymorph or solvate thereof, modulates
the activity of a molecular target if it stimulates or inhibits the
activity of the molecular target by at least 5-fold, at least
10-fold, at least 20-fold, at least 50-fold, at least 100-fold
relative to the activity of the molecular target under the same
conditions but lacking only the presence of said compound. The
activity of a molecular target may be measured by any reproducible
means. The activity of a molecular target may be measured in vitro
or in vivo. For example, the activity of a molecular target may be
measured in vitro by an enzymatic activity assay or a DNA binding
assay, or the activity of a molecular target may be measured in
vivo by assaying for expression of a reporter gene.
[0885] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
does not significantly modulate the activity of a molecular target
if the addition of the compound does not stimulate or inhibit the
activity of the molecular target by greater than 10% relative to
the activity of the molecular target under the same conditions but
lacking only the presence of said compound.
[0886] As used herein, the term "isozyme selective" means
preferential inhibition or stimulation of a first isoform of an
enzyme in comparison to a second isoform of an enzyme (e.g.,
preferential inhibition or stimulation of a protein
methyltransferase isozyme alpha in comparison to a protein
methyltransferase isozyme beta). Preferably, a compound of the
present invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, demonstrates a minimum of
a fourfold differential, preferably a tenfold differential, more
preferably a fifty fold differential, in the dosage required to
achieve a biological effect. Preferably, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, demonstrates this
differential across the range of inhibition, and the differential
is exemplified at the IC.sub.50, i.e., a 50% inhibition, for a
molecular target of interest.
[0887] Administering a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, to a cell or a subject in need thereof may result
in or can result in modulation (i.e., stimulation or inhibition) of
an activity of a protein methyltransferase of interest.
[0888] The present invention provides methods to assess biological
activity of a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof or methods of identifying a test compound as an
inhibitor of a Y641 mutant of EZH2. In one embodiment the method
includes combining an isolated Y641 mutant of EZH2 with a histone
substrate, a methyl group donor (such as S-adenosylmethionine
(SAM)), and a test compound, wherein the histone substrate
comprises a form of H3-K27 selected from the group consisting of
unmethylated H3-K27, monomethylated H3-K27, dimethylated H3-K27,
and any combination thereof; and performing an assay to detect
methylation of H3-K27 in the histone substrate, thereby identifying
the test compound as an inhibitor of the Y641 mutant of EZH2 when
methylation of H3-K27 in the presence of the test compound is less
than methylation of H3-K27 in the absence of the test compound. The
assay to detect methylation of H3-K27 can be selected to measure
the rate of methylation, the extent of methylation, or both the
rate and extent of methylation.
[0889] The Y641 mutant of EZH2 is isolated as a PRC2 complex or
functional equivalent thereof. As used herein, the term "isolated"
means substantially separated from other components with which the
complex may be found as it occurs in nature. A compound can be
isolated without necessarily being purified. In one embodiment the
mutant of EZH2 is isolated as a complex of a Y641 mutant of EZH2
together with EED and SUZ12. In another embodiment the mutant of
EZH2 is isolated as a complex of a Y641 mutant of EZH2 together
with EED, SUZ12, and RbAp48. Under appropriate conditions, a PRC2
complex or functional equivalent thereof exhibits histone
methyltransferase activity for H3-K27. In one embodiment the
complex is composed of recombinantly expressed component
polypeptides, e.g., EZH2, EED, SUZ12, with or without RbAp48.
[0890] The isolated Y641 mutant of EZH2 is combined with a histone
substrate. A histone substrate includes any suitable source of
histone polypeptides or fragments thereof that can serve as
substrate for EZH2. In one embodiment the histone substrate
includes histones isolated from a subject. The histones can be
isolated from cells of a subject using any suitable method; such
methods are well known to persons skilled in the art and need not
be further specified here. See, for example, Fang et al. (2004)
Methods Enzymol 377:213-26. In accordance with the Examples below,
in one embodiment the histone substrate is provided as nucleosomes.
In accordance with the Examples below, in one embodiment the
histone substrate is provided as avian (chicken) erythrocyte
nucleosomes.
[0891] Histone substrate so provided may include an admixture of
states of histone modification, including various states of H3-K27
methylation as judged by Western blotting with H3-K27 methylation
state-specific antibodies. In one embodiment the histone substrate
may be provided as purified full-length histone H3. Such purified
full-length histone H3 may be provided as a homogeneous preparation
in respect of states of H3-K27 methylation, or as an admixture of
various states of H3-K27 methylation. Homogeneous preparations of
isolated histone H3 in respect of states of H3-K27 methylation may
be prepared in part by passage over an immunoaffinity column loaded
with suitable H3-K27 methylation state-specific antibodies or by
immunoprecipitation using magnetic beads coated with suitable
H3-K27 methylation state-specific antibodies. Alternatively or in
addition, the methylation state of H3-K27 can be characterized as
part of performing the assay. For example, the starting material
histone substrate might be characterized as containing 50 percent
unmethylated H3-K27, 40 percent monomethylated H3-K27, 10 percent
dimethylated H3-K27, and 0 percent trimethylated H3-K27.
[0892] In one embodiment the histone substrate includes a peptide
library or a suitable peptide comprising one or more amino acid
sequences related to histone H3, including, in particular, a
sequence that encompasses H3-K27. For example, in one embodiment,
the histone substrate is a peptide fragment that corresponds to
amino acid residues 21-44 of histone H3. The peptide library or
peptide can be prepared by peptide synthesis according to
techniques well known in the art and optionally modified so as to
incorporate any desired degree of methylation of lysine
corresponding to H3-K27. As described in the Examples below, such
peptides can also be modified to incorporate a label, such as
biotin, useful in performing downstream assays. In one embodiment
the label is appended to the amino (N)-terminus of the peptide(s).
In one embodiment the label is appended to the carboxy (C)-terminus
of the peptide(s).
[0893] Detection of methylation of H3-K27 can be accomplished using
any suitable method. In one embodiment, the source of donor methyl
groups includes methyl groups that are labeled with a detectable
label. The detectable label in one embodiment is an isotopic label,
e.g., tritium. Other types of labels may include, for example,
fluorescent labels.
[0894] Detection of formation of trimethylated H3-K27 can be
accomplished using any suitable method. For example, detection of
formation of trimethylated H3-K27 can be accomplished using an
assay to detect incorporation of labeled methyl groups, such as
described above, optionally combined with a chromatographic or
other method to separate labeled products by size, e.g.,
polyacrylamide gel electrophoresis (PAGE), capillary
electrophoresis (CE), or high pressure liquid chromatography
(HPLC). Alternatively or in addition, detection of formation of
trimethylated H3-K27 can be accomplished using antibodies that are
specific for trimethylated H3-K27.
[0895] Detection of conversion of monomethylated H3-K27 to
dimethylated H3-K27 can be accomplished using any suitable method.
In one embodiment the conversion is measured using antibodies
specific for monomethylated H3-K27 and dimethylated H3-K27. For
example, starting amounts or concentrations of monomethylated
H3-K27 and dimethylated H3-K27 may be determined using appropriate
antibodies specific for monomethylated H3-K27 and dimethylated
H3-K27. Following the combination of enzyme, substrate, methyl
group donor, and test compound, resulting amounts or concentrations
of monomethylated H3-K27 and dimethylated H3-K27 may then be
determined using appropriate antibodies specific for monomethylated
H3-K27 and dimethylated H3-K27. The beginning and resulting amounts
or concentrations of monomethylated H3-K27 and dimethylated H3-K27
can then be compared. Alternatively or in addition, beginning and
resulting amounts or concentrations of monomethylated H3-K27 and
dimethylated H3-K27 can then be compared to corresponding amounts
of concentrations from a negative control. A negative control
reaction, in which no test agent is included in the assay, can be
run in parallel or as a historical control. Results of such control
reaction can optionally be subtracted from corresponding results of
the experimental reaction prior to or in conjunction with making
the comparison mentioned above.
[0896] Because the dimethylated form of H3-K27 may be further
methylated in the same assay, a reduction in the amount or
concentration of monomethylated H3-K27 may not appear to correspond
directly to an increase in dimethylated H3-K27. In this instance,
it may be presumed, however, that a reduction in the amount or
concentration of monomethylated H3-K27 is, by itself, reflective of
conversion of monomethylated H3-K27 to dimethylated H3-K27.
[0897] Detection of conversion of dimethylated H3-K27 to
trimethylated H3-K27 can be accomplished using any suitable method.
In one embodiment the conversion is measured using antibodies
specific for dimethylated H3-K27 and trimethylated H3-K27. For
example, starting amounts or concentrations of dimethylated H3-K27
and trimethylated H3-K27 may be determined using appropriate
antibodies specific for dimethylated H3-K27 and trimethylated
H3-K27. Following the combination of enzyme, substrate, and test
compound, resulting amounts or concentrations of dimethylated
H3-K27 and trimethylated H3-K27 may then be determined using
appropriate antibodies specific for dimethylated H3-K27 and
trimethylated H3-K27. The beginning and resulting amounts or
concentrations of dimethylated H3-K27 and trimethylated H3-K27 can
then be compared. Alternatively or in addition, beginning and
resulting amounts or concentrations of dimethylated H3-K27 and
trimethylated H3-K27 can then be compared to corresponding amounts
of concentrations from a negative control. A negative control
reaction, in which no test agent is included in the assay, can be
run in parallel or as a historical control. Results of such control
reaction can optionally be subtracted from corresponding results of
the experimental reaction prior to or in conjunction with making
the comparison mentioned above.
[0898] A test agent is identified as an inhibitor of the Y641
mutant of EZH2 when methylation of H3-K27 with the test compound is
less than methylation of H3-K27 without the test compound. In one
embodiment, a test agent is identified as an inhibitor of the Y641
mutant of EZH2 when formation of trimethylated H3-K27 in the
presence of the test compound is less than formation of
trimethylated H3-K27 in the absence of the test compound.
[0899] The present invention also provides a method for identifying
a selective inhibitor of a Y641 mutant of EZH2. In one embodiment
the method includes combining an isolated Y641 mutant of EZH2 with
a histone substrate, a methyl group donor (e.g., SAM), and a test
compound, wherein the histone substrate comprises a form of H3-K27
selected from the group consisting of monomethylated H3-K27,
dimethylated H3-K27, and a combination of monomethylated H3-K27 and
dimethylated H3-K27, thereby forming a test mixture; combining an
isolated wild-type EZH2 with a histone substrate, a methyl group
donor (e.g., SAM), and a test compound, wherein the histone
substrate comprises a form of H3-K27 selected from the group
consisting of monomethylated H3-K27, dimethylated H3-K27, and a
combination of monomethylated H3-K27 and dimethylated H3-K27,
thereby forming a control mixture; performing an assay to detect
trimethylation of the histone substrate in each of the test mixture
and the control mixture; calculating the ratio of (a)
trimethylation with the Y641 mutant of EZH2 and the test compound
(M+) to (b) trimethylation with the Y641 mutant of EZH2 without the
test compound (M-); calculating the ratio of (c) trimethylation
with wild-type EZH2 and the test compound (WT+) to (d)
trimethylation with wild-type EZH2 without the test compound (WT-);
comparing the ratio (a)/(b) with the ratio (c)/(d); and identifying
the test compound as a selective inhibitor of the Y641 mutant of
EZH2 when the ratio (a)/(b) is less than the ratio (c)/(d). In one
embodiment the method further includes taking into account a
negative control without test compound for either or both of the
test mixture and the control mixture.
[0900] In some assays, immunological reagents, e.g., antibodies and
antigens, are employed. Fluorescence can be utilized in the
measurement of enzymatic activity in some assays. As used herein,
"fluorescence" refers to a process through which a molecule emits a
photon as a result of absorbing an incoming photon of higher energy
by the same molecule. Specific methods for assessing the biological
activity of the disclosed compounds are described in the
examples.
[0901] Administering a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, to a cell or a subject in need thereof may or can
result in modulation (i.e., stimulation or inhibition) of an
activity of an intracellular target (e.g., substrate). Several
intracellular targets may or can be modulated with the compounds of
the present invention, including, but not limited to, protein
methyltransferase.
[0902] Activating refers to placing a composition of matter (e.g.,
protein or nucleic acid) in a state suitable for carrying out a
desired biological function. A composition of matter capable of
being activated also has an unactivated state. An activated
composition of matter may have an inhibitory or stimulatory
biological function, or both.
[0903] Elevation refers to an increase in a desired biological
activity of a composition of matter (e.g., a protein or a nucleic
acid). Elevation may occur through an increase in concentration of
a composition of matter.
[0904] As used herein, "a cell cycle checkpoint pathway" refers to
a biochemical pathway that is involved in modulation of a cell
cycle checkpoint. A cell cycle checkpoint pathway may have
stimulatory or inhibitory effects, or both, on one or more
functions comprising a cell cycle checkpoint. A cell cycle
checkpoint pathway is comprised of at least two compositions of
matter, preferably proteins, both of which contribute to modulation
of a cell cycle checkpoint. A cell cycle checkpoint pathway may be
activated through an activation of one or more members of the cell
cycle checkpoint pathway. Preferably, a cell cycle checkpoint
pathway is a biochemical signaling pathway.
[0905] As used herein, "cell cycle checkpoint regulator" refers to
a composition of matter that can function, at least in part, in
modulation of a cell cycle checkpoint. A cell cycle checkpoint
regulator may have stimulatory or inhibitory effects, or both, on
one or more functions comprising a cell cycle checkpoint. A cell
cycle checkpoint regulator can be a protein or not a protein.
[0906] Treating cancer or a cell proliferative disorder may result
in or can result in cell death, and preferably, cell death would
result in a decrease of at least 10% in number of cells in a
population. More preferably, cell death means a decrease of at
least 20%; more preferably, a decrease of at least 30%; more
preferably, a decrease of at least 40%; more preferably, a decrease
of at least 50%; most preferably, a decrease of at least 75%.
Number of cells in a population may be measured by any reproducible
means. A number of cells in a population can be measured by
fluorescence activated cell sorting (FACS), immunofluorescence
microscopy and light microscopy. Methods of measuring cell death
are as shown in Li et al., Proc Natl Acad Sci USA. 100(5): 2674-8,
2003. In an aspect, cell death occurs by apoptosis.
[0907] Preferably, an effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, would not be
significantly cytotoxic to normal cells. A therapeutically
effective amount of a compound is not significantly cytotoxic to
normal cells if administration of the compound in a therapeutically
effective amount does not induce cell death in greater than 10% of
normal cells. A therapeutically effective amount of a compound does
not significantly affect the viability of normal cells if
administration of the compound in a therapeutically effective
amount does not induce cell death in greater than 10% of normal
cells. In an aspect, cell death occurs by apoptosis.
[0908] Contacting a cell with a compound of the present invention,
or a pharmaceutically acceptable salt, prodrug, metabolite,
polymorph or solvate thereof, may or can induce or activate cell
death selectively in cancer cells. Administering to a subject in
need thereof a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, may or can induce or activate cell death
selectively in cancer cells. Contacting a cell with a compound of
the present invention, or a pharmaceutically acceptable salt,
prodrug, metabolite, polymorph or solvate thereof, may or can
induce cell death selectively in one or more cells affected by a
cell proliferative disorder. Preferably, administering to a subject
in need thereof a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, would induce cell death selectively in one or more
cells affected by a cell proliferative disorder.
[0909] One aspect of the present invention relates to a method of
treating or preventing cancer (e.g., the course of which can be
influenced by modulating EZH2-mediated protein methylation) by
administering a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, polymorph or
solvate thereof, to a subject in need thereof, where administration
of the compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, polymorph or solvate thereof,
results in one or more of the following: prevention of cancer cell
proliferation by accumulation of cells in one or more phases of the
cell cycle (e.g. G1, G1/S, G2/M), or induction of cell senescence,
or promotion of tumor cell differentiation; promotion of cell death
in cancer cells via cytotoxicity, necrosis or apoptosis, without a
significant amount of cell death in normal cells, antitumor
activity in animals with a therapeutic index of at least 2. As used
herein, "therapeutic index" is the maximum tolerated dose divided
by the efficacious dose. The present invention also relates to a
method used to identify suitable candidates for treating or
preventing cancer.
[0910] One skilled in the art may refer to general reference texts
for detailed descriptions of known techniques discussed herein or
equivalent techniques. These texts include Ausubel et al., Current
Protocols in Molecular Biology, John Wiley and Sons, Inc. (2005);
Sambrook et al., Molecular Cloning, A Laboratory Manual (3.sup.rd
edition), Cold Spring Harbor Press, Cold Spring Harbor, N.Y.
(2000); Coligan et al., Current Protocols in Immunology, John Wiley
& Sons, N.Y.; Enna et al., Current Protocols in Pharmacology,
John Wiley & Sons, N.Y.; Fingl et al., The Pharmacological
Basis of Therapeutics (1975), Remington's Pharmaceutical Sciences,
Mack Publishing Co., Easton, Pa., 18.sup.th edition (1990). These
texts can, of course, also be referred to in making or using an
aspect of the invention.
[0911] As used herein, "combination therapy" or "co-therapy"
includes the administration of a compound of the present invention,
or a pharmaceutically acceptable salt, prodrug, metabolite,
polymorph or solvate thereof, and at least a second agent as part
of a specific treatment regimen intended to provide the beneficial
effect from the co-action of these therapeutic agents. The
beneficial effect of the combination may include, but is not
limited to, pharmacokinetic or pharmacodynamic co-action resulting
from the combination of therapeutic agents. Administration of these
therapeutic agents in combination typically is carried out over a
defined time period (usually minutes, hours, days or weeks
depending upon the combination selected). "Combination therapy" may
be, but generally is not, intended to encompass the administration
of two or more of these therapeutic agents as part of separate
monotherapy regimens that incidentally and arbitrarily result in
the combinations of the present invention.
[0912] "Combination therapy" is intended to embrace administration
of these therapeutic agents in a sequential manner, wherein each
therapeutic agent is administered at a different time, as well as
administration of these therapeutic agents, or at least two of the
therapeutic agents, in a substantially simultaneous manner.
Substantially simultaneous administration can be accomplished, for
example, by administering to the subject a single capsule having a
fixed ratio of each therapeutic agent or in multiple, single
capsules for each of the therapeutic agents. Sequential or
substantially simultaneous administration of each therapeutic agent
can be effected by any appropriate route including, but not limited
to, oral routes, intravenous routes, intramuscular routes, and
direct absorption through mucous membrane tissues. The therapeutic
agents can be administered by the same route or by different
routes. For example, a first therapeutic agent of the combination
selected may be administered by intravenous injection while the
other therapeutic agents of the combination may be administered
orally. Alternatively, for example, all therapeutic agents may be
administered orally or all therapeutic agents may be administered
by intravenous injection. The sequence in which the therapeutic
agents are administered is not narrowly critical.
[0913] "Combination therapy" also embraces the administration of
the therapeutic agents as described above in further combination
with other biologically active ingredients and non-drug therapies
(e.g., surgery or radiation treatment). Where the combination
therapy further comprises a non-drug treatment, the non-drug
treatment may be conducted at any suitable time so long as a
beneficial effect from the co-action of the combination of the
therapeutic agents and non-drug treatment is achieved. For example,
in appropriate cases, the beneficial effect is still achieved when
the non-drug treatment is temporally removed from the
administration of the therapeutic agents, perhaps by days or even
weeks.
[0914] A compound of the present invention, or a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof,
may be administered in combination with a second chemotherapeutic
agent. The second chemotherapeutic agent (also referred to as an
anti-neoplastic agent or anti-proliferative agent) can be an
alkylating agent; an antibiotic; an anti-metabolite; a detoxifying
agent; an interferon; a polyclonal or monoclonal antibody; an EGFR
inhibitor; a HER2 inhibitor; a histone deacetylase inhibitor; a
hormone; a mitotic inhibitor; an MTOR inhibitor; a multi-kinase
inhibitor; a serine/threonine kinase inhibitor; a tyrosine kinase
inhibitors; a VEGF/VEGFR inhibitor; a taxane or taxane derivative,
an aromatase inhibitor, an anthracycline, a microtubule targeting
drug, a topoisomerase poison drug, an inhibitor of a molecular
target or enzyme (e.g., a kinase or a protein methyltransferase), a
cytidine analogue drug or any chemotherapeutic, anti-neoplastic or
anti-proliferative agent listed in
www.cancer.org/docroot/cdg/cdg_0.asp.
[0915] Exemplary alkylating agents include, but are not limited to,
cyclophosphamide (Cytoxan; Neosar); chlorambucil (Leukeran);
melphalan (Alkeran); carmustine (BiCNU); busulfan (Busulfex);
lomustine (CeeNU); dacarbazine (DTIC-Dome); oxaliplatin (Eloxatin);
carmustine (Gliadel); ifosfamide (Ifex); mechlorethamine
(Mustargen); busulfan (Myleran); carboplatin (Paraplatin);
cisplatin (CDDP; Platinol); temozolomide (Temodar); thiotepa
(Thioplex); bendamustine (Treanda); or streptozocin (Zanosar).
[0916] Exemplary antibiotics include, but are not limited to,
doxorubicin (Adriamycin); doxorubicin liposomal (Doxil);
mitoxantrone (Novantrone); bleomycin (Blenoxane); daunorubicin
(Cerubidine); daunorubicin liposomal (DaunoXome); dactinomycin
(Cosmegen); epirubicin (Ellence); idarubicin (Idamycin); plicamycin
(Mithracin); mitomycin (Mutamycin); pentostatin (Nipent); or
valrubicin (Valstar).
[0917] Exemplary anti-metabolites include, but are not limited to,
fluorouracil (Adrucil); capecitabine (Xeloda); hydroxyurea
(Hydrea); mercaptopurine (Purinethol); pemetrexed (Alimta);
fludarabine (Fludara); nelarabine (Arranon); cladribine (Cladribine
Novaplus); clofarabine (Clolar); cytarabine (Cytosar-U); decitabine
(Dacogen); cytarabine liposomal (DepoCyt); hydroxyurea (Droxia);
pralatrexate (Folotyn); floxuridine (FUDR); gemcitabine (Gemzar);
cladribine (Leustatin); fludarabine (Oforta); methotrexate (MTX;
Rheumatrex); methotrexate (Trexall); thioguanine (Tabloid); TS-1 or
cytarabine (Tarabine PFS).
[0918] Exemplary detoxifying agents include, but are not limited
to, amifostine (Ethyol) or mesna (Mesnex).
[0919] Exemplary interferons include, but are not limited to,
interferon alfa-2b (Intron A) or interferon alfa-2a
(Roferon-A).
[0920] Exemplary polyclonal or monoclonal antibodies include, but
are not limited to, trastuzumab (Herceptin); ofatumumab (Arzerra);
bevacizumab (Avastin); rituximab (Rituxan); cetuximab (Erbitux);
panitumumab (Vectibix); tositumomaModine131 tositumomab (Bexxar);
alemtuzumab (Campath); ibritumomab (Zevalin; In-111; Y-90 Zevalin);
gemtuzumab (Mylotarg); eculizumab (Soliris) ordenosumab.
[0921] Exemplary EGFR inhibitors include, but are not limited to,
gefitinib (Iressa); lapatinib (Tykerb); cetuximab (Erbitux);
erlotinib (Tarceva); panitumumab (Vectibix); PKI-166; canertinib
(CI-1033); matuzumab (Emd7200) or EKB-569.
[0922] Exemplary HER2 inhibitors include, but are not limited to,
trastuzumab (Herceptin); lapatinib (Tykerb) or AC-480.
[0923] Histone Deacetylase Inhibitors include, but are not limited
to, vorinostat (Zolinza).
[0924] Exemplary hormones include, but are not limited to,
tamoxifen (Soltamox; Nolvadex); raloxifene (Evista); megestrol
(Megace); leuprolide (Lupron; Lupron Depot; Eligard; Viadur);
fulvestrant (Faslodex); letrozole (Femara); triptorelin (Trelstar
LA; Trelstar Depot); exemestane (Aromasin); goserelin (Zoladex);
bicalutamide (Casodex); anastrozole (Arimidex); fluoxymesterone
(Androxy; Halotestin); medroxyprogesterone (Provera; Depo-Provera);
estramustine (Emcyt); flutamide (Eulexin); toremifene (Fareston);
degarelix (Firmagon); nilutamide (Nilandron); abarelix (Plenaxis);
or testolactone (Teslac).
[0925] Exemplary mitotic inhibitors include, but are not limited
to, paclitaxel (Taxol; Onxol; Abraxane); docetaxel (Taxotere);
vincristine (Oncovin; Vincasar PFS); vinblastine (Velban);
etoposide (Toposar; Etopophos; VePesid); teniposide (Vumon);
ixabepilone (Ixempra); nocodazole; epothilone; vinorelbine
(Navelbine); camptothecin (CPT); irinotecan (Camptosar); topotecan
(Hycamtin); amsacrine or lamellarin D (LAM-D).
[0926] Exemplary MTOR inhibitors include, but are not limited to,
everolimus (Afinitor) or temsirolimus (Torisel); rapamune,
ridaforolimus; or AP23573.
[0927] Exemplary multi-kinase inhibitors include, but are not
limited to, sorafenib (Nexavar); sunitinib (Sutent); BIBW 2992;
E7080; Zd6474; PKC-412; motesanib; or AP24534.
[0928] Exemplary serine/threonine kinase inhibitors include, but
are not limited to, ruboxistaurin; eril/easudil hydrochloride;
flavopiridol; seliciclib (CYC202; Roscovitrine); SNS-032
(BMS-387032); Pkc412; bryostatin; KAI-9803; SF1126; VX-680;
Azd1152; Arry-142886 (AZD-6244); SCIO-469; GW681323; CC-401;
CEP-1347 or PD 332991.
[0929] Exemplary tyrosine kinase inhibitors include, but are not
limited to, erlotinib (Tarceva); gefitinib (Iressa); imatinib
(Gleevec); sorafenib (Nexavar); sunitinib (Sutent); trastuzumab
(Herceptin); bevacizumab (Avastin); rituximab (Rituxan); lapatinib
(Tykerb); cetuximab (Erbitux); panitumumab (Vectibix); everolimus
(Afinitor); alemtuzumab (Campath); gemtuzumab (Mylotarg);
temsirolimus (Torisel); pazopanib (Votrient); dasatinib (Sprycel);
nilotinib (Tasigna); vatalanib (Ptk787; ZK222584); CEP-701; SU5614;
MLN518; XL999; VX-322; Azd0530; BMS-354825; SKI-606 CP-690; AG-490;
WHI-P154; WHI-P131; AC-220; or AMG888.
[0930] Exemplary VEGF/VEGFR inhibitors include, but are not limited
to, bevacizumab (Avastin); sorafenib (Nexavar); sunitinib (Sutent);
ranibizumab; pegaptanib; or vandetinib.
[0931] Exemplary microtubule targeting drugs include, but are not
limited to, paclitaxel, docetaxel, vincristin, vinblastin,
nocodazole, epothilones and navelbine.
[0932] Exemplary topoisomerase poison drugs include, but are not
limited to, teniposide, etoposide, adriamycin, camptothecin,
daunorubicin, dactinomycin, mitoxantrone, amsacrine, epirubicin and
idarubicin.
[0933] Exemplary taxanes or taxane derivatives include, but are not
limited to, paclitaxel and docetaxol.
[0934] Exemplary general chemotherapeutic, anti-neoplastic,
anti-proliferative agents include, but are not limited to,
altretamine (Hexalen); isotretinoin (Accutane; Amnesteem; Claravis;
Sotret); tretinoin (Vesanoid); azacitidine (Vidaza); bortezomib
(Velcade) asparaginase (Elspar); levamisole (Ergamisol); mitotane
(Lysodren); procarbazine (Matulane); pegaspargase (Oncaspar);
denileukin diftitox (Ontak); porfimer (Photofrin); aldesleukin
(Proleukin); lenalidomide (Revlimid); bexarotene (Targretin);
thalidomide (Thalomid); temsirolimus (Torisel); arsenic trioxide
(Trisenox); verteporfin (Visudyne); mimosine (Leucenol); (1M
tegafur-0.4 M 5-chloro-2,4-dihydroxypyrimidine-1 M potassium
oxonate) or lovastatin.
[0935] In another aspect, the second chemotherapeutic agent can be
a cytokine such as G-CSF (granulocyte colony stimulating factor).
In another aspect, a compound of the present invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or
derivative thereof, may be administered in combination with
radiation therapy. Radiation therapy can also be administered in
combination with a compound of the present invention and another
chemotherapeutic agent described herein as part of a multiple agent
therapy. In yet another aspect, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or derivative thereof, may be administered in
combination with standard chemotherapy combinations such as, but
not restricted to, CMF (cyclophosphamide, methotrexate and
5-fluorouracil), CAF (cyclophosphamide, adriamycin and
5-fluorouracil), AC (adriamycin and cyclophosphamide), FEC
(5-fluorouracil, epirubicin, and cyclophosphamide), ACT or ATC
(adriamycin, cyclophosphamide, and paclitaxel), rituximab, Xeloda
(capecitabine), Cisplatin (CDDP), Carboplatin, TS-1 (tegafur,
gimestat and otastat potassium at a molar ratio of 1:0.4:1),
Camptothecin-11 (CPT-11, Irinotecan or Camptosar.TM.), CHOP
(cyclophosphamide, hydroxydaunorubicin, oncovin, and prednisone or
prednisolone), R-CHOP (rituximab, cyclophosphamide,
hydroxydaunorubicin, oncovin, prednisone or prednisolone), or CMFP
(cyclophosphamide, methotrexate, 5-fluorouracil and
prednisone).
[0936] In preferred embodiments, a compound of the present
invention, or a pharmaceutically acceptable salt, prodrug,
metabolite, polymorph or solvate thereof, may be administered with
an inhibitor of an enzyme, such as a receptor or non-receptor
kinase. Receptor and non-receptor kinases are, for example,
tyrosine kinases or serine/threonine kinases. Kinase inhibitors
described herein are small molecules, polynucleic acids,
polypeptides, or antibodies.
[0937] Exemplary kinase inhibitors include, but are not limited to,
Bevacizumab (targets VEGF), BMW 2992 (targets EGFR and Erb2),
Cetuximab/Erbitux (targets Erb1), Imatinib/Gleevic (targets
Bcr-Abl), Trastuzumab (targets Erb2), Gefitinib/Iressa (targets
EGFR), Ranibizumab (targets VEGF), Pegaptanib (targets VEGF),
Erlotinib/Tarceva (targets Erb1), Nilotinib (targets Bcr-Abl),
Lapatinib (targets Erb1 and Erb2/Her2), GW-572016/lapatinib
ditosylate (targets HER2/Erb2), Panitumumab/Vectibix (targets
EGFR), Vandetinib (targets RET/VEGFR), E7080 (multiple targets
including RET and VEGFR), Herceptin (targets HER2/Erb2), PKI-166
(targets EGFR), Canertinib/CI-1033 (targets EGFR),
Sunitinib/SU-11464/Sutent (targets EGFR and FLT3),
Matuzumab/Emd7200 (targets EGFR), EKB-569 (targets EGFR), Zd6474
(targets EGFR and VEGFR), PKC-412 (targets VEGR and FLT3),
Vatalanib/Ptk787/ZK222584 (targets VEGR), CEP-701 (targets FLT3),
SU5614 (targets FLT3), MLN518 (targets FLT3), XL999 (targets FLT3),
VX-322 (targets FLT3), Azd0530 (targets SRC), BMS-354825 (targets
SRC), SKI-606 (targets SRC), CP-690 (targets JAK), AG-490 (targets
JAK), WHI-P154 (targets JAK), WHI-P131 (targets JAK),
sorafenib/Nexavar (targets RAF kinase, VEGFR-1, VEGFR-2, VEGFR-3,
PDGFR-.beta., KIT, FLT-3, and RET), Dasatinib/Sprycel (BCR/ABL and
Src), AC-220 (targets Flt3), AC-480 (targets all HER proteins,
"panHER"), Motesanib diphosphate (targets VEGF1-3, PDGFR, and
c-kit), Denosumab (targets RANKL, inhibits SRC), AMG888 (targets
HER3), and AP24534 (multiple targets including Flt3).
[0938] Exemplary serine/threonine kinase inhibitors include, but
are not limited to, Rapamune (targets mTOR/FRAP1), Deforolimus
(targets mTOR), Certican/Everolimus (targets mTOR/FRAP1), AP23573
(targets mTOR/FRAP1), Eril/Fasudil hydrochloride (targets RHO),
Flavopiridol (targets CDK), Seliciclib/CYC202/Roscovitrine (targets
CDK), SNS-032/BMS-387032 (targets CDK), Ruboxistaurin (targets
PKC), Pkc412 (targets PKC), Bryostatin (targets PKC), KAI-9803
(targets PKC), SF1126 (targets PI3K), VX-680 (targets Aurora
kinase), Azd1152 (targets Aurora kinase), Arry-142886/AZD-6244
(targets MAP/MEK), SCIO-469 (targets MAP/MEK), GW681323 (targets
MAP/MEK), CC-401 (targets JNK), CEP-1347 (targets JNK), and PD
332991 (targets CDK).
[0939] The disorder in which EZH2-mediated protein methylation
plays a part can be a neurological disease. The compounds of this
invention may thus also be used for treating or studying neurologic
diseases such as epilepsy, schizophrenia, bipolar disorder or other
psychological and/or psychiatric disorders, neuropathies, skeletal
muscle atrophy, and neurodegenerative diseases, e.g., a
neurodegenerative disease. Exemplary neurodegenerative diseases
include: Alzheimer's, Amyotrophic Lateral Sclerosis (ALS), and
Parkinson's disease. Another class of neurodegenerative diseases
includes diseases caused at least in part by aggregation of
poly-glutamine. Diseases of this class include: Huntington's
Diseases, Spinalbulbar Muscular Atrophy (SBMA or Kennedy's Disease)
Dentatorubropallidoluysian Atrophy (DRPLA), Spinocerebellar Ataxia
1 (SCA1), Spinocerebellar Ataxia 2 (SCA2), Machado-Joseph Disease
(MJD; SCA3), Spinocerebellar Ataxia 6 (SCA6), Spinocerebellar
Ataxia 7 (SCAT), and Spinocerebellar Ataxia 12 (SCA12).
[0940] Any other disease in which epigenetic methylation, which is
mediated by EZH2, plays a role may be treatable or preventable
using compounds and methods described herein, or such diseases and
potential treatments thereof may be studied with the compounds
described herein.
4. PHARMACEUTICAL COMPOSITIONS
[0941] The present invention also provides pharmaceutical
compositions comprising a compound of any of the Formulae described
herein in combination with at least one pharmaceutically acceptable
excipient or carrier.
[0942] A "pharmaceutical composition" is a formulation containing
the compounds of the present invention in a form suitable for
administration to a subject. In one embodiment, the pharmaceutical
composition is in bulk or in unit dosage form. The unit dosage form
is any of a variety of forms, including, for example, a capsule, an
IV bag, a tablet, a single pump on an aerosol inhaler or a vial.
The quantity of active ingredient (e.g., a formulation of the
disclosed compound or salt, hydrate, solvate or isomer thereof) in
a unit dose of composition is an effective amount and is varied
according to the particular treatment involved. One skilled in the
art will appreciate that it is sometimes necessary to make routine
variations to the dosage depending on the age and condition of the
patient. The dosage will also depend on the route of
administration. A variety of routes are contemplated, including
oral, pulmonary, rectal, parenteral, transdermal, subcutaneous,
intravenous, intramuscular, intraperitoneal, inhalational, buccal,
sublingual, intrapleural, intrathecal, intranasal, and the like.
Dosage forms for the topical or transdermal administration of a
compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants. In
one embodiment, the active compound is mixed under sterile
conditions with a pharmaceutically acceptable carrier, and with any
preservatives, buffers, or propellants that are required.
[0943] As used herein, the phrase "pharmaceutically acceptable"
refers to those compounds, anions, cations, materials,
compositions, carriers, and/or dosage forms which are, within the
scope of sound medical judgment, suitable for use in contact with
the tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other problem or complication,
commensurate with a reasonable benefit/risk ratio.
[0944] "Pharmaceutically acceptable excipient" means an excipient
that is useful in preparing a pharmaceutical composition that is
generally safe, non-toxic and neither biologically nor otherwise
undesirable, and includes excipient that is acceptable for
veterinary use as well as human pharmaceutical use. A
"pharmaceutically acceptable excipient" as used in the
specification and claims includes both one and more than one such
excipient.
[0945] A pharmaceutical composition of the invention is formulated
to be compatible with its intended route of administration.
Examples of routes of administration include parenteral, e.g.,
intravenous, intradermal, subcutaneous, oral (e.g., inhalation),
transdermal (topical), and transmucosal administration. Solutions
or suspensions used for parenteral, intradermal, or subcutaneous
application can include the following components: a sterile diluent
such as water for injection, saline solution, fixed oils,
polyethylene glycols, glycerine, propylene glycol or other
synthetic solvents; antibacterial agents such as benzyl alcohol or
methyl parabens; antioxidants such as ascorbic acid or sodium
bisulfite; chelating agents such as ethylenediaminetetraacetic
acid; buffers such as acetates, citrates or phosphates, and agents
for the adjustment of tonicity such as sodium chloride or dextrose.
The pH can be adjusted with acids or bases, such as hydrochloric
acid or sodium hydroxide. The parenteral preparation can be
enclosed in ampoules, disposable syringes or multiple dose vials
made of glass or plastic.
[0946] A compound or pharmaceutical composition of the invention
can be administered to a subject in many of the well-known methods
currently used for chemotherapeutic treatment. For example, for
treatment of cancers, a compound of the invention may be injected
directly into tumors, injected into the blood stream or body
cavities or taken orally or applied through the skin with patches.
The dose chosen should be sufficient to constitute effective
treatment but not so high as to cause unacceptable side effects.
The state of the disease condition (e.g., cancer, precancer, and
the like) and the health of the patient should preferably be
closely monitored during and for a reasonable period after
treatment.
[0947] The term "therapeutically effective amount", as used herein,
refers to an amount of a pharmaceutical agent to treat, ameliorate,
or prevent an identified disease or condition, or to exhibit a
detectable therapeutic or inhibitory effect. The effect can be
detected by any assay method known in the art. The precise
effective amount for a subject will depend upon the subject's body
weight, size, and health; the nature and extent of the condition;
and the therapeutic or combination of therapeutics selected for
administration. Therapeutically effective amounts for a given
situation can be determined by routine experimentation that is
within the skill and judgment of the clinician. In a preferred
aspect, the disease or condition to be treated is cancer. In
another aspect, the disease or condition to be treated is a cell
proliferative disorder.
[0948] For any compound, the therapeutically effective amount can
be estimated initially either in cell culture assays, e.g., of
neoplastic cells, or in animal models, usually rats, mice, rabbits,
dogs, or pigs. The animal model may also be used to determine the
appropriate concentration range and route of administration. Such
information can then be used to determine useful doses and routes
for administration in humans. Therapeutic/prophylactic efficacy and
toxicity may be determined by standard pharmaceutical procedures in
cell cultures or experimental animals, e.g., ED.sub.50 (the dose
therapeutically effective in 50% of the population) and LD.sub.50
(the dose lethal to 50% of the population). The dose ratio between
toxic and therapeutic effects is the therapeutic index, and it can
be expressed as the ratio, LD.sub.50/ED.sub.50. Pharmaceutical
compositions that exhibit large therapeutic indices are preferred.
The dosage may vary within this range depending upon the dosage
form employed, sensitivity of the patient, and the route of
administration.
[0949] Dosage and administration are adjusted to provide sufficient
levels of the active agent(s) or to maintain the desired effect.
Factors which may be taken into account include the severity of the
disease state, general health of the subject, age, weight, and
gender of the subject, diet, time and frequency of administration,
drug combination(s), reaction sensitivities, and tolerance/response
to therapy. Long-acting pharmaceutical compositions may be
administered every 3 to 4 days, every week, or once every two weeks
depending on half-life and clearance rate of the particular
formulation.
[0950] The pharmaceutical compositions containing active compounds
of the present invention may be manufactured in a manner that is
generally known, e.g., by means of conventional mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping, or lyophilizing processes. Pharmaceutical compositions
may be formulated in a conventional manner using one or more
pharmaceutically acceptable carriers comprising excipients and/or
auxiliaries that facilitate processing of the active compounds into
preparations that can be used pharmaceutically. Of course, the
appropriate formulation is dependent upon the route of
administration chosen.
[0951] Pharmaceutical compositions suitable for injectable use
include sterile aqueous solutions (where water soluble) or
dispersions and sterile powders for the extemporaneous preparation
of sterile injectable solutions or dispersion. For intravenous
administration, suitable carriers include physiological saline,
bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or
phosphate buffered saline (PBS). In all cases, the composition must
be sterile and should be fluid to the extent that easy
syringeability exists. It must be stable under the conditions of
manufacture and storage and must be preserved against the
contaminating action of microorganisms such as bacteria and fungi.
The carrier can be a solvent or dispersion medium containing, for
example, water, ethanol, polyol (for example, glycerol, propylene
glycol, and liquid polyethylene glycol, and the like), and suitable
mixtures thereof. The proper fluidity can be maintained, for
example, by the use of a coating such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. Prevention of the action of
microorganisms can be achieved by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
ascorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include isotonic agents, for example, sugars,
polyalcohols such as manitol and sorbitol, and sodium chloride in
the composition. Prolonged absorption of the injectable
compositions can be brought about by including in the composition
an agent which delays absorption, for example, aluminum
monostearate and gelatin.
[0952] Sterile injectable solutions can be prepared by
incorporating the active compound in the required amount in an
appropriate solvent with one or a combination of ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the active
compound into a sterile vehicle that contains a basic dispersion
medium and the required other ingredients from those enumerated
above. In the case of sterile powders for the preparation of
sterile injectable solutions, methods of preparation are vacuum
drying and freeze-drying that yields a powder of the active
ingredient plus any additional desired ingredient from a previously
sterile-filtered solution thereof.
[0953] Oral compositions generally include an inert diluent or an
edible pharmaceutically acceptable carrier. They can be enclosed in
gelatin capsules or compressed into tablets. For the purpose of
oral therapeutic administration, the active compound can be
incorporated with excipients and used in the form of tablets,
troches, or capsules. Oral compositions can also be prepared using
a fluid carrier for use as a mouthwash, wherein the compound in the
fluid carrier is applied orally and swished and expectorated or
swallowed. Pharmaceutically compatible binding agents, and/or
adjuvant materials can be included as part of the composition. The
tablets, pills, capsules, troches and the like can contain any of
the following ingredients, or compounds of a similar nature: a
binder such as microcrystalline cellulose, gum tragacanth or
gelatin; an excipient such as starch or lactose, a disintegrating
agent such as alginic acid, Primogel, or corn starch; a lubricant
such as magnesium stearate or Sterotes; a glidant such as colloidal
silicon dioxide; a sweetening agent such as sucrose or saccharin;
or a flavoring agent such as peppermint, methyl salicylate, or
orange flavoring.
[0954] For administration by inhalation, the compounds are
delivered in the form of an aerosol spray from pressured container
or dispenser, which contains a suitable propellant, e.g., a gas
such as carbon dioxide, or a nebulizer.
[0955] Systemic administration can also be by transmucosal or
transdermal means. For transmucosal or transdermal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the art,
and include, for example, for transmucosal administration,
detergents, bile salts, and fusidic acid derivatives. Transmucosal
administration can be accomplished through the use of nasal sprays
or suppositories. For transdermal administration, the active
compounds are formulated into ointments, salves, gels, or creams as
generally known in the art.
[0956] The active compounds can be prepared with pharmaceutically
acceptable carriers that will protect the compound against rapid
elimination from the body, such as a controlled release
formulation, including implants and microencapsulated delivery
systems. Biodegradable, biocompatible polymers can be used, such as
ethylene vinyl acetate, polyanhydrides, polyglycolic acid,
collagen, polyorthoesters, and polylactic acid. Methods for
preparation of such formulations will be apparent to those skilled
in the art. The materials can also be obtained commercially from
Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal
suspensions (including liposomes targeted to infected cells with
monoclonal antibodies to viral antigens) can also be used as
pharmaceutically acceptable carriers. These can be prepared
according to methods known to those skilled in the art, for
example, as described in U.S. Pat. No. 4,522,811.
[0957] It is especially advantageous to formulate oral or
parenteral compositions in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit form as used
herein refers to physically discrete units suited as unitary
dosages for the subject to be treated; each unit containing a
predetermined quantity of active compound calculated to produce the
desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
of the invention are dictated by and directly dependent on the
unique characteristics of the active compound and the particular
therapeutic effect to be achieved.
[0958] In therapeutic applications, the dosages of the
pharmaceutical compositions used in accordance with the invention
vary depending on the agent, the age, weight, and clinical
condition of the recipient patient, and the experience and judgment
of the clinician or practitioner administering the therapy, among
other factors affecting the selected dosage. Generally, the dose
should be sufficient to result in slowing, and preferably
regressing, the growth of the tumors and also preferably causing
complete regression of the cancer. Dosages can range from about
0.01 mg/kg per day to about 5000 mg/kg per day. In preferred
aspects, dosages can range from about 1 mg/kg per day to about 1000
mg/kg per day. In an aspect, the dose will be in the range of about
0.1 mg/day to about 50 g/day; about 0.1 mg/day to about 25 g/day;
about 0.1 mg/day to about 10 g/day; about 0.1 mg to about 3 g/day;
or about 0.1 mg to about 1 g/day, in single, divided, or continuous
doses (which dose may be adjusted for the patient's weight in kg,
body surface area in m.sup.2, and age in years). An effective
amount of a pharmaceutical agent is that which provides an
objectively identifiable improvement as noted by the clinician or
other qualified observer. For example, regression of a tumor in a
patient may be measured with reference to the diameter of a tumor.
Decrease in the diameter of a tumor indicates regression.
Regression is also indicated by failure of tumors to reoccur after
treatment has stopped. As used herein, the term "dosage effective
manner" refers to amount of an active compound to produce the
desired biological effect in a subject or cell.
[0959] The pharmaceutical compositions can be included in a
container, pack, or dispenser together with instructions for
administration.
[0960] The compounds of the present invention are capable of
further forming salts. All of these forms are also contemplated
within the scope of the claimed invention.
[0961] As used herein, "pharmaceutically acceptable salts" refer to
derivatives of the compounds of the present invention wherein the
parent compound is modified by making acid or base salts thereof.
Examples of pharmaceutically acceptable salts include, but are not
limited to, mineral or organic acid salts of basic residues such as
amines, alkali or organic salts of acidic residues such as
carboxylic acids, and the like. The pharmaceutically acceptable
salts include the conventional non-toxic salts or the quaternary
ammonium salts of the parent compound formed, for example, from
non-toxic inorganic or organic acids. For example, such
conventional non-toxic salts include, but are not limited to, those
derived from inorganic and organic acids selected from
2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic,
benzene sulfonic, benzoic, bicarbonic, carbonic, citric, edetic,
ethane disulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic,
gluconic, glutamic, glycolic, glycollyarsanilic, hexylresorcinic,
hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxymaleic,
hydroxynaphthoic, isethionic, lactic, lactobionic, lauryl sulfonic,
maleic, malic, mandelic, methane sulfonic, napsylic, nitric,
oxalic, pamoic, pantothenic, phenylacetic, phosphoric,
polygalacturonic, propionic, salicyclic, stearic, subacetic,
succinic, sulfamic, sulfanilic, sulfuric, tannic, tartaric, toluene
sulfonic, and the commonly occurring amine acids, e.g., glycine,
alanine, phenylalanine, arginine, etc.
[0962] Other examples of pharmaceutically acceptable salts include
hexanoic acid, cyclopentane propionic acid, pyruvic acid, malonic
acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid,
3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic
acid, muconic acid, and the like. The present invention also
encompasses salts formed when an acidic proton present in the
parent compound either is replaced by a metal ion, e.g., an alkali
metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, tromethamine, N-methylglucamine,
and the like. In the salt form, it is understood that the ratio of
the compound to the cation or anion of the salt can be 1:1, or any
ration other than 1:1, e.g., 3:1, 2:1, 1:2, or 1:3.
[0963] It should be understood that all references to
pharmaceutically acceptable salts include solvent addition forms
(solvates) or crystal forms (polymorphs) as defined herein, of the
same salt.
[0964] The compounds of the present invention can also be prepared
as esters, for example, pharmaceutically acceptable esters. For
example, a carboxylic acid function group in a compound can be
converted to its corresponding ester, e.g., a methyl, ethyl or
other ester. Also, an alcohol group in a compound can be converted
to its corresponding ester, e.g., acetate, propionate or other
ester.
[0965] The compounds of the present invention can also be prepared
as prodrugs, for example, pharmaceutically acceptable prodrugs. The
terms "pro-drug" and "prodrug" are used interchangeably herein and
refer to any compound which releases an active parent drug in vivo.
Since prodrugs are known to enhance numerous desirable qualities of
pharmaceuticals (e.g., solubility, bioavailability, manufacturing,
etc.), the compounds of the present invention can be delivered in
prodrug form. Thus, the present invention is intended to cover
prodrugs of the presently claimed compounds, methods of delivering
the same and compositions containing the same. "Prodrugs" are
intended to include any covalently bonded carriers that release an
active parent drug of the present invention in vivo when such
prodrug is administered to a subject. Prodrugs in the present
invention are prepared by modifying functional groups present in
the compound in such a way that the modifications are cleaved,
either in routine manipulation or in vivo, to the parent compound.
Prodrugs include compounds of the present invention wherein a
hydroxy, amino, sulfhydryl, carboxy or carbonyl group is bonded to
any group that may be cleaved in vivo to form a free hydroxyl, free
amino, free sulfhydryl, free carboxy or free carbonyl group,
respectively.
[0966] Examples of prodrugs include, but are not limited to, esters
(e.g., acetate, dialkylaminoacetates, formates, phosphates,
sulfates and benzoate derivatives) and carbamates (e.g.,
N,N-dimethylaminocarbonyl) of hydroxy functional groups, esters
(e.g., ethyl esters, morpholinoethanol esters) of carboxyl
functional groups, N-acyl derivatives (e.g., N-acetyl)N-Mannich
bases, Schiff bases and enaminones of amino functional groups,
oximes, acetals, ketals and enol esters of ketone and aldehyde
functional groups in compounds of the invention, and the like, See
Bundegaard, H., Design of Prodrugs, p1-92, Elesevier, New
York-Oxford (1985).
[0967] The compounds, or pharmaceutically acceptable salts, esters
or prodrugs thereof, are administered orally, nasally,
transdermally, pulmonary, inhalationally, buccally, sublingually,
intraperintoneally, subcutaneously, intramuscularly, intravenously,
rectally, intrapleurally, intrathecally and parenterally. In one
embodiment, the compound is administered orally. One skilled in the
art will recognize the advantages of certain routes of
administration.
[0968] The dosage regimen utilizing the compounds is selected in
accordance with a variety of factors including type, species, age,
weight, sex and medical condition of the patient; the severity of
the condition to be treated; the route of administration; the renal
and hepatic function of the patient; and the particular compound or
salt thereof employed. An ordinarily skilled physician or
veterinarian can readily determine and prescribe the effective
amount of the drug required to prevent, counter, or arrest the
progress of the condition.
[0969] Techniques for formulation and administration of the
disclosed compounds of the invention can be found in Remington: the
Science and Practice of Pharmacy, 19.sup.th edition, Mack
Publishing Co., Easton, Pa. (1995). In an embodiment, the compounds
described herein, and the pharmaceutically acceptable salts
thereof, are used in pharmaceutical preparations in combination
with a pharmaceutically acceptable carrier or diluent. Suitable
pharmaceutically acceptable carriers include inert solid fillers or
diluents and sterile aqueous or organic solutions. The compounds
will be present in such pharmaceutical compositions in amounts
sufficient to provide the desired dosage amount in the range
described herein.
[0970] All percentages and ratios used herein, unless otherwise
indicated, are by weight. Other features and advantages of the
present invention are apparent from the different examples. The
provided examples illustrate different components and methodology
useful in practicing the present invention. The examples do not
limit the claimed invention. Based on the present disclosure the
skilled artisan can identify and employ other components and
methodology useful for practicing the present invention.
[0971] In the synthetic schemes described herein, compounds may be
drawn with one particular configuration for simplicity. Such
particular configurations are not to be construed as limiting the
invention to one or another isomer, tautomer, regioisomer or
stereoisomer, nor does it exclude mixtures of isomers, tautomers,
regioisomers or stereoisomers; however, it will be understood that
a given isomer, tautomer, regioisomer or stereoisomer may have a
higher level of activity than another isomer, tautomer, regioisomer
or stereoisomer.
[0972] Compounds designed, selected and/or optimized by methods
described above, once produced, can be characterized using a
variety of assays known to those skilled in the art to determine
whether the compounds have biological activity. For example, the
molecules can be characterized by conventional assays, including
but not limited to those assays described below, to determine
whether they have a predicted activity, binding activity and/or
binding specificity.
[0973] Furthermore, high-throughput screening can be used to speed
up analysis using such assays. As a result, it can be possible to
rapidly screen the molecules described herein for activity, using
techniques known in the art. General methodologies for performing
high-throughput screening are described, for example, in Devlin
(1998) High Throughput Screening, Marcel Dekker; and U.S. Pat. No.
5,763,263. High-throughput assays can use one or more different
assay techniques including, but not limited to, those described
below.
[0974] All publications and patent documents cited herein are
incorporated herein by reference as if each such publication or
document was specifically and individually indicated to be
incorporated herein by reference. Citation of publications and
patent documents is not intended as an admission that any is
pertinent prior art, nor does it constitute any admission as to the
contents or date of the same. The invention having now been
described by way of written description, those of skill in the art
will recognize that the invention can be practiced in a variety of
embodiments and that the foregoing description and examples below
are for purposes of illustration and not limitation of the claims
that follow.
5. EXAMPLES
General Experimental
NMR
[0975] .sup.1H-NMR spectra were taken using CDCl.sub.3 unless
otherwise stated and were recorded at 400 or 500 MHz using a Varian
or Oxford instruments magnet (500 MHz) instruments. Multiplicities
indicated are s=singlet, d=doublet, t=triplet, q=quartet,
quint=quintet, sxt=sextet, m=multiplet, dd=doublet of doublets,
dt=doublet of triplets; br indicates a broad signal.
LCMS and HPLC
[0976] Shimadzu LC-Q, Shimadzu LCMS-2010EV or Waters Acquity Ultra
Performance LC. HPLC: Products were analyzed by Shimadzu SPD-20A
with 150.times.4.5 mm YMC ODS-M80 column or 150.times.4.6 mm
YMC-Pack Pro C18 column at 1.0 ml/min.
[0977] Mobile phase was MeCN:H2O=3:2 (containing 0.3% SDS and 0.05%
H.sub.3PO.sub.4),
[0978] 0.05% TFA in water, 0.05% TFA in acetonitrile (gradient
Initial 20%, then 0.05% TFA/MeCN to conc. to 95% in 3 min. holds
for 0.5 min. at 3.51 to 4.50 min then 0.05% TFA/MeCN conc.
20%).
[0979] Alternatively the LCMS, 2 different methods were used; the
one we use the most is the high pH (METCR1600) and the other one
for more standard compounds (METCR1416).
[0980] 0.1% Formic acid in water--Mobile phase "A" 0.1% Formic acid
in acetonitrile--Mobile phase "B" utilizing Waters Atlantis dC18,
2.1 mm.times.100 mm, 3 .mu.m column, with a flow rate=0.6 ml/min
Column temperature=40.degree. C.; Time (mins) % B 0.00 min 5% B.
5.0 mins 100% B, 5.4 mins 100% B and 0.42 mins 5% B
[0981] 3.5 minute method refers to Atlantis dC18, 2.1 mm.times.50
mm, 3 .mu.m column, flow rate of 1 ml/min at 40 C. Mobile phase A
Formic acid (aq.) 0.1% mobile phase B formic acid (MeCN) 0.1%,
injection 3 .mu.L, gradient 0 mins (5% organic), 2.5 min (100%
organic), 2.7 mins (100% organic), 2.71 min (5% organic), 3.5 min
(5% organic)
[0982] 7.0 minute method refers to Atlantis dC18, 2.1 mm.times.100
mm, 3 .mu.m column, flow rate of 0.6 ml/min at 40 C. Mobile phase A
Formic acid (aq.) 0.1% mobile phase B formic acid (MeCN) 0.1%,
injection 3 .mu.L, gradient 0 mins (5% organic), 5 min (100%
organic), 5.4 mins (100% organic), 5.42 min (5% organic), 7 min (5%
organic)
[0983] Both the 3, 5 and 7 minute methods were performed on a MS18
Shimadzu LCMS-2010EV or a MS19 Shimadzu LCMS-2010EV system
utilizing LC-20AB pumps and SPD-M20A PDA detectors.
[0984] Products were purified by HPLC/MS using Waters
AutoPurification System with 3100 Mass Detector.
[0985] HPLC analyses may also be performed on a Shimdazu LC-2010
CHT using an YMC ODS-A, C18, (150.times.4.6.times.5 .mu.m) column
at ambient temperature with a flow Rate of 1.4 ml/min. An injection
volume of 10 .mu.l is utilized and detection occurs via UV/PDA.
Mobile Phase A is 0.05% TFA in water and Mobile Phase B is 0.05 TFA
in acetonitrile with a gradient program of Initial 5% B to 95% B in
8 min, hold for 1.5 min, at 9.51 to 12 min B. conc. 0.5%. The
diluent is the mobile phase
Other
[0986] Automated flash column chromatography was performed on a
Biotage Isolera version 4. 10 g SNAP cartridge running at 12 ml/min
or a 25 g SNAP cartridge running at 25 ml/min and detecting at 254
nm and 280 nm.
[0987] Select Nitrile reductions may be performed on a ThalesNano
H-Cube.RTM. according to the conditions described in the
experimental procedure.
General Procedure for the Synthesis of the Pyridone Amines
##STR00447##
[0989] To a solution of the cyano compound (1 eq.) in MeOH, a
catalytic amount of Raney Nickel and ammonia solution were added.
The mixture was stirred at RT under hydrogen (balloon pressure) for
1 hr. On completion of the reaction, the mixture was filtered
through a celite bed washing with MeOH. The filtrate was
concentrated under reduced pressure to afford the desired amine.
This procedure is applicable to a wide variety of R groups
described within the examples. In general the cyano pyridines care
available from commercial vendors or can be synthesized via methods
known to those of ordinary skill in the art.
Syntheses of Starting Materials or Intermediates
3-(Aminomethyl)-6-methyl-4-propyl-1,2-dihydropyridin-2-one HCl
salt
##STR00448##
[0991] To a stirred solution of t-BuOK (20.0 g, 179 mmol) and
cyanoacetamide (16.5 g, 196 mmol) in DMSO (300 mL) was added
(3E)-3-hepten-2-one (20.0 g, 178 mmol). The reaction mixture was
stirred at 23.degree. C. for 30 minutes and then additional t-BuOK
(60.0 g, 712 mmol) was added to the reaction mixture. The reaction
mixture was placed under oxygen atmosphere and stirred for 16 h.
The reaction mixture was then purged with argon and was cooled to
0.degree. C. The mixture was diluted with aq. HCl and the resulting
precipitate was collected. The solid was washed with water and
dried to give
6-methyl-2-oxo-4-propyl-1,2-dihydropyridine-3-carbonitrile (15.0 g,
47% yield).
[0992] To a stirred solution of
6-methyl-2-oxo-4-propyl-1,2-dihydropyridine-3-carbonitrile (15.0 g,
85.1 mmol) in methanol (600 mL) and concentrated HCl (15 mL) was
added Pd(OH).sub.2 (15.0 g). The mixture was stirred for 48 hours
under H.sub.2 atmosphere. The reaction mixture was filtered and
filtrate was concentrated in vacuo. Ethanol was added to the
residue, the resulting precipitate was collected and dried to give
the titled compound as a white solid (13.0 g, 60% yield).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 11.86 (br. s., 1H),
6.00 (s, 1H), 3.78 (q, J=5.5 Hz, 2H), 3.61 (br. s., 2H), 2.46 (m,
2H), 2.17 (s, 3H), 1.50 (sxt, J=7.4 Hz, 2H), 0.91 (t, J=7.4 Hz,
3H).
6-Methyl-2-oxo-4-(propan-2-yl)-1,2-dihydropyridine-3-carbonitrile
##STR00449##
[0994] To a solution of 2-cyanoacetamide (35.1 g, 417 mmol) and
t-BuOK (42.5 g, 379 mmol) in DMSO (631 ml) was added
5-methyl-3-hexen-2-one (50.0 ml, 379 mmol) under N.sub.2
atmosphere. The mixture was stirred at 23.degree. C. for 30 min and
then additional t-BuOK (127 g, 1137 mmol) was added. The N.sub.2
gas was displaced by O.sub.2 gas and the mixture was stirred for 45
h at 23.degree. C. under oxygen. The mixture was cooled to
0.degree. C., diluted with H.sub.2O (200 ml) and HCl (5N, 227 ml,
slowly added). The mixture was stirred for 15 min at 0.degree. C.
and the solid was filtered with Buchner funnel. The solid was
washed with H.sub.2O (1500 ml) and dried with hot-air (55.degree.
C., 16 h) to give
6-methyl-2-oxo-4-(propan-2-yl)-1,2-dihydropyridine-3-carbonitrile
(26.6 g, 40% yield). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta.
6.14 ppm (s, 1H), 3.25-3.29 (m, 1H), 2.45 (s, 3H), 1.26 (d, J=6.8
Hz, 6H); LC-MS: m/z 177.1 [M+1], 198.9 [M+23].
3-(Aminomethyl)-6-methyl-4-(propan-2-yl)-1,2-dihydropyridin-2-one
HCl salt
##STR00450##
[0996] To a solution of
6-methyl-2-oxo-4-(propan-2-yl)-1,2-dihydropyridine-3-carbonitrile
(5.00 g, 28.4 mmol) and in MeOH (400 ml) and HCl (8.8 ml, 12 M) was
added 10% Pd(OH).sub.2 (5.17 g, 3.68 mmol) under N.sub.2
atmosphere. The N.sub.2 gas was displaced by H.sub.2 gas and the
mixture was stirred for 24 h at 23.degree. C. under hydrogen. The
H.sub.2 gas was displaced by N.sub.2 gas and the mixture was
filtered through celite, washed with MeOH and concentrated. The
residue was triturated with EtOH-TBME, the solid was collected with
Buchner funnel and dried in vacuo to give the titled compound (6.15
g, 100%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 11.9
(br-s, 1H), 8.03 (br-s, 2H), 6.12 (s, 1H), 3.82-3.84 (m, 2H),
3.08-3.12 (m, 1H), 2.19 (s, 3H), 1.12 (d, J=6.8 Hz, 6H).
6-Methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-3-carbonitrile
##STR00451##
[0998] To a solution of 2-cyanoacetamide (14.0 g, 166 mmol) and
trifluoroacetylacetone (20.0 ml, 166 mmol) in H.sub.2O (332 ml) was
added K.sub.2CO.sub.3 (6.60 g, 47.9 mmol). The mixture was stirred
at 23.degree. C. for 15 h. The precipitated solid was filtered with
Buchner funnel, washed with ice cold H.sub.2O, and dried with hot
air (60.degree. C., 16 h) to give the titled compound (17.6 g,
52%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 2.38 (s,
3H), 6.66 (s, 1H).
3-(Aminomethyl)-6-methyl-4-(trifluoromethyl)-1,2-dihydropyridin-2-one
hydrochloride HCl salt
##STR00452##
[1000] To a solution of
6-methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-3-carbonitrile
(400 mg, 1.98 mmol) in MeOH (19.8 ml) and HCl (436 ul, 12M) was
added 10% Pd(OH).sub.2 (361 mg, 0.257 mmol) under N.sub.2
atmosphere. The N.sub.2 gas was displaced by H.sub.2 gas and the
mixture was stirred for 18 h at 23.degree. C. under hydrogen. The
H.sub.2 gas was displaced by N.sub.2 gas. The mixture was filtered
through Celite, washed with MeOH and concentrated. The residue was
triturated with MeOH-Et.sub.2O, collected with Buchner funnel, and
dried in vacuo to give the titled compound (433 mg, 100%). .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. ppm 2.31 (s, 3H), 3.88 (s,
2H), 6.43 (s, 1H).
5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
##STR00453##
[1002] To a stirred solution of
6-methyl-2-oxo-4-(propan-2-yl)-1,2-dihydropyridine-3-carbonitrile
(225 mg, 1.277 mmol) in MeCN (6 mL) was added Selectfluor (620 mg,
1.75 mmol). The reaction mixture was stirred at 50.degree. C. for 3
h. After cooling to 23.degree. C., the reaction mixture was
concentrated in vacuo. The residue was purified by column
chromatography (50% to 100% EtOAc-heptane to obtain the titled
compound (90 mg, 36%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 3.39 (m, 1H), 2.44 (d, J=3.1 Hz, 3H), 1.41 (dd, J=7.0, 3.1 Hz,
6H); LCMS E-S (M+H)=195.2.
5-Fluoro-6-methyl-2-oxo-4-propyl-1,2-dihydropyridine-3-carbonitrile
##STR00454##
[1004] The title compound was prepared in the same manner as
described for
5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
(20% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 2.80
(dt, J=7.9, 1.9 Hz 2H), 2.31 (d, J=2.1 Hz, 3H), 1.73 (tq, J=7.4,
7.4 Hz, 2H), 1.06 (t, J=7.3 Hz, 3H); LCMS E-S (M+H)=195.2.
5-Fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
##STR00455##
[1006] The title compound was prepared in the same manner as
described for
5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
(15% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 2.46 (d,
J=3.1 Hz, 3H), 2.44 (d, J=7.4, 3.1 Hz, 3H); LCMS E-S
(M+H)=167.2.
3-(Aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one
##STR00456##
[1008]
5-Fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitr-
ile (100 mg, 0.515 mmol) in 100 mL flask was dissolved in a mixture
of MeOH (6 mL) and 2 mL NH.sub.3aq (2 mL, 25%). Reduction was
conducted using H-Cube with Raney-Ni as a catalyst at room
temperature for 3-4 h. On completion of reaction (monitored by
TLC), reaction was concentrated under reduced pressure to afford
titled compound as a grey solid (90 mg, 90%). .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. ppm 4.04 (s, 2H), 3.22 (m, 1H), 2.24 (d, J=3.4
Hz, 3H), 1.32 (dd, J=7.0, 1.8 Hz, 6H); LCMS E-S (M+H)=199.2.
3-(Aminomethyl)-5-fluoro-6-methyl-4-propylpyridin-2(1H)-one
##STR00457##
[1010] The titled compound was prepared in the same manner as
described for
3-(aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one (92%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.76 (s, 2H),
2.61 (dt, J=7.2, 1.8 Hz 2H), 2.31 (d, J=2.8 Hz, 3H), 1.57 (tq,
J=7.6, 7.6 Hz, 2H), 1.00 (t, J=7.3 Hz, 3H); LCMS E-S
(M+H)=199.2.
3-(Aminomethyl)-5-fluoro-4,6-dimethylpyridin-2(1H)-one
##STR00458##
[1012] The title compound was prepared in the same manner as
described for
3-(Aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one (92%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.79 (s, 2H),
2.29 (d, J=3.3 Hz, 3H), 2.24 (d, J=2.1 Hz, 3H); LCMS E-S
(M+H)=171.2.
2-Methyl-3-nitro-5-(trifluoromethyl)benzoic acid
##STR00459##
[1014] To a stirred solution of 2-methyl-5-(trifluoromethyl)benzoic
acid (2.00 g, 9.80 mmol) in H.sub.2SO.sub.4 (20 mL, 12 N) was added
HNO.sub.3 (2 mL, 12N) at 0.degree. C. After stirring for 3 hours,
water was added and resulting precipitate was collected. The solid
was washed with water and dried to give the titled compound as a
white solid (1.90 g, 76% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 8.43 (s, 1H), 8.15 (s, 1H), 2.77 (s, 3H); MS (ES)
[M-H] 247.8.
Methyl 2-methyl-3-nitro-5-(trifluoromethyl)benzoate
##STR00460##
[1016] To a solution of 2-methyl-3-nitro-5-(trifluoromethyl)benzoic
acid (21.0 g, 84.3 mmol) in DMF (200 mL) was added potassium
carbonate (35.0 g, 253 mmol) and methyl iodide (24.0 g, 169 mmol).
The mixture was stirred at 60.degree. C. and after stirring for 6 h
the reaction mixture was cooled to 23.degree. C. and was filtered.
The filtrate was concentrated in vacuo and the residue was
dissolved in EtOAc and water. The aqueous phase was extracted with
EtOAc (2.times.250 mL and the combined organic layers were
concentrated in vacuo. The residue was purified by column
chromatography (SiO.sub.2 heptane-EtOac=4/1) to give the titled
compound (20.0 g, 90% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. (ppm); 8.27 (s, 1H), 8.11 (s, 1H), 3.99 (s, 3H), 2.71 (s,
3H); MS (ES) [M-H] 261.9.
Methyl 3-amino-2-methyl-5-(trifluoromethyl)benzoate
##STR00461##
[1018] To a stirred solution of methyl
2-methyl-3-nitro-5-(trifluoromethyl)benzoate (20.0 g, 76.0 mmol) in
MeOH (400 ml) was added Pd(OH).sub.2 (2.00 g). The reaction mixture
was stirred for 3 hours under hydrogen atmosphere. After purging
with nitrogen, the mixture was filtered and the filtrate was
concentrated in vacuo to give the titled compound as an orange
solid (10.2 g, 58% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.47 (brs, 1H), 7.01 (d, J=2.0 Hz, 1H), 3.92 (m, 2H),
3.91 (s, 3H), 2.39 (s, 3H).
tert-Butyl-4-{[3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl]amin-
o}piperidine-1-carboxylate
##STR00462##
[1020] To a stirred solution of methyl
3-amino-2-methyl-5-(trifluoromethyl)benzoate (6.00 g, 25.7 mmol) in
CH.sub.2Cl.sub.2 (120 mL) and AcOH (6 mL) was added N-Boc
piperidinone (6.70 g, 33.4 mmol) and sodium triacetoxyborohydride
(13.6 g, 64.3 mmol). The reaction mixture was stirred at 23.degree.
C. for 20 hours. Then saturated NaHCO.sub.3 was added and the
mixture was separated. The aqueous layer was extracted with
CH.sub.2Cl.sub.2 (3.times.50 mL) and the combined organic layer
were concentrated in vacuo. The residue was purified by silica gel
column chromatography (SiO.sub.2 heptane/EtOAc=4/1) to give the
titled compound (10.6 g, 98% yield). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.37 (s, 1H), 6.88 (s, 1H), 4.05 (m, 2H),
3.90 (s, 3H), 3.01 (m, 3H), 2.32 (s, 3H), 1.71 (m, 2H), 1.48-1.61
(m, 2H), 1.45 (s, 9H); MS (ES) [M-H] 415.1.
tert-Butyl-4-{ethyl[3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl-
]amino}piperidine-1-carboxylate
##STR00463##
[1022] To a stirred solution of tert-butyl
4-{[3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl]amino}piperidi-
ne-1-carboxylate (10.6 g, 25.4 mmol) in CH.sub.2Cl.sub.2 (200 mL)
and AcOH (10 mL) was added acetaldehyde (2.80 g, 63.5 mmol) and
sodium triacetoxyborohydride (13.4 g, 63.5 mmol). The reaction
mixture was stirred at 23.degree. C. for 24 hours. Then saturated
NaHCO.sub.3 was added and the mixture was separated. The aqueous
layer was extracted with CH.sub.2Cl.sub.2 (3.times.50 mL) and the
combined organic layer was concentrated in vacuo. The residue was
purified by silica gel column chromatography
(SiO.sub.2Heptane/EtOAc=5/1) to give the titled compound as oil
(4.50 g, 40% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm;
7.82 (br.s., 1H), 7.45 (d, J=1.6 Hz, 1H), 3.96-4.18 (m, 2H), 3.92
(s, 3H), 3.08 (q, J=7.0 Hz, 2H), 2.89 (tt, J=11.0, 3.7 Hz, 1H),
2.69 (m, 2H), 2.55 (s, 3H), 1.73 (m, 2H), 1.49-1.56 (m, 2H), 1.45
(s, 9H), 0.86 (t, J=7.0 Hz, 3H); MS (ES) [M+Na] 467.0.
Methyl
3-[ethyl(piperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoate
TFA salt
##STR00464##
[1024] To a stirred solution of tert-butyl
4-{ethyl[3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl]amino}pip-
eridine-1-carboxylate (4.50 g, 10.1 mmol) in CH.sub.2Cl.sub.2 (50
mL) was added TFA (5 mL). The reaction mixture was stirred at rt
for 18 hours. Then reaction mixture was concentrated in vacuo to
give the titled compound as a crude product (7.70 g).
Methyl
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)-
benzoate
##STR00465##
[1026] To a stirred solution of methyl
3-[ethyl(piperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoate
TFA salt (crude material, 5.30 g, 12.0 mmol) in CH.sub.2Cl.sub.2
(150 mL) and AcOH (10 mL) was added formaldehyde (15 mL, 44%
aqueous solution) and sodium triacetoxyborohydride (6.40 g, 30.0
mmol). The reaction mixture was stirred at 23.degree. C. for 2.5
days. Then saturated NaHCO.sub.3 was added and the mixture was
separated. The aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.50 mL) and the combined organic layers were concentrated
in vacuo. The residue was purified by silica gel column
chromatography (NH--SiO.sub.2 heptane/EtOAc=2/1) to give the titled
compound as oil (1.20 cg, 28% yield). .sup.1H-NMR (400 cMhz,
CDCl.sub.3) .delta. ppm; 7.81 (br.s., 1H), 7.45 (d, J=2.0 Hz, 1H),
3.92 (s, 3H), 3.10 (q, J=7.0 Hz, 2H), 2.83 (m, 2H), 2.66-2.77 (m,
1H), 2.54 (s, 3H), 2.24 (s, 3H), 1.84-1.94 (m, 2H), 1.63-1.78 (m,
4H), 0.86 (t, J=7.0 Hz, 3H); MS (ES) [M+H] 359.1.
Methyl
3-{ethyl[1-(propan-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoro-
methyl)benzoate
##STR00466##
[1028] To a stirred solution of methyl
3-[ethyl(piperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoate
TFA salt (crude material, 1.30 g, 3.36 mmol) in CH.sub.2Cl.sub.2
(20 mL) and AcOH (5 mL) was added acetone (590 mg, 10.1 mmol) and
sodium triacetoxyborohydride (1.80 g, 8.40 mmol). The reaction
mixture was stirred at 23.degree. C. for 6 days and then saturated
NaHCO.sub.3 was added and the mixture was separated. The aqueous
layer was extracted with CH.sub.2Cl.sub.2 (3.times.30 mL) and the
combined organic layer was concentrated in vacuo. The residue was
purified by silica gel column chromatography (NH--SiO.sub.2
heptane/ethylacetate=5/1) to give the titled compound as oil (220
mg, 17% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.79
(br.s., 1H), 7.44 (d, J=1.6 Hz, 1H), 3.91 (s, 3H), 3.10 (q, J=7.0
Hz, 2H), 2.86 (m, 2H), 2.61-2.75 (m, 2H), 2.53 (s, 3H), 2.06 (m,
2H), 1.70-1.80 (m, 2H), 1.58-1.69 (m, 2H), 1.00 (s.times.2, 6H),
0.84 (t, J=7.0 Hz, 3H); MS (ES) [M+H] 387.2.
3-[Ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoic
acid
##STR00467##
[1030] To a stirred solution of methyl
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoa-
te (700 mg, 1.95 mmol) in methanol (15 mL) was added aq. NaOH (5M,
590 ul, 2.93 mmol). The reaction mixture was stirred at 50.degree.
C. for 3 hours. After cooling to 23.degree. C., HCl (2 M) was added
to the mixture. The mixture was acidified to pH=5 and was
concentrated in vacuo to give the titled compound as crude products
(1.26 g).
[1031] Following the same preparation method for
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoi-
c acid, the following titled compounds were prepared from methyl
3-amino-2-methyl-5-(trifluoromethyl)benzoate and tert-butyl
3-oxoazetidine-1-carboxylate.
t-butyl
3-((3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl)amino)a-
zetidine-1-carboxylate
##STR00468##
[1033] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.49 (s, 1H),
6.60 (s, 1H), 4.39 (dd, J=7.2 Hz, 8.8 Hz, 2H), 4.26 (m, 1H), 4.19
(br, d, J=5.6 Hz, 1H), 3.93 (s, 3H), 3.78 (m, 2H), 2.41 (s, 3H),
1.47 (s, 9H).
tert-butyl
3-(ethyl(3-(methoxycarbonyl)-2-methyl-5-(trifluoromethyl)phenyl-
)amino)azetidine-1-carboxylate
##STR00469##
[1035] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.86 (s, 1H),
7.20 (s, 1H), 4.15 (m, 1H), 3.99 (dd, J=8.4 Hz, J=8.4 Hz, 2H), 3.94
(s, 3H), 3.65 (br, 2H), 3.00 (q, J=7.2 Hz, 2H), 2.60 (s, 3H), 1.43
(s, 9H), 0.90 (t, J=6.8 Hz, 3H); MS (ES) (M+1) 417.41.
Methyl
3-(azetidin-3-yl(ethyl)amino)-2-methyl-5-(trifluoromethyl)benzoate
##STR00470##
[1037] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.89 (s, 1H),
7.20 (s, 1H), 4.39 (m, 1H), 3.94 (s, 3H), 3.87 (m, 2H), 3.68 (br,
2H), 2.99 (q, J=6.8 Hz, 2H), 2.64 (s, 3H), 0.92 (t, J=6.8 Hz, 3H);
MS (ES) (M+1) 317.32.
Methyl
3-(ethyl(1-methylazetidin-3-yl)amino)-2-methyl-5-(trifluoromethyl)b-
enzoate
##STR00471##
[1039] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 7.83 (s, 1H),
7.24 (s, 1H), 4.03 (m, 1H), 3.94 (s, 3H), 3.60 (br, dd, J=7.2, 7.2
Hz, 2H), 2.94 (q, J=7.6 Hz, 2H), 2.72 (br, t, J=6.8 Hz, 2H), 2.59
(s, 3H), 2.33 (s, 3H), 0.90 (t, J=7.2 Hz, 3H); MS (ES) (M+1)
331.09.
3-(ethyl(1-methylazetidin-3-yl)amino)-2-methyl-5-(trifluoromethyl)benzoic
acid
##STR00472##
[1041] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 7.83 (s, 1H),
7.37 (s, 1H), 4.44 (br, 1H), 3.86 (br, 2H), 3.33 (s, 2H), 3.04 (q,
J=6.8 Hz, 2H), 3.03 (s, 3H), 2.64 (s, 3H), 0.90 (t, J=7.2 Hz, 3H);
MS (ES) (M+1) 317.32.
tert-butyl
3-((5-Chloro-3-(methoxycarbonyl)-2-methylphenyl)amino)azetidine-
-1-carboxylate
##STR00473##
[1043] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 7.05 (d, J=2.0
Hz, 1H), 6.48 (d, J=2.0 Hz, 1H), 4.31 (dd, J=7.2, 7.8 Hz, 2H),
4.26-4.21 (m, 1H), 3.87 (s, 3H), 3.82 (dd, J=4.4, 8.3 Hz, 2H), 2.29
(s, 3H), 1.45 (s, 9H).
tert-butyl
3-((5-Chloro-3-(methoxycarbonyl)-2-methylphenyl)(ethyl)amino)az-
etidine-1-carboxylate
##STR00474##
[1045] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 7.57 (d, J=2.0
Hz, 1H), 7.18 (d, J=2.0 Hz, 1H), 4.19-4.14 (m, 1H), 3.98 (dd,
J=8.0, 7.5 Hz, 2H), 3.89 (s, 3H), 3.62-3.56 (br, 2H), 2.98 (q,
J=6.7 Hz, 2H), 2.49 (s, 3H), 1.42 (s, 9H), 0.88 (t, J=7.2 Hz,
3H).
Methyl
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methylbenzoate
##STR00475##
[1047] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 7.53 (d, J=2.0
Hz, 1H), 7.17 (d, J=2.0 Hz, 1H), 4.01 (m, 1H), 3.89 (s, 3H), 3.57
(dd, J=6.5, 8.0 Hz, 2H), 2.93 (q, J=7.5 Hz, 2H), 2.83 (bt, J=7.0
Hz, 2H), 2.49 (s, 3H), 2.33 (s, 3H), 0.90 (t, J=7.0 Hz, 3H); LCMS
(M+H)=297.30.
3-{Ethyl[1-(propan-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoromethyl)-
benzoic acid
##STR00476##
[1049] To a stirred solution of methyl
3-{ethyl[1-(propan-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoromethyl-
)benzoate (220 mg, 0.569 mmol) in methanol (4 mL) was added aq.
NaOH (5M, 170 ul, 0.854 mmol). The reaction mixture was stirred at
50.degree. C. for 2 hours. After cooling to 23.degree. C. HCl (2 M)
was added to the mixture. The mixture was acidified to pH=5 and was
concentrated in vacuo to give the titled compound as crude products
(510 mg).
Methyl
3-{[(2R*,6R*)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-2-methyl-5-
-(trifluoromethyl)benzoate
##STR00477##
[1051] To a stirred solution of methyl
3-amino-2-methyl-5-(trifluoromethyl)benzoate (400 mg, 1.72 mmol) in
THF (15 mL) and TFA (491 uL) was added (2R,
6S)-1-benzyl-2,6-dimethylpiperidin-4-one (410 mg, 1.89 mmol) at
0.degree. C. The mixture was stirred at 60.degree. C. for 1 hour
under nitrogen atmosphere. Then the mixture was cooled to 0.degree.
C. in an ice bath, sodium triacetoxyborohydride (765 mg, 3.43 mmol)
was added. The reaction mixture was stirred at 23.degree. C. for 2
hours. Then aq. NaHCO.sub.3 was added at 0.degree. C. until pH 8-9,
and the content was extracted with ethylacetate. The organic layer
was dried over MgSO.sub.4 and filtered. The filtrate was
concentrated in vacuo. The residue was purified by silica gel
column chromatography (NH--SiO.sub.2; heptane/ethylacetate=3/1) to
give the titled compound as a colorless oil. (278 mg, 37% yield).
.sup.1H-NMR (400M Hz, CDCl.sub.3) .delta. ppm; 7.21-7.40 (m, 6H),
6.90 (s, 1H), 3.95 (d, J=14.0 Hz, 1H), 3.90 (s, 3H), 3.60-3.73 (m,
2H), 3.46 (d, J=14.0 Hz, 1H), 3.08-3.12 (m, 1H), 2.98-3.03 (m, 1H),
2.30 (s, 3H), 2.02-2.08 (m, 1H), 1.83-1.88 (m, 1H), 1.52-1.60 (m,
1H), 1.17-1.26 (m, 1H), 1.11 (d, J=6.8 Hz, 3H), 1.13 (d, J=6.4 Hz,
3H).
Methyl
3-{[(2R*,6S*)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-2-methyl-5-
-(trifluoromethyl)benzoate (diastereomeric mixture)
##STR00478##
[1053] To a stirred mixture of MS3A (1.0 g), methyl
3-amino-2-methyl-5-(trifluoromethyl) benzoate (350 mg, 1.50 mmol)
and (2R, 6S)-1-benzyl-2,6-dimethylpiperidin-4-one (359 mg, 1.65
mmol) in chloroform (10 mL) was added acetic acid (500 uL) at
0.degree. C. and stirred at r.t. for 7 hours. To the reaction
mixture was added sodium triacetoxyborohydride (670 mg, 3.00 mmol)
at 0.degree. C. The reaction mixture was stirred at 23.degree. C.
for 1.5 hours. Then the reaction mixture was quenched with aq.
NaHCO.sub.3 until pH 8-9. The MS 3 .ANG. was removed by Celite
filtration and the content was extracted with ethylacetate, dried
over MgSO.sub.4, filtered and the filtrate was concentrated in
vacuo. The residue was purified by silica gel column chromatography
(heptane/ethylacetate=5/1 to 3/1). Target fractions were collected
and concentrated in vacuo. The residue was purified by silica gel
column chromatography (NH--SiO.sub.2; heptane/ethylacetate=5/1) to
give the titled compound as a light yellow oil (223 mg, 62%
yield.). For major isomer; .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.40-7.21 (m, 6H), 6.85 (s, 1H), 4.00-3.95 (m, 1H),
3.91 (s, 3H), 3.88 (d, 2H), 3.84-3.76 (m, 1H), 2.83-2.73 (m, 2H),
2.25 (s, 3H), 1.83-1.69 (m, 4H), 1.13 (d, J=6.4 Hz, 6H).
Methyl
3-{[(2R*,6R*)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-2-m-
ethyl-5-(trifluoromethyl)benzoate
##STR00479##
[1055] To a stirred solution of methyl 3-{[(2R,
6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-2-methyl-5-(trifluoromethy-
l)benzoate (278 mg, 0.640 mmol) in CH.sub.2Cl.sub.2 (5 mL) and AcOH
(500 uL) was added acetaldehyde (359 mg, 6.40 mmol) and sodium
triacetoxyborohydride (428 mg, 1.92 mmol) at 0.degree. C. The
reaction mixture was stirred at 23.degree. C. for 7 hours. Then the
reaction mixture was quenched with aq. NaHCO.sub.3 until pH 8-9.
The content was extracted with ethylacetate, dried over MgSO.sub.4
and filtered. The filtrate was concentrated in vacuo. The residue
was purified by silica gel column chromatography (NH--SiO.sub.2;
heptane/ethylacetate=5/1) to give the titled compound as a light
yellow oil (236 mg, 80% yield.). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.79 (s, 1H), 7.43 (s, 1H), 7.21-7.36 (m, 5H), 3.92
(s, 3H), 3.87 (d, J=14.0 Hz, 1H), 3.38 (d, J=14.0 Hz, 1H),
3.04-3.15 (m, 4H), 2.73-2.80 (m, 1H), 2.53 (s, 3H), 1.76-1.82 (m,
1H), 1.51-1.63 (m, 2H), 1.34-1.42 (m, 1H), 1.06 (d, J=6.0 Hz, 3H),
0.93 (d, J=6.8 Hz, 3H), 0.86 (t, J=6.8 Hz, 3H).
Methyl
3-{[(2R*,6S*)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-2-m-
ethyl-5-(trifluoromethyl)benzoate(diastereomeric mixture)
##STR00480##
[1057] To a stirred solution of methyl 3-{[(2R,
6S)-1-benzyl-2,6-dimethylpiperidin-4-yl]
amino}-2-methyl-5-(trifluoromethyl)benzoate (223 mg, 0.513 mmol) in
chloroform (4 mL) and acetic acid (500 uL) was added acetaldehyde
(1.0 ml) and stirred for 30 min at 0.degree. C. Then sodium
triacetoxyborohydride (343 mg, 1.54 mmol) was added to the reaction
mixture at 0.degree. C. The reaction mixture was stirred at
23.degree. C. for 4 hours. Then the reaction mixture was quenched
with aq. NaHCO.sub.3 until pH 8-9. The content was extracted with
ethylacetate. The organic layer was washed with brine. The organic
layer was dried over MgSO.sub.4 and filtered. The filtrate was
concentrated in vacuo. The residue was purified by silica gel
column chromatography (heptane/ethylacetate=4/1 to 1/2) to give the
titled compound as a colorless oil. (105.5 mg, 44% yield). MS (ES)
[M+H] 463.3.
3-{[(2R*,4S*,6R*)-1-Benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-N-[(4,-
6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-5-(trifluorometh-
yl)benzamide
##STR00481##
[1059] To a stirred solution of methyl 3-{[(2R,
6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-2-methyl-5-(trifluoromethyl)benzoate (236 mg, 0.510
mmol) in ethanol (5 mL) was added aq. NaOH (5 M, 269 ul, 1.02
mmol). The reaction mixture was stirred at 80.degree. C. for 2
hours. After cooling to rt, the reaction mixture was concentrated
in vacuo and dried under reduced pressure to give crude carboxylic
acid sodium salt.
[1060] To a stirred solution of crude carboxylic acid sodium salt
and 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt
(125 mg, 0.664 mmol) in DMSO (5 mL) was added PyBOP (398 mg, 0.766
mmol) and Hunig's base (445 ul, 2.55 mmol). The reaction mixture
was stirred at 23.degree. C. for 19 hours. The reaction mixture was
quenched with water and extracted with ethylacetate. The organic
layer was washed with water (twice) and brine. The organic layer
was dried over MgSO.sub.4 and filtered. The combined organic layer
was concentrated in vacuo. The residue was purified by silica gel
column chromatography (NH--SiO.sub.2; ethylacetate/MeOH=50/1-8/1)
to give the titled compound as a white amorphous (267 mg, 90%
yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.14-7.36
(m, 8H), 5.94 (s, 1H), 4.53-4.56 (m, 2H), 3.85 (d, J=14.0 Hz, 1H),
3.58 (d, J=14.0 Hz, 1H), 3.02-3.14 (m, 4H), 2.72-2.80 (m, 1H), 2.40
(s, 3H), 2.32 (s, 3H), 2.21 (s, 3H), 1.32-1.82 (m, 4H), 1.05 (d,
J=6.4 Hz, 3H), 0.92 (d, J=6.8 Hz, 3H), 0.84 (t, J=6.8 Hz, 3H).
3-{[(2R*,4R*,6S*)-1-Benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-N-[(4,-
6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-5-(trifluorometh-
yl)benzamide
##STR00482##
[1062] To a stirred solution of methyl 3-{[(2R,
6S)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-2-methyl-5-(trifluo-
romethyl)benzoate (106 mg, 0.228 mmol) in ethanol (2.0 mL) was
added aq. NaOH (5 M, 91.2 ul, 0.456 mmol). The reaction mixture was
stirred at 80.degree. C. for 1.5 hours. After cooling to rt,
solvent was removed in vacuo and dried under reduced pressure. To a
stirred solution of this residue and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt
(55.9 mg, 0.297 mmol) in DMSO (2 mL) was added Hunig's base (199
ul, 1.14 mmol) and PyBOP (178 mg, 0.342 mmol). The reaction mixture
was stirred at rt for 4 hours. The reaction mixture was quenched
with water. The content was extracted with ethylacetate. The
organic layer was washed with water twice and brine. The organic
layer was dried over MgSO.sub.4 and filtered. The filtrate was
concentrated in vacuo. The residue was purified by silica gel
column chromatography (NH SiO.sub.2, ethylacetate/MeOH=50/1 to
40/1) to give the titled compound as a colorless oil (35.6 mg, 27%
yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.34-7.08
(m, 7H), 5.96 (s, 1H), 4.54 (d, J=6.0 Hz, 2H), 3.81 (s, 2H), 3.53
(m, 1H), 2.99-2.91 (m, 2H), 2.81 (m, 2H), 2.41 (s, 3H), 2.24 (s,
3H), 2.20 (s, 3H), 1.82-1.47 (m, 4H), 1.00 (d, J=6.4 Hz, 6H), 0.81
(t, J=6.8 Hz, 3H); MS(ES) [M+H] 583.0.
tert-Butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl]amino}piperidin-
e-1-carboxylate
##STR00483##
[1064] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1.40 g, 7.04 mmol) in
CH.sub.2Cl.sub.2 (30 mL) and AcOH (2.50 g, 42.2 mmol) was added
N-Boc piperidinone (1.96 g, 9.85 mmol) and sodium
triacetoxyborohydride (4.40 g, 21.1 mmol). The reaction mixture was
stirred at 23.degree. C. for 18 hours and then saturated
NaHCO.sub.3 was added and the mixture was separated. The aqueous
layer was extracted with CH.sub.2Cl.sub.2 (3.times.20 mL) and the
combined organic layers were concentrated in vacuo. The residue was
purified by silica gel column chromatography (SiO.sub.2
heptane/ethylacetate=10/1 to 1/1) to give the titled compound as
oil (2.40 g, 89% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
ppm; 7.09 (s, 1H), 6.69 (s, 1H), 4.05 (m, 2H), 3.88 (s, 3H), 3.63
(m, 1H), 3.42 (m, 1H), 2.97 (m, 2H), 2.23 (s, 3H), 2.05 (m, 2H),
1.45 (s, 9H), 1.27 (m, 2H).
tert-Butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl](ethyl)amino}pi-
peridine-1-carboxylate
##STR00484##
[1066] To a stirred solution of tert-butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl]
amino}piperidine-1-carboxylate (2.40 g, 6.28 mmol) in
dichloromethane (50 mL) and acetic acid (1.8 mL) was added
acetaldehyde (830 ul, 12.6 mmol) and sodium triacetoxyborohydride
(4.00 g, 18.8 mmol). The reaction mixture was stirred at rt for 18
hours. Then aq. NaHCO.sub.3 was added and the mixture was
separated. The aqueous layer was extracted with dichloromethane.
The combined organic layer was concentrated in vacuo. The residue
was purified by silica gel column chromatography (SiO.sub.2
heptane/ethylacetate=10/1) to give the titled compound as oil (2.40
g, 89% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.56
(s, 1H), 7.21 (d, J=1.6 Hz, 1H), 4.04 (m, 2H), 3.89 (s, 3H), 3.02
(q, J=7.0 Hz, 2H), 2.88 (m, 1H), 2.68 (m, 2H), 2.44 (s, 3H), 1.71
(m, 2H), 1.48-1.61 (m, 2H), 1.45 (s, 9H), 0.83-0.89 (t, J=7.0 Hz,
3H); MS(ES) [M+Na] 433.0.
Methyl
5-chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methylbenzoate
##STR00485##
[1068] To a stirred solution of tert-butyl
4-{ethyl[3-(methoxycarbonyl)-2-methyl-5-chlorophenyl]amino}piperidine-1-c-
arboxylate (2.20 g, 5.35 mmol) in dichloromethane (20 mL) was added
TFA (4 mL). The reaction mixture was stirred at rt for 2 hours.
Then reaction mixture was concentrated in vacuo to give TFA salt as
a crude product.
[1069] To a stirred solution of TFA salt in THF (20 mL) and acetic
acid (305 ul) was added formaldehyde (3 mL, 44% aqueous solution)
and sodium triacetoxyborohydride (3.40 g, 16.1 mmol) at 0.degree.
C. The reaction mixture was stirred at 23.degree. C. for 19 hours
and then saturated NaHCO.sub.3 was added and the mixture was
separated. The aqueous layer was extracted with EtOAc (50 mL) and
the combined organic layer was washed with brine, dried over
Na.sub.2SO.sub.4 and filtered and the filtrate concentrated in
vacuo. The residue was purified by silica gel column chromatography
(NH--SiO.sub.2 heptane/ethylacetate=1/1) to give title compound as
oil (1.54 g, 89% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
ppm; 7.54 (d, J=2.4 Hz, 1H), 7.21 (d, J=2.4 Hz, 1H), 3.89 (s, 3H),
3.07 (q, J=6.8 Hz, 2H), 2.81-2.85 (m, 2H), 2.66-2.70 (m, 1H), 2.44
(s, 3H), 2.24 (s, 3H), 1.86-1.93 (m, 2H), 1.65-1.75 (m, 4H), 0.86
(t, J=6.8 Hz, 3H).
5-Chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methylbenzoic
acid
##STR00486##
[1071] To a stirred solution of methyl
5-chloro-3-[ethyl(1-methylpiperidin-4-yl) amino]-2-methylbenzoate
(1.06 g, 3.26 mmol) in methanol (10 mL) was added aq. NaOH (5 M,
780 ul, 3.92 mmol). The reaction mixture was stirred at 50.degree.
C. for 18 hours. After cooling to 23.degree. C., HCl (2 M, 2 mL))
was added to the mixture and the mixture was acidified to pH=5 and
was concentrated in vacuo to give the titled compound as crude
products (1.80 g).
Methyl
3-{[(2R,6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-5-fluoro-2-m-
ethylbenzoate
##STR00487##
[1073] To a stirred solution of methyl
3-amino-5-fluoro-2-methylbenzoate (520 mg, 2.84 mmol) in THF (20
mL) and TFA (813 uL) was added
(2R,6S)-1-benzyl-2,6-dimethylpiperidin-4-one (679 mg, 3.12 mmol) at
0.degree. C. The mixture was stirred at 60.degree. C. for 1 hour
under nitrogen atmosphere. Then the mixture was cooled to 0.degree.
C. in an ice bath, sodium triacetoxyborohydride (1.27 g, 5.68 mmol)
was added. The reaction mixture was stirred at rt for 2 hours. Then
aq. NaHCO.sub.3 was added at 0.degree. C. until pH 8-9, and the
content was extracted with ethylacetate. The organic layer was
dried over MgSO.sub.4 and filtered. The filtrate was concentrated
in vacuo. The residue was purified by silica gel column
chromatography (NH--SiO.sub.2; heptane/ethylacetate=5/1) to give
titled compound as a colorless oil (331 mg, 30% yield.).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.35-7.39 (m, 2H),
7.27-7.30 (m, 2H), 7.21-7.26 (m, 1H), 6.76 (dd, J=2.4, 9.2 Hz, 1H),
6.45 (dd, J=2.4, 11.2 Hz, 1H), 3.94 (d, J=14.4 Hz, 1H), 3.87 (s,
3H), 3.58-3.65 (m, 2H), 3.46 (d, J=14.4 Hz, 1H), 3.06-3.12 (m, 1H),
2.94-3.01 (m, 1H), 2.21 (s, 3H), 2.02-2.07 (m, 1H), 1.82-1.89 (m,
1H), 1.15-1.22 (m, 1H), 1.12 (d, J=6.0 Hz, 3H), 1.09 (d, J=7.2 Hz,
3H).
Methyl
3-{[(2R,4R,6S)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-5-fluoro--
2-methylbenzoate
##STR00488##
[1075] To a stirred solution of methyl
3-amino-5-fluoro-2-methylbenzoate (171 mg, 0.931 mmol) in
chloroform (6 mL) and AcOH (266 uL) was added
(2R,6S)-1-benzyl-2,6-dimethylpiperidin-4-one (212 mg, 0.980 mmol)
and MS3A (500 mg) at 0.degree. C. The mixture was stirred at rt for
2 hour under nitrogen atmosphere. Then the mixture was cooled to
0.degree. C. in an ice bath and sodium triacetoxyborohydride (415
mg, 1.86 mmol) was added. The reaction mixture was stirred at rt
for 5 hours. Then aq. NaHCO.sub.3 was added at 0.degree. C. until
pH 8-9, and the content was extracted with ethylacetate. The
organic layer was dried over MgSO.sub.4 and filtered. The filtrate
was concentrated in vacuo. The residue was purified by silica gel
column chromatography (1. SiO.sub.2; heptane/ethylacetate=5/1 to
3/1, 2. NH--SiO.sub.2; heptane/ethylacetate=5/1, 3. NH--SiO.sub.2;
heptane/ethylacetate=10/1 to 6/1) to give the titled compound as a
colorless oil (85.0 mg, 24% yield.). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.38-7.20 (m, 5H), 6.77 (dd, J=2.4, 9.2
Hz, 1H), 6.40 (dd, J=2.4, 11.6 Hz, 1H), 3.95-3.90 (m, 1H), 3.88 (s,
3H), 3.87 (s, 2H), 2.84-2.74 (m, 2H), 2.16 (s, 3H), 1.81-1.66 (m,
4H), 1.11 (d, J=6.0 Hz, 6H).
Methyl
3-{[(2R,6R)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-5-flu-
oro-2-methylbenzoate
##STR00489##
[1077] To a stirred solution of methyl
3-{[(2R,6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]
amino}-5-fluoro-2-methylbenzoate (269 mg, 0.701 mmol) in
dichloromethane (5 mL) and acetic acid (500 uL) was added
acetaldehyde (124 mg, 2.80 mmol) and sodium triacetoxyborohydride
(469 mg, 2.10 mmol) at 0.degree. C. The reaction mixture was
stirred at rt for 12 hours. Then the reaction mixture was quenched
with aq. NaHCO.sub.3 until pH 8-9. The content was extracted with
ethylacetate. The organic layer was dried over MgSO.sub.4 and
filtered. The filtrate was concentrated in vacuo. The residue was
purified by silica gel column chromatography (NH--SiO.sub.2;
heptane/ethylacetate=5/1) to give the titled compound as a light
yellow oil (277 mg, 96% yield). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.19-7.36 (m, 6H), 6.97 (dd, J=2.4, 10.4 Hz, 1H), 3.89
(s, 3H), 3.86 (d, J=10.4 Hz, 1H), 3.39 (d, J=10.4 Hz, 1H),
3.02-3.10 (m, 4H), 2.74-2.80 (m, 1H), 2.43 (s, 3H), 1.74-1.82 (m,
1H), 1.51-1.63 (m, 2H), 1.32-1.40 (m, 1H), 1.05 (d, J=6.4 Hz, 3H),
0.93 (d, J=7.2 Hz, 3H), 0.86 (t, J=6.8 Hz, 3H); MS (ES) [M+H]
413.0.
Methyl
3-{[(2R,4R,6S)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-5--
fluoro-2-methylbenzoate
##STR00490##
[1079] To a stirred solution of methyl
3-{[(2R,4R,6S)-1-benzyl-2,6-dimethylpiperidin-4-yl]amino}-5-fluoro-2-meth-
ylbenzoate (84.9 mg, 0.221 mmol) and acetaldehyde (124 mg, 2.21
mmol) in chloroform (2 mL), was added acetic acid (200 uL) at
0.degree. C. and stirred at 23.degree. C. for 1 hour. To the
reaction mixture was added sodium triacetoxyborohydride (148 mg,
0.662 mmol) at 0.degree. C. The reaction mixture was stirred at RT
for 3.5 hours. To the reaction mixture were added MS 3 .ANG. (200
mg) and acetaldehyde (500 ul) at 0.degree. C. and stirred at r.t.
for 25 minutes. To the reaction mixture was added sodium
triacetoxyborohydride (148 mg, 0.662 mmol) at 0.degree. C. and
stirred at rt for 12 hours. Then the reaction mixture was then
quenched with saturated NaHCO.sub.3 until pH=8-9. The content was
extracted with EtOAc. The organic layer was washed with brine. The
organic layer was dried over MgSO.sub.4 and filtered. The filtrate
was concentrated in vacuo. The residue was purified by silica gel
column chromatography (NH--SiO2; heptane/ethylacetate=5/1) to give
title compound as a colorless oil (48.5 mg, 53% yield). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm; 7.36-7.19 (m, 6H), 6.96 (dd,
J=2.8, 9.6 Hz, 1H), 3.90 (s, 3H), 3.82 (s, 2H), 3.44 (m, 1H),
2.98-2.91 (m, 2H), 2.88-2.87 (m, 2H), 2.31 (s, 3H), 1.82-1.70 (m,
2H), 1.57-1.47 (m, 2H), 0.99 (d, J=6.4 Hz, 6H), 0.83 (t, J=6.8 Hz,
3H).
3-{[(2R,6R)-1-Benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-N-[(4,6-dime-
thyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-fluoro-2-methylbenzamide
##STR00491##
[1081] To a stirred solution of methyl
3-{[(2R,6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-5-fluoro-2-methylbenzoate (277 mg, 0.672 mmol) in
ethanol (5 mL) was added aq. NaOH (5 M, 269 ul, 1.35 mmol). The
reaction mixture was stirred at 80.degree. C. for 2 hours. After
cooling to rt, the reaction mixture was concentrated in vacuo and
dried under reduced pressure to give crude carboxylic acid sodium
salt.
[1082] To a stirred solution of crude carboxylic acid sodium salt
and 3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt
(165 mg, 0.874 mmol) in DMSO (5 mL) was added PyBOP (525 mg, 1.01
mmol) and Hunig's base (586 ul, 3.36 mmol). The reaction mixture
was stirred at 23.degree. C. for 5 hours. The reaction mixture was
quenched with water and extracted with ethylacetate. The organic
layer was washed with water (2.times.10 mL) and brine (1.times.10
mL). The organic layer was dried over MgSO.sub.4 and filtered and
the filtrate concentrated in vacuo. The residue was purified by
silica gel column chromatography (NH--SiO.sub.2; ethylacetate only
to ethylacetate/MeOH=50/1-8/1) to give titled compound as a white
solid (378 mg, quantitative yield). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.20-7.38 (m, 5H), 7.02-7.10 (m, 1H),
6.80-6.84 (m, 1H), 6.73-6.78 (m, 1H), 5.93 (s, 1H), 4.51 (d, J=6.0
Hz, 2H), 3.85 (d, J=10.0 Hz, 1H), 3.38 (d, J=10.0 Hz, 1H),
3.01-3.05 (m, 4H), 2.67-2.81 (m, 1H), 2.40 (s, 3H), 2.23 (s, 3H),
2.23 (s, 3H), 1.52-1.83 (m, 3H), 1.32-1.39 (m, 1H), 1.05 (d, J=6.4
Hz, 3H), 0.93 (d, J=6.8 Hz, 3H), 0.84 (t, J=6.8 Hz, 3H).
3-{[(2R,4R,6S)-1-Benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-N-[(4,6-d-
imethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-fluoro-2-methylbenzamide
##STR00492##
[1084] To a stirred solution of methyl
3-{[(2R,4R,6S)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-5-fluoro-2-methylbenzoate (48.5 mg, 0.118 mmol) in
ethanol (1.5 mL) was added aq. NaOH (5 M, 47.0 ul, 0.235 mmol). The
reaction mixture was stirred at 80.degree. C. for 1.5 hours. After
cooling to rt, solvent was removed in vacuo and dried under reduced
pressure. To a stirred solution of this residue and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt
(28.8 mg, 0.153 mmol) in DMSO (1 mL) was added PyBOP (91.8 mg,
0.176 mmol) and Hunig's base (102 ul, 0.588 mmol). The reaction
mixture was stirred at 23.degree. C. for 13.5 hours. The reaction
mixture was quenched with water, and the mixture was extracted with
ethylacetate. The organic layer was washed with water (2.times.10
mL) and brine (1.times.10 mL). The organic layer was dried over
MgSO.sub.4, filtered and the filtrate was concentrated in vacuo.
The residue was purified by silica gel column chromatography (NH
SiO.sub.2, ethylacetate/MeOH=50/1 to 8/1) to give title compound as
a white solid (50.1 mg, 79% yield). MS(ES) [M+H] 533.3.
Example 1
Synthesis of Compound 1:
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methylbenzamide
##STR00493##
[1085] Step 1: Synthesis of methyl
5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate
##STR00494##
[1087] To a stirred solution of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate (0.5 g, 1.75 mmol) in
acetonitrile (10 mL), Cs.sub.2CO.sub.3 (1.02 g, 2.63 mmol) and
methyl iodide (1.45 g, 3.5 mmol) were added to it. The resulting
reaction mixture was stirred at 80.degree. C. for 4 h. Upon
completion, the solvent was removed under reduced pressure and
residue dissolved in water and extracted with ethyl acetate. Crude
material obtained was purified by column chromatography over silica
gel affording the desired compound product without further
purification (0.39 g).
Step 2: Synthesis of
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methylbenzamide
##STR00495##
[1089] To a stirred solution of methyl
5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (1 equiv.) in
ethanol (5 mL), aqueous NaOH solution (1 equiv.) was added and
reaction stirred at 60.degree. C. for 4 h. On completion, ethanol
was removed under reduced pressure and residue acidified with 1N
HCl to pH 6. The aqueous phase was extracted with 10% MeOH/DCM.
Combined organic layers were dried over sodium sulfate and solvent
removed under reduced pressure affording pure acid (yield 50-60%).
To a solution of this acid (1 equiv.) in DMSO (1.5 mL), PyBOP (1.5
equiv.) was added and reaction stirred at room temperature for 15
min. Then 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.)
was added and reaction stirred overnight. On completion, water was
added and the resulting solid precipitate filtered and washed with
water. Then this solid was stirred with acetonitrile for 10 min and
filtered again to obtain pure target molecule (yield 50-60%). LCMS:
446.15 (M+1).sup.+; HPLC: 94.48% (@254 nm) (R.sub.t; 5.257);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.20 (t,
1H, J=4.8 Hz), 7.25 (s, 1H), 7.04 (s, 1H), 5.85 (s, 1H), 4.23 (d,
2H, J=4.8 Hz), 3.37-3.45 (m, 1H), 2.49 (3H merged in solvent peak),
2.17 (s, 3H), 2.13 (s, 3H), 2.10 (s, 3H), 1.53-1.67 (m, 4H),
1.38-1.50 (m, 4H).
Example 2
Synthesis of Compound 2:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(piperidine-4-yl)amino)benzamide
##STR00496##
[1090] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00497##
[1092] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mass was poured on ice cold water, solid precipitated was filtered,
resulting residue was washed with water and dried under vacuum
giving the desired compound (71.7 g, 99.9%) which was used for
further reaction.
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00498##
[1094] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g,
4415 mmol) and methyl iodide (626.63 g, 4415 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 8 h. On
completion, solid precipitated was filtered, residue washed with
diethyl ether (5 times). Combined organic layers were dried,
concentrated under reduced pressure giving the desired compound
(302 g, 99%) which was used for further reaction.
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00499##
[1096] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 7 h. On
completion, reaction mass was filtered through celite giving
washing of water and ethyl acetate, filtrate was extracted with
ethyl acetate. The combined organic layers were dried, concentrated
under reduced pressure giving the desired compound.
Step 4: tert-butyl
4-((5-bromo-3-(methoxycarbonyl)amino)piperidine-1-carboxylate
##STR00500##
[1098] To a stirred solution of above crude methyl
3-amino-5-bromo-2-methylbenzoate (5 g, 20.57 mmol) and tert-butyl
4-oxopiperidine-1-carboxylate (8.2 g, 41.15 mmol) in methanol (50
mL), acetic acid (1.2 g, 20.57 mmol) was added and reaction stirred
at room temperature for 3 h. Then sodium cyanoborohydride (1.55 g,
24.6 mmol) was added at 0.degree. C. and reaction stirred overnight
at room temperature. On completion, solvent was removed under
reduced pressure and crude material was purified by column
chromatography to afford compound 5 (5 g, 57%).
Step 5: tert-butyl 4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl
(methyl)amino)piperidine-1-carboxylate
##STR00501##
[1100] To a stirred solution of tert-butyl
4-((5-bromo-3-(methoxycarbonyl)amino)piperidine-1-carboxylate (3 g,
7.06 mmol) in acetonitrile (25 mL), cesium carbonate (4.57 g, 14.11
mmol) and methyl iodide (5 g, 35.2 mmol) were added; resulting
reaction mass was heated at 80.degree. C. for 7 h. On completion,
reaction mass was cooled to room temperature and filtered, residue
was washed with ethyl acetate and filtrate was concentrated and
then purified by column chromatography to afford the desired
compound (2.5 g, 81%).
Step 6: tert-butyl
4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbam-
oyl)-2-methylphenyl)(methyl)amino)piperidin-1-carboxylate
##STR00502##
[1102] Aqueous NaOH (0.37 g, 9.38 mmol) was added to a solution of
tert-butyl 4-((5-bromo-3-(methoxycarbonyl)-2-methylphenyl
(methyl)amino)piperidine-1-carboxylate (2 g, 4.69 mmol) in MeOH (20
mL) and stirred at 60.degree. C. for 1 h. After completion of the
reaction, ethanol was removed under reduced pressure and acidified
using dilute HCl up to pH 6 and pH 4 was adjusted using citric
acid. Extraction was carried out using ethyl acetate. Combined
organic layers were dried concentrated giving respective acid (1.7
g, 90%).
[1103] The acid (1.7 g, 4.22 mmol) was then dissolved in DMSO (10
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (1.42 g,
9.38 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (3.66 g, 7.04 mmol) was
added to it and stirring was continued for overnight. After
completion, reaction mass was poured into ice to obtain solid, this
was filtered and washed with acetonitrile followed by purification
with column chromatography to afford the desired compound (1.3 g,
50%).
Step 7:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-m-
ethyl-3-(methyl(piperidine-4-yl)amino)benzamide
##STR00503##
[1105] A stirred solution of tert-butyl
4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbam-
oyl)-2-methylphenyl)(methyl)amino)piperidin-1-carboxylate (1.3 g,
2.39 mmol) in DCM (10 mL) was cooled to 0.degree. C. and TFA (2 mL)
was added to it. Reaction mass was stirred at room temperature for
1 h. On completion, reaction was concentrated to dryness. Residue
was basified with aqueous sodium bicarbonate till pH 8 and aqueous
layer extracted with 20% MeOH/DCM. Combined organic layers were
dried over sodium sulfate and concentrated to afford the desired
compound (0.9 g, 85%). LCMS: 461.15 (M+1).sup.+; HPLC: 98.52% (@254
nm) (R.sub.t; 4.673; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
8.22 (t, 1H), 7.19 (s, 1H), 7.01 (s, 1H), 5.85 (s, 1H), 4.23 (d,
2H, J=4 Hz), 2.91 (d, 2H, J=11.6 Hz), 2.78 (b s, 1H), 2.55 (s, 3H),
2.32-2.35 (m, 2H), 2.18 (s, 3H), 2.10 (s, 6H), 1.44-1.50 (m,
4H).
Example 3
Synthesis of Compound 3:
5-chloro-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2-methylbenzamide
##STR00504##
[1106] Step 1: 5-chloro-2-methyl-3-nitrobenzoic acid
##STR00505##
[1108] 5-Chloro-2-methylbenzoic acid (4 g, 23.39 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C. lot wise.
After 10 minutes nitrating mixture {prepared as mixing Conc. HNO3
(3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4 (4.4 mL)} was added
drop wise at -10.degree. C. Resulting reaction mass was stirred at
-10.degree. C. for 30 minutes. On completion, reaction mixture was
poured on ice cold water, solid precipitated was filtered, washed
with water and dried under vacuum giving desired compound (4.95 g,
99%).
Step 2: methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00506##
[1110] To stirred solution of 5-chloro-2-methyl-3-nitrobenzoic acid
(6.75 g, 31.25 mmol) in DMF (33 mL), sodium carbonate (13.23 g,
125.18 mmol) and methyl iodide (17.77 g, 125.2 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 4 h. On
completion, water was added to the reaction mass and extraction was
carried out using DCM. Combined organic layers were dried,
concentrated under reduced pressure and purified by column
chromatography over silica (60-120 mesh size) giving desired
compound (6 g, 83%).
Step 3: methyl 3-amino-5-chloro-2-methyl benzoate
##STR00507##
[1112] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 26.13 mmol) in ethanol (60
mL), ammonium chloride (6 g, 112.1 mmol) dissolved in water (60 mL)
and iron powder (11.88 g, 208.4 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 1 h. On
completion, water was added to reaction mass and reaction mixture
was filtered through celite, filtrate was extracted with ethyl
acetate. Combined organic layers were washed with water, dried,
concentrated under reduced pressure giving desired compound.
Step 4: methyl 5-chloro-3-(cyclohexylamino)-2-methylbenzoate
##STR00508##
[1114] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (1 g, 5.025 mmol) and cyclohexanone (0.739 g, 7.75 mmol)
in methanol (10 mL), acetic acid (0.301 g, 5.02 mmol) was added and
reaction stirred at room temperature for 3 h. Then sodium
cyanoborohydride (390 mg, 0.00621 mmol) was added and reaction
stirred overnight. On completion, solvent was removed under reduced
pressure and crude material was purified by column chromatography
to afford the desired compound (1.4 g, contaminated with
cyclohexanone).
Step 5: methyl
5-chloro-3-(cyclohexyl(methyl)amino)-2-methylbenzoate (6)
##STR00509##
[1116] To a stirred solution of methyl
5-chloro-3-(cyclohexylamino)-2-methylbenzoate (1.4 g, 0.0049 mole)
in acetonitrile (10 mL), cesium carbonate (3.2 g, 0.0099 mole) and
methyl iodide (3.5 g, 0.0024 mole) were added; resulting reaction
mass was heated at 80.degree. C. for 13 h. On completion, reaction
mass was cooled to room temperature and filtered, residue was
washed with ethyl acetate and filtrate was concentrated and then
purified by column chromatography to afford desired compound (800
mg, 55%).
Step 6:
5-chloro-3-(cyclohexyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00510##
[1118] Aqueous NaOH (0.16 g, 4.06 mmol) was added to a solution of
compound 6 (0.8 g, 2.71 mmol) in EtOH (5 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.7 g, 91%).
[1119] The acid (0.7 g, 2.49 mmol) was then dissolved in DMSO (2
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.752 g,
4.98 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.9 g, 3.73 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed column purification to provide the title compound (0.250
g, 25%).
[1120] LCMS: 416.35 (M+1).sup.+; HPLC: 99.01% (@254 nm) (R.sub.t;
5.408; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (s, 1H), 8.20 (t, 1H, J=4.8 Hz), 7.05 (d, 1H, J=2 Hz), 6.88
(d, 1H, J=2 Hz), 5.85 (s, 1H), 4.24 (d, 2H, J=4.8 Hz), 2.70 (m,
1H), 2.56 (s, 3H), 2.18 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H),
1.75-1.35 (m, 7H), 1.25-1.05 (m, 3H).
Example 4
Synthesis of Compound 4:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(1-pivaloylpiperidin-4-yl)amino)benzamide
##STR00511##
[1122] To a stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(piperidine-4-yl)amino)benzamide (0.05 g, 0.112 mmol) in
DMF (2 mL), EDCI.HCl (0.064 g, 0.336 mmol), HOBt (0.03 g, 0.22
mmol), triethyl amine (0.03 g, 0.336 mmol) and pivalic acid (0.023
g, 0.224 mmol) were added at room temperature and stirred at same
temperature for 18 h. On completion, water was added and extracted
with 10% MeOH/DCM. Combined organic layers were dried, concentrated
giving crude material; which then purified by column chromatography
to afford the desired compound (0.03 g, 56%). LCMS: 545.20
(M+1).sup.+; HPLC: 98.60% (@254 nm) (R.sub.t; 6.355; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.20 (t,
1H, J=4.4 Hz), 7.25 (d, 1H, J=2.8 Hz), 7.05 (d, 1H, J=1.2 Hz), 5.85
(s, 1H), 4.23-4.25 (m, 4H), 3.00 (m, 1H), 2.76 (t, 2H, J=12.4 Hz),
2.54 (s, 3H), 2.18 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 1.62-1.65
(m, 2H), 1.42-1.48 (m, 2H), 1.18 (s, 9H).
Example 5
Synthesis of Compound 5:
5-bromo-3-(cyclopentyl(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methylbenzamide
##STR00512##
[1123] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00513##
[1125] To a mixture of 2-methyl-3-nitrobenzoic acid (15 g, 82.80
mmol) in conc. H.sub.2SO.sub.4 (60 mL), 1,3-dibromo-5,5-dimethyl
2,4-imidazolidinedione (13.07 g, 45.71 mmol) was added and reaction
mixture was stirred at room temperature for 5 h. Then reaction
mixture was slowly poured in to 400 mL of ice cold water. Solid
that precipitated out was filtered, washed and dried under vacuum
the desired compound (21 g, 98.22%).
Step 2: methyl 3-bromo-5-nitrobenzoate (3)
##STR00514##
[1127] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (16 g, 61.54 mmol) in DMF (160 mL), iodomethane (35.72 g, 248
mmol) and sodium carbonate (26.28 g, 248 mmol) were added.
Resulting reaction mass was stirred at 60.degree. C. for 8 h. On
completion, reaction mass was filtered and inorganic solid residue
washed with ethyl acetate. Combined filtrates were concentrated
under vacuum till dryness. The residue was re-dissolved in ethyl
acetate and washed with 5% sodium bicarbonate solution (700 mL)
followed by 5M HCl solution (300 mL). Organic layer was finally
washed with brine, dried over sodium sulfate and concentrated to
afford desired compound (16 g, 95%).
Step 3: ethyl 3-amino-5-bromobenzoate
##STR00515##
[1129] To a stirred solution of methyl 3-bromo-5-nitrobenzoate (17
g, 62.04 mmol) in ethanol (85 mL), was added NH.sub.4Cl solution
(17 g in 85 mL water, 317.8 mmol) followed by Fe powder (27.82 g,
498.12 mmol). Resulting reaction mass was stirred at 90.degree. C.
for 1 h. On completion, reaction mass was filtered and filtrate
concentrated till dryness to get solid which was dissolved in sat.
sodium bicarbonate solution. Aqueous layer was extracted with ethyl
acetate (3.times.50 mL). Combined organic layers were dried over
sodium sulfate and concentrated to afford the desired compound (15
g, 99.14%).
Step 4: methyl 3-bromo-5-(cyclopentylamino)benzoate
##STR00516##
[1131] To a stirred solution of ethyl 3-amino-5-bromobenzoate (2 g,
8.73 mmol) and cyclopentanone (2.2 g, 26.1 mmol) in methanol (20
mL), acetic acid (1.04 g, 17.4 mmol) was added and reaction stirred
at room temperature for 3 h. Then sodium cyanoborohydride (1.37 g,
21.83 mmol) was added and reaction stirred overnight. On
completion, solvent was removed under reduced pressure and crude
material was purified by column chromatography to afford desired
compound (1.6 g, 61%).
Step 5: methyl
5-bromo-3-(cyclopentyl(ethyl)amino)-2-methylbenzoate
##STR00517##
[1133] To a stirred solution of methyl
3-bromo-5-(cyclopentylamino)benzoate (0.3 g, 0.96 mmol) in DMF (5
mL), cesium carbonate (0.628 g, 1.92 mmol) and ethyl iodide (0.748
g, 4.8 mmol) were added; resulting reaction mass was heated at
80.degree. C. for 24 h. On completion, reaction mass was cooled to
room temperature and filtered, residue was washed with ethyl
acetate and filtrate was concentrated and then purified by column
chromatography to afford desired compound (0.150 g, 46%).
Step 6:
5-bromo-3-(cyclopentyl(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dih-
ydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00518##
[1135] Aqueous NaOH (0.03 g, 0.75 mmol) was added to a solution of
methyl 5-bromo-3-(cyclopentyl(ethyl)amino)-2-methylbenzoate (0.170
g, 0.75 mmol) in EtOH (10 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6. Extraction was
carried out using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.15 g, 92.59%).
[1136] The acid (0.15 g, 0.461 mmol) was then dissolved in DMSO (5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.14 g,
0.923 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.35 g, 0.692 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed by HPLC purification to provide the title compound (0.050
g, 24%) as the TFA salt.: LCMS: 460.10 (M+1).sup.+; HPLC: 99.22%
(@254 nm) (R.sub.t; 5.115; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.24 (s, 1H), 7.32
(s, 1H), 7.09 (s, 1H), 5.86 (s, 1H), 4.24 (d, 2H, J=4.8 Hz),
3.47-3.48 (m, 1H), 2.96 (m, 2H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10
(s, 3H), 1.33-1.63 (m, 8H), 0.78 (t, 3H, J=6.4 Hz).
Example 6
Synthesis of Compound 7:
5-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydrop-
yridin-3-yl)methyl)-2-methylbenzamide
##STR00519##
[1137] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00520##
[1139] 5-chloro-2-methylbenzoic acid (4 g, 23.4 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C. in small
portions and stirred for 10 minutes. Nitrating mixture {prepared by
mixing Conc. HNO3 (3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4
(4.4 mL)} was added drop wise at -10.degree. C. The resulting
mixture was stirred at -10.degree. C. for 30 minutes. On
completion, the reaction mixture was poured into ice cold water,
the solid was collected by filtration, washed with water and dried
under vacuum to give 5-chloro-2-methyl-3-nitrobenzoic acid (4.95 g,
99%).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00521##
[1141] To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic
acid, prepared analogously to above, (6.75 g, 31.3 mmol) in DMF (33
mL) was added sodium carbonate (13.2 g, 125 mmol) and methyl iodide
(17.8 g, 125 mmol). The resulting mixture was heated at 60.degree.
C. for 4 h. On completion, water was added to the mixture and
extraction was carried out using DCM. Combined organic layers were
dried, concentrated under reduced pressure and purified by column
chromatography over silica to give methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 83.6%).
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR00522##
[1143] To a stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 26.1 mmol) in ethanol (60
mL), was added ammonium chloride (6 g, 112 mmol) dissolved in water
(60 mL) and iron powder (11.9 g, 208 mmol). The resulting mixture
was heated at 80.degree. C. for 1 h. On completion, water was added
to mixture and reaction mixture was filtered through celite, and
the filtrate was extracted with ethyl acetate. Combined organic
layers were washed with water, dried, and concentrated under
reduced pressure to give methyl 3-amino-5-chloro-2-methyl
benzoate.
Step 4: Synthesis of methyl
5-chloro-3-(cyclopentylamino)-2-methylbenzoate
##STR00523##
[1145] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (1 g, 5.0 mmol) and cyclopentanone (2.1 g, 25.0 mmol) in
methanol (10 mL) was added acetic acid (0.6 g, 5.0 mmol), and the
mixture was stirred at room temperature for 3 h. Then sodium
cyanoborohydride (0.78 g, 12.5 mmol) was added and the reaction
stirred overnight. On completion, solvent was removed under reduced
pressure and crude material was purified by column silica
chromatography to afford methyl
5-chloro-3-(cyclopentylamino)-2-methylbenzoate (1.2 g, 89%).
Step 5: Synthesis of methyl
5-chloro-3-(cyclopentyl(methyl)amino)-2-methylbenzoate
##STR00524##
[1147] To a stirred solution of methyl
5-chloro-3-(cyclopentylamino)-2-methylbenzoate (0.5 g, 1.86 mmol)
in acetonitrile (5 mL) was added cesium carbonate (1.22 g, 3.7
mmol) and methyl iodide (1.32 g, 9.33 mmol). The resulting mixture
was heated at 80.degree. C. for 12 h. On completion, the mixture
was cooled to room temperature and filtered, the residue was washed
with ethyl acetate and the filtrate was concentrated and then
purified by column chromatography to afford methyl
5-chloro-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (0.5 g,
95%).
Step 6: Synthesis of
5-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydrop-
yridin-3-yl)methyl)-2-methylbenzamide
[1148] Aqueous NaOH (0.106 g, 2.66 mmol) was added to a solution of
methyl 5-chloro-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (0.5
g, 1.77 mmol) in MeOH (5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, methanol was removed under
reduced pressure and acidified using dilute HCl up to pH 6 and pH 4
was adjusted using citric acid. Extraction was carried out using
ethyl acetate. Combined organic layers were dried concentrated
giving respective acid (0.4 g, 84%).
[1149] The acid (0.1 g, 0.37 mmol) was then dissolved in DMSO (0.5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.114 g,
0.74 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.29 g, 0.56 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, mixture was poured into ice to obtain a
solid, this was filtered and washed with acetonitrile followed
column purification to provide
5-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydrop-
yridin-3-yl)methyl)-2-methylbenzamide (100 mg, 66%). LCMS: 402.15
(M+1).sup.+; HPLC: 96.46% (@254 nm) (R.sub.t; 5.289; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.19 (t,
1H, J=4.8 Hz), 7.13 (d, 1H, J=2 Hz), 6.91 (d, 1H, J=2 Hz), 5.85 (s,
1H), 4.23 (d, 2H, J=5.2 Hz), 3.41-3.45 (m, 1H), 2.50 (3H merged in
solvent peak), 2.18 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 1.38-1.67
(m, 8H).
Example 7
Synthesis of Compound 8:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(methyl(piperidin-4-yl)amino)benzamide
##STR00525##
[1150] Step 1: 5-chloro-2-methyl-3-nitrobenzoic acid
##STR00526##
[1152] 5-Chloro-2-methylbenzoic acid (4 g, 23.39 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C. lot wise.
After 10 minutes nitrating mixture {prepared as mixing Conc. HNO3
(3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4 (4.4 mL)} was added
drop wise at -10.degree. C. Resulting reaction mass was stirred at
-10.degree. C. for 30 minutes. On completion, reaction mixture was
poured on ice cold water, solid precipitated was filtered, washed
with water and dried under vacuum giving desired compound (4.95 g,
99%).
Step 2: methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00527##
[1154] To stirred solution of 5-chloro-2-methyl-3-nitrobenzoic acid
(6.75 g, 31.25 mmol) in DMF (33 mL), sodium carbonate (13.23 g,
125.18 mmol) and methyl iodide (17.77 g, 125.2 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 4 h. On
completion, water was added to the reaction mass and extraction was
carried out using DCM. Combined organic layers were dried,
concentrated under reduced pressure and purified by column
chromatography over silica (60-120 mesh size) giving desired
compound (6 g, 84%).
Step 3: methyl 3-amino-5-chloro-2-methyl benzoate
##STR00528##
[1156] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 26.13 mmol) in ethanol (60
mL), ammonium chloride (6 g, 112.1 mmol) dissolved in water (60 mL)
and iron powder (11.88 g, 208.4 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 1 h. On
completion, water was added to reaction mass and reaction mixture
was filtered through celite, filtrate was extracted with ethyl
acetate. Combined organic layers were washed with water, dried,
concentrated under reduced pressure giving the desired
compound.
Step 4: tert-butyl
4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl)amino)
piperidine-1-carboxylate
##STR00529##
[1158] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (6 g, 30.15 mmol) and tert-butyl
4-oxopiperidine-1-carboxylate (29 g, 150 mmol) in methanol (10 mL),
acetic acid (1.8 g, 30.1 mmol) was added and reaction stirred at
room temperature for 3 h. Then sodium cyanoborohydride (4.7 g, 75.3
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford the desired compound (4
g, 30%).
Step 5: tert-butyl 4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl)
(methyl)amino)piperidine-1-carboxylate
##STR00530##
[1160] To a stirred solution of tert-butyl
4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl)amino)
piperidine-1-carboxylate (4 g, 9.09 mmol) in acetonitrile (50 mL),
cesium carbonate (5.9 g, 18.09 mmol) and methyl iodide (6.95 g,
48.94 mmol) were added; resulting reaction mass was heated at
80.degree. C. for 12 h. On completion, reaction mass was cooled to
room temperature and filtered, residue was washed with ethyl
acetate and filtrate was concentrated and then purified by column
chromatography to afford the desired compound (2.2 g, 53%).
Step 6: tert-butyl
4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate
##STR00531##
[1162] Aqueous NaOH (0.75 g, 1.86 mmol) was added to a solution of
tert-butyl 4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl)
(methyl)amino)piperidine-1-carboxylate (0.5 g, 1.26 mmol) in MeOH
(5 mL) and stirred at 60.degree. C. for 1 h. After completion of
the reaction, ethanol was removed under reduced pressure and
acidified using dilute HCl up to pH 6 and pH 4 was adjusted using
citric acid. Extraction was carried out using ethyl acetate.
Combined organic layers were dried concentrated giving respective
acid (0.4 g, 84%).
[1163] The acid (0.4 g, 1.04 mmol) was then dissolved in DMSO (1
mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.316 g,
2.09 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.816 g, 1.56 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed column purification to provide the desired compound (0.225
g, 41%).
Step 7:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methyl-3-(methyl(piperidin-4-yl)amino)benzamide
##STR00532##
[1165] A stirred solution of tert-butyl
4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)piperidine-1-carboxylate (0.2 g,
0.632 mmol) in DCM (5 mL) was cooled to 0.degree. C. and TFA (0.15
mL) was added to it. Reaction mass was stirred at room temperature
for 1 h. On completion, reaction was concentrated to dryness.
Residue was purified by solvent washings to afford the desired
compound (0.14 g, 53%) as its TFA salt. LCMS: 417.25 (M+1).sup.+;
HPLC: 96.16% (@254 nm) (R.sub.t; 4.677; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.49 (bs, 1H), 8.20
(t, 1H), 8.18 (bs, 1H), 7.19 (s, 1H), 6.96 (s, 1H), 5.86 (s, 1H),
4.25 (d, 2H, J=4.4 Hz), 3.25 (d, 2H, J=12 Hz), 3.08 (m, 1H), 2.88
(bs, 2H), 2.56 (s, 3H), 2.18 (s, 3H), 2.17 (s, 3H), 2.10 (s, 3H),
1.76 (bs, 4H).
Example 8
Synthesis of Compound 10:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00533##
[1166] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00534##
[1168] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 152 mmol)
was added portion wise at room temperature, and stirring was
continued at room temperature for 5 h. On completion, the mixture
was poured into ice cold water, the resulting solid was filtered,
washed with water and dried under vacuum to give
5-bromo-2-methyl-3-nitrobenzoic acid (71.7 g, 99.9%).
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00535##
[1170] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid,
prepared analogously to above, (287 g, 1103 mmol) in DMF (150 mL),
sodium carbonate (468 g, 4415 mmol) and methyl iodide (626.6 g,
4415 mmol) were added. Resulting mixture was heated at 60.degree.
C. for 8 h. On completion, solid was remove by filtration, and the
residue washed with diethyl ether (5 times). The combined organic
layers were dried, concentrated under reduced pressure to give
methyl 5-bromo-2-methyl-3-nitrobenzene (302 g, 99%).
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00536##
[1172] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzene
(150 g, 544 mmol) in ethanol (750 mL) was added a solution of
ammonium chloride (150 g, 2777 mmol) in water (750 mL) and iron
powder (93.3 g, 1636 mmol). The resulting mixture was heated at
80.degree. C. for 7 h. On completion, mixture was filtered through
celite; the celite was washed with water and ethyl acetate and the
filtrate was extracted with ethyl acetate. Combined organic layers
were dried, concentrated under reduced pressure to give methyl
3-amino-5-bromo-2-methylbenzoate.
Step 4: Synthesis of methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate
##STR00537##
[1174] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (2 g, 8.23 mmol) and
dihydro-2H-pyran-4(3)-one (1.06 g, 10.6 mmol) in methanol (20 mL),
acetic acid (0.5 g, 8.23 mmol) was added and the reaction stirred
at room temperature for 3 h. Then sodium cyanoborohydride (0.622 g,
9.87 mmol) was added at 0.degree. C. and reaction stirred overnight
at room temperature. On completion, the solvent was removed under
reduced pressure and the crude material was purified by column
chromatography to afford methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (1.6
g, 61%).
Step 5: Synthesis of methyl
5-bromo-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzoate
##STR00538##
[1176] To a stirred solution of methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (0.4
g, 1.26 mmol) in acetonitrile (15 mL), cesium carbonate (0.79 g,
2.44 mmol) and methyl iodide (0.86 g, 6.11 mmol) were added;
resulting mixture was heated at 80.degree. C. for 7 h. On
completion, the mixture was cooled to room temperature and
filtered, the residue was washed with ethyl acetate and the
filtrate was concentrated and then purified by column
chromatography to afford Synthesis of methyl
5-bromo-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzoate
(0.33 g, 80%).
Step 6: Synthesis of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00539##
[1178] Aqueous NaOH (0.19 g, 4.89 mmol) was added to a solution of
compound 6 (0.8 g, 2.4 mmol) in MeOH (20 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.70 g, 92%).
[1179] The acid (0.7 g, 2.24 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.74 g,
4.89 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.9 g, 3.6 mmol) was
added to it and stirring was continued for overnight. After
completion, mixture was poured into ice to obtain a solid. The
solid was filtered and washed with acetonitrile followed by
purification with column chromatography to afford
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide (0.53 g, 54%).
LCMS: 462.23 (M+1).sup.+; HPLC: 98.57% (@254 nm) (R.sub.t; 5.276;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.20
(t, 1H, J=4.8 Hz), 7.24 (d, 1H, J=2 Hz), 7.04 (d, 1H, J=2 Hz), 5.85
(s, 1H), 4.24 (d, 2H, J=4.8 Hz), 3.84 (d, 2H, J=10.8 Hz), 3.22-3.28
(m, 2H), 2.96 (m, 1H), 2.56 (s, 3H), 2.18 (s, 3H), 2.13 (s, 3H),
2.10 (s, 3H), 1.52-1.61 (m, 4H).
Example 9
Synthesis of Compound 13: 2, 5-dichloro-3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)
benzamide
##STR00540##
[1180] Step 1: Synthesis of 2,5-dichloro-3-nitrobenzoic acid
##STR00541##
[1182] 2,5-dichlorobenzoic acid (5 g, 26.2 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (50 mL) at -10.degree. C. in small
portions. After 10 minutes nitrating mixture {prepared by mixing
Conc. HNO.sub.3 (2.5 mL) with conc. H.sub.2SO.sub.4 (10 mL)} was
added drop wise at -10.degree. C. The resulting reaction mixture
was stirred at -10.degree. C. for 2 h. On completion, the reaction
mixture was poured into ice cold water and a solid precipitated.
The solid was collected by filtration, washed with water and dried
under vacuum to give 2,5-dichloro-3-nitrobenzoic acid (3.4 g,
65%).
Step 2: Synthesis of methyl 2,5-dichloro-3-nitrobenzoate
##STR00542##
[1184] To stirred solution of 2,5-dichloro-3-nitrobenzoic acid (3.2
g, 13.67 mmol) in DMF (30 mL) was added sodium carbonate (4.3 g,
40.56 mmol) and methyl iodide (9.7 g, 68.30 mmol). The reaction
mixture was heated at 60.degree. C. for 18 h. On completion, water
was added to the reaction mixture, and the product was extracted
using DCM. The combined organic layers were dried and concentrated
under reduced pressure to give methyl 2,5-dichloro-3-nitrobenzoate
(3 g, 90%).
Step 3: Synthesis of methyl 3-amino-2,5-dichlorobenzoate
##STR00543##
[1186] To stirred solution of methyl 2,5-dichloro-3-nitrobenzoate
(3.3 g, 13.1 mmol) in ethanol (20 mL) was added ammonium chloride
(3.5 g, 64.81 mmol) dissolved in water (20 mL) and iron powder (7
g, 82.35 mmol). The reaction mixture was heated at 80.degree. C.
for 3 h. On completion, water was added to the reaction mixture and
the solids removed by filtration through celite. The filtrate was
extracted with ethyl acetate. The combined organic layers were
washed with water, dried, and concentrated under reduced pressure
to give methyl 3-amino-2,5-dichlorobenzoate (2.5 g, 66%).
Step 4: Synthesis of methyl 2, 5-dichloro-3-(cyclopentylamino)
benzoate
##STR00544##
[1188] To a stirred solution of methyl 3-amino-2,5-dichlorobenzoate
(2.5 g, 11.5 mmol) and cyclopentanone (4.8 g, 57.1 mmol) in
methanol (5 mL) was added acetic acid (0.6 g, 11.3 mmol) and
stirring was continued at room temperature for 3 h. Then sodium
cyanoborohydride (1.4 g, 22.2 mmol) was added and the reaction
stirred overnight. On completion, the solvent was removed under
reduced pressure and the product was purified by column
chromatography to afford methyl 2, 5-dichloro-3-(cyclopentylamino)
benzoate (0.275 g, 8%).
Step 5: Synthesis of methyl 2, 5-dichloro-3-(cyclopentyl (methyl)
amino) benzoate
##STR00545##
[1190] To a stirred solution of methyl 2,
5-dichloro-3-(cyclopentylamino) benzoate (0.275 g, 0.958 mmol) in
acetonitrile (5 mL) was added cesium carbonate (0.622 g, 1.9 mmol)
and methyl iodide (0.689 g, 4.78 mmol). The reaction mixture was
heated at 80.degree. C. for 18 h. On completion, the reaction was
cooled to room temperature and filtered. The residue was washed
with ethyl acetate and the filtrate was concentrated to afford
methyl 2, 5-dichloro-3-(cyclopentyl (methyl) amino) benzoate (0.27
g, 93%).
Step 6: Synthesis of 2, 5-dichloro-3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)
benzamide
##STR00546##
[1192] Aqueous NaOH (0.50 g, 1.24 mmol) was added to a solution of
methyl 2, 5-dichloro-3-(cyclopentyl (methyl) amino) benzoate (0.25
g, 0.83 mmol) in MeOH (5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, methanol was removed under
reduced pressure and acidified using dilute HCl up to pH 6 and to
pH 4 using citric acid. The product was extracted using ethyl
acetate. The combined organic layers were dried and concentrated to
give the desired acid (0.21 g, 88%).
[1193] The acid (0.21 g, 0.755 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.3 g, 1.97
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.757 g, 1.45 mmol) was added
to it and stirring was continued for overnight. After completion of
the reaction, reaction mixture was poured into ice to obtain a
solid. The solid was filtered and washed with acetonitrile. The
compound was further purified by prep. HPLC to afford 2,
5-dichloro-3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)
benzamide (0.1 g, 36%) as TFA salt. LCMS: 422.10 (M+1).sup.+; HPLC:
97.58% (@254 nm) (R.sub.t; 6.973; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.35 (t, 1H), 7.23
(s, 1H), 6.98 (s, 1H), 5.85 (s, 1H), 4.24 (d, 2H, J=4.4 Hz), 3.66
(m, 1H), 2.59 (s, 3H), 2.18 (s, 3H), 2.10 (s, 3H), 1.70 (bs, 2H),
1.61 (bs, 2H), 1.48 (bs, 4H).
Example 10
Synthesis of Compound 14:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00547##
[1194] Step 1: 5-chloro-2-methyl-3-nitrobenzoic acid
##STR00548##
[1196] 2-Methyl-3-nitrobenzoic acid (4 g, 23.39 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C. lot wise.
After 10 minutes nitrating mixture {prepared as mixing Conc. HNO3
(3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4 (4.4 mL)} was added
drop wise at -10.degree. C. Resulting reaction mass was stirred at
-10.degree. C. for 30 minutes. On completion, reaction mixture was
poured on ice cold water, solid precipitated was filtered, washed
with water and dried under vacuum giving desired compound (4.95 g,
99%).
Step 2: methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00549##
[1198] To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic
acid (6.75 g, 31.25 mmol) in DMF (33 mL), sodium carbonate (13.23
g, 125.18 mmol) and methyl iodide (17.77 g, 125.2 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 4 h. On
completion, water was added to the reaction mass and extraction was
carried out using DCM. Combined organic layers were dried,
concentrated under reduced pressure and purified by column
chromatography over silica (60-120 mesh size) giving desired
compound (6 g, 84%).
Step 3: methyl 3-amino-5-chloro-2-methyl benzoate
##STR00550##
[1200] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 26.13 mmol) in ethanol (60
mL), ammonium chloride (6 g, 112.1 mmol) dissolved in water (60 mL)
and iron powder (11.88 g, 208.4 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 1 h. On
completion, water was added to reaction mass and reaction mixture
was filtered through celite, filtrate was extracted with ethyl
acetate. Combined organic layers were washed with water, dried,
concentrated under reduced pressure giving desired compound which
was used as is.
Step 4: methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino) benzoate
##STR00551##
[1202] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (1 g, 5.025 mmol) and dihydro-2H-pyran-4(3H)-one (753 mg,
7.537 mmol) in methanol (10 mL), acetic acid (301 mg, 5.025 mmol)
was added and reaction stirred at room temperature for 3 h. Then
sodium cyanoborohydride (373 mg, 6.03 mmol) was added and reaction
stirred overnight. On completion, solvent was removed under reduced
pressure and crude material was purified by column chromatography
to afford the desired compound (700 mg, 49%).
Step 5: methyl
5-chloro-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)
benzoate
##STR00552##
[1204] To a stirred solution of methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino) benzoate (700
mg, 2.473 mmol) in acetonitrile (10 mL), cesium carbonate (1.61 g,
4.946 mmol) and methyl iodide (1.82 g, 0.8 ml, 12.36 mmol) were
added; resulting reaction mass was heated at 80.degree. C. for 13
h. On completion, reaction mass was cooled to room temperature and
filtered, residue was washed with ethyl acetate and filtrate was
concentrated and then purified by column chromatography to afford
desired compound (480 mg, 65%).
Step 6:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00553##
[1206] Aqueous NaOH (96 mg, 2.424 mmol) was added to a solution of
methyl 5-chloro-2-methyl-3-(methyl(tetrahydro-2H-pyran-4-yl)amino)
benzoate (0.48 g, 1.616 mmol) in EtOH (2 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.4 g, 87%).
[1207] The acid (0.4 g, 1.41 mmol) was then dissolved in DMSO (1
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.429 g,
2.82 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.10 g, 2.12 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed column purification to provide the title compound (0.17 g,
29%): LCMS: 418.25 (M+1).sup.+; HPLC: 97.72% (@254 nm) (R.sub.t;
5.022; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.48 (s, 1H), 8.21 (t, 1H, J=4.4 Hz), 7.13 (s, 1H), 6.92 (s, 1H),
5.85 (s, 1H), 4.24 (d, 2H, J=4.4 Hz), 3.84 (d, 2H, J=10.8 Hz), 3.25
(t, 2H, J=11.2 Hz), 2.96 (m, 1H), 2.56 (s, 3H), 2.18 (s, 3H), 2.15
(s, 3H), 2.10 (s, 3H), 1.65-1.52 (m, 4H).
Example 11
Synthesis of Compound 15: tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate
##STR00554##
[1208] Step 1: Synthesis of methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino) cyclohexyl)
amino)-2-methylbenzoate
##STR00555##
[1210] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (5 g, 20.6 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (5.6 g, 26.7 mmol) in methanol (50 mL)
was added acetic acid (1.2 g, 20.6 mmol) and the mixture was
stirred at room temperature for 3 h. Then sodium cyanoborohydride
(1.6 g, 26.7 mmol) was added and the reaction stirred overnight. On
completion, the solvent was removed under reduced pressure and the
product purified by column chromatography to afford compound methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino) cyclohexyl)
amino)-2-methylbenzoate (3 g, 33.3%).
Step 2: Synthesis of methyl 5-bromo-3-((4-((tert-butoxycarbonyl)
amino) cyclohexyl) (methyl) amino)-2-methylbenzoate
##STR00556##
[1212] To a stirred solution of methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino) cyclohexyl)
amino)-2-methylbenzoate (1.5 g, 4.6 mmol) in acetonitrile (15 mL)
was added cesium carbonate (2.63 g, 8.1 mmol) and methyl iodide
(2.86 g, 20.32 mmol). The mixture was heated at 80.degree. C. for 7
h. On completion, the reaction was cooled to room temperature,
filtered, and the residue was washed with ethyl acetate. The
filtrate was concentrated to afford methyl
5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl) (methyl)
amino)-2-methylbenzoate (1.5 g).
Step 3: Synthesis of tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate
##STR00557##
[1214] Aqueous NaOH (0.26 g, 6.62 mmol) was added to a solution of
methyl 5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl)
(methyl) amino)-2-methylbenzoate (1.5 g, 3.3 mmol) in MeOH (20 mL)
and stirred at 60.degree. C. for 1 h. After completion of the
reaction, methanol was removed under reduced pressure and the
mixture was acidified using dilute HCl up to pH 6 and to pH 4 using
citric acid. The product was extracted with ethyl acetate. Combined
organic layers were dried and concentrated to give the desired acid
(1.33 g, 92%).
[1215] The acid (1.3 g, 2.96 mmol) was then dissolved in DMSO (10
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.34 g, 2.2
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.88 g, 1.7 mmol) was added to
it and stirring was continued for overnight. After completion of
the reaction, the mixture was poured onto ice to obtain solid. The
solid was filtered and washed with acetonitrile followed by
purification with column chromatography to afford tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate. (0.5 g,
26%). LCMS: 575.80 (M+1).sup.+; HPLC: 99.99% (@254 nm) (R.sub.t;
6.094, 7.065 [mixture of diastereomers]; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.20 (d, 1H, J=3.6
Hz), 7.21 (s, 1H), 7.18 (s, 1H), 7.01 (s, 1H), 6.66 (d, 1H, J=7.2
Hz), 5.85 (s, 1H), 4.23 (d, 2H, J=3.2 Hz), 3.44 (bs, 1H), 3.15 (bs,
1H), 2.55-2.66 (m, 3H), 2.18 (s, 3H), 2.10 (s, 6H), 1.76-1.79 (m,
2H), 1.51-1.63 (m, 3H), 1.36-1.38 (m, 11H), 1.07-1.16 (m, 1H).
Example 12
Synthesis of Compound 16:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methyl-5-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)benzamide
##STR00558##
[1216] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00559##
[1218] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mass was poured on ice cold water, solid precipitated was filtered,
resulting residue was washed with water and dried under vacuum
giving the desired compound (71.7 g, 100%)
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00560##
[1220] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid
(287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415
mmol) and methyl iodide (626.63 g, 4415 mmol) were added. Resulting
reaction mass was heated at 60.degree. C. for 8 h. On completion,
solid precipitated was filtered, residue washed with diethyl ether
(5 times). Combined organic layers were dried, concentrated under
reduced pressure giving the desired crude compound (302 g, 99%)
which was used without further purification.
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00561##
[1222] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 7 h. On
completion, reaction mass was filtered through celite giving
washing of water and ethyl acetate, filtrate was extracted with
ethyl acetate. Combined organic layers were dried, concentrated
under reduced pressure giving the desired compound which was used
further as is.
Step 4: methyl 5-bromo-3-(cyclopentylamino)-2-methylbenzoate
##STR00562##
[1224] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (0.3 g, 1.33 mmol) and
cyclopentanone (0.56 g, 6.6 mmol) in methanol (3 mL), acetic acid
(0.159 g, 2.6 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.208 g, 3.3
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
used as is in the next step.
Step 5: Synthesis of methyl
5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate
##STR00563##
[1226] A solution of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate (1 g, 3.22 mmol),
Cs.sub.2CO.sub.3 (2.10 g, 6.45 mmol) and methyl iodide (2.3 g,
16.12 mmol) in acetonitrile (15 mL) was heated at 80.degree. C. for
14 h. After complete consumption of starting material reaction
mixture was filtered. The filtrate was concentrated and the
obtained residue was purified by column chromatography to provide
the desired compound (0.9 g, 86%).
Step 6:
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methylbenzamide
##STR00564##
[1228] Aqueous NaOH (0.166 g, 4.15 mmol) was added to a solution of
methyl 5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (0.9
g, 2.76 mmol) in EtOH (9 mL) and water (2.2 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.85 g, 98%).
[1229] The acid (0.85 g, 2.73 mmol) was then dissolved in DMSO (10
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.83 g,
5.46 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (2.13 g, 4.09 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed by ether to provide the desired compound (0.450 g,
37%).
Step 7: tert-butyl
4-(3-(cyclopentyl(methyl)amino)-5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)carbamoyl)-4-methylphenyl)-5,6-dihydropyridine-1(2H)-carboxy-
late
##STR00565##
[1231] To a degassed solution of
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methylbenzamide (1.0 g, 2.24 mmol) and boronic acid
(0.833 g, 2.69 mmol) in dioxane (20 mL) was added
Pd(PPh.sub.3).sub.4 (0.260 g, 0.224 mmol, 0.1 eq) and the solution
was again purged with argon for 20 min. To the reaction mixture Aq
Na.sub.2CO.sub.3 (0.857, 8.08 mmol) solution in 1 mL water was
added and reaction mixture was again purged with argon for 20 min.
The reaction was heated at 100.degree. C. for 90 min. The reaction
mixture was then cooled to room temperature and poured in to cold
water and extracted with dichloromethane. Crude compound purified
by column chromatography to provide the desired compound (0.900 g,
64%).
Step 8:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methyl-5-(1,2,3,6-tetrahydropyridin-4-yl)benzamide
##STR00566##
[1233] To a stirred solution of tert-butyl
4-(3-cyclopentyl)methyl)amino)-5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-
-3-yl)methyl)carbamoyl)-4-methylphenyl)-5,6-dihydropyridine-1(2H)-carboxyl-
ate (0.800 g, 1.45 mmol) in DCM was added TFA (3 mL) and the
reaction mixture was stirred at RT for 2 h. On completion, reaction
was concentrated to dryness. Residue was basified with aqueous
sodium bicarbonate till pH 8 and aqueous layer extracted with 20%
MeOH/DCM. Combined organic layers were dried over sodium sulfate
and concentrated to afford crude material which was purified by
preparative HPLC to afford the desired compound (0.620 g, 95%).
Step 9:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methyl-5-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)benz-
amide
##STR00567##
[1235]
3-(Cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyri-
din-3-yl)methyl)-2-methyl-5-(1,2,3,6-tetrahydropyridin-4-yl)benzamide
(0.100 g, 0.223 mmol) was dissolved in methanol (5 mL) and cooled
to 0.degree. C., formalin (0.067 g, 0.19 mL, 2.23 mmol) was added.
Resulting reaction mass was stirred at same temperature for 30
minutes. Sodium cyanoborohydride (0.041 g, 0.66 mmol) was added to
above reaction mass and stirred at room temperature for 4 h. After
completion, solvent were removed under reduced pressure and water
was added to the residue, extraction was carried out using DCM.
Combined organic layers were dried, concentrated and purified by
column chromatography to provide the desired compound (0.065 g,
63%): LCMS: 463.40 (M+1).sup.+; HPLC: 98.79% (@254 nm) (R.sub.t;
3.911; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.43 (s, 1H), 8.04 (t, 1H, J=4.8 Hz), 7.17 (s, 1H), 6.95 (s, 1H),
6.07 (s, 1H), 5.85 (s, 1H), 4.25 (d, 2H, J=4.8 Hz), 3.45 (m, 1H),
2.98 (bs, 2H), 2.53 (m, 2H), 2.42 (bs, 2H), 2.26 (s, 3H), 2.18 (s,
3H), 2.17 (s, 3H), 2.10 (s, 3H), 1.67 (m, 2H), 1.59 (m, 2H), 1.49
(m, 2H), 1.40 (m, 2H).
Example 13
Synthesis of Compound 17:
3-((1-acetylpiperidin-4-yl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo--
1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00568##
[1237] The
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)--
2-methyl-3-(methyl(piperidine-4-yl)amino)benzamide (0.2 g, 0.447
mmol) dissolved in DMSO (2 mL) and acetic acid (0.054 g, 0.896
mmol) were added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.342 g, 0.672 mmol) was added
to it and stirring was continued for overnight. After completion,
reaction mass was poured into ice, and extracted using 10%
MeOH/DCM. Combined organic layers were dried, concentrated to
provide crude material, which then purified with column
chromatography using silica (100-200 mesh size) to afford the
desired compound (0.15 g, 17%). LCMS: 503.20 (M+1).sup.+; HPLC:
96.06% (@254 nm) (R.sub.t; 5.143; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.48 (s, 1H), 8.21 (t, 1H, J=4.8
Hz), 7.23 (d, 1H, J=1.6 Hz), 7.04 (d, 1H, J=1.6 Hz), 5.85 (s, 1H),
4.32 (d, 1H, J=12.4 Hz), 4.23 (d, 2H, J=4.8 Hz), 3.77 (d, 1H,
J=13.2 Hz), 2.98 (t, 2H, J=11.2 Hz), 2.54 (s, 3H), 2.18 (s, 3H),
2.13 (s, 3H), 2.10 (s, 3H), 1.97 (s, 3H), 1.53-1.60 (m, 4H),
1.38-1.41 (m, 1H).
Example 14
Synthesis of Compound 18:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methyl-5-(1-methylpiperidin-4-yl)benzamide
##STR00569##
[1239] To a solution of
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methyl-5-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)benzamide
(0.045 g, 0.097 mmol) in ethanol added 10% Pd/C and the reaction
was stirred under H.sub.2 at bladder pressure. After complete
consumption of the starting material reaction mixture was filtered
through celite. The filtrate was concentrated and triturated in
acetonitrile and filtered to provide the desired compound (0.030 g,
66%): LCMS: 465.40 (M+1).sup.+; HPLC: 95.39% (@254 nm) (R.sub.t;
3.884; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.44 (s, 1H), 7.98 (t, 1H), 7.00 (s, 1H), 6.77 (s, 1H), 5.85 (s,
1H), 4.25 (d, 2H, J=4.4 Hz), 3.42 (m, 1H), 2.84 (d, 2H, J=11.2 Hz),
2.47 (s, 3H), 2.37 (m, 1H), 2.18 (s, 6H), 2.15 (s, 3H), 2.10 (s,
3H), 1.93 (t, 2H, J=10.8 Hz), 1.70-1.55 (m, 8H), 1.48 (m, 2H), 1.38
(m, 2H).
Example 15
Synthesis of Compound 20: 3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(tri fluoromethyl) benzamide
##STR00570##
[1240] Step 1: Synthesis of 2-methyl-3-nitro-5-(trifluoromethyl)
benzoic acid
##STR00571##
[1242] 2-methyl-5-(trifluoromethyl) benzoic acid (4 g, 19.6 mmol)
was added to cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C.
in small portions. After 10 minutes nitrating mixture {prepared by
mixing conc. HNO.sub.3 (2.77 g, 44.44 mmol) with conc.
H.sub.2SO.sub.4 (3.6 mL)} was added drop wise at -10.degree. C. The
resulting solution was stirred at -10.degree. C. for 30 minutes. On
completion, the reaction mixture was poured into ice cold water.
The solid was filtered, washed with water and dried under vacuum to
give 2-methyl-3-nitro-5-(trifluoromethyl) benzoic acid (4.95 g,
99%).
Step 2: Synthesis of methyl 2-methyl-3-nitro-5-(trifluoromethyl)
benzoate
##STR00572##
[1244] To stirred solution of 2-methyl-3-nitro-5-(trifluoromethyl)
benzoic acid (1 g, 4.01 mmol) in DMF (3 mL) was added sodium
carbonate (0.63 g, 6.02 mmol) and methyl iodide (1.14 g, 8.03
mmol). The resulting mixture was heated at 60.degree. C. for 4 h.
On completion, water was added to the reaction, and the product
extracted using DCM. Combined organic layers were dried,
concentrated under reduced pressure and purified by column
chromatography over silica to give methyl
2-methyl-3-nitro-5-(trifluoromethyl) benzoate (1 g, 95.2%).
Step 3: Synthesis of methyl 3-amino-2-methyl-5-(trifluoromethyl)
benzoate
##STR00573##
[1246] To a stirred solution of methyl
2-methyl-3-nitro-5-(trifluoromethyl) benzoate (5.6 g, 20.97 mmol)
in ethanol (40 mL) was added ammonium chloride (5.6 g, 104.6 mmol)
dissolved in water (30 mL) and iron powder (4.67 g, 85.16 mmol).
The resulting mixture was heated at 80.degree. C. for 1 h. On
completion, water was added to the reaction then the mixture was
filtered through celite. The filtrate was extracted with ethyl
acetate. The combined organic layers were dried then concentrated
under reduced pressure to give methyl
3-amino-2-methyl-5-(trifluoromethyl) benzoate.
Step 4: Synthesis of methyl
3-(cyclopentylamino)-2-methyl-5-(trifluoromethyl)benzoate
##STR00574##
[1248] To a stirred solution of methyl
3-amino-2-methyl-5-(trifluoromethyl) benzoate (2.5 g, 10.7 mmol)
and cyclopentanone (4.5 g, 53.49 mmol) in methanol (25 mL) was
added acetic acid (1.98 g, 21.44 mmol). After 3 h, sodium
cyanoborohydride (1.68 g, 24.3 mmol) was added and the reaction
stirred overnight. On completion, the solvent was removed under
reduced pressure and crude material was purified by column
chromatography to afford methyl
3-(cyclopentylamino)-2-methyl-5-(trifluoromethyl)benzoate (1.8 g,
55.9%).
Step 5: Synthesis of methyl 3-(cyclopentyl (methyl)
amino)-2-methyl-5-(trifluoromethyl) benzoate
##STR00575##
[1250] To a stirred solution of methyl
3-(cyclopentylamino)-2-methyl-5-(trifluoromethyl)benzoate (1.6 g,
5.38 mmol) in acetonitrile (25 mL) was added cesium carbonate (3.5
g, 10.7 mmol) and methyl iodide (3.8 g, 26.8 mmol). The mixture was
heated at 80.degree. C. for 8 h. On completion, the reaction was
cooled to room temperature and filtered. The residue was washed
with ethyl acetate and the filtrate was concentrated to give methyl
3-(cyclopentyl (methyl) amino)-2-methyl-5-(trifluoromethyl)
benzoate.
Step 6: Synthesis of 3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(trifluoromethyl) benzamide
##STR00576##
[1252] Aqueous NaOH (0.762 g, 15.4 mmol) was added to a solution of
methyl 3-(cyclopentyl (methyl) amino)-2-methyl-5-(trifluoromethyl)
benzoate (2 g, 6.34 mmol) in MeOH (20 mL) and the mixture was
stirred at 60.degree. C. for 1 h. After completion of the reaction,
methanol was removed under reduced pressure and the residue
acidified using dilute HCl up to pH 6 and to pH 4 using citric
acid. The product was extracted using ethyl acetate. The combined
organic layers were dried and concentrated to give the desired acid
(1.5 g, 78%).
[1253] The acid (0.5 g, 1.69 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.505 g,
3.32 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.29 g, 2.49 mmol) was
added and stirring was continued overnight. After completion of the
reaction, the mixture was poured onto ice to obtain a solid. The
solid was filtered and washed with acetonitrile followed by ether
to provide compound 3-(cyclopentyl
(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(trifluoromethyl) benzamide (100 mg, 14%).
Analytical Data: LCMS: 436.20 (M+1).sup.+; HPLC: 89.22% (@254 nm)
(R.sub.t; 6.094; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.47 (s, 1H), 8.30 (t, 1H), 7.37 (s, 1H), 7.18 (s, 1H), 5.86 (s,
1H), 4.26 (d, 2H, J=4.4 Hz), 3.51 (m, 1H), 2.53 (s, 3H), 2.25 (s,
3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.41-1.69 (m, 8H).
Example 16
Synthesis of Compound 21: 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide
##STR00577##
[1254] Step 1: Synthesis of methyl 3-amino-2-methylbenzoate
##STR00578##
[1256] To stirred solution of methyl 3-amino-2-methylbenzoic acid
(5 g, 33.1 mmol) in methanol (50 mL), was added conc.
H.sub.2SO.sub.4 (5 mL) at 0.degree. C. and the mixture was heated
at 70.degree. C. for 22 h. On completion, methanol was removed
under reduced pressure, and then sat. sodium bicarbonate solution
was added and the product extracted with 10% MeOH/DCM. Combined
organic layers were dried, concentrated to yield methyl
3-amino-2-methylbenzoate (5 g, 91%).
Step 2: Synthesis of methyl
3-(cyclopentylamino)-2-methylbenzoate
##STR00579##
[1258] To a stirred solution of methyl 3-amino-2-methylbenzoate (2
g, 12.1 mmol) and cyclopentanone (5 g, 60.6 mmol) in methanol (20
mL) was added acetic acid (0.72 g, 12.12 mmol) and the reaction
stirred at room temperature for 3 h. Sodium cyanoborohydride (1.13
g, 18.2 mmol) was added and the reaction stirred overnight. On
completion, the solvent was removed under reduced pressure and the
product was purified by column chromatography to afford methyl
3-(cyclopentylamino)-2-methylbenzoate (2.2 g, 78%).
Step 3: Synthesis of methyl 3-(cyclopentyl (methyl)
amino)-2-methylbenzoate
##STR00580##
[1260] To a stirred solution of methyl
3-(cyclopentylamino)-2-methylbenzoate (2.2 g, 9.44 mmol) in
acetonitrile (20 mL) was added cesium carbonate (6.13 g, 18.8 mmol)
and methyl iodide (6.7 g, 47.2 mmol). The resulting mixture was
heated at 80.degree. C. for 12 h. On completion, the mixture was
cooled to room temperature and filtered, washing with ethyl
acetate. The filtrate was concentrated and then purified by column
chromatography to afford methyl 3-(cyclopentyl (methyl)
amino)-2-methylbenzoate (2 g, 85%).
Step 4: Synthesis of 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide
##STR00581##
[1262] Aqueous NaOH (0.48 g, 12.14 mmol) was added to a solution of
methyl 3-(cyclopentyl (methyl) amino)-2-methylbenzoate (2 g, 8.09
mmol) in MeOH (20 mL) and stirred at 60.degree. C. for 1 h. After
completion of the reaction, methanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate. Combined organic layers were dried and concentrated to
give the desired acid (1.3 g, 69%).
[1263] The acid (0.3 g, 1.28 mmol) was then dissolved in DMSO (5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.39 g, 2.5
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.99 g, 1.92 mmol) was added
to it and stirring was continued for overnight. After completion of
the reaction, mixture was poured into ice to obtain solid, this was
filtered and washed with acetonitrile followed column purification
to provide 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methylbenzamide (0.020 g, 4%).
[1264] Analytical Data: LCMS: 368.15 (M+1).sup.+; HPLC: 99.49%
(@254 nm) (R.sub.t; 4.186; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.43 (s, 1H), 8.00 (t, 1H, J=4
Hz), 7.09-7.16 (m, 2H), 6.90 (d, 1H, J=6.8 Hz), 5.84 (s, 1H), 4.25
(d, 2H, J=5.2 Hz), 3.41 (t, 1H, J=6.8 Hz), 2.47 (s, 3H), 2.18 (d,
6H, J=4.4 Hz), 2.10 (s, 3H), 1.58-1.67 (m, 4H), 1.37-1.52 (m,
4H).
Example 17
Synthesis of Compound 23:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(methyl(1-methylpiperidin-4-yl)amino)benzamide
##STR00582##
[1266]
5-Chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-m-
ethyl-3-(methyl(piperidin-4-yl)amino)benzamide (0.08 g, 0.192 mmol)
was dissolved in methanol (5 mL) and cooled to 0.degree. C.,
formalin (0.028 g, 0.965 mmol) was added. Resulting reaction mass
was stirred at same temperature for 30 minutes. Sodium
cyanoborohydride (0.023 g, 0.38 mmol) was added to above reaction
mass and stirred at room temperature for 4 h. After completion,
solvent were removed under reduced pressure and water was added to
the residue, extraction was carried out using DCM. Combined organic
layers were dried, concentrated and purified by column
chromatography giving the desired compound (0.02 g, 24%): LCMS:
431.25 (M+1).sup.+; HPLC: 98.80% (@254 nm) (R.sub.t; 4.757; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.20 (t,
1H), 7.15 (d, 1H, J=1.2 Hz), 6.94 (d, 1H, J=2 Hz), 5.85 (s, 1H),
4.25 (d, 2H, J=4.8 Hz), 3.05 (bs, 2H), 2.88 (bs, 1H), 2.55 (s, 3H),
2.44 (bs, 2H), 2.18 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 1.68 (bs,
4H).
Example 18
Synthesis of Compound 25:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methyl-5-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-
benzamide
##STR00583##
[1267] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00584##
[1269] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mass was poured on ice cold water, solid precipitated was filtered,
resulting residue was washed with water and dried under vacuum to
give the desired compound (71.7 g, 100%).
Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00585##
[1271] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g,
4415 mmol) and methyl iodide (626.63 g, 4415 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 8 h. On
completion, solid precipitated was filtered, residue washed with
diethyl ether (5 times). Combined organic layers were dried,
concentrated under reduced pressure giving the desired crude
compound (302 g, 99%).
Step 2: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00586##
[1273] To stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 7 h. On
completion, reaction mass was filtered through celite giving
washing of water and ethyl acetate, filtrate was extracted with
ethyl acetate. Combined organic layers were dried, concentrated
under reduced pressure giving the desired compound.
Step 3: methyl 5-bromo-3-(cyclopentylamino)-2-methylbenzoate
##STR00587##
[1275] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (0.3 g, 1.33 mmol) and
cyclopentanone (0.56 g, 6.6 mmol) in methanol (3 mL), acetic acid
(0.159 g, 2.6 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.208 g, 3.3
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure to give the desired
compound.
Step 4: methyl
5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate
##STR00588##
[1277] To a stirred solution of the crude methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate (0.7 g, 2.25 mmol) in
acetonitrile (15 mL), cesium carbonate (1.47 g, 4.50 mmol) and
methyl iodide (1.6 g, 11.26 mmol) were added; resulting reaction
mass was heated at 80.degree. C. for 7 h. On completion, reaction
mass was cooled to room temperature and filtered, residue was
washed with ethyl acetate and filtrate was concentrated and then
purified by column chromatography to afford the desired compound
(0.6 g, 82%).
Step 5:
5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00589##
[1279] Aqueous NaOH (0.11 g, 2.75 mmol) was added to a solution of
methyl 5-bromo-3-(cyclopentyl(methyl)amino)-2-methylbenzoate (0.6
g, 1.8 mmol) in MeOH (1.5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate. The combined organic layers were dried and concentrated to
give the respective acid (0.5 g, 87%).
[1280] The acid (0.5 g, 1.60 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.49 g,
3.22 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.25 g, 2.41 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed by ether to provide the desired compound (0.315 g,
44%).
Step 6:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methyl-5-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridi-
n-4-yl)benzamide
##STR00590##
[1282] A solution of
5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2-methylbenzamide (1 equiv.),
(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)boronic acid
(1.2 equiv.) and Pd (PPh.sub.3).sub.4 (0.1 equiv.) in 1,4-dioxane
(4 mL) was purged with argon for 10 min. Then, 2 M Na.sub.2CO.sub.3
solution (3.6 equiv.) was added to it and argon was purged again
for 10 min. The reaction mixture was stirred at 100.degree. C. for
2 h. After completion of the reaction, water was added to it and
extraction was carried out using DCM. The combined organic layers
were washed with water, dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated under reduced pressure to afford crude
material which was purified preparative HPLC to give the desired
compound as its TFA salt (0.045, 23%). LCMS: 505.39 (M+1).sup.+;
HPLC: 99.95% (@254 nm) (R.sub.t; 4.091; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR (MeOD, 400
MHz) .delta. 7.70 (s, 1H), 7.46 (s, 1H), 6.21 (s, 1H), 6.13 (s,
1H), 4.48 (s, 2H), 4.23 (bs, 1H), 3.14 (s, 3H), 2.69 (s, 2H), 2.42
(s, 3H), 2.39 (s, 3H), 2.24 (s, 3H), 1.92 (, 2H), 1.81 (m, 2H),
1.67 (m, 4H), 1.60 (s, 6H), 1.53 (s, 6H).
Example 19
Synthesis of Compound 26: tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)amino)cyclohexyl)carbamate
##STR00591##
[1283] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00592##
[1285] To a mixture of 2-methyl-3-nitrobenzoic acid (15 g, 82.80
mmol) in conc. H.sub.2SO.sub.4 (60 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (13.07 g, 45.71
mmol) was added and reaction mixture was stirred at room
temperature for 5 h. After completion of reaction, reaction mixture
was slowly poured onto ice cold water (400 mL). The precipitate was
collected by filtration to give the desired compound (21 g,
98.22%).
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzoate
##STR00593##
[1287] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (16 g, 61.54 mmol) in DMF (160 mL), iodomethane (35.72 g, 248
mmol) and sodium carbonate (26.28 g, 248 mmol) were added.
Resulting reaction mass was stirred at 60.degree. C. for 8 h. On
completion, reaction mass was filtered and inorganic solid residue
washed with ethyl acetate. Combined filtrate was concentrated under
vacuum till dryness. The residue was re-dissolved in ethyl acetate
and washed with 5% sodium bicarbonate solution (700 mL) followed by
5M HCl solution (300 mL). Organic layer was finally washed with
brine, dried over sodium sulfate, filtered and concentrated to
afford the desired compound (16 g, 94.50%).
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00594##
[1289] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzoate (17 g, 62.04 mmol) in ethanol (85
mL), was added NH.sub.4Cl solution (17 g in 85 mL water, 317.8
mmol) followed by Fe powder (27.82 g, 498.11 mmol). Resulting
reaction mass was stirred at 90.degree. C. for 1 h. On completion,
reaction mass was filtered and filtrate was concentrated till
dryness to get solid which was dissolved in saturated sodium
bicarbonate solution. The aqueous layer was extracted with ethyl
acetate. Combined organic layers were dried over sodium sulfate and
concentrated to afford the desired compound 4 (15 g, 99%).
Step 4: methyl 5-bromo-3-((4-((tert-butoxycarbonyl)
amino)cyclohexyl)amino)-2-methylbenzoate
##STR00595##
[1291] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (5 g, 20.57 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (5.67 g, 26.74 mmol) in methanol (50
mL), acetic acid (1.2 g, 20.57 mmol) was added and reaction stirred
at room temperature for 4 h. Then sodium cyanoborohydride (1.68 g,
26.74 mmol) was added and reaction stirred 18 h. On completion,
solvent was removed under reduced pressure; water was added and
extracted with DCM. Combined organic were dried, concentrated and
purified by column chromatography using silica (100-200 mesh size)
to afford the desired compound (3 g, 33%).
Step 5: tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)amino)cyclohexyl)carbamate
##STR00596##
[1293] Aqueous NaOH (0.247 g, 6.83 mmol) was added to a solution of
methyl 5-bromo-3-((4-((tert-butoxycarbonyl)
amino)cyclohexyl)amino)-2-methylbenzoate (1.5 g, 3.41 mmol) in MeOH
(20 mL) and stirred at 60.degree. C. for 1 h. After completion of
the reaction, ethanol was removed under reduced pressure and
acidified using dilute HCl up to pH 6 and pH 4 was adjusted using
citric acid. Extraction was carried out using DCM. Combined organic
layers were dried concentrated giving respective acid (1.28 g,
89%).
[1294] The acid (1.28 g, 3.01 mmol) was then dissolved in DMSO (10
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (1.03 g,
6.83 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (2.6 g, 5.12 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice and
extracted with 5% MeOH/DCM. The combined organic layers were dried
and concentrated to obtain a crude solid, which was purified by
column chromatography to afford the desired compound (0.75 g, 45%).
LCMS: 561.15 (M+1).sup.+; HPLC: 99.99% (@254 nm) (R.sub.t; 6.876,
7.080 [mixture of diastereomers]; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.44 (s, 1H), 8.07 (t, 1H, J=4.8
Hz), 6.77 (d, 1H, J=7.6 Hz), 6.66 (s, 1H), 6.51-6.54 (m, 1H), 5.84
(s, 1H), 4.71 (d, 1H, J=7.6 Hz), 4.21-4.22 (m, 2H), 3.44 (bs, 1H),
3.19 (bs, 1H), 2.16 (s, 3H), 2.10 (s, 3H), 1.93-2.03 (m, 5H), 1.78
(s, 1H), 1.56-1.72 (m, 3H), 1.37 (s, 9H), 1.26-1.31 (m, 2H).
Example 20
Synthesis of Compound 28:
5-bromo-3-(cyclohexylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-2-methylbenzamide
##STR00597##
[1295] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00598##
[1297] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mass was poured on ice cold water, solid precipitated was filtered,
resulting residue was washed with water and dried under vacuum
giving the desired compound (71.7 g, 100%).
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00599##
[1299] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid
(287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415
mmol) and methyl iodide (626.63 g, 4415 mmol) were added. Resulting
reaction mass was heated at 60.degree. C. for 8 h. On completion,
solid precipitated was filtered, residue washed with diethyl ether
(5 times). Combined organic layers were dried, concentrated under
reduced pressure giving the desired compound (302 g, 99%).
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00600##
[1301] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 7 h. On
completion, reaction mass was filtered through celite giving
washing of water and ethyl acetate, filtrate was extracted with
ethyl acetate. Combined organic layers were dried, concentrated
under reduced pressure to give the desired compound which was used
directly in the next step.
Step 4: methyl 5-bromo-3-(cyclohexylamino)-2-methylbenzoate
##STR00601##
[1303] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (0.3 g, 1.33 mmol) and
cyclohexanone (0.64 g, 6.6 mmol) in methanol (3 mL), acetic acid
(0.159 g, 2.6 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.208 g, 3.3
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and was used directly in
the next step.
Step 5:
5-bromo-3-(cyclohexylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyri-
din-3-yl)methyl)-2-methylbenzamide
##STR00602##
[1305] Aqueous NaOH (0.049 g, 1.23 mmol) was added to a solution of
methyl 5-bromo-3-(cyclohexylamino)-2-methylbenzoate (0.2 g, 0.61
mmol) in MeOH (5 mL) and stirred at 60.degree. C. for 1 h. After
completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate. Combined organic layers were dried concentrated giving
respective acid (0.17 g, 89%).
[1306] The acid (0.17 g, 0.54 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.099 g,
0.65 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.42 g, 0.817 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was purified by preparative HPLC to provide the
desired compound as its TFA salt (0.042 g, 15%). LCMS: 446.16
(M+1).sup.+; HPLC: 99.85% (@254 nm) (R.sub.t; 6.918; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.08 (s,
1H), 6.66 (s, 1H), 6.51 (d, 1H, J=0.8 Hz), 5.84 (s, 1H), 4.70 (s,
1H), 4.21 (d, 2H, J=4.4 Hz), 3.24 (s, 1H), 2.16 (s, 3H), 2.10 (s,
3H), 1.94 (s, 3H), 1.88 (d, 2H, J=10 Hz), 1.69-1.72 (m, 3H),
1.22-1.36 (m, 5H).
Example 21
Synthesis of Compound 30:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3-
-(methyl(1-methylpiperidin-4-yl)amino)benzamide
##STR00603##
[1307] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
##STR00604##
[1309] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mass was poured on ice cold water, solid precipitated was filtered,
resulting residue was washed with water and dried under vacuum
giving the desired compound (71.7 g, 99.9%) which was used for
further reaction.
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00605##
[1311] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid
(287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415
mmol) and methyl iodide (626.63 g, 4415 mmol) were added. Resulting
reaction mass was heated at 60.degree. C. for 8 h. On completion,
solid precipitated was filtered, residue washed with diethyl ether
(5 times). Combined organic layers were dried, concentrated under
reduced pressure giving the desired compound (302 g, 99).
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00606##
[1313] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
Resulting reaction mass was heated at 80.degree. C. for 7 h. On
completion, reaction mass was filtered through celite giving
washing of water and ethyl acetate, filtrate was extracted with
ethyl acetate. The combined organic layers were dried, concentrated
under reduced pressure giving the desired compound.
Step 4: methyl
5-bromo-2-methyl-3-((1-methylpiperidin-4-yl)amino)benzoate
##STR00607##
[1315] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (2 g, 8.23 mmol) and
1-methylpiperidin-4-one (1.86 g, 16.46 mmol) in methanol (20 mL),
acetic acid (0.5 g, 8.23 mmol) was added and reaction stirred at
room temperature for 3 h. Then sodium cyanoborohydride (0.622 g,
9.87 mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford the desired compound
(0.9 g, 33%).
Step 5: methyl
5-bromo-2-methyl-3-(methyl(1-methylpiperidin-4-yl)amino)benzoate
##STR00608##
[1317] To a stirred solution of methyl
5-bromo-2-methyl-3-((1-methylpiperidin-4-yl)amino)benzoate (0.6 g,
1.76 mmol) in acetonitrile (15 mL), cesium carbonate (1.14 g, 3.52
mmol) and methyl iodide (1.2 g, 8.8 mmol) were added; resulting
reaction mass was heated at 80.degree. C. for 7 h. On completion,
reaction mass was cooled to room temperature and filtered, residue
was washed with ethyl acetate and filtrate was concentrated and
then purified by column chromatography to afford the desired
compound (0.5 g, 80%).
Step 6:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-m-
ethyl-3-(methyl(1-methylpiperidin-4-yl)amino)benzamide
##STR00609##
[1319] Aqueous NaOH (0.11 g, 2.75 mmol) was added to a solution of
methyl
5-bromo-2-methyl-3-(methyl(1-methylpiperidin-4-yl)amino)benzoate
(0.5 g, 1.4 mmol) in MeOH (20 mL) and stirred at 60.degree. C. for
1 h. After completion of the reaction, ethanol was removed under
reduced pressure and acidified using dilute HCl up to pH 6 and pH 4
was adjusted using citric acid. Extraction was carried out using
ethyl acetate. Combined organic layers were dried concentrated
giving respective acid (0.42 g, 90%).
[1320] The acid (0.5 g, 0.974 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.45 g, 2.8
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (1.1 g, 2.1 mmol) was added to
it and stirring was continued for overnight. After completion of
the reaction, reaction mass was poured into ice to obtain solid,
this was filtered and washed with acetonitrile followed by
purification with column chromatography to afford the desired
compound (0.2 g, 30%). LCMS: 475.02 (M+1).sup.+; HPLC: 94.25% (@254
nm) (R.sub.t; 4.570; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (s, 1H), 8.13 (t, 1H, J=4.8 Hz), 6.78 (d, 1H, J=0.8 Hz), 6.60
(d, 1H, J=1.6 Hz), 5.85 (s, 1H), 4.94 (d, 1H, J=8.8 Hz), 4.22 (d,
2H, J=4.8 Hz), 3.62-3.64 (m, 1H), 3.37-3.46 (m, 3H), 3.11 (s, 3H),
3.09 (s, 3H), 2.17 (s, 3H), 2.10 (s, 3H), 1.99-2.09 (m, 2H), 1.98
(s, 3H), 1.86-1.93 (m, 2H).
Example 22
Synthesis of Compound 31:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isoprop-
yl(methyl)amino)-2-methylbenzamide
##STR00610##
[1321] Step 1: Synthesis of methyl
5-bromo-3-(isopropylamino)-2-methylbenzoate
##STR00611##
[1323] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (1 equiv.) in methanol (5 mL),
acetone (5 equiv.) and acetic acid (2 equiv.) were added and
reaction mixture stirred at room temperature for 2 h. Then
NaBH.sub.3CN (3 equiv.) was added at 0.degree. C. The resulting
reaction mixture was stirred further for 16 h at room temperature.
On completion, solvent was evaporated and water added to the
residue and the extracted with ethyl acetate. The combined organic
layers were dried over sodium sulfate and concentrated to afford
crude product which was used in next step without further
purification (yield 80-90%).
Step 2: Synthesis of methyl
5-bromo-3-(isopropyl(methyl)amino)-2-methylbenzoate
##STR00612##
[1325] To a stirred solution of methyl
5-bromo-3-(isopropylamino)-2-methylbenzoate (0.5 g, 1.75 mmol) in
acetonitrile (10 mL), Cs.sub.2CO.sub.3 (1.02 g, 2.63 mmol) and
methyl iodide (1.45 g, 3.5 mmol) were added to it. The resulting
reaction mixture was stirred at 80.degree. C. for 4 h. Upon
completion, the solvent was removed under reduced pressure and
residue dissolved in water and extracted with ethyl acetate. Crude
material obtained was purified by column chromatography over silica
gel affording the desired compound product without further
purification (0.39 g)
Step 3:
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(-
isopropyl(methyl)amino)-2-methylbenzamide
##STR00613##
[1327] To a stirred solution of methyl
5-bromo-3-(isopropyl(methyl)amino)-2-methylbenzoate (1 equiv.) in
ethanol (5 mL), aqueous NaOH solution (1 equiv.) was added and
reaction stirred at 60.degree. C. for 4 h. On completion, ethanol
was removed under reduced pressure and residue acidified with 1N
HCl to pH 6. The aqueous phase was extracted with 10% MeOH/DCM.
Combined organic layers were dried over sodium sulfate and solvent
removed under reduced pressure affording pure acid (yield 50-60%).
To a solution of this acid (1 equiv.) in DMSO (1.5 mL), PyBOP (1.5
equiv.) was added and reaction stirred at room temperature for 15
min. Then 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.)
was added and reaction stirred overnight. On completion, water was
added and the resulting solid precipitate filtered and washed with
water. Then this solid was stirred with acetonitrile for 10 min and
filtered again to obtain pure target molecule (yield 50-60%).
[1328] LCMS: 420.15 (M+1).sup.+; HPLC: 93.04% (@254 nm) (R.sub.t;
4.791); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H),
8.19 (t, 1H, J=4.8 Hz), 7.14 (d, 1H, J=1.6 Hz), 7.00 (d, 1H, J=1.2
Hz), 5.85 (s, 1H), 4.23 (d, 2H, J=4.4 Hz), 3.16-3.20 (m, 1H), 2.52
(s, 3H), 2.17 (s, 3H), 2.10 (s, 6H), 1.01 (d, 6H, J=6.4 Hz).
Example 23
Synthesis of Compound 32:
5-bromo-N-((4,6-dihydro-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-3--
((tetrahydro-2H-pyran-4-yl)amino)benzoate
##STR00614##
[1329] Step 1: 5-bromo-2-methyl-3-nitrobenzoic acid
[1330] To a mixture of 2-methyl-3-nitrobenzoic acid (15 g, 82.80
mmol) in conc. H.sub.2SO.sub.4 (60 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (13.07 g, 45.71
mmol) was added and reaction mixture was stirred at room
temperature for 5 h. After completion of reaction, reaction mixture
was slowly poured onto ice cold water (400 mL). Solid that
precipitated out was filtered and dried under vacuum to obtain the
desired compound (21 g, 98%).
Step 2: methyl 5-bromo-2-methyl-3-nitrobenzoate
##STR00615##
[1332] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (16 g, 61.54 mmol) in DMF (160 mL), iodomethane (35.72 g, 248
mmol) and sodium carbonate (26.28 g, 248 mmol) were added.
Resulting reaction mass was stirred at 60.degree. C. for 8 h. On
completion, reaction mass was filtered and inorganic solid residue
washed with ethyl acetate. Combined filtrate was concentrated under
vacuum till dryness. The residue was re-dissolved in ethyl acetate
and washed with 5% sodium bicarbonate solution (700 mL) followed by
5M HCl solution (300 mL). Organic layer was finally washed with
brine, dried over sodium sulfate and concentrated to afford the
desired compound (16 g, 95%).
Step 3: methyl 3-amino-5-bromo-2-methylbenzoate
##STR00616##
[1334] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzoate (17 g, 62.04 mmol) in ethanol (85
mL), was added NH.sub.4Cl solution (17 g in 85 mL water, 317.8
mmol) followed by Fe powder (27.82 g, 498.11 mmol). Resulting
reaction mass was stirred at 90.degree. C. for 1 h. On completion,
reaction mass was filtered and filtrate was concentrated till
dryness to get solid which was dissolved in saturated sodium
bicarbonate solution. Aqueous layer was extracted with ethyl
acetate. Combined organic layers were dried over sodium sulfate and
concentrated to afford the desired compound (15 g, 99%).
Step 4: methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate
##STR00617##
[1336] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (2 g, 8.23 mmol) and tetra hydro
pyran-4-one (1.06 g, 10.66 mmol) in methanol (20 mL), acetic acid
(0.5 g, 8.23 mmol) was added and reaction stirred at room
temperature for 18 h. Then sodium cyanoborohydride (0.62 g, 9.83
mmol) was added and reaction stirred 3 h. On completion, solvent
was removed under reduced pressure; water was added and extracted
with DCM. Combined organic were dried, concentrated and purified by
column chromatography using silica (100-200 mesh size) to afford
the desired compound (1.6 g, 62%).
Step 5:
5-bromo-N-((4,6-dihydro-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-me-
thyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate
##STR00618##
[1338] Aqueous NaOH (0.146 g, 3.66 mmol) was added to a solution of
methyl 5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate
(0.6 g, 1.88 mmol) in MeOH (8 mL) and stirred at 60.degree. C. for
1 h. After completion of the reaction, ethanol was removed under
reduced pressure and acidified using dilute HCl up to pH 6 and pH 4
was adjusted using citric acid. Extraction was carried out using
DCM. Combined organic layers were dried concentrated giving
respective acid (0.515 g, 90%).
[1339] The acid (0.515 g, 1.67 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.50 g, 3.3
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (1 g, 1.98 mmol) was added to
it and stirring was continued for overnight. After completion of
the reaction, reaction mass was poured into ice and extracted with
5% MeOH/DCM. Combined organic layers were dried and concentrated to
obtain crude solid, this was purified by column chromatography to
afford the desired compound (0.30 g, 37%). LCMS: 447.84
(M+1).sup.+; HPLC: 99.78% (@254 nm) (R.sub.t; 5.753; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.09 (t,
1H, J=4.8 Hz), 6.76 (s, 1H), 6.53 (d, 1H, J=1.6 Hz), 5.84 (s, 1H),
4.82 (d, 1H, J=8 Hz), 4.42 (d, 2H, J=4.8 Hz), 3.85 (d, 2H, J=11.2
Hz), 3.37-3.53 (m, 3H), 2.16 (s, 3H), 2.10 (s, 3H), 1.95 (s, 3H),
1.81 (d, 2H, J=12.8 Hz), 1.44-1.54 (m, 2H).
Example 24
Synthesis of Compound 33:
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methylbenzamide
##STR00619##
[1340] Step 1: Synthesis of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate
##STR00620##
[1342] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (1 equiv.) in methanol (5 mL),
cyclopentanone (5 equiv.) and acetic acid (2 equiv.) were added and
reaction mixture stirred at room temperature for 2 h. Then
NaBH.sub.3CN (3 equiv.) was added at 0.degree. C. The resulting
reaction mixture was stirred further for 16 h at room temperature.
On completion, solvent was evaporated and water added to the
residue and the extracted with ethyl acetate. The combined organic
layers were dried over sodium sulfate and concentrated to afford
crude product which was used in next step without further
purification (yield 80-90%).
Step 2: Synthesis of
5-bromo-3-(cyclopentylamino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methylbenzamide
##STR00621##
[1344] To a stirred solution of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate (1 equiv.) in ethanol
(5 mL), aqueous NaOH solution (1 equiv.) was added and reaction
stirred at 60.degree. C. for 4 h. On completion, ethanol was
removed under reduced pressure and residue acidified with 1N HCl to
pH 6. Aqueous phase was extracted with 10% MeOH/DCM. Combined
organic layers were dried over sodium sulfate and solvent removed
under reduced pressure affording pure acid (yield 50-60%). To a
solution of this acid (1 equiv.) in DMSO (1.5 mL), PyBOP (1.5
equiv.) was added and reaction stirred at room temperature for 15
min. Then 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.)
was added and reaction stirred overnight. On completion, water was
added and the solid precipitate filtered and washed with water.
This solid was then stirred with acetonitrile for 10 min and
filtered again to obtain the pure target molecule (50-60%
yield).
[1345] LCMS: 432.10 (M+1).sup.+; HPLC: 97.15% (@254 nm) (R.sub.t;
6.834); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H),
8.09 (t, 1H, J=4.4&5.2 Hz), 6.65 (d, 1H, J=1.6 Hz), 6.54 (d,
1H, J=1.6 Hz), 5.84 (s, 1H), 4.85 (d, 1H, J=6 Hz), 4.22 (d, 2H,
J=5.2 Hz), 3.73 (m, 1H), 2.16 (s, 3H), 2.10 (s, 3H), 1.95 (s, 3H),
1.91 (m 2H), 1.70-1.62 (m, 2H), 1.60-1.45 (m, 4H).
Example 25
Synthesis of Compound 36:
5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2,4-dimethylbenzamide
##STR00622##
[1346] Step 1: 5-bromo-2,4-dimethylbenzoic acid
##STR00623##
[1348] To the stirred solution of NaOH (60 g) in water (1152 ml)
was added bromine (36 ml) dropwise at 0.degree. C. and stirred the
reaction mixture for 45 min. at same temperature.
2,4-dimethylbenzoic acid (12 g, 79.9 mmol) was added in above
reaction mixture at 0.degree. C. portion wise and stirred the
reaction at room temperature for 3 h. After completion of reaction
of reaction (TLC) reaction mixture was acidified using conc. HCl,
solid was filtered through Buchner funnel and wash with water and
dried to afford the desired compound (12 g, 65%) which was used
without further purification.
Step 2: 5-bromo-2,4-dimethyl-3-nitrobenzoic acid
##STR00624##
[1350] 5-Bromo-2,4-dimethylbenzoic acid (12 g, 52.17 mmol) was
added to cooled conc. H.sub.2SO.sub.4 (48 mL) at -10.degree. C. lot
wise. After 10 minutes nitrating mixture {prepared as mixing Conc.
HNO.sub.3 (6 mL,) with conc. H.sub.2SO.sub.4 (24 mL)} was added
drop wise at -10.degree. C. Resulting reaction mass was stirred at
-10.degree. C. for 30 minutes. On completion, reaction mixture was
poured on ice cold water, solid precipitated was filtered, washed
with water and dried under vacuum giving desired compound (13 g,
91%).
Step 3: methyl 3-bromo-5-nitrobenzoate
##STR00625##
[1352] To stirred solution of 5-bromo-2,4-dimethyl-3-nitrobenzoic
acid (13 g, 47.44 mmol) in DMF (120 mL), sodium carbonate (15 g,
142.2 mmol) and methyl iodide (4.4 ml, 71.16 mmol) were added.
Resulting reaction mass was heated at 60.degree. C. for 8 h. On
completion, water was added to the reaction mass and extraction was
carried out using ethyl acetate. Combined organic layers were
washed with sat. bicarbonate solution and 5 N HCl, dried over
sodium sulphate, concentrated under reduced pressure giving desired
compound (12.9 g, 86%).
Step 4: methyl 3-amino-5-bromo-2,4-dimethylbenzoate
##STR00626##
[1354] To stirred solution of methyl 3-bromo-5-nitrobenzoate (12.9
g, 44.79 mmol) in ethanol (65 mL), ammonium chloride (13 g, 223.95
mmol) dissolved in water (65 mL) and iron powder (20 g, 358.33
mmol) were added under stirring. Resulting reaction mass was heated
at 80.degree. C. for 1 h. On completion, reaction mixture was
filtered through celite bed, celite bed was washed with ethanol,
filtrate was extracted with ethyl acetate. Combined organic layers
were washed with water, dried, concentrated under reduced pressure
giving desired compound (11.0 g, 95%).
Step 5: methyl
5-bromo-3-(cyclopentylamino)-2,4-dimethylbenzoate
##STR00627##
[1356] To a stirred solution of methyl
3-amino-5-bromo-2,4-dimethylbenzoate (1.4 g, 5.42 mmol) and
cyclopentanone (2.4 mL, 27.13 mmol) in methanol (20 mL), acetic
acid (0.650 g, 10.84 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.85 g, 13.56
mmol) was added and reaction was stirred for overnight. On
completion, solvent was removed water was added and compound was
extracted with ethyl acetate. Combined organic layer was collected,
dried over Na.sub.2SO.sub.4 and concentrate under reduced pressure
and crude material was purified by column chromatography to afford
the desired compound (1.2 g, 68%).
Step 6:methyl
5-bromo-3-(cyclopentyl(methyl)amino)-2,4-dimethylbenzoate
##STR00628##
[1358] To a stirred solution of methyl
5-bromo-3-(cyclopentylamino)-2,4-dimethylbenzoate (0.3 g, 1.07
mmol) in DMF (5 mL), cesium carbonate (0.697 g, 2.14 mmol) and
ethyl iodide (0.3 mL, 5.38 mmol) were added; resulting reaction
mass was heated at 80.degree. C. for 12 h. On completion, reaction
mass was cooled to room temperature and filtered, residue was
washed with ethyl acetate and filtrate was concentrated and then
purified by column chromatography to afford desired compound (0.3
g, 82%).
Step 7:
5-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2,4-dimethylbenzamide
##STR00629##
[1360] Aqueous NaOH (0.022 g, 0.553 mmol) was added to a solution
of methyl 5-bromo-3-(cyclopentyl(methyl)amino)-2,4-dimethylbenzoate
(0.125 g, 0.368 mmol) in EtOH (10 mL) and H.sub.2O (1 mL) and
stirred at 60.degree. C. for 1 h. After completion of the reaction,
ethanol was removed under reduced pressure and acidified using
dilute HCl up to pH 6. Extraction was carried out using ethyl
acetate. Combined organic layers were dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure to give respective acid (0.1 g,
84.03%).
[1361] The acid (0.1 g, 0.308 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.07 g,
0.462 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.320 g, 0.616 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed by HPLC purification to provide the title compound (0.018
g, 12.7%) as its TFA salt: LCMS: 460.10 (M+1).sup.+; HPLC: 98.10%
(@254 nm) (R.sub.t; 8.291; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.18 (t, 1H, J=4.4
Hz), 7.20 (s, 1H), 5.85 (s, 1H), 4.26 (d, 2H, J=4.8 Hz), 3.51-3.52
(m, 1H), 2.64 (s, 3H), 2.30 (s, 3H), 2.17 (s, 3H), 2.14 (s, 3H),
2.10 (s, 3H), 1.48-1.59 (m, 6H), 1.31-1.32 (m, 2H).
Example 26
Synthesis of Compound 48:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(N-methylcyclopentanecarboxamido)benzamide
##STR00630##
[1362] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00631##
[1364] To a stirred ice cold (-10 to -15.degree. C.) conc.
H.sub.2SO.sub.4 (136 mL) was added 5-chloro-2-methylbenzoic acid
(20 g, 0.117 mmol) in portions. The nitrating mixture [Conc.
H.sub.2SO.sub.4 (22 mL) and HNO.sub.3 (11.05 mL] was added dropwise
at -10 to -15.degree. C. and resulting mixture was allowed stir for
30 min. The obtained precipitate was filtered and dried. The solid
was the dissolved in ethyl acetate and dried (Na.sub.2SO.sub.4) and
concentrated to provide desired product (23.6 g, 93%) as a white
solid.
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00632##
[1366] To a solution of 5-chloro-2-methyl-3-nitrobenzoic acid (23.5
g, 0.109 mmol) in DMF (120 mL) was added sodium carbonate (46.21,
0.436 mmol) and methyl iodide (27.2 mL, 0.43 mmol) at room
temperature. The reaction mixture was heated at 60.degree. C. for 3
h. After complete consumption of starting material the reaction
mixture was filtered and the residue washed with ethyl acetate. The
filtrate was concentrated under reduced pressure and the crude
product purified by column chromatography to provide the desired
compound (16 g, 64%) as thick oil which solidified on standing.
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methylbenzoate
##STR00633##
[1368] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (16 g, 69 mmol) in ethanol (160
mL), ammonium chloride (16 g, 53.4 mmol) dissolved in water (160
mL) and iron powder (31.2 g, 55.85 mmol) were added under stirring.
The resulting reaction mixture was heated at 80.degree. C. for 1 h.
On completion, water was added to and the reaction mixture filtered
through celite. The filtrate was extracted with ethyl acetate and
the combined organic layers washed with water, dried and
concentrated under reduced pressure giving desired compound (12.10
g, 86%) without further purification.
Step 4: Synthesis of methyl
5-chloro-3-(cyclopentanecarboxamido)-2-methylbenzoate
##STR00634##
[1370] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (0.4 g, 2.0 mmol) in pyridine (2
mL), cyclopentyl chloride (0.39 mL, 3.00 mmol) was added at
0.degree. C. The reaction mixture was stirred at room temperature
for 2 h. After complete consumption of the starting material the
reaction was quenched with ice water. The resulting precipitate was
filtered of to provide desired product (0.5 g, 84%).
Step 5: Synthesis of methyl
5-chloro-2-methyl-3-(N-methylcyclopentanecarboxamido)benzoate
##STR00635##
[1372] To a stirred solution of methyl
5-chloro-3-(cyclopentanecarboxamido)-2-methylbenzoate (0.3 g, 1.01
mmol) in DMF, sodium hydride (0.060 g, 0.0015 mmol) was added at
0.degree. C. and reaction mixture was allowed to stir for 15 min.
Methyl iodide (0.32 mL, 0.50 mmol) was added and reaction mixture
and was stirred at room temperature. After complete consumption of
starting material, the reaction was quenched with addition of water
and extracted with ethyl acetate. The combined organic layer was
the washed with brine, dried (Na.sub.2SO.sub.4) and concentrated.
The crude compound was used for the next step.
Step 6: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(N-methylcyclopentanecarboxamido)benzamide
##STR00636##
[1374] Aq. NaOH (0.077 g, 1.94 mmol) was added to a solution of
methyl
5-chloro-2-methyl-3-(N-methylcyclopentanecarboxamido)benzoate (0.4
g, 1.2 mmol) in 4:1 EtOH: Water (15 mL) and stirred at 65.degree.
C. for 1 h. After completion of the reaction, ethanol was removed
under reduced pressure and acidified with 1N HCl. The reaction
mixture was extracted with ethyl acetate. Combined organic layers
were washed with brine, dried and concentrated to provide
respective acid (0.35 g, 91%).
[1375] The acid (0.3 g, 0.10 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.309 g,
0.20 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.826 g, 0.155 mmol) was
added to it and stirring was continued overnight. After completion
the reaction mixture was poured into ice to obtain a solid which
was filtered and washed with acetonitrile. Final purification was
performed using prep. HPLC purification to the desired product as
the TFA salt (0.120 g, 27%).
[1376] LCMS: 430.20 (M+1).sup.+; HPLC: 99.79% (@254 nm) (R.sub.t;
6.086); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.55 (bs, 1H),
8.43 (t, 1H), 7.46 (s, 1H), 7.29 (s, 1H), 5.87 (s, 1H), 4.25 (s,
2H), 3.01 (s, 3H), 2.35 (m, 1H), 2.19 (s, 3H), 2.10 (s, 3H), 2.06
(s, 3H), 1.70-1.45 (m, 6H), 1.35 (m, 2H).
Example 27
Synthesis of Compound 49:
3-((2-aminoethyl)(methyl)amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihyd-
ropyridin-3-yl)methyl)-2-methylbenzamide
##STR00637##
[1378] A stirred solution of tert-butyl
(2-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carb-
amoyl)-2-methylphenyl)(methyl)amino)ethyl)carbamate (0.2 g, 4.2
mmol) in DCM (5 mL) was cooled to 0.degree. C. and TFA (1 mL) was
added to it. The reaction mixture was stirred at room temperature
for 1 h. On completion, the reaction was concentrated to dryness.
Half of the material was purified by solvent washings giving pure
3-((2-aminoethyl)(methyl)amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihyd-
ropyridin-3-yl)methyl)-2-methylbenzamide as the TFA salt (0.060 g,
76%).
[1379] LCMS: 377.15 (M+1).sup.+; HPLC: 97.57% (@254 nm) (R.sub.t;
4.611); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H),
8.21 (t, 1H), 7.69 (bs, 3H), 7.16 (s, 1H), 6.97 (s, 1H), 5.86 (s,
1H), 4.25 (d, 2H, J=4.4 Hz), 3.07 (t, 2H), 2.96 (m, 2H), 2.59 (s,
3H), 2.18 (s, 6H), 2.10 (s, 3H).
[1380] Remaining material was basified with aqueous sodium
bicarbonate till pH 8 and aqueous layer extracted with 20%
MeOH/DCM. Combined organic layers were dried over sodium sulfate
and concentrated to afford desired compound as free base which used
for further reaction (0.075 g).
Example 28
Synthesis of Compound 50: tert-butyl
(2-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carb-
amoyl)-2-methylphenyl)(methyl)amino)ethyl)carbamate
##STR00638##
[1381] Step 1: Synthesis of methyl
3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-5-chloro-2-methylbenzoate
##STR00639##
[1383] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1 g, 4.6 mmol) and tert-butyl
(2-oxoethyl)carbamate (1.4 g, 8.8 mmol) in methanol (10 mL), acetic
acid (0.027 g, 4.6 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.352 g, 4.68
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford the desired product
(0.62 g, 38%).
Step 2: Synthesis of methyl
3-((2-((tert-butoxycarbonyl)amino)ethyl)(methyl)amino)-5-chloro-2-methylb-
enzoate
##STR00640##
[1385] To a stirred solution of methyl
3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-5-chloro-2-methylbenzoate
(0.5 g, 1.46 mmol) in acetonitrile (10 mL), cesium carbonate (0.95
g, 2.92 mmol) and methyl iodide (1 g, 7.3 mmol) were added and the
resulting reaction mixture heated at 80.degree. C. for 12 h. On
completion, the reaction mixture was cooled to room temperature and
filtered, the residue washed with ethyl acetate and the filtrate
concentrated before purification by column chromatography to afford
desired product (0.325 g, 62%).
Step 3: Synthesis of tert-butyl
(2-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carb-
amoyl)-2-methylphenyl)(methyl)amino)ethyl)carbamate
##STR00641##
[1387] Aqueous NaOH (0.071 g, 1.79 mmol) was added to a solution of
methyl
3-((2-((tert-butoxycarbonyl)amino)ethyl)(methyl)amino)-5-chloro-2-methylb-
enzoate (0.425 g, 1.19 mmol) in MeOH (5 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 and adjusted using citric acid. Extraction was
carried out using ethyl acetate and the combined organic layers
dried and concentrated giving respective acid (0.34 g, 84%).
[1388] The acid (0.34 g, 0.99 mmol) was then dissolved in DMSO (1.5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.300 g,
1.98 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.77 g, 1.48 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, the reaction mixture was poured into
ice to obtain solid, this was filtered and washed with acetonitrile
followed column purification to provide tert-butyl
(2-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carb-
amoyl)-2-methylphenyl)(methyl)amino)ethyl)carbamate (0.27 g,
58%).
[1389] LCMS: 477.25 (M+1).sup.+; HPLC: 97.92% (@254 nm) (R.sub.t;
6.229); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H),
8.17 (t, 1H), 7.07 (s, 1H), 6.88 (s, 1H), 6.78 (bs, 1H), 5.85 (s,
1H), 4.24 (d, 2H, J=4.4 Hz), 3.07 (t, 2H), 2.84 (t, 2H), 2.62 (s,
3H), 2.17 (s, 3H), 2.13 (s, 3H), 2.10 (s, 3H), 1.35 (s, 9H).
Example 29
Synthesis of Compound 51:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((2-(di-
methylamino)ethyl)(methyl)amino)-2-methylbenzamide
##STR00642##
[1391]
3-((2-Aminoethyl)(methyl)amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-
-dihydropyridin-3-yl)methyl)-2-methylbenzamide (0.075 g, 0.199
mmol) was dissolved in methanol (5 mL) and cooled to 0.degree. C.,
formaldehyde (0.056 g, 1.86 mmol) was added. The resulting reaction
mixture was stirred at the same temperature for 30 minutes then
sodium cyanoborohydride (0.023 g, 0.366 mmol) was added to above
reaction mixture and stirred at room temperature for 4 h. After
completion, solvent was removed under reduced pressure and water
added before extraction with DCM. The combined organic layers were
dried, concentrated and purified by column chromatography to
provide
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((2-(di-
methylamino)ethyl)(methyl)amino)-2-methylbenzamide (0.040 g, 50%).
LCMS: 405.25 (M+1).sup.+; HPLC: 89.93% (@254 nm) (R.sub.t; 4.634);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.21 (s,
1H), 7.08 (s, 1H), 5.85 (s, 1H), 4.24 (bs, 2H), 2.91 (t, 2H), 2.63
(s, 3H), 2.33-2.35 (m, 2H), 2.11-2.18 (m, 15H).
Example 30
Synthesis of Compound 52:
3-(allyl(cyclopentyl)amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2-methylbenzamide
##STR00643##
[1392] Step 1: Synthesis of methyl
3-(allyl(cyclopentyl)amino)-5-chloro-2-methylbenzoate
##STR00644##
[1394] Methyl 5-chloro-3-(cyclopentylamino)-2-methylbenzoate (1.2
g, 4.46 mmol) was dissolved in DMF (12 mL) and cooled to 0.degree.
C. NaH (0.21 g, 8.92 mmol) was added after 10 minutes and allyl
bromide (1.07 g, 8.9 mmol) was added. The reaction mixture was
stirred at room temperature for 4 h and then heated at 80.degree.
C. for 18 h. The reaction was quenched with ice water and extracted
with ethyl acetate. Combined organic layers were dried,
concentrated giving crude methyl
3-(allyl(cyclopentyl)amino)-5-chloro-2-methylbenzoate (0.4 g,
29.4%).
Step 2: Synthesis of 3-(allyl (cyclopentyl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00645##
[1396] Aqueous NaOH (0.078 g, 1.95 mmol) was added to a solution of
methyl 3-(allyl(cyclopentyl)amino)-5-chloro-2-methylbenzoate (0.4
g, 1.30 mmol) in MeOH (5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate and the combined organic layers concentrated giving
respective acid (0.37 g, 97.6%).
[1397] The acid (0.25 g, 0.85 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.259 g,
1.74 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.66 g, 1.27 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mixture was poured into ice to
obtain a solid, which was filtered and washed with acetonitrile
followed column purification to provide the desired 3-(allyl
(cyclopentyl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide (0.175 g, 48%).
[1398] LCMS: 428.30 (M+1).sup.+; HPLC: 96.37% (@254 nm) (R.sub.t;
6.357); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H),
8.21 (t, 1H), 7.16 (s, 1H), 6.92 (s, 1H), 5.85 (s, 1H), 5.63-5.69
(m, 1H), 4.98 (m, 2H), 4.23 (d, 2H, J=4 Hz), 3.54 (d, 2H, J=5.6
Hz), 3.46-3.50 (m, 1H), 2.18 (s, 3H), 2.16 (s, 3H), 2.10 (s, 3H),
1.67 (m, 2H), 1.59 (m, 2H), 1.47 (m, 2H), 1.36 (m, 2H).
Example 31
Synthesis of Compound 53: 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(piperazin-1-yl) benzamide
##STR00646##
[1399] Step 1: Synthesis of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate
##STR00647##
[1401] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (3 g, 13.3 mmol) and
cyclopentanone (5.6 g, 66 mmol) in methanol (30 mL), acetic acid
(1.59 g, 26.6 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (2.08 g, 29.4
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
used without further purification.
Step 2: Synthesis of methyl 5-bromo-3-(cyclopentyl (methyl)
amino)-2-methylbenzoate
##STR00648##
[1403] To a stirred solution of methyl
5-bromo-3-(cyclopentylamino)-2-methylbenzoate (1.6 g, 5.38 mmol) in
acetonitrile (20 mL), cesium carbonate (3.5 g, 10.73 mmol) and
methyl iodide (3.87 g, 27.25 mmol) were added and the resulting
reaction mixture heated at 80.degree. C. for 12 h. On completion,
the reaction mixture was cooled to room temperature and filtered.
The residue was washed with ethyl acetate and filtrate concentrated
and purified by column chromatography to afford desired compound
(1.6 g, 95%).
Step 3: Synthesis of tert-butyl 4-(3-(cyclopentyl (methyl)
amino)-5-(methoxycarbonyl)-4-methylphenyl)piperazine-1-carboxylate
##STR00649##
[1405] A solution of methyl 5-bromo-3-(cyclopentyl (methyl)
amino)-2-methylbenzoate (1 g, 3.07 mmol), tert-butyl
piperazine-1-carboxylate (0.85 g, 4.61 mmol) and Cs.sub.2CO.sub.3
(0.5 g, 1.53 mmol) in toluene (25 mL) was purged with argon for 10
min. Then, Pd.sub.2(dba).sub.3 (0.31 g, 0.307 mmol) and BINAP
(0.038 g, 0.061 mmol) was added to it and argon purged again for 10
min. The reaction mixture was stirred at 100.degree. C. for 8 h.
After completion, water was added to it and extracted with ethyl
acetate. The combined organic layers were washed with water, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford crude material which was purified by
column chromatography to afford the title compound (0.44 g,
33.3%).
Step 4: Synthesis tert-butyl 4-(3-(cyclopentyl (methyl)
amino)-5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)--
4-methylphenyl)piperazine-1-carboxylate
##STR00650##
[1407] Aqueous NaOH (0.061 g, 1.53 mmol) was added to a solution of
tert-butyl 4-(3-(cyclopentyl (methyl)
amino)-5-(methoxycarbonyl)-4-methylphenyl)piperazine-1-carboxylate
(0.44 g, 1.02 mmol) in MeOH (10 mL) and stirred at 60.degree. C.
for 1 h. After completion of the reaction, ethanol was removed
under reduced pressure and acidified using dilute HCl up to pH 6
and pH 4 was adjusted using citric acid. Extraction was carried out
using ethyl acetate. Combined organic layers were dried
concentrated giving respective acid (0.3 g, 70%).
[1408] The acid (0.3 g, 0.71 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.215 g,
1.43 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.56 g, 1.07 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mixture was poured into ice to
obtain solid, which was filtered and washed with acetonitrile
followed by ether to provide tert-butyl 4-(3-(cyclopentyl (methyl)
amino)-5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)--
4-methylphenyl)piperazine-1-carboxylate (0.30 g, 75.7%).
Step 5: Synthesis of 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(piperazin-1-yl) benzamide
##STR00651##
[1410] A stirred solution of tert-butyl 4-(3-(cyclopentyl (methyl)
amino)-5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)--
4-methylphenyl)piperazine-1-carboxylate (0.30, 0.54 mmol) in DCM
(10 mL) was cooled to 0.degree. C. and TFA (3 mL) was added to it.
The reaction mixture was stirred at room temperature for 1 h. Upon
completion, the mixture was concentrated to dryness. Half crude was
purified by solvent washing giving the title compound as TFA salt
(0.01 g, 8.33%).
[1411] LCMS: 452.39 (M+1).sup.+; HPLC: 83.92% (@254 nm) (R.sub.t;
3.825); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H),
8.71 (s, 2H), 7.98 (s, 1H), 6.79 (s, 1H), 6.55 (s, 1H), 5.85 (s,
1H), 4.24 (d, 2H, J=3.2 Hz), 3.46 (1H in solvent peak), 3.22-3.26
(m, 8H), 2.50 (3H in solvent peak), 2.18 (s, 3H), 2.10 (s, 6H),
1.41-1.67 (m, 8H).
[1412] Half of the crude was basified with aqueous sodium
bicarbonate till pH 8 and aqueous layer extracted with 20%
MeOH/DCM. Combined organic layers were dried over sodium sulfate
and concentrated to afford the title compound as free base.
Example 32
Synthesis of Compound 54:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-methyl-5-(4-methylpiperazin-1-yl)benzamide
##STR00652##
[1414] 3-(Cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(piperazin-1-yl) benzamide (free base) (0.100 g,
0.223 mmol) was dissolved in methanol (5 mL) and cooled to
0.degree. C. before formalin (0.033 g, 1.1 mL, 2.23 mmol) was
added. The resulting reaction mixture was stirred at same
temperature for 30 minutes. Sodium cyanoborohydride (0.013 g, 0.22
mmol) was added to above reaction mixture and stirred at room
temperature for 4 h. After completion, solvent was removed under
reduced pressure and water added to the residue, with extraction
carried out using DCM. The combined organic layers were dried,
concentrated and purified by column chromatography to provide
desired 3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-5-(4-methylpiperazin-1-yl) benzamide (0.037 g,
36%).
[1415] LCMS: 466.40 (M+1).sup.+; HPLC: 99.18% (@254 nm) (R.sub.t;
3.871); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H),
7.94 (t, 1H), 6.71 (s, 1H), 6.48 (s, 1H), 5.85 (s, 1H), 4.24 (d,
2H, J=3.2 Hz), 3.39-3.43 (m, 1H), 3.07 (bs, 4H), 2.46 (s, 3H), 2.31
(s, 3H), 2.18 (s, 3H), 2.10 (s, 3H), 2.08 (s, 3H), 1.66 (m, 2H),
1.59 (m, 2H), 1.39-1.48 (m, 4H).
Example 33
Synthesis of Compound 55:
5-chloro-3-(cyclohexyl(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyr-
idin-3-yl)methyl)-2-methylbenzamide
##STR00653##
[1416] Step 1: Synthesis of methyl
5-chloro-3-(cyclohexylamino)-2-methylbenzoate
##STR00654##
[1418] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1 g, 5.02 mmol) and
cyclohexanone (2.45 g, 25 mmol) in methanol (10 mL), acetic acid
(0.3 g, 5.02 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.63 g, 10.05
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford methyl
5-chloro-3-(cyclohexylamino)-2-methylbenzoate (1.2 g, 89.92%).
Step 2: Synthesis of methyl 5-chloro-3-(cyclohexyl (ethyl)
amino)-2-methylbenzoate
##STR00655##
[1420] To a stirred solution of methyl
5-chloro-3-(cyclohexylamino)-2-methylbenzoate (1.2 g, 4.3 mmol) in
dry DMF (15 mL), cesium carbonate (2.78 g, 8.5 mmol) and ethyl
iodide (3.35 g, 21.47 mmol) were added. The resulting reaction
mixture was heated at 80.degree. C. for 18 h. On completion, the
reaction mixture was cooled to room temperature and filtered, and
the residue washed with ethyl acetate. The filtrate was
concentrated and purified by column chromatography to afford methyl
5-chloro-3-(cyclohexyl (ethyl) amino)-2-methylbenzoate (0.25 g,
22.7%).
Step 3: Synthesis of 5-chloro-3-(cyclohexyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00656##
[1422] Aqueous NaOH (0.065 g, 1.61 mmol) was added to a solution of
methyl 5-chloro-3-(cyclohexyl (ethyl) amino)-2-methylbenzoate (0.25
g, 0.809 mmol) in EtOH (5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate. Combined organic layers were dried concentrated giving
respective acid (0.22 g, 92%).
[1423] The acid (0.22 g, 0.745 mmol) was then dissolved in DMSO (2
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.246 g,
1.49 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.58 g, 1.11 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, the mixture was poured into ice to
obtain a solid which was filtered and washed with acetonitrile.
Final purification by column purification gave
5-chloro-3-(cyclohexyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide (0.200 g, 46.6%). LCMS: 430.20
(M+1).sup.+; HPLC: 92.49% (@254 nm) (R.sub.t; 5.264); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.21 (t, 1H), 7.12
(s, 1H), 6.91 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4.4 Hz),
3.01-3.03 (m, 2H), 2.64-2.66 (m, 1H), 2.18 (s, 3H), 2.13 (s, 3H),
2.10 (s, 3H), 1.68 (m, 4H), 1.51-1.53 (m, 1H), 1.07-1.34 (m, 5H),
0.78 (t, 3H, J=6.8 Hz).
Example 34
Synthesis of Compound 56:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide
##STR00657##
[1424] Step 1: Synthesis of methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino) benzoate
[1425] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1 g, 5.02 mmol) and
tetrahydropyran-4-one (2.5 g, 25 mmol) in methanol (10 mL), acetic
acid (0.3 g, 5.02 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.63 g, 10.05
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (0.5
g, 35.5%).
Step 2: Synthesis of methyl 5-chloro-3-(ethyl
(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate
##STR00658##
[1427] To a stirred solution of methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl)amino)benzoate (0.5
g, 1.76 mmol) in dry CAN (15 mL), cesium carbonate (1.2 g, 3.68
mmol) and ethyl iodide (2.7 g, 17.3 mmol) were added and the
resulting reaction mixture was heated at 80.degree. C. for 18 h. On
completion, the mixture was cooled to room temperature and filtered
with the residue washed with ethyl acetate. The filtrate was
concentrated and then purified by column chromatography to afford
desired methyl 5-chloro-3-(ethyl (tetrahydro-2H-pyran-4-yl)
amino)-2-methylbenzoate (0.180 g, 34%).
Step 3: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide
##STR00659##
[1429] Aqueous NaOH (0.05 g, 1.22 mmol) was added to a solution of
methyl 5-chloro-3-(ethyl (tetrahydro-2H-pyran-4-yl)
amino)-2-methylbenzoate (0.18 g, 0.608 mmol) in EtOH (5 mL) and
stirred at 60.degree. C. for 1 h. After completion of the reaction,
ethanol was removed under reduced pressure and acidified using
dilute HCl up to pH 6 and pH 4 and adjusted using citric acid.
Extraction was carried out using ethyl acetate and the combined
organic layers were dried and concentrated giving respective acid
(0.15 g, 87%).
[1430] The acid (0.15 g, 0.530 mmol) was then dissolved in DMSO (1
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.160 g,
1.06 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.413 g, 0.79 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mixture was poured into ice
and extraction was carried out using 10% MeOH/DCM. Combined organic
layers were dried, concentrated and purified by silica gel column
to afford
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
tetrahydro-2H-pyran-4-yl)amino)-2-methylbenzamide (0.100 g,
43.88%). LCMS: 432.25 (M+1).sup.+; HPLC: 90.46% (@254 nm) (R.sub.t;
4.833); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H),
8.22 (t, 1H), 7.19 (s, 1H), 6.96 (s, 1H), 5.85 (s, 1H), 4.23 (d,
2H, J=4 Hz), 3.81 (d, 2H, J=10 Hz), 3.20-3.21 (m, 2H), 2.94-3.02
(m, 3H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 1.48-1.61 (m,
4H), 0.78 (t, 3H, J=6.4 Hz).
Example 35
Synthesis of Compound 57:
5-chloro-3-(cycloheptyl(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2-methylbenzamide
##STR00660##
[1431] Step 1: Synthesis of methyl
5-chloro-3-(cycloheptylamino)-2-methylbenzoate
##STR00661##
[1433] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1 g, 5.02 mmol) and
cycloheptanone (2.81 g, 25 mmol) in methanol (10 mL), acetic acid
(0.3 g, 5.02 mmol) was added and reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (0.65 g, 10.05
mmol) was added and reaction stirred overnight. On completion,
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford
5-chloro-3-(cycloheptylamino)-2-methylbenzoate (0.8 g, 56.2%).
Step 2: Synthesis of methyl 5-chloro-3-(cycloheptyl (ethyl)
amino)-2-methylbenzoate
##STR00662##
[1435] To a stirred solution of
5-chloro-3-(cycloheptylamino)-2-methylbenzoate (0.8 g, 2.70 mmol)
in dry DMF (10 mL), cesium carbonate (1.76 g, 5.4 mmol) and ethyl
iodide (2.11 g, 13.5 mmol) were added and the resulting reaction
mixture heated at 80.degree. C. for 18 h. On completion, the
mixture was cooled to room temperature and filtered, the residue
washed with ethyl acetate and the filtrate concentrated before
purification by column chromatography to afford the desired methyl
5-chloro-3-(cycloheptyl (ethyl) amino)-2-methylbenzoate (0.220 g,
25%).
Step 3: Synthesis of 5-chloro-3-(cycloheptyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00663##
[1437] Aqueous NaOH (0.04 g, 1.02 mmol) was added to a solution of
methyl 5-chloro-3-(cycloheptyl (ethyl) amino)-2-methylbenzoate
(0.22 g, 0.68 mmol) in EtOH (5 mL) and stirred at 60.degree. C. for
1 h. After completion of the reaction, ethanol was removed under
reduced pressure and acidified using dilute HCl up to pH 6 and pH 4
and adjusted using citric acid. Extraction was carried out using
ethyl acetate. With the combined organic layers dried and
concentrated giving the respective acid (0.18 g, 85%).
[1438] The acid (0.18 g, 0.585 mmol) was then dissolved in DMSO
(1.5 mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.177
g, 1.16 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.45 g, 0.87 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, the reaction mixture was poured into
ice to obtain solid, which was filtered and washed with
acetonitrile before column purification to 5-chloro-3-(cycloheptyl
(ethyl) amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide (0.100 g, 38.7%). LCMS: 444.25
(M+1).sup.+; HPLC: 89.74% (@254 nm) (R.sub.t; 5.933); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.21 (t, 1H), 7.07
(s, 1H), 6.88 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4 Hz),
3.02-3.03 (m, 2H), 2.77 (bs, 1H), 2.18 (s, 3H), 2.13 (s, 3H), 2.10
(s, 3H), 1.76 (m, 2H), 1.59-1.62 (m, 4H), 1.46 (m, 4H), 1.28 (m,
2H), 0.78 (t, 3H, J=6 Hz).
Example 36
Synthesis of Compound 58:
Cis-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1-
,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00664##
[1439] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00665##
[1441] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276.2 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) was added portion wise at room temperature and reaction mass
was stirred at room temperature for 5 h. On completion, reaction
mixture was poured onto ice cold water, solid precipitated was
filtered, resulting residue was washed with water and dried under
vacuum giving desired compound (71.7 g, 100%) which was used as is
in subsequent steps.
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00666##
[1443] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid
(287 g, 1103 mmol) in DMF (150 mL), sodium carbonate (468 g, 4415
mmol) and methyl iodide (626.63 g, 4415 mmol) were added. The
resulting reaction mixture was heated at 60.degree. C. for 8 h. On
completion, a solid precipitated was filtered, residue washed with
diethyl ether (5 times). The combined organic layers were dried,
concentrated under reduced pressure giving desired compound (302 g,
99%) which was used as is in subsequent steps.
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00667##
[1445] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
The resulting reaction mixture was heated at 80.degree. C. for 7 h.
On completion, reaction mass was filtered through celite; giving
washings of water and ethyl acetate to the residue, filtrate was
extracted with ethyl acetate. Combined organic layers were dried,
concentrated under reduced pressure giving desired compound which
was used in subsequent steps without further purification.
Step 4: Synthesis of cis and trans methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino) cyclohexyl)
amino)-2-methylbenzoate
##STR00668##
[1447] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (5 g, 20.57 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (5.6 g, 26.7 mmol) in methanol (50 mL),
acetic acid (1.2 g, 20.57 mmol) was added and reaction stirred at
room temperature for 8 h. Then sodium cyanoborohydride (1.6 g,
26.74 mmol) was added at 0.degree. C. and reaction stirred
overnight. On completion, solvent was removed under reduced
pressure and crude material was purified by column chromatography
twice eluting with ethyl acetate: hexane to afford (4 g, 44%) of
less polar cis isomer and (3 g, 33%) of the pure more polar trans
isomer.
Step 5: Synthesis of Cis-methyl
5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl) (methyl)
amino)-2-methylbenzoate
##STR00669##
[1449] To a stirred solution of mixture of cis isomer of methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino) cyclohexyl)
amino)-2-methylbenzoate (4 g, 9.09 mmol) in acetonitrile (50 mL)
was added cesium carbonate (5.9 g, 18.18 mmol) and methyl iodide
(6.45 g, 45.45 mmol). The resulting reaction mixture was heated at
80.degree. C. for 7 h. On completion, the reaction mixture was
cooled to room temperature and filtered, the residue was washed
with ethyl acetate and filtrate was concentrated to afford desired
crude compound which then purified by column chromatography giving
desired compound (1.4 g, 34%).
Step 6: Synthesis of Cis-tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino)cyclohexyl)carbamate
##STR00670##
[1451] Aqueous NaOH (0.23 g, 5.72 mmol) was added to a solution of
cis-methyl 5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl)
(methyl) amino)-2-methylbenzoate (1.3 g, 2.86 mmol) in MeOH (20 mL)
and stirred at 60.degree. C. for 1 h. After completion of the
reaction, ethanol was removed under reduced pressure and acidified
using dilute HCl up to pH 6 and pH 4 was adjusted using citric
acid. Extraction was carried out using ethyl acetate. Combined
organic layers were dried concentrated giving respective acid (1.13
g, 90.1%).
[1452] The acid (1.13 g, 2.57 mmol) was then dissolved in DMSO (10
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.87 g,
5.72 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (2.23 g, 4.28 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, reaction mass was poured into ice to
obtain solid, this was filtered and washed with acetonitrile
followed by purification with column chromatography to afford the
desired compound (0.8 g, 49%).
Step 7: Synthesis of
Cis-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1-
,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00671##
[1454] A stirred solution of cis-tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl) (methyl)amino)cyclohexyl)carbamate (0.8 g,
1.39 mmol) in DCM (25 mL) was cooled to 0.degree. C. and TFA (5 mL)
was added to it. The reaction mixture was stirred at room
temperature for 1 h. Upon completion, reaction was concentrated to
dryness. Residue was basified with aqueous sodium bicarbonate till
pH 8 and aqueous layer extracted with 20% MeOH/DCM. The combined
organic layers were dried over sodium sulfate, filtered and
concentrated to afford the desired compound (600 mg, 91% yield).
LCMS: 475.15 (M+1).sup.+; HPLC % 95.88 (@254 nm) (R.sub.t; 4.832;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 8.22 (t, 1H), 7.20
(s, 1H), 7.01 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=3.6 Hz),
2.82-2.86 (m, 2H), 2.53 (s, 3H), 2.18 (s, 3H), 2.13 (s, 3H), 2.10
(s, 3H), 1.82-1.84 (m, 2H), 1.34-1.44 (m, 6H).
[1455] 100 mg of
cis-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1-
,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (free base) was
then dissolved in methanolic HCl (5 mL) and stirred at room
temperature for 1 h. On completion of salt formation solvent was
removed under reduced pressure and resulting solid was purified by
washing with diethyl ether to afford the corresponding HCl salt.
Analytical Data of HCl salt: LCMS: 475.20 (M+1).sup.+; HPLC % 96.09
(@254 nm) (R.sub.t; 4.818; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.51 (s, 1H), 8.28 (t, 1H), 8.08
(s, 3H), 7.31 (s, 1H), 7.08 (s, 1H), 5.90 (s, 1H), 4.24 (d, 2H,
J=4.4 Hz), 3.05-3.13 (m, 2H), 2.54 (s, 3H), 2.19 (s, 6H), 2.12 (s,
3H), 1.83 (m, 2H), 1.66 (m, 2H), 1.59-1.60 (m, 2H), 1.46 (m,
2H).
Example 37
Synthesis of Compounds 59:
Trans-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-
-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00672##
[1456] Step 1: Synthesis of Trans-methyl
5-bromo-3-((4-((tert-butoxycarbonyl) amino) cyclohexyl) (methyl)
amino)-2-methylbenzoate
##STR00673##
[1458] To a stirred solution of trans-methyl
5-bromo-3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methylbenzo-
ate (3 g, 6.81 mmol) in acetonitrile (40 mL), cesium carbonate (4.4
g, 13.62 mmol) and methyl iodide (4.83 g, 34.05 mmol) were added;
the resulting reaction mixture was heated at 80.degree. C. for 7 h.
Upon completion, the reaction mixture was cooled to room
temperature and filtered, the residue was washed with ethyl acetate
and filtrate was concentrated to afford desired crude compound
which then purified by column chromatography giving the desired
compound (1.3 g, 43%).
Step 2: Synthesis of Trans-tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino) cyclohexyl)carbamate
##STR00674##
[1460] Aqueous NaOH (0.23 g, 5.72 mmol) was added to a solution of
trans-methyl 5-bromo-3-((4-((tert-butoxycarbonyl) amino)
cyclohexyl) (methyl) amino)-2-methylbenzoate (1.3 g, 2.86 mmol) in
MeOH (20 mL) and was stirred at 60.degree. C. for 1 h. After
completion of the reaction, ethanol was removed under reduced
pressure and the residue was acidified using dilute HCl up to pH 6
and to pH 4 using citric acid. The mixture was extracted was with
ethyl acetate. The combined organic layers were dried, filtered and
concentrated to give the respective acid (1 g, 83%).
[1461] The acid (1 g, 2.27 mmol) was then dissolved in DMSO (5 mL)
and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.65 g, 4.54
mmol) was added to it. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (1.7 g, 3.4 mmol) was added to
it and stirring was continued for overnight. After completion of
the reaction, reaction mass was poured into ice to obtain solid,
this was filtered and washed with acetonitrile followed by
purification with column chromatography to afford the desired
compound (0.7 g, 54%).
Step 3: Synthesis of
Trans-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-
-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00675##
[1463] A stirred solution of trans-tert-butyl
(4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methylphenyl)(methyl)amino) cyclohexyl)carbamate (0.7 g,
1.21 mmol) in DCM (8 mL) was cooled to 0.degree. C. and TFA (2.5
mL) was added to it. Reaction mass was stirred at room temperature
for 1 h. On completion, reaction was concentrated to dryness.
Residue was basified with aqueous sodium bicarbonate till pH 8 and
aqueous layer extracted with 20% MeOH/DCM. The combined organic
layers were dried over sodium sulfate, filtered and concentrated to
afford crude material which was then purified by solvent washings
giving desired compound (0.5 g, 86%). LCMS: 475.20 (M+1).sup.+;
HPLC % 92.35 (@254 nm) (R.sub.t; 4.416; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 8.21 (t, 1H), 7.18 (s, 1H), 7.01
(s, 1H), 5.85 (s, 1H), 4.23 (d, 2H), 2.63-2.66 (m, 2H), 2.55 (s,
3H), 2.17 (s, 3H), 2.10 (s, 6H), 1.79-1.82 (m, 2H), 1.60 (m, 2H),
1.46-1.49 (m, 2H), 1.06-1.09 (m, 2H).
[1464] 100 mg of
trans-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-
-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (free base) was
then dissolved in methanolic HCl (5 mL) and stirred at room
temperature for 1 h. On completion of salt formation, the solvent
was removed under reduced pressure and resulting solid was purified
by washing with diethyl ether to afford the corresponding HCl salt.
Analytical data of HCl salt: LCMS: 475.20 (M+1).sup.+; HPLC % 91.40
(@254 nm) (R.sub.t; 4.408; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.51 (s, 1H), 8.28 (t, 1H), 8.06
(s, 3H), 7.31 (s, 1H), 7.08 (s, 1H), 5.89 (s, 1H), 4.24 (d, 2H,
J=4.4 Hz), 2.92 (bs, 1H), 2.62-2.91 (m, 4H), 2.19 (s, 3H), 2.13 (s,
3H), 2.11 (s, 3H), 1.97 (d, 2H, J=10.8 Hz), 1.70 (m, 2H), 1.53-1.56
(m, 2H), 1.31-1.34 (m, 2H).
Example 38
Synthesis of Compound 60:
Cis-3-((4-acetamidocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00676##
[1466] To a stirred solution of compound
cis-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-1-
,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (free base)
(0.225, 0.474 mmol) in DMF (3 mL), EDCI.HCl (0.138 g, 0.718 mmol),
HOBt (0.064 g, 0.47 mmol), triethyl amine (0.1 g, 0.99 mmol) and
acetic acid (0.057 g, 0.949 mmol) were added at room temperature
and stirred at same temperature for 18 h. On completion, water was
added and extracted with 10% MeOH/DCM. Combined organic layers were
dried, concentrated giving crude material; which then purified by
column chromatography to afford the desired compound (0.17 g, 72%).
LCMS: 517.25 (M+1).sup.+; HPLC % 95.83 (@254 nm) (R.sub.t; 4.894;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.22
(t, 1H), 7.76 (d, 1H, J=7.2 Hz), 7.23 (s, 1H), 7.03 (s, 1H), 5.85
(s, 1H), 4.23 (d, 2H, J=3.6 Hz), 3.71 (bs, 1H), 2.89 (m, 1H), 2.53
(s, 3H), 2.18 (s, 3H), 2.16 (s, 3H), 2.10 (s, 3H), 1.81 (s, 3H),
1.74-1.76 (m, 2H), 1.53 (m, 2H), 1.39 (m, 4H).
Example 39
Synthesis of Compounds 61:
Trans-3-((4-acetamidocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-
-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00677##
[1468] To a stirred solution of
trans-3-((4-aminocyclohexyl)(methyl)amino)-5-bromo-N-((4,6-dimethyl-2-oxo-
-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (0.225, 0.474
mmol) in DMF (3 mL), EDCI.HCl (0.138 g, 0.718 mmol), HOBt (0.064 g,
0.47 mmol), triethyl amine (0.1 g, 0.99 mmol) and acetic acid
(0.057 g, 0.949 mmol) were added at room temperature and stirred at
same temperature for 18 h. On completion, water was added and
extracted with 10% MeOH/DCM. Combined organic layers were dried,
concentrated giving crude material; which then purified by column
chromatography to afford desired compound (0.13 g, 53%).
[1469] LCMS: 517.20 (M+1).sup.+; HPLC % 93.92 (@254 nm) (R.sub.t;
4.713; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (s, 1H), 8.20 (t, 1H), 7.65 (d, 1H, J=7.6 Hz), 7.18 (s, 1H),
7.01 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=3.6 Hz), 3.42-3.44 (m,
1H), 2.68-2.71 (m, 1H), 2.56 (s, 3H), 2.18 (s, 3H), 2.10 (s, 6H),
1.77-1.80 (m, 2H), 1.74 (s, 3H), 1.62-1.65 (m, 2H), 1.46-1.54 (m,
2H), 1.07-1.23 (m, 2H).
Example 40
Synthesis of Compound 62:
2-bromo-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)benzamide
##STR00678##
[1470] Step 1: methyl 2-bromo-3-nitrobenzoate
##STR00679##
[1472] To a stirred solution of 2-bromo-3-nitrobenzoate (3 g, 12.19
mmol) in DMF (33 mL), sodium carbonate (5.16 g, 48.67 mmol) and
methyl iodide (6.92 g, 48.67 mmol) were added. Resulting reaction
mixture was heated at 60.degree. C. for 4 h. On completion, water
was added to the mixture and extraction carried out using DCM. The
combined organic layers were dried and concentrated under reduced
pressure to obtain crude methyl 2-bromo-3-nitrobenzoate (4 g,
crude).
Step 2: Synthesis of methyl 3-amino-2-bromobenzoate
##STR00680##
[1474] To a stirred solution of methyl 2-bromo-3-nitrobenzoate (4
g, 15.38 mmol) in ethanol (20 mL), ammonium chloride (4 g, 74.07
mmol) dissolved in water (20 mL) and iron powder (6.88 g, 123 mmol)
were added under stirring. Resulting reaction mixture was heated at
80.degree. C. for 1 h. On completion, water was added to the
mixture and filtered through celite. The filtrate was extracted
with ethyl acetate and the combined organic layers washed with
water and sodium bicarbonate solution, dried and concentrated under
reduced pressure giving the desired methyl 3-amino-2-bromobenzoate
(3 g, 85%) which was used without further purification.
Step 3: Synthesis of methyl 2-bromo-3-(cyclopentylamino)
benzoate
##STR00681##
[1476] To a stirred solution of methyl 3-amino-2-bromobenzoate (3
g, 13.0 mmol) and cyclopentanone (5.4 g, 64.28 mmol) in methanol
(20 mL), acetic acid (1.56 g, 26 mmol) was added and reaction
stirred at room temperature for 3 h. Then sodium cyanoborohydride
(2 g, 31.7 mmol) was added and reaction stirred overnight. On
completion, solvent was removed under reduced pressure and crude
material purified by column chromatography to afford methyl
2-bromo-3-(cyclopentylamino) benzoate (1.4 g, 36%).
Step 4: Synthesis of methyl 2-bromo-3-(cyclopentyl (methyl) amino)
benzoate
##STR00682##
[1478] To a stirred solution of methyl 2-bromo-3-(cyclopentylamino)
benzoate (1.4 g, 4.69 mmol) in acetonitrile (10 mL), cesium
carbonate (3 g, 9.2 mmol) and methyl iodide (3.38 g, 23.3 mmol)
were added and the resulting reaction mixture heated at 80.degree.
C. for 12 h. On completion, the mixture was cooled to room
temperature and filtered and the residue was washed with ethyl
acetate. The filtrate was concentrated and purified by column
chromatography to afford methyl 2-bromo-3-(cyclopentyl (methyl)
amino) benzoate (1.1 g, 75%).
Step 5: Synthesis of 2-bromo-3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide
##STR00683##
[1480] Aqueous NaOH (0.064 g, 1.6 mmol) was added to a solution of
methyl 2-bromo-3-(cyclopentyl (methyl) amino) benzoate (0.25 g,
0.801 mmol) in MeOH (5 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate. Combined organic layers were dried concentrated giving
respective acid (0.2 g, 84%).
[1481] The acid (0.2 g, 0.67 mmol) was then dissolved in DMSO (1.5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.204 g,
1.34 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.516 g, 1.00 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, the mixture was poured into ice to
obtain a solid, which was filtered and washed with acetonitrile.
Final purification by preparative HPLC provided
2-bromo-3-(cyclopentyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)benzamide
as the TFA salt (0.02 g, 6.89%). LCMS: 432.10 (M+1).sup.+; HPLC %
96.05 (@254 nm) (R.sub.t; 4.908); .sup.1H NMR (DMSO-d.sub.6, 400
MHz) .delta. 11.47 (s, 1H), 8.21 (t, 1H), 7.26-7.29 (m, 2H), 6.94
(d, 1H, J=8 Hz), 5.86 (s, 1H), 4.25 (d, 2H), 3.59 (m, 1H), 2.56 (s,
3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.61-1.67 (m, 4H), 1.46 (bs,
4H).
Example 41
Synthesis of Compound 63:
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-vinylbenzamide
##STR00684##
[1482] Step 1: Synthesis of methyl 2-bromo-3-nitrobenzoate
##STR00685##
[1484] To a stirred solution of 2-bromo-3-nitrobenzoic acid (3 g,
12.19 mmol) in DMF (30 mL), sodium carbonate (5.16 g, 48.6 mmol)
and methyl iodide (6.92 g, 48.7 mmol) were added. Resulting
reaction mixture was heated at 60.degree. C. for 8 h. On
completion, solid precipitated was filtered, residue washed with
ethyl acetate (5 times). Combined organic layers were dried,
concentrated under reduced pressure giving desired crude
2-bromo-3-nitrobenzoate (4 g, 99%) which was used without further
purification.
Step 2: Synthesis of methyl 3-amino-2-bromobenzoate
##STR00686##
[1486] To stirred solution of above crude 2-bromo-3-nitrobenzoate
(4 g, 15.38 mmol) in ethanol (20 mL), ammonium chloride (4 g, 74.0
mmol) dissolved in water (20 mL) and iron powder (6.87 g, 123 mmol)
were added under stirring. The resulting reaction mixture was
heated at 80.degree. C. for 7 h. On completion, the reaction
mixture was filtered through celite, washing with water and ethyl
acetate and the filtrate extracted with ethyl acetate. The combined
organic layers were dried and concentrated under reduced pressure
giving the desired crude product (3 g).
Step 3: Synthesis of methyl 2-bromo-3-(cyclopentylamino)
benzoate
##STR00687##
[1488] To a stirred solution of methyl 3-amino-2-bromobenzoate (3
g, 13.01 mmol) and cyclopentanone (5.6 g, 66 mmol) in methanol (30
mL), acetic acid (1.59 g, 26.6 mmol) was added and the reaction
stirred at room temperature for 3 h. Then sodium cyanoborohydride
(2.08 g, 29.4 mmol) was added and the mixture stirred overnight. On
completion, solvent was removed under reduced pressure and crude
material was purified by column chromatography to afford methyl
2-bromo-3-(cyclopentylamino) benzoate (1.4 g, 36%).
Step 4: Synthesis of methyl 2-bromo-3-(cyclopentyl (methyl) amino)
benzoate
##STR00688##
[1490] To a stirred solution of methyl 2-bromo-3-(cyclopentylamino)
benzoate (1.4 g, 4.69 mmol) in acetonitrile (20 mL), cesium
carbonate (3.0 g, 9.2 mmol) and methyl iodide (3.38 g, 23.4 mmol)
were added and the resulting reaction mixture heated at 80.degree.
C. for 12 h. On completion, the mixture was cooled to room
temperature and filtered and the residue washed with ethyl acetate.
The filtrate was concentrated and then purified by column
chromatography to afford desired methyl 2-bromo-3-(cyclopentyl
(methyl) amino) benzoate (1.1 g, 75%).
Step 5: Synthesis of methyl
3-(cyclopentyl(methyl)amino)-2-vinylbenzoate
##STR00689##
[1492] To a stirred solution of methyl 2-bromo-3-(cyclopentyl
(methyl) amino) benzoate (1 equiv.) and
4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.2 equiv.) in
dioxane/water mixture (5 mL+1 mL), Na.sub.2CO.sub.3 (3.6 equiv.)
was added and solution purged with argon for 15 min. Then,
Pd(PPh.sub.3).sub.4 (0.1 equiv.) was added and argon purged for 10
min. and the reaction mixture heated at 100.degree. C. for 2 h. On
completion, the reaction mixture was diluted with water and
extracted with 10% MeOH/DCM. The combined organic layers were dried
over Na.sub.2SO.sub.4 and solvent removed under reduced pressure to
afford crude material which was purified by column chromatography
over silica gel to afford the desired methyl
3-(cyclopentyl(methyl)amino)-2-vinylbenzoate.
Step 6: Synthesis of
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-vinylbenzamide
##STR00690##
[1494] To a stirred solution of methyl
3-(cyclopentyl(methyl)amino)-2-vinylbenzoate (1 equiv.) in ethanol
(10 mL), aqueous NaOH solution (1.5 equiv. in 10 mL water) was
added and reaction stirred at 60.degree. C. for 4 h. On completion,
ethanol was removed under reduced pressure and residue acidified
with 1N HCl to pH 6. The precipitate was filtered, washed with
water and dried to obtain pure corresponding acid.
[1495] To a solution of this respective acid (1 equiv.) in DMSO (10
mL), PyBOP (1.5 equiv.) was added and reaction stirred at rt for 15
min. Then 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.)
was added and reaction stirred overnight. On completion, water was
added and the resulting solid filtered and washed with water. This
solid was stirred with acetonitrile for 10 min and filtered again
to obtain
3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3--
yl)methyl)-2-vinylbenzamide which was purified by preparative HPLC
yielding the TFA salt (0.045 g, 10.39%). LCMS: 380.25 (M+1).sup.+;
HPLC % 98.00 (@254 nm) (R.sub.t; 4.323); .sup.1H NMR (DMSO-d.sub.6,
400 MHz) .delta. 11.51 (s, 1H), 8.01 (s, 1H), 7.25 (bs, 2H),
6.81-6.88 (m, 2H), 5.85 (s, 1H), 5.49 (d, 1H, J=18 Hz), 5.40 (bs,
1H), 4.34 (3H merged in solvent peak), 4.19 (d, 3H, J=4.8 Hz), 2.17
(s, 3H), 2.10 (s, 3H), 1.61 (m, 4H), 1.47 (bs, 4H).
Example 42
Synthesis of Compound 64:
5-chloro-3-(cyclopentyl(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropy-
ridin-3-yl)methyl)-2-methylbenzamide
##STR00691##
[1496] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00692##
[1498] 5-Chloro-2-methylbenzoic acid (4 g, 23.39 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree. C. lot wise.
After 10 minutes nitrating mixture [Prepared by mixing Conc. HNO3
(3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4 (4.4 mL)] was added
drop wise at -10.degree. C. Resulting reaction mixture was stirred
at -10.degree. C. for 30 minutes. On completion, the mixture was
poured on ice cold water and the resulting solid precipitated was
filtered, washed with water and dried under vacuum giving the
desired chloro-2-methyl-3-nitrobenzoic acid (4.95 g, 99%).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00693##
[1500] To a stirred solution of chloro-2-methyl-3-nitrobenzoic acid
(6.75 g, 31.25 mmol) in DMF (33 mL), sodium carbonate (13.23 g,
125.18 mmol) and methyl iodide (17.77 g, 125.2 mmol) were added.
The resulting reaction mixture was heated at 60.degree. C. for 4 h.
On completion, water was added and extracted out using DCM.
Combined organic layers were dried, concentrated under reduced
pressure and purified by column chromatography over silica (60-120
mesh size) giving methyl 5-chloro-2-methyl-3-nitrobenzoate (6 g,
83.65%).
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR00694##
[1502] To a stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6 g, 26.13 mmol) in ethanol (60
mL), ammonium chloride (6 g, 112.1 mmol) dissolved in water (60 mL)
and iron powder (11.88 g, 208.4 mmol) were added under stirring.
The resulting reaction mixture was heated at 80.degree. C. for 1 h.
On completion, water was added to reaction and the mixture was
filtered through celite. The filtrate was extracted with ethyl
acetate and the combined organic layers were washed with water,
dried, concentrated under reduced pressure giving methyl
3-amino-5-chloro-2-methyl benzoate which was used without further
purification.
Step 4: Synthesis of methyl
5-chloro-3-(cyclopentylamino)-2-methylbenzoate
##STR00695##
[1504] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (1 g, 5.02 mmol) and cyclopentanone (2.1 g, 25 mmol) in
methanol (10 mL), acetic acid (0.3 g, 5.02 mmol) was added and
reaction stirred at room temperature for 3 h. Then sodium
cyanoborohydride (0.37 g, 5.90 mmol) was added and reaction stirred
overnight. On completion, solvent was removed under reduced
pressure and crude material was purified by column chromatography
to afford methyl 5-chloro-3-(cyclopentylamino)-2-methylbenzoate
(0.62 g, 46.2%).
Step 5: Synthesis of methyl 5-chloro-3-(cyclopentyl (ethyl)
amino)-2-methylbenzoate
##STR00696##
[1506] To a stirred solution of methyl
5-chloro-3-(cyclopentylamino)-2-methylbenzoate (0.62 g, 2.32 mmol)
in dry DMF (10 mL), cesium carbonate (3.78 g, 11.6 mmol) and ethyl
iodide (5.43 g, 34.8 mmol) were added and the resulting reaction
mass was heated at 80.degree. C. for 18 h. On completion, reaction
mixture was cooled to room temperature and filtered. The residue
was washed with ethyl acetate and filtrate concentrated and
purified by column chromatography to afford desired methyl
5-chloro-3-(cyclopentyl (ethyl) amino)-2-methylbenzoate (0.101 g,
15%).
Step 6: Synthesis of 5-chloro-3-(cyclopentyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00697##
[1508] Aqueous NaOH (0.1 g, 2.5 mmol) was added to a solution of
methyl 5-chloro-3-(cyclopentyl (ethyl) amino)-2-methylbenzoate (0.5
g, 1.69 mmol) in EtOH (10 mL) and stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using ethyl
acetate and the combined organic layers were dried concentrated
giving respective acid (0.41 g, 86%).
[1509] The acid (0.10 g, 0.355 mmol) was then dissolved in DMSO
(1.5 mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.104
g, 0.77 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.277 g, 0.533 mmol) was
added to it and stirring was continued for overnight. After
completion of the reaction, the mixture was poured into ice and the
resulting solid filtered and washed with acetonitrile followed by
additional solvent washings to yield 5-chloro-3-(cyclopentyl
(ethyl) amino)-N-((4, 6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide (0.107 g, 72%). LCMS: 416.25 (M+1).sup.+;
HPLC % 91.83 (@254 nm) (R.sub.t; 5.021); .sup.1H NMR (DMSO-d.sub.6,
400 MHz) .delta. 11.45 (s, 1H), 8.21 (t, 1H), 7.19 (s, 1H), 6.95
(s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4 Hz), 3.45 (t, 1H),
2.94-2.96 (m, 2H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H),
1.58-1.64 (m, 4H), 1.47 (m, 2H), 1.32 (m, 2H), 0.77 (t, 3H, J=6.8
Hz).
Example 43
Synthesis of Compound 19:
5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)benzamide
##STR00698##
[1510] Step 1: Synthesis of 5-bromo-2-chloro-3-nitrobenzoic
acid
##STR00699##
[1512] 5-bromo-2-chlorobenzoic acid: (5 g, 27.3 mmol) was added to
cooled conc. H.sub.2SO.sub.4 (20 mL) at -10.degree. C. in small
portions. After 10 minutes nitrating mixture {prepared by mixing
conc. HNO.sub.3 (2.5 mL) with conc. H.sub.2SO.sub.4 (10 mL)} was
added drop wise at -10.degree. C. The reaction mixture was stirred
at -10.degree. C. for 30 minutes. On completion, the reaction
mixture was poured into ice cold water and the solid collected by
filtration. The solid was washed with water and dried under vacuum
to give 5-bromo-2-chloro-3-nitrobenzoic acid: (3.6 g, 60.5%).
Step 2: Synthesis of methyl 5-bromo-2-chloro-3-nitrobenzoate
##STR00700##
[1514] To stirred solution of -bromo-2-chloro-3-nitrobenzoic acid
(3.6 g, 12.9 mmol) in DMF (40 mL) was added sodium carbonate (4.1
g, 38.6 mmol) and methyl iodide (4 mL, 64.3 mmol). The reaction
mixture was heated at 60.degree. C. for 8 h. On completion, water
was added to the reaction mixture and the product was extracted
with ethyl acetate. The combined organic layers were washed with
sat. bicarbonate solution and 5 N HCl. The organics were dried and
concentrated under reduced pressure to afford methyl
5-bromo-2-chloro-3-nitrobenzoate (3.4 g, 89.9%).
Step 3: Synthesis of methyl 3-amino-5-bromo-2-chlorobenzoate
##STR00701##
[1516] To stirred solution of methyl
5-bromo-2-chloro-3-nitrobenzoate (3.4 g, 11.6 mmol) in ethanol (19
mL) was added ammonium chloride (3.4 g, 57.8 mmol) dissolved in
water (30 mL) and iron powder (5.16 g, 92.5 mmol). The reaction
mixture was heated at 80.degree. C. for 1 h. On completion, the
reaction mixture was filtered through celite washing with ethanol
and ethylacetate. The filtrate was extracted with ethyl acetate and
the combined organic layers were washed with water, dried and
concentrated under reduced pressure to give methyl
3-amino-5-bromo-2-chlorobenzoate (2.8 g, 92.1%).
Step 4: Synthesis of methyl
5-bromo-2-chloro-3-(cyclopentylamino)benzoate
##STR00702##
[1518] To a stirred solution of methyl
3-amino-5-bromo-2-chlorobenzoate (2.8 g, 10.6 mmol) and
cyclopentanone (4.47 g, 53.2 mmol) in methanol (20 mL) was added
acetic acid (1.3 mL, 21.2 mmol) and the reaction stirred at room
temperature for 3 h. Then sodium cyanoborohydride (1.7 g, 26.6
mmol) was added and the reaction stirred overnight. On completion,
the solvent was removed under reduced pressure and the product was
purified by column chromatography to afford methyl
5-bromo-2-chloro-3-(cyclopentylamino)benzoate (0.5 g, 14.2%).
Step 5: Synthesis of methyl
5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)benzoate
##STR00703##
[1520] To a stirred solution of methyl
5-bromo-2-chloro-3-(cyclopentylamino)benzoate (0.550 g, 1.65 mmol)
in acetonitrile (20 mL) was added cesium carbonate (1.08 g, 3.31
mmol) and methyl iodide (1.17 g, 8.25 mmol). The reaction mixture
was heated at 80.degree. C. for 12 h. On completion, the reaction
mixture was cooled to room temperature and filtered. The residue
was washed with ethyl acetate and the filtrate was concentrated and
purified by column chromatography to afford methyl
5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)benzoate (0.350 g,
61%).
Step 6: Synthesis of
5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)benzamide
##STR00704##
[1522] Aqueous NaOH (0.06 g, 1.52 mmol) was added to a solution of
methyl 5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)benzoate (0.350
g, 1.01 mmol) in EtOH (4 mL) and water (1 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and to pH 4 with citric acid. The product was extracted
with ethyl acetate. The combined organic layers were dried and
concentrated to give the desired acid (0.266 g, 79.4%).
[1523] The acid (0.265 g, 0.80 mmol) was then dissolved in DMSO (3
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (0.243 g,
1.60 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.624 g, 1.20 mmol) was
added and stirring was continued overnight. After completion of the
reaction, the reaction mixture was poured into ice to obtain a
solid. The solid was filtered and washed with acetonitrile followed
by purification by column chromatography and prep. HPLC to give
5-bromo-2-chloro-3-(cyclopentyl(methyl)amino)-N-((4,6-dimethyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)benzamide (0.012 g, 3.22%).
[1524] Analytical Data: LCMS: 466.05 (M+1).sup.+; HPLC: 99.28%
(@254 nm) (R.sub.t; 6.917; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR (CD3OD, 400
MHz) .delta. 7.42 (d, 1H, J=2 Hz), 7.24 (d, 1H, J=1.6 Hz), 6.16 (s,
1H), 4.66 (s, 2H), 3.71 (m, 1H), 2.70 (s, 3H), 2.37 (s, 3H), 2.25
(s, 3H), 1.79 (m, 2H), 1.70 (m, 2H), 1.56 (m, 4H).
Example 44
Syntheses of Compounds 91, 92, 97, 98, 102, 104, 105, 117-123, 126,
127, 137, 144, 157, 191, 192, 205-209, 212, 213, 222, 243-245, 268,
269, 273, 276-279, 282-287, 290, 301, 302, 306, 308-311, 313, 315,
319-322, 328, 332-336, 339, 340, 344, 347-349, 356, 386, 387, 389,
390, and 392-417
[1525] Compounds 91, 92, 97, 98, 102, 104, 105, 117-123, 126, 127,
137, 144, 157, 191, 192, 205-209, 212, 213, 222, 243-245, 268, 269,
273, 276-279, 282-287, 290, 301, 302, 306, 308-311, 313, 315,
319-322, 328, 332-336, 339, 340, 344, 347-349, 356, 386, 387, 389,
390, and 392-417 were synthesized as described below.
Compound 91: 3-(allyl (piperidin-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00705##
[1526] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00706##
[1528] 5-Chloro-2-methylbenzoic acid (4.0 g, 23 mmol) was added
portionwise to cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree.
C. After 10 minutes the nitrating mixture {consisting of
concentrated HNO3 (3.3 g, 52 mmol) and concentrated H.sub.2SO.sub.4
(4.4 mL)} was added dropwise at -10.degree. C. The mixture was
stirred at -10.degree. C. for 30 minutes and poured on ice cold
water. The solid precipitate was filtered, washed with water and
dried under vacuum to give the title compound (4.95 g, 99%).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00707##
[1530] To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic
acid (6.75 g, 31.3 mmol) in DMF (33 mL), were added sodium
carbonate (13.23 g, 125 mmol) and methyl iodide (17.77 g, 125
mmol). The mixture was heated at 60.degree. C. for 4 h. On
completion, water was added to the reaction mass and extraction was
carried out using DCM. The combined organic layers were dried,
concentrated under reduced pressure, and purified by column
chromatography over silica gel (to afford the title compound (6.0
g, 83%).
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR00708##
[1532] To a stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6.0 g, 26 mmol) in ethanol (60
mL) were added ammonium chloride (6.0 g, 110 mmol) dissolved in
water (60 mL) and iron powder (11.9 g, 208 mmol) under stirring.
The mixture was heated at 80.degree. C. for 1 h. On completion,
water was added and the reaction mixture was filtered through
celite. The filtrate was extracted with ethyl acetate. The combined
organic layers were washed with water, dried, concentrated under
reduced pressure to afford the title compound which was used
without further purification.
Step 4: Synthesis of tert-butyl
4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl) amino)
piperidine-1-carboxylate
##STR00709##
[1534] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (5.0 g, 25 mmol) and tert-butyl
4-oxopiperidine-1-carboxylate (25.3 g, 127 mmol) in methanol (50
mL), acetic acid (1.5 g, 25 mmol) was added and the reaction
stirred at room temperature for 8 h. Then sodium cyanoborohydride
(1.89 g, 30.1 mmol) was added at 0.degree. C. and the reaction
stirred overnight at room temperature. On completion, the solvent
was removed under reduced pressure and the crude material was
purified by column chromatography to afford the title compound (5.0
g, 52%).
Step 5: Synthesis of tert-butyl
4-(allyl(5-chloro-3-(methoxycarbonyl)-2-methylphenyl)
amino)piperidine-1-carboxylate
##STR00710##
[1536] To a stirred solution of tert-butyl
4-((5-chloro-3-(methoxycarbonyl)-2-methylphenyl) amino)
piperidine-1-carboxylate (0.70 g, 1.82 mmol) in DMF (7 mL), was
added NaH (0.052 g, 2.19 mmol) at 0.degree. C. After stirring for
20 minutes, 3-bromoprop-1-ene (0.44 g, 3.64 mmol) was added and the
mixture was heated at 80.degree. C. for 15 h. On completion, the
reaction mass was quenched with ice cold water and extraction was
carried out using DCM. The combined organic layers were dried,
concentrated under reduced pressure and the residue was purified by
column chromatography to afford the title compound (0.3 g,
39%).
Step 6: Synthesis of tert-butyl 4-(allyl
(5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)
carbamoyl)-2-methylphenyl) amino) piperidine-1-carboxylate
##STR00711##
[1538] Aqueous NaOH (0.113 g, 2.83 mmol) was added to a solution of
tert-butyl 4-(allyl(5-chloro-3-(methoxycarbonyl)-2-methylphenyl)
amino)piperidine-1-carboxylate (0.60 g, 1.41 mmol) in ethanol (15
mL). After stirring at 60.degree. C. for 1 h ethanol was removed
under reduced pressure and the mixture acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using DCM. The combined organic layers were dried and
concentrated giving respective acid (0.5 g, 86%). The above acid
(0.5 g, 1.22 mmol) was then dissolved in DMSO (5 mL) and 3-(amino
methyl)-4, 6-dimethylpyridin-2(1H)-one (0.37 g, 2.44 mmol) was
added to it. The reaction mixture was stirred at room temperature
for 15 min before PYBOP (0.95 g, 1.83 mmol) was added to it and
stirring was continued overnight. The mixture was poured into ice
and extracted with 10% MeOH/DCM. The combined organic layers were
dried, concentrated, and purified by column chromatography to
afford the title compound (0.2 g, 30%).
Step 7: Synthesis of 3-(allyl (piperidin-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00712##
[1540] A stirred solution of compound tert-butyl 4-(allyl
(5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl) methyl)
carbamoyl)-2-methylphenyl) amino) piperidine-1-carboxylate (0.2 g,
0.36 mmol) in DCM (5 mL) was cooled to 0.degree. C. and TFA (0.5
mL) was added. The mixture was stirred at room temperature for 18
h. and concentrated to dryness. The residue was washed with diethyl
ether and then purified by preparative HPLC to afford the title
compound as TFA salt (0.06 g, 37%). LCMS: 443.25 (M+1).sup.+; HPLC:
97.14% (@254 nm) (R.sub.t; 5.074; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.38 (bs, 1H), 8.21
(t, 1H), 8.05 (d, 1H), 7.21 (d, 1H, J=4.4 Hz), 6.97 (d, 1H, J=1.6
Hz), 5.86 (s, 1H), 5.56-5.64 (m, 1H), 4.97-5.09 (m, 2H), 4.24 (d,
2H, J=4.4 Hz), 3.62 (d, 2H, J=5.2 Hz), 3.23 (m, 2H), 3.10-3.16 (m,
1H), 2.85-2.88 (m, 2H), 2.19 (s, 3H), 2.18 (s, 3H), 2.10 (s, 3H),
1.83-1.86 (m, 2H), 1.68-1.73 (m, 2H).
Compound 97:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-3-(piperidin-4-yl (propyl) amino) benzamide
##STR00713##
[1542] To a stirred solution of 3-(allyl (piperidin-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide (0.04 g, 0.09 mmol) in MeOH (3 mL) was
added 10% Pd/C (0.01 g) and reaction stirred at room temperature
under hydrogen (balloon pressure) for 2 h. The mixture was filtered
through celite and the filtrate was concentrated under reduced
pressure to obtain a crude solid which was purified by preparative
HPLC to afford the title compound as TFA salt (0.012 g, 40%). LCMS:
445.25 (M+1).sup.+; HPLC: 95.52% (@254 nm) (R.sub.t; 5.102; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (s, 1H), 8.39
(bs, 1H), 8.22 (t, 1H), 8.03 (bs, 1H), 7.25 (s, 1H), 6.98 (s, 1H),
5.86 (s, 1H), 4.24 (d, 2H, J=4.8 Hz), 3.24-3.27 (m, 2H), 3.00 (m,
1H), 2.92 (m, 2H), 2.83-2.86 (m, 2H), 2.18 (s, 3H), 2.17 (s, 3H),
2.10 (s, 3H), 1.79-1.82 (m, 2H), 1.67-1.71 (m, 2H), 1.18-1.23 (m,
2H), 0.75 (t, 3H, J=8 Hz).
Compound 144:
3,6-dichloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-[me-
thyl(piperidin-4-yl)amino]pyridine-2-carboxamide
##STR00714##
[1543] Step 1: Synthesis of methyl
3,4,6-trichloropyridine-2-carboxylate
##STR00715##
[1545] To a stirred solution of methyl
3,6-dichloropyridine-2-carboxylate (0.96 g, 4.66 mmol) in TFA (5
ml) was added hydrogen peroxide (30% w/w aqueous solution, 435
.mu.l, 2.5 mmol) and the reaction mixture was heated at 60.degree.
C. for 20 h. The reaction mixture was then cooled and slowly poured
onto saturated K.sub.2CO.sub.3 solution (100 ml), followed by
extraction of the aqueous layer with EtOAc (2.times.200 ml),
washing of the combined organic phases with brine (2.times.50 ml),
drying (Na.sub.2SO.sub.4) and evaporation. The desired
3,6-dichloro-2-(methoxycarbonyl)pyridin-1-ium-1-olate was used
crude in the next stage of the synthesis without any further
purification. To the crude
3,6-dichloro-2-(methoxycarbonyl)pyridin-1-ium-1-olate (.about.70%
purity, 2.40 g, 7.7 mmol) was added POCl.sub.3 (3.5 ml, 38 mmol)
and the solution was heated to 100.degree. C. for 4 h. After
cooling the POCl.sub.3 was removed in vacuo to give a white solid
which was chromatographed over silica gel eluting with 0% to 10% of
EtOAc in heptane to afford the title compound as colourless needles
(340 mg, 30% over two steps). LC-MS 100%, 2.20 min (3.5 minute
LC-MS method), m/z=239.8/241.7, .sup.1H NMR (500 MHz, Chloroform-d)
.delta. 7.51 (1H, s), 3.92 (3H, s).
Step 2: Synthesis of methyl
4-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(methyl)amino)-3,6-dichloropy-
ridine-2-carboxylate
##STR00716##
[1547] To a stirred solution of methyl
3,4,6-trichloropyridine-2-carboxylate (310 mg, 1.29 mmol) in DMF (5
ml) was added TEA (359 .mu.l, 2.58 mmol) followed by tert-butyl
4-(methylamino)piperidine-1-carboxylate (276 mg, 1.29 mmol) and the
reaction mixture was heated at 100.degree. C. for 4 h. The reaction
mixture was then cooled to room temperature and poured onto water
(50 ml), followed by extraction of the product into TBME
(3.times.50 ml), washing of the combined organics with brine (50
ml), drying with Na.sub.2SO.sub.4 and evaporation. The crude
product was then purified over a 10 g silica Isolute column eluting
with a gradient of 0% to 50% EtOAc in heptane to afford the title
compound as a white solid (95 mg, 18%). LC-MS 95%, 2.29 min (3.5
minute LC-MS method), m/z=418.1/419.8, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 6.87 (s, 1H) 4.12-4.34 (m, 2H) 3.99 (s, 3H)
3.69-3.80 (m, 1H) 2.79 (s, 3H) 2.64-2.77 (m, 2H) 1.75 (br. s., 4H)
1.48 (s, 9H).
Step 3: Synthesis of
4-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(methyl)amino)-3,6-dichloropy-
ridine-2-carboxylic acid
##STR00717##
[1549] To a stirred solution of methyl
4-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(methyl)amino)-3,6-dichloropy-
ridine-2-carboxylate (95 mg, 0.23 mmol) in THF (5 ml) was added 2M
aqueous NaOH (2.3 ml, 4.54 mmol) and the reaction mixture was left
to stir at room temperature for 16 h after which time the THF was
evaporated in vacuo. The aqueous phase was then treated with an
aqueous 10% citric acid solution to pH 5-6 and then extracted with
EtOAc (3.times.50 ml), the combined organic phases were then dried
with Na.sub.2SO.sub.4 and evaporated to give the title compound as
a white powder (77 mg, 84%). LC-MS 100%, 2.02 min (3.5 minute LC-MS
method), m/z=403.9/405.6, .sup.1H NMR (500 MHz, Methanol-d4)
.delta. 7.09 (s, 1H) 4.14-4.22 (m, 2H) 3.80-3.89 (m, 1H) 2.84 (s,
5H) 1.74-1.81 (m, 4H) 1.46 (s, 9H).
Step 4: Synthesis of tert-butyl
4-[(3,6-dichloro-2-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]c-
arbamoyl} pyridin-4-yl)(methyl)amino]piperidine-1-carboxylate
##STR00718##
[1551] To a stirred solution of
4-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(methyl)amino)-3,6-dichloropy-
ridine-2-carboxylic acid (75 mg, 0.19 mmol) in DMF (2 ml) cooled to
0.degree. C. h was added DIPEA (48 .mu.l, 0.28 mmol) followed by
PyBOP (116 mg, 0.22 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89% purity,
35 mg, 0.2 mmol). The reaction mixture was then stirred at room
temperature for 16 h after which the reaction was poured onto water
(30 ml) and extracted with EtOAc (3.times.50 ml) after which the
combined organics were washed with brine (50 ml) and dried
(Na.sub.2SO.sub.4) and evaporated. The crude product was purified
using a 5 g silica Isolute column eluting with a gradient of 0% to
100% EtOAc in heptane to afford the title compound as a white solid
which contained .about.45% w/w tripyrrolidinophosphene oxide by
NMR--this compound was used in the next stage of the synthesis
without any further purification. LC-MS 93%, 1.97 min (3.5 minute
LC-MS method), m/z=538.0/539.45/542.05, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 10.03 (br. s, 1H), 8.13-7.97 (m, 1H), 6.79
(s, 1H), 5.87 (s, 1H), 4.50 (d, J=6.1 Hz, 2H), 4.16 (br. m, 2H),
3.78 3.62 (m, 1H), 2.72 (s, 3H), 2.65 (br. m, 2H), 2.35 (s, 3H),
2.22 (s, 3H), 1.73-1.63 (m, 4H), 1.44 (s, 9H).
Step 5: Synthesis of
3,6-dichloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-[me-
thyl(piperidin-4-yl)amino]pyridine-2-carboxamide hydrochloride
##STR00719##
[1553]
Tert-butyl4-[(3,6-dichloro-2-{[(4,6-dimethyl-2-oxo-1,2-dihydropyrid-
in-3-yl)methyl]carbamoyl}pyridin-4-yl)
(methyl)amino]piperidine-1-carboxylate (310 mg, 1.29 mmol) was
dissolved in 4M HCl in dioxane solution (3 ml) and stirred at room
temperature for 16 h after which the solvent was evaporated and the
compound purified by preparative HPLC to afford the title compound
as a beige powder (13 mg, 14%). LC-MS 100%, 2.41 min (7 minute
LC-MS method), m/z=438.1/439.8, .sup.1H NMR (500 MHz, Methanol-d4)
.delta. 7.12 (s, 1H), 6.11 (s, 1H), 4.46 (s, 2H), 4.01-3.81 (m,
1H), 3.45 (d, J=12.9 Hz, 2H), 3.12-2.97 (m, 2H), 2.82 (s, 3H), 2.35
(s, 3H), 2.22 (s, 3H), 2.13-1.93 (m, 4H).
Compound 386:
3,6-dichloro-n-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-[me-
thyl (oxan-4-yl)amino]pyridine-2-carboxamide
##STR00720##
[1554] Step 1: Synthesis of methyl
3,6-dichloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylate
[1555] To a stirred solution of methyl
3,4,6-trichloropyridine-2-carboxylate (600 mg, 2.50 mmol) in DMF
(12 ml) was added TEA (696 .mu.l, 4.99 mmol) followed by
N-methyloxan-4-amine (287 mg, 2.50 mmol) and the reaction mixture
was heated at 100.degree. C. for 20 h. The reaction mixture was
then cooled to room temperature and poured onto water (100 ml),
followed by extraction of the product into EtOAc (3.times.100 ml),
washing of the combined organics with brine (50 ml), drying with
Na.sub.2SO.sub.4 and evaporation. The crude product was then
purified over a 10 g silica Isolute column eluting with a gradient
of 0% to 60% EtOAc in heptane to afford the title compound as a
white solid (114 mg, 14%). LC-MS 100%, 1.91 min (3.5 minute LC-MS
method), m/z=319.3/320.9, .sup.1H NMR (500 MHz, Chloroform-d)
.delta. 6.87 (s, 1H), 4.15-4.00 (m, 2H), 3.98 (s, 3H), 3.93-3.71
(m, 1H), 3.54-3.24 (m, 2H), 2.82 (s, 3H), 1.94 (dd, J=12.1, 4.7 Hz,
2H), 1.70 (d, J=10.2 Hz, 2H). (br. s., 4H) 1.48 (s, 9H)
Step 2: Synthesis of
3,6-dichloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylic
acid
##STR00721##
[1557] To a stirred solution of methyl
3,6-dichloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylate (114
mg, 0.36 mmol) in THF (5 ml) was added 2M aqueous NaOH (0.89 ml,
1.79 mmol) and the reaction mixture was left to stir at room
temperature for 20 h after which time the THF was evaporated in
vacuo. The aqueous phase was then treated with an aqueous 10%
citric acid solution to pH 5-6 and then extracted with EtOAc
(3.times.50 ml) followed by a solution of 1:1 IPA/CHCl.sub.3
(2.times.50 ml), the combined organic phases were then washed with
brine (50 ml) dried with Na.sub.2SO.sub.4 and evaporated to give
the title compound as a white solid (85 mg, 78%). LC-MS 100%, 1.47
min (3.5 minute LC-MS method), m/z=305.5/306.9, .sup.1H NMR (500
MHz, Methanol-d4) .delta. 7.10 (s, 1H), 4.03 (dd, J=11.5, 4.4 Hz,
2H), 3.99-3.86 (m, 1H), 3.47 (t, J=11.0 Hz, 2H), 2.90 (s, 3H),
2.05-1.90 (m, 2H), 1.74 (d, J=12.4 Hz, 2H).
Step 3: Synthesis of
3,6-dichloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-[me-
thyl (oxan-4-yl)amino]pyridine-2-carboxamide
##STR00722##
[1559] To a stirred solution of
3,6-dichloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylic acid
(85 mg, 0.28 mmol) in DMF (2 ml) was added DIPEA (73 .mu.l, 0.42
mmol) and HATU (127 mg, 0.33 mmol). The reaction was stirred at
room temperature for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 52 mg,
0.31 mmol) was added and the reaction was stirred at room
temperature for 16 h. The reaction mixture was then poured onto 50
ml of water and the aqueous phase was extracted with EtOAc
(3.times.50 ml), washed with brine (50 ml) dried (Na.sub.2SO.sub.4)
and evaporated to give an oil. The product was then purified using
a 5 g silica Isolute column eluting with 0% to 5% MeOH in DCM and
evaporated to give a glassy solid which was triturated with diethyl
ether and filtered to afford the title compound as a white powder
(50 mg, 41%). LC-MS 95%, m/z=439.0/440.8, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 11.33 (br. s, 1H), 8.20 (br. s, 1H), 6.79 (s,
1H), 5.93 (s, 1H), 4.51 (s, 2H), 4.02 (dd, J=11.4, 3.8 Hz, 2H),
3.92-3.72 (m, 1H), 3.39 (t, J=11.3 Hz, 2H), 2.78 (s, 3H), 2.38 (s,
3H), 2.27 (s, 3H), 1.89 (tt, J=12.0, 6.2 Hz, 2H), 1.83-1.52 (m,
2H).
Compound 387:
3-bromo-6-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-
-[methyl(oxan-4-yl)amino]pyridine-2-carboxamide
##STR00723##
[1560] Step 1: Synthesis of methyl
3-bromo-6-chloropyridine-2-carboxylate
##STR00724##
[1562] To a stirred solution of
3-bromo-6-chloropyridine-2-carboxylic acid (2.00 g, 8.46 mmol) in
MeOH (40 ml) was added H.sub.2SO.sub.4 (189 .mu.l, 3.55 mmol) and
the solution was heated to reflux for 16 h. The reaction was cooled
and the MeOH removed under reduced pressure, the resulting solid
was dissolved in EtOAc (100 ml) and washed with saturated
NaHCO.sub.3 solution (2.times.50 ml) followed by brine (50 ml),
dried (Na.sub.2SO.sub.4) and evaporated under reduced pressure to
afford the title compound as an off-white solid (1.97 g, 93%) which
was used without any further purification. LC-MS 98%,
m/z=249.8/251.7/253.7
Step 2: Synthesis of methyl
3-bromo-4,6-dichloropyridine-2-carboxylate
##STR00725##
[1564] To a stirred solution of methyl
3-bromo-6-chloropyridine-2-carboxylate (1.92 g, 7.67 mmol) in TFA
(18 ml) was added hydrogen peroxide (30% w/w aqueous solution, 5.22
ml, 53.7 mmol) and the reaction mixture was heated at 60.degree. C.
for 21 h. The reaction mixture was then cooled and slowly poured
onto saturated K.sub.2CO.sub.3 solution (100 ml), followed by
extraction of the aqueous layer with EtOAc (3.times.100 ml),
washing of the combined organic phases with brine (2.times.50 ml),
drying (Na.sub.2SO.sub.4) and evaporation. The desired
3-bromo-6-chloro-2-(methoxycarbonyl)pyridin-1-ium-1-olate (2.61 g,
.about.75% purity) was used crude in the next stage of the
synthesis without any further purification. To the crude
3-bromo-6-chloro-2-(methoxycarbonyl)pyridin-1-ium-1-olate
(.about.75% purity, 2.61 g, 7.35 mmol) was added POCl.sub.3 (3.42
ml, 36.7 mmol) and the solution was heated to 100.degree. C. for 4
h. After cooling the POCl.sub.3 was remove in vacuo to give a white
solid which was columned over silica eluting with 0% to 10% of
EtOAc in heptane to afford the title compound as a pale yellow
powder (1.07 g, 49% over two steps). LC-MS 99%, 2.02 min (3.5
minute LC-MS method), m/z=283.7/285.7/287.7, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.56 (s, 1H) 4.00 (s, 3H).
Step 3: Synthesis of methyl
3-bromo-6-chloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylate
##STR00726##
[1566] To a stirred solution of methyl
3-bromo-4,6-dichloropyridine-2-carboxylate (600 mg, 2.1 mmol) in
DMF (5 ml) was added TEA (587 .mu.l, 4.21 mmol) followed by
N-methyloxan-4-amine (240 mg, 2.1 mmol) and the reaction mixture
was heated at 80.degree. C. for 20 h. The reaction mixture was then
cooled to room temperature and poured onto water (100 ml), followed
by extraction of the product into EtOAc (3.times.100 ml), washing
of the combined organics with brine (50 ml), drying with
Na.sub.2SO.sub.4 and evaporation. The crude product was then
purified over a 10 g silica Isolute column eluting with a gradient
of 0% to 100% EtOAc in heptane to afford the title compound as a
white solid (110 mg, 14%). LC-MS 100%, 1.94 min (3.5 minute LC-MS
method), m/z=362.8/365.2/346.8, .sup.1H NMR (250 MHz, Chloroform-d)
.delta. 6.87 (s, 1H), 4.17-3.99 (m, 2H), 3.98 (s, 3H), 3.93-3.70
(m, 1H), 3.41 (t, J=10.9 Hz, 2H), 2.81 (s, 3H), 1.93 (dd, J=11.9,
4.6 Hz, 2H), 1.71 (d, J=10.3 Hz, 2H).
Step 4: Synthesis of
3-bromo-6-chloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylic
acid
##STR00727##
[1568] To a stirred solution of methyl
3-bromo-6-chloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylate
(102 mg, 0.28 mmol) in THF (2 ml) was added 2M aqueous NaOH (0.70
ml, 1.40 mmol) and the reaction mixture was left to stir at room
temperature for 18 h after which time the THF was evaporated in
vacuo. The aqueous phase was then treated with an aqueous 10%
citric acid solution to pH 5-6 and then extracted with EtOAc
(3.times.50 ml) followed by a solution of 1:1 IPA/CHCl.sub.3
(2.times.50 ml), the combined organic phases were then washed with
brine (50 ml) dried with Na.sub.2SO.sub.4 and evaporated to give
the title compound as a white solid (72 mg, 73%). LC-MS 100%, 1.49
min (3.5 minute LC-MS method), m/z=349.0/351.0/352.9, .sup.1H NMR
(500 MHz, Methanol-d4) .delta. 7.07 (s, 1H), 4.02 (dd, J=11.3, 4.3
Hz, 2H), 3.96-3.87 (m, 1H), 3.46 (t, J=11.0 Hz, 2H), 2.85 (s, 3H),
1.96 (dd, J=12.3, 4.3 Hz, 2H), 1.77 (s, 2H).
Step 5: Synthesis of
3-bromo-6-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-4-
-[methyl(oxan-4-yl)amino]pyridine-2-carboxamide
##STR00728##
[1570] To a stirred solution of
3-bromo-6-chloro-4-[methyl(oxan-4-yl)amino]pyridine-2-carboxylic
acid (73 mg, 0.21 mmol) in DMF (2 ml) were added DIPEA (560, 0.31
mmol) and HATU (95 mg, 0.25 mmol) at 0.degree. C. The reaction was
stirred at 0.degree. C. for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 52 mg,
0.31 mmol) was added and the reaction was stirred at room
temperature for 16 h. The reaction mixture was then poured onto 50
ml of water, the aqueous phase was extracted with EtOAc (3.times.50
ml), washed with brine (50 ml) dried (Na.sub.2SO.sub.4) and
evaporated to give an oil. The product was then purified using a 5
g silica Isolute column eluting with 0% to 4% MeOH in DCM and
evaporated to give a glassy solid which was triturated with diethyl
ether and filtered and dried in a vacuum oven at 40.degree. C. for
24 h to afford the title compound as a white powder (50 mg, 41%).
LC-MS 99%, m/z=483.1/485.0/486.9, .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 10.94 (br. s, 1H), 8.14-7.98 (m, 1H), 6.83 (s, 1H), 5.92
(s, 1H), 4.55 (d, J=6.1 Hz, 2H), 4.15-3.94 (m, 2H), 3.93-3.70 (m,
1H), 3.40 (t, J=11.0 Hz, 2H), 2.78 (s, 3H), 2.39 (s, 3H), 2.28 (s,
3H), 1.99-1.82 (m, 2H), 1.79-1.64 (m, 2H).
Compound 213:
5-(azetidin-3-yl)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
3-[ethyl(oxan-4-yl)amino]-2-methylbenzamide
##STR00729##
[1571] Step 1: Synthesis of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate
##STR00730##
[1573] To a stirred solution of methyl
3-amino-2-methy-5-bromobenzoate (4.06 g, 16.6 mmol) in DCE (60 ml)
under a nitrogen atmosphere was added oxan-4-one (3.07 ml, 33.3
mmol) followed by acetic acid (5.71 ml, 99.8 mmol) and the reaction
was left to stir for 5 min before the addition of sodium
triacetoxyborohydride (10.6 g, 49.9 mmol). The reaction was stirred
for 3 h at room temperature whereupon distilled water (50 ml) was
added and the solution was neutralised with solid NaHCO3. The
phases were separated and then the aqueous phase was washed with
EtOAc (2.times.100 ml), the combined organics were then dried using
Na2SO4, filtered and evaporated. The residue was purified by FCC
using silica and eluting with a 10% to 40% EtOAc in heptane
gradient to afford the title compound as an off-white solid (3.88
g, 71%). LC-MS 100%, 2.10 min (3.5 minute LC-MS method),
m/z=327.9/329.8, .sup.1H NMR (500 MHz, Chloroform-d) .delta. 7.23
(d, J=1.9 Hz, 1H), 6.84 (d, J=1.7 Hz, 1H), 4.02 (dt, J=11.7, 3.5
Hz, 2H), 3.87 (s, 3H), 3.65 (d, J=7.2 Hz, 1H), 3.59-3.45 (m, 3H),
2.23 (s, 3H), 2.05 (d, J=13.8 Hz, 2H), 1.51 (ddd, J=24.1, 10.8, 4.2
Hz, 2H).
Step 2: Synthesis of methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate
##STR00731##
[1575] To a stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (2.0 g, 6.1 mmol) in
DCE (10 ml) under a nitrogen atmosphere was added acetaldehyde (1.0
ml, 18 mmol) followed by acetic acid (2.1 ml, 37 mmol) and the
reaction was left to stir for 5 min before the addition of sodium
triacetoxyborohydride (6.6 g, 31 mmol). The reaction was stirred
for 16 h at room temperature after which time additional
acetaldehyde (1.0 ml, 22 mmol) and sodium triacetoxyborohydride
(3.0 g, 14 mmol) were added and the reaction was stirred for a
further 6 h whereupon distilled water (100 ml) was added and the
phases were separated. The aqueous phase was washed with EtOAc
(3.times.100 ml), the combined organics were then dried using
Na.sub.2SO.sub.4, filtered and evaporated to afford the title
compound as a colourless oil (2.10 g, 93%) which was suitable for
use without any further purification. LC-MS 96%, 2.38 min (3.5
minute LC-MS method), m/z=356.1/357.9, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 7.70 (d, J=2.0 Hz, 1H), 7.36 (d, J=2.0 Hz,
1H), 3.95 (d, J=11.3 Hz, 2H), 3.88 (d, J=9.1 Hz, 3H), 3.32 (td,
J=11.5, 2.5 Hz, 2H), 3.04 (q, J=7.1 Hz, 2H), 2.98-2.87 (m, 1H),
2.44 (s, 3H), 1.77-1.55 (m, 4H), 0.86 (t, J=7.1 Hz, 3H).
Step 3: Synthesis of tert-butyl
3-{3-[ethyl(oxan-4-yl)amino]-5-(methoxycarbonyl)-4-methylphenyl}azetidine-
-1-carboxylate
##STR00732##
[1577] To a dry flask was added zinc dust (40 mg, 0.61 mmol)
followed by anhydrous DMA (2 ml) and the vessel was flushed with
nitrogen whilst stirring vigorously and heating to 65.degree. C.
TMS-Cl (9 .mu.l, 0.07 mmol) and 1,2-dibromoethane (6 .mu.l, 0.07
mmol) were added and the reaction was stirred at 65.degree. C. for
30 mins, followed by the dropwise addition of N-Boc-3-iodoazetidine
(133 mg, 0.47 mmol) as a solution in anhydrous DMA (1 ml). The
reaction was then cooled to room temperature and methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate (100 mg, 0.28
mmol) was added as a solution in anhydrous DMA (2 ml). The
resulting solution was degassed with nitrogen for 5 mins after
which Pd(dppf)Cl.sub.2.DCM (7 mg, 0.01 mmol) and copper (I) iodide
(3 mg, 0.02 mmol) were added as solids. The reaction was heated to
80.degree. C. for 16 h and then cooled to room temperature followed
by the addition of saturated NH.sub.4Cl solution (50 ml). The
aqueous phase was extracted with EtOAc (3.times.100 ml), and then
the combined organic phases were washed with brine (2.times.50 ml),
dried with Na.sub.2SO.sub.4, filtered and evaporated. The residue
was purified over a 5 g Isolute column using 10-30% EtOAc in
heptane as the eluent to afford the title compound as a colourless
oil (77 mg, 62%). LC-MS 98%, 2.19 min (3.5 minute LC-MS method),
m/z=433.2, .sup.1H NMR (500 MHz, CDCl3) .delta. 7.49 (d, J=1.6 Hz,
1H), 7.16 (d, J=1.6 Hz, 1H), 4.31 (t, J=8.7 Hz, 2H), 3.94 (dd,
J=14.4, 5.5 Hz, 4H), 3.89 (s, 3H), 3.77-3.64 (m, 1H), 3.31 (td,
J=11.4, 2.7 Hz, 2H), 3.06 (q, J=7.0 Hz, 2H), 2.99-2.87 (m, 1H),
2.46 (s, 3H), 1.65 (ddd, J=11.1, 8.6, 6.0 Hz, 4H), 1.46 (s, 9H),
0.85 (dd, J=13.1, 6.1 Hz, 3H).
Step 4: Synthesis of
5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-3-[ethyl(oxan-4-yl)amino]-2-me-
thylbenzoic acid
##STR00733##
[1579] To a stirred solution of tert-butyl
3-{3-[ethyl(oxan-4-yl)amino]-5-(methoxycarbonyl)-4-methylphenyl}azetidine-
-1-carboxylate (78 mg, 0.18 mmol) in THF (2 ml) and MeOH (0.1 ml)
was added 2M NaOH solution (0.9 ml, 1.8 mmol) and the reaction was
stirred at room temperature for 16 h followed by heating at
50.degree. C. for 22 h. The reaction was cooled to room temperature
and the solvent removed in vacuo after which the aqueous solution
was adjusted to pH 5 using 1M HCl. The product was extracted into
DCM (2.times.50 ml), dried with Na.sub.2SO.sub.4 and filtered to
afford the title compound as a colourless oil (54 mg, 72%). LC-MS
100%, 1.70 min (3.5 minute LC-MS method), m/z=419.2, 1H NMR (500
MHz, MeOD) .delta. 7.77 (s, 1H), 7.55 (s, 1H), 4.36 (t, J=8.2 Hz,
2H), 3.90 (d, J=27.0 Hz, 5H), 3.71 (t, J=6.5 Hz, 1H), 3.36 (t,
J=11.3 Hz, 3H), 2.55 (s, 3H), 1.92-1.81 (m, 1H), 1.64 (d, J=55.8
Hz, 3H), 1.49-1.42 (m, 10H), 0.93 (s, 3H).
Step 5: Synthesis of
5-(azetidin-3-yl)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
3-[ethyl(oxan-4-yl)amino]-2-methylbenzamide
##STR00734##
[1581] To a stirred solution of
5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-3-[ethyl(oxan-4-yl)amino]-2-me-
thylbenzoic acid (54 mg, 0.13 mmol) in DMF (2 ml) and the reaction
was cooled using an ice bath followed by the addition of DIPEA (45
.mu.l, 0.26 mmol) and HATU (59 mg, 0.15 mmol)). The reaction was
stirred at 0.degree. C. for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 24 mg,
0.14 mmol) was added and the reaction was warmed room temperature
for 16 h. The reaction mixture was then poured onto 50 ml of water
and the aqueous phase was extracted with EtOAc (3.times.50 ml),
washed with brine (50 ml) dried (Na.sub.2SO.sub.4), filtered and
evaporated to give an oil. The product was then purified using a 5
g silica Isolute column eluting with 0% to 2.5% MeOH in DCM and
evaporated to afford tert-butyl
3-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-5-[e-
thyl(oxan-4-yl)amino]-4-methylphenyl)azetidine-1-carboxylate as a
colourless oil (29 mg). LC-MS 90%, m/z=553.3. This material was
dissolved in 4M HCl in dioxane solution (3 ml) and stirred at room
temperature for 90 min after which the solvent was evaporated to
dryness and the compound purified by preparative HPLC to afford the
title compound as a white solid (11 mg, 8% over 2 steps). LC-MS
97%, 2.17 min (7 minute LC-MS method), m/z=453.2, 1H NMR (500 MHz,
MeOD) .delta. 7.26 (s, 1H), 7.18 (s, 1H), 6.14 (s, 1H), 4.48 (s,
2H), 4.39 (s, 2H), 4.22 (s, 3H), 3.93 (d, J=10.5 Hz, 2H), 3.40-3.33
(m, 2H), 3.29-2.90 (m, 3H), 2.40 (s, 3H), 2.31 (s, 3H), 2.26 (s,
3H), 1.68 (d, J=26.3 Hz, 4H), 0.87 (s, 3H).
Compound 245:
5-(azetidin-3-yl)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
2-methyl-3-[methyl(oxan-4-yl)amino]benzamide
##STR00735##
[1582] Step 1: Synthesis of methyl
5-bromo-2-methyl-3-[methyl(oxan-4-yl)amino]benzoate
##STR00736##
[1584] To a stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (1.20 g, 3.66 mmol)
in DCE (10 ml) under a nitrogen atmosphere was added
paraformaldehyde (659 mg, 21.9 mmol) followed by acetic acid (1.26
ml, 21.9 mmol) and the reaction was left to stir for 5 min before
the addition of sodium triacetoxyborohydride (4.65 g, 21.9 mmol).
The reaction was stirred for 16 h at room temperature after which
time additional paraformaldehyde (325 mg, 10.8 mmol) and sodium
triacetoxyborohydride (2.40 g, 11.3 mmol) was added and the
reaction was stirred for a further 18 h whereupon distilled water
(30 ml) was added and solid NaHCO.sub.3 was added until the aqueous
phase was pH 7 and the phases were separated, the aqueous phase was
washed with EtOAc (3.times.50 ml), the combined organics were then
washed with brine (50 ml), dried using Na.sub.2SO.sub.4, filtered
and evaporated. The crude residue was purified using FCC over
silica eluting with 0-10% EtOAc in heptane to afford the title
compound as a colourless oil (728 mg, 52%). LC-MS 87%,
m/z=341.9/343.9, 1H NMR (500 MHz, Chloroform-d) .delta. 7.60 (d,
J=2.0 Hz, 1H), 7.25 (d, J=1.9 Hz, 1H), 3.92 (d, J=11.1 Hz, 2H),
3.82 (s, 3H), 3.27 (td, J=11.5, 2.0 Hz, 2H), 2.94-2.79 (m, 1H),
2.57 (s, 3H), 2.36 (s, 3H), 1.74-1.52 (m, 4H).
Step 2: Synthesis of
5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-methyl-3-[methyl(oxan-4-yl)a-
mino] benzoic acid
##STR00737##
[1586] To a dry flask was added zinc dust (436 mg, 6.67 mmol) and
the flask was heated using a heat gun for a few minutes. Then
anhydrous DMA (20 ml) was added under nitrogen whilst stirring
vigorously and heating to 65.degree. C. TMS-Cl (102 .mu.l, 0.8
mmol) and 1,2-dibromoethane (70 .mu.l, 0.8 mmol) were added and the
reaction was stirred at 65.degree. C. for 30 min followed by the
dropwise addition of N-Boc-3-iodoazetidine (1.46 g, 5.16 mmol) as a
solution in anhydrous DMA (15 ml). The reaction was then cooled to
room temperature and methyl
5-bromo-2-methyl-3-[methyl(oxan-4-yl)amino]benzoate (1.06 g, 3.09
mmol) was added as a solution in anhydrous DMA (15 ml). The
resulting solution was degassed with nitrogen for 10 mins after
which Pd(dppf)Cl.sub.2.DCM (76 mg, 0.09 mmol) and copper (I) iodide
(35 mg, 0.19 mmol) were added as solids. The reaction was heated to
80.degree. C. for 90 min and then cooled to room temperature
followed by the addition of distilled water (100 ml) and saturated
NH.sub.4Cl solution (10 ml). The aqueous phase was extracted with
TBME (3.times.50 ml), and then the combined organic phases were
dried with Na.sub.2SO.sub.4, filtered and evaporated. The residue
was purified over a 25 g Isolute column using 0%-40% EtOAc in
heptane as the eluent to afford the title compound as a pale brown
oil (507 mg). LC-MS 97%, m/z=420.0. To a stirred solution of
tert-butyl
3-[3-(methoxycarbonyl)-4-methyl-5-[methyl(oxan-4-yl)amino]phenyl]azetidin-
e-1-carboxylate (85 mg, 0.2 mmol) in THF (2 ml) and MeOH (0.1 ml)
was added 2M NaOH solution (1.02 ml, 2.03 mmol) and the reaction
was stirred at 50.degree. C. for 16 h after which time 4M NaOH
solution (0.5 ml, 2.0 mmol) was added and the reaction was heated
for a further 24 h. The reaction was cooled to room temperature and
the solvent removed in vacuo after which the aqueous solution was
adjusted to pH 3 using 1M HCl. The product was extracted into DCM
(2.times.50 ml), washed with brine (20 ml), dried with
Na.sub.2SO.sub.4 and filtered to afford the title compound as an
off-white solid (47 mg, 21% over two steps). LC-MS 95%, m/z=405.1,
1H NMR (500 MHz, Chloroform-d) .delta. 7.66 (t, J=7.0 Hz, 1H), 7.19
(d, J=1.3 Hz, 1H), 4.34 (t, J=8.7 Hz, 2H), 3.98 (dd, J=16.6, 10.2
Hz, 4H), 3.79-3.69 (m, 2H), 3.35 (dd, J=11.5, 10.0 Hz, 2H), 2.97
(td, J=10.8, 5.3 Hz, 1H), 2.67 (s, 3H), 2.55 (d, J=6.6 Hz, 3H),
1.74 (ddd, J=15.5, 12.1, 4.3 Hz, 2H), 1.66 (d, J=11.1 Hz, 2H), 1.48
(s, 9H).
Step 3: Synthesis of tert-butyl
3-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-4-me-
thyl-5-[methyl(oxan-4-yl)amino]phenyl)azetidine-1-carboxylate
##STR00738##
[1588] To a stirred solution of
5-{1-[(tert-butoxy)carbonyl]azetidin-3-yl}-2-methyl-3-[methyl(oxan-4-yl)a-
mino]benzoic acid (47 mg, 0.12 mmol) in DMF (2 ml) were added DIPEA
(40 .mu.l, 0.23 mmol) and HATU (53 mg, 0.14 mmol) at 0.degree. C.
The reaction was stirred at 0.degree. C. for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 22 mg,
0.13 mmol) was added and the reaction was warmed room temperature
for 16 h. The reaction mixture was then poured onto distilled water
(50 ml) and the aqueous phase was extracted with DCM (3.times.50
ml), washed with brine (50 ml) dried (Na.sub.2SO.sub.4), filtered
and evaporated to give an oil. The product was then purified using
a 5 g silica Isolute column eluting with 0% to 5% MeOH in DCM and
evaporated to afford the title compound as a colourless oil (44 mg,
67%). LC-MS 96%, m/z=539.4, .sup.1H NMR (500 MHz, Chloroform-d)
.delta. 11.68 (s, 1H), 6.95 (t, J=5.8 Hz, 1H), 6.92 (d, J=4.2 Hz,
2H), 5.88 (s, 1H), 4.47 (d, J=5.8 Hz, 2H), 4.20 (t, J=8.7 Hz, 2H),
3.90 (d, J=11.2 Hz, 2H), 3.86-3.73 (m, 2H), 3.62-3.50 (m, 1H), 3.26
(t, J=10.8 Hz, 2H), 2.89 (ddd, J=14.6, 10.8, 3.7 Hz, 1H), 2.55 (s,
3H), 2.33 (s, 3H), 2.20 (s, 3H), 2.14 (s, 3H), 1.75-1.47 (m, 4H),
1.39 (d, J=5.7 Hz, 9H).
Step 4: Synthesis of
5-(azetidin-3-yl)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]--
2-methyl-3-[methyl(oxan-4-yl)amino]benzamide
##STR00739##
[1590] To a stirred solution of tert-butyl
3-(3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carbamoyl}-4-me-
thyl-5-[methyl(oxan-4-yl)amino]phenyl)azetidine-1-carboxylate (29
mg, 1.29 mmol) in DCM (4 ml) was added TFA (1 ml) and the solution
was stirred at room temperature for 90 min after which the solvent
was evaporated and saturated aqueous NaHCO.sub.3 was added until pH
7-8. The aqueous phase was extracted with a 1:1 mixture
IPA:CHCl.sub.3 (2.times.50 ml), and the combined organic phases
washed with brine (30 ml, dried with Na.sub.2SO.sub.4, filtered and
evaporated. The residue was purified by preparative HPLC to afford
the title compound as a white solid (15 mg, 42%). LC-MS 100%,
m/z=439.3, 1H NMR (500 MHz, MeOD) .delta. 7.19 (d, J=1.4 Hz, 1H),
7.10 (d, J=1.5 Hz, 1H), 6.12 (s, 1H), 4.46 (s, 2H), 4.41-4.27 (m,
2H), 4.26-4.13 (m, 3H), 3.93 (d, J=11.4 Hz, 2H), 3.36 (td, J=11.3,
3.3 Hz, 2H), 3.06 (dq, J=14.6, 5.0 Hz, 1H), 2.65 (s, 3H), 2.39 (s,
3H), 2.26 (s, 3H), 2.24 (s, 3H), 1.75-1.60 (m, 4H).
Compound 269:
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-[methyl-
(oxan-4-yl)amino]-5-(1-methylazetidin-3-yl)benzamide
##STR00740##
[1591] Step 1: Synthesis of methyl
5-(azetidin-3-yl)-2-methyl-3-[methyl(oxan-4-yl)amino]benzoate
##STR00741##
[1593] To a stirred solution of tert-butyl
3-[3-(methoxycarbonyl)-4-methyl-5-[methyl(oxan-4-yl)amino]phenyl]azetidin-
e-1-carboxylate (400 mg, 0.96 mmol) in DCM (4 ml) was added TFA (1
ml) and the solution was stirred at room temperature for 45 min
after which the solvent was evaporated to dryness and saturated
aqueous NaHCO.sub.3 was added until pH 7-8. The aqueous phase was
extracted with DCM (3.times.30 ml), washed with brine (30 ml),
dried with MgSO.sub.4, filtered and evaporated to afford the title
compound as an orange oil (370 mg, 95%). LC-MS 96%, m/z=319.1,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. 7.49 (d, J=1.4 Hz, 1H),
7.20 (d, J=1.3 Hz, 1H), 4.32-4.10 (m, 5H), 3.98 (d, J=11.0 Hz, 2H),
3.90 (d, J=8.4 Hz, 3H), 3.34 (t, J=10.8 Hz, 2H), 2.99 (ddd, J=14.7,
10.9, 3.8 Hz, 1H), 2.65 (d, J=8.3 Hz, 3H), 2.51-2.41 (m, 3H), 1.73
(ddd, J=15.7, 12.1, 4.3 Hz, 2H), 1.64 (d, J=11.2 Hz, 2H).
Step 2: Synthesis of methyl
2-methyl-3-[methyl(oxan-4-yl)amino]-5-(1-methylazetidin-3-yl)benzoate
##STR00742##
[1595] To a stirred solution of methyl
5-(azetidin-3-yl)-2-methyl-3-[methyl(oxan-4-yl)amino]benzoate (320
mg, 1.01 mmol) in DCE (10 ml) under a nitrogen atmosphere was added
paraformaldehyde (151 mg, 5.03 mmol) followed by acetic acid (0.115
ml, 2.01 mmol) and the reaction was left to stir for 5 min before
the addition of sodium triacetoxyborohydride (852 mg, 4.02 mmol).
The reaction was stirred for 16 h at room temperature after which
time additional paraformaldehyde (21 mg) and sodium
triacetoxyborohydride (98 mg) was added and the reaction was
stirred for a further 4 h whereupon distilled water (20 ml) was
added and solid NaHCO.sub.3 was added until the aqueous phase was
pH 7. The phases were separated, the aqueous phase was washed with
DCM (3.times.20 ml), the combined organic phases were then washed
with brine (50 ml), dried using MgSO.sub.4, filtered and evaporated
to afford the title compound as an oil (296 mg, 89%). LC-MS 100%,
m/z=333.1, 1H NMR (500 MHz, Chloroform-d) .delta. 7.42 (d, J=1.4
Hz, 1H), 7.09 (s, 1H), 3.96 (d, J=11.2 Hz, 2H), 3.92-3.82 (m, 5H),
3.74 (d, J=7.2 Hz, 1H), 3.30 (dt, J=22.2, 15.0 Hz, 4H), 3.00-2.89
(m, 1H), 2.62 (s, 3H), 2.49-2.39 (m, 6H), 1.75-1.59 (m, 4H).
Step 3: Synthesis of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-[methyl-
(oxan-4-yl)amino]-5-(1-methylazetidin-3-yl)benzamide
##STR00743##
[1597] To a stirred solution of methyl
2-methyl-3-[methyl(oxan-4-yl)amino]-5-(1-methylazetidin-3-yl)benzoate
(296 mg, 0.89 mmol) in THF (10 ml) and MeOH (1 ml) was added 2M
NaOH solution (4.5 ml, 8.9 mmol) and the reaction was stirred at
50.degree. C. for 16 h after which time 4M NaOH solution (1 ml, 4.5
mmol) was added and the reaction was heated for a further 24 h. The
reaction was cooled to room temperature and the product was
neutralised with 1M HCl and evaporated to dryness followed by
azeotrope with toluene to remove residual water. The crude acid was
taken through to the next stage without any purification. The crude
residue of
2-methyl-3-[methyl(oxan-4-yl)amino]-5-(1-methylazetidin-3-yl)benzoic
acid was suspended in DMF (10 ml) and the solution was cooled using
an ice bath followed by the addition of DIPEA (433 .mu.l, 2.49
mmol) and HATU (567 mg, 1.49 mmol). The reaction was stirred at
0.degree. C. for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 221 mg,
1.29 mmol) was added and the reaction was warmed room temperature
for 16 h. The reaction mixture was then poured onto distilled water
(50 ml) and the aqueous phase was extracted with DCM (3.times.50
ml), washed with brine (50 ml) dried (Na.sub.2SO.sub.4), filtered
and evaporated to give an oil. The product was then purified using
preparative HPLC method (and evaporated to dryness followed by
trituration with ether and filtration to afford the title compound
as an off-white solid (51 mg, 10%). LC-MS 91%, m/z=453.2, 1H NMR
(500 MHz, MeOD) .delta. 7.14 (d, J=1.7 Hz, 1H), 7.09 (d, J=1.7 Hz,
1H), 6.12 (s, 1H), 4.46 (s, 2H), 4.41 (t, J=9.2 Hz, 2H), 4.22 (t,
J=9.3 Hz, 2H), 4.14 (dd, J=17.4, 8.5 Hz, 1H), 3.98-3.89 (m, 2H),
3.36 (td, J=11.3, 3.4 Hz, 2H), 3.06 (ddd, J=14.6, 9.8, 4.9 Hz, 1H),
2.97 (s, 3H), 2.64 (s, 3H), 2.39 (s, 3H), 2.26 (s, 3H), 2.24 (s,
3H), 1.73-1.62 (m, 4H).
Compound 389:
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[ethyl(oxan-4-yl-
) amino]-4-methylpyridine-3-carboxamide
##STR00744##
[1598] Step 1: Synthesis of methyl
5-amino-4-methylpyridine-3-carboxylate
##STR00745##
[1600] To a stirred solution of
5-amino-4-methylpyridine-3-carboxylic acid (0.94 g, 6.18 mmol) in
MeOH (20 mlat 0.degree. C. was added thionyl chloride drop-wise.
The solution was then heated to 50.degree. C. for 3.5 h, a further
10 ml of MeOH was added and the reaction heated at 50.degree. C.
for 5 h followed by stirring at room temperature for 64 h. The
solvent was then evaporated under reduced pressure and saturated
NaHCO.sub.3 solution was added until pH 7-8. The product was
extracted into EtOAc (2.times.50 ml) and the combined organics were
washed with brine (50 ml) dried using Na.sub.2SO.sub.4, filtered
and evaporated to afford the title compound as an off-white powder
(824 mg, 80%) which was suitable for use in the next stage without
any purification. LC-MS m/z=167.0, 1H NMR (500 MHz, Chloroform-d)
.delta. 8.52 (d, J=34.2 Hz, 1H), 8.16 (s, 1H), 3.95 (d, J=4.8 Hz,
3H), 3.78 (s, 2H), 2.42 (s, 3H).
Step 2: Synthesis of methyl
4-methyl-5-[(oxan-4-yl)amino]pyridine-3-carboxylate
##STR00746##
[1602] To a stirred solution of methyl
5-amino-4-methylpyridine-3-carboxylate (805 mg, 4.84 mmol) in DCE
(20 ml) under a nitrogen atmosphere was added oxan-4-one (537
.mu.l, 5.80 mmol) followed by trifluoroacetic acid (742 .mu.l, 9.69
mmol) and the reaction was left to stir for 5 min before the
addition of sodium triacetoxyborohydride (1.54 g, 7.27 mmol). The
reaction was stirred for 16 h at room temperature after which time
more oxan-4-one (100 .mu.l) and sodium triacetoxyborohydride (300
mg) were added and the reaction stirred for a further 6 h.
Distilled water (20 ml) was added and solid NaHCO.sub.3 was added
until pH 8. The phases were separated and then the aqueous phase
was washed with EtOAc (2.times.50 ml), the combined organics were
washed with brine (50 ml), dried using Na.sub.2SO.sub.4, filtered
and evaporated to afford the title compound as a pale yellow solid
(1.11 g, 88%) which was used in the next stage without any further
purification. LC-MS 96%, m/z=251.0, 1H NMR (500 MHz, Chloroform-d)
.delta. 8.40 (s, 1H), 8.10 (s, 1H), 4.03 (dt, J=11.9, 3.5 Hz, 2H),
3.92 (s, 3H), 3.68-3.58 (m, 1H), 3.54 (tt, J=8.1, 4.1 Hz, 3H), 2.35
(s, 3H), 2.12-2.05 (m, 2H), 1.53 (ddd, J=23.8, 11.0, 4.3 Hz,
2H).
Step 3: Synthesis of methyl
5-[ethyl(oxan-4-yl)amino]-4-methylpyridine-3-carboxylate
##STR00747##
[1604] To a stirred solution of methyl
4-methyl-5-[(oxan-4-yl)amino]pyridine-3-carboxylate (600 mg, 2.40
mmol) in DCE (20 ml) under a nitrogen atmosphere was added
acetaldehyde (1.07 ml, 19.2 mmol) followed by trifluoroacetic acid
(367 .mu.l, 4.79 mmol) and the reaction was left to stir for 5 min
before the addition of sodium triacetoxyborohydride (3.05 g, 14.4
mmol). The reaction was stirred for 16 h at room temperature after
which time more acetaldehyde (500 .mu.l) and sodium
triacetoxyborohydride (1.0 g) were added and the reaction stirred
for a further 16 h. Distilled water (20 ml) was added and solid
NaHCO.sub.3 was added until pH 8. The phases were separated and
then the aqueous phase was washed with DCM (2.times.50 ml), the
combined organics were washed with brine (50 ml), dried using
Na.sub.2SO.sub.4, filtered and evaporated. The residue was purified
using a 25 g Isolute silica column using a gradient of 0% to 50%
EtOAc in heptane to afford the title compound as a pale yellow oil
(418 mg, 60%). LC-MS 96%, m/z=279.1, 1H NMR (500 MHz, Chloroform-d)
.delta. 8.76 (s, 1H), 8.44 (s, 1H), 3.97 (d, J=11.0 Hz, 2H), 3.93
(s, 3H), 3.33 (td, J=11.6, 2.1 Hz, 2H), 3.13 (q, J=7.1 Hz, 2H),
3.04 (tt, J=10.9, 4.0 Hz, 1H), 2.54 (s, 3H), 1.76-1.59 (m, 4H),
0.89 (t, J=7.1 Hz, 3H).
Step 4: Synthesis of
5-[ethyl(oxan-4-yl)amino]-4-methylpyridine-3-carboxylic acid
##STR00748##
[1606] To a stirred solution of methyl
5-[ethyl(oxan-4-yl)amino]-4-methylpyridine-3-carboxylate (415 mg,
1.49 mmol) in THF (7 ml) and MeOH (3 ml) was added 4M NaOH solution
(932 .mu.l, 3.73 mmol). The reaction was stirred at room
temperature for 4.5 h after which the solution was neutralised via
the addition of 1M HCl and the solvent was removed under reduced
pressure, followed by the addition of saturated brine (10 ml). The
aqueous phase was extracted with 1:1 IPA:CHCl.sub.3 (3.times.50
ml), the combined organics washed with brine (30 ml), dried using
Na.sub.2SO.sub.4, filtered and evaporated to afford the title
compound as an off-white solid (305 mg, 77%) which was suitable for
use in the next stage without any further purification. LC-MS 99%,
m/z=265.1, 1H NMR (500 MHz, DMSO-d6) .delta. 8.57 (s, 1H), 8.46 (s,
1H), 3.82 (d, J=11.6 Hz, 2H), 3.25 (t, J=10.9 Hz, 2H), 3.16-2.99
(m, 3H), 2.45 (s, 3H), 1.64 (d, J=11.0 Hz, 2H), 1.48 (dt, J=11.9,
5.7 Hz, 2H), 0.81 (t, J=7.0 Hz, 3H).
Step 5: Synthesis of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-[ethyl(oxan-4-yl-
) amino]-4-methylpyridine-3-carboxamide
##STR00749##
[1608] To a stirred solution of
5-[ethyl(oxan-4-yl)amino]-4-methylpyridine-3-carboxylic acid (300
mg, 1.13 mmol) in DMF (10 ml) was added DIPEA (395 .mu.l, 2.27
mmol) and HATU (518 mg, 1.36 mmol). The reaction was stirred at
room temperature for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 214 mg,
1.25 mmol) was added and the reaction was stirred at room
temperature for 18 h. The reaction mixture was then poured onto 500
ml of water and the product was extracted into EtOAc (3.times.200
ml), the combined organic phase was washed with brine (2.times.100
ml), dried (Na.sub.2SO.sub.4) and evaporated. The crude product was
then purified using preparative-HPLC and the resulting solid was
triturated with diethyl ether and isolated by filtration to afford
the title compound as an off white solid (115 mg, 25%). LC-MS 100%,
m/z=399.2, .sup.1H NMR (500 MHz, Acetone) .delta. 10.97 (s, 1H),
8.32 (s, 1H), 8.22 (s, 1H), 7.73 (s, 1H), 5.92 (s, 1H), 4.41 (d,
J=5.3 Hz, 2H), 3.85 (dd, J=11.3, 3.0 Hz, 2H), 3.28 (td, J=11.7, 1.9
Hz, 2H), 3.15 (q, J=7.1 Hz, 2H), 3.09 (ddd, J=11.1, 7.2, 3.9 Hz,
1H), 2.32 (d, J=1.5 Hz, 6H), 2.23 (s, 3H), 1.69 (dd, J=12.6, 1.7
Hz, 2H), 1.55 (ddd, J=24.3, 12.2, 4.4 Hz, 2H), 0.85 (t, J=7.1 Hz,
3H).
Compound 322:
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(oxan-4-yl-
) amino]-2-methyl-5-(1-methylazetidin-3-yl)benzamide
##STR00750##
[1609] Step 1: Synthesis of methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate
##STR00751##
[1611] To a stirred solution of methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate (622 mg, 1.90
mmol) in DCE (10 ml) under a nitrogen atmosphere was added
acetaldehyde (318 .mu.l, 5.69 mmol) followed by acetic acid (650
.mu.l, 11.4 mmol) and the reaction was left to stir for 5 min
before the addition of sodium triacetoxyborohydride (2.00 g, 9.48
mmol). The reaction was stirred for 16 h at room temperature after
which time additional acetaldehyde (300 .mu.l) and sodium
triacetoxyborohydride (1.2 g) were added. The reaction was stirred
for a further 6 h whereupon distilled water (100 ml) was added and
solid NaHCO.sub.3 was added until the aqueous phase was pH 7. The
phases were separated, the aqueous phase was washed with DCM
(2.times.50 ml), the combined organic phases were then washed with
brine (100 ml), dried using Na.sub.2SO.sub.4, filtered and
evaporated. The crude residue was purified using FCC over silica
eluting with 0-5% EtOAc in heptane to afford the title compound as
a pale yellow oil (47 0 mg, 70%). LC-MS 100 m/z=356.0/357.9. 1H NMR
(500 MHz, Chloroform-d) .delta. 7.64 (d, J=2.0 Hz, 1H), 7.29 (d,
J=2.0 Hz, 1H), 3.89 (d, J=11.3 Hz, 2H), 3.83 (s, 3H), 3.26 (td,
J=11.5, 2.6 Hz, 2H), 2.98 (q, J=7.1 Hz, 2H), 2.86 (dd, J=9.7, 5.2
Hz, 1H), 2.38 (s, 3H), 1.67-1.52 (m, 4H), 0.80 (t, J=7.1 Hz,
3H).
Step 2: Synthesis of methyl
5-(azetidin-3-yl)-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate
##STR00752##
[1613] To a dry flask was added zinc dust (186 mg, 2.85 mmol) and
the flask was heated using a heat gun for a few minutes. Then
anhydrous DMA (15 ml) was added under nitrogen whilst stirring
vigorously and heating to 65.degree. C. TMSCl (44 .mu.l, 0.34 mmol)
and 1,2-dibromoethane (30 .mu.l, 0.34 mmol) were added and the
reaction was stirred at 65.degree. C. for 30 mins followed by the
dropwise addition of N-Boc-3-iodoazetidine (624 mg, 2.20 mmol) as a
solution in anhydrous DMA (5 ml). The reaction was then cooled to
room temperature methyl
5-bromo-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate (470 mg, 1.32
mmol) was added as a solution in anhydrous DMA (10 ml). The
resulting solution was degassed with nitrogen for 10 mins after
which Pd(dppf)Cl.sub.2.DCM (32 mg, 0.04 mmol) and copper (I) iodide
(15 mg, 0.08 mmol) were added as solids. The reaction was heated to
80.degree. C. for 60 min and then cooled to room temperature
followed by the addition of distilled water (50 ml) and saturated
NH.sub.4Cl solution (10 ml). The aqueous phase was extracted with
EtOAc (3.times.50 ml) and then the combined organics were washed
with brine (20 ml), dried with Na.sub.2SO.sub.4, filtered and
evaporated to dryness followed by heptane azeotrope to remove any
residual DMA. The residue was purified over a 10 g Isolute column
using 0%-25% EtOAc in heptane as the eluent to afford the title
compound as a pale brown oil (565 mg). LC-MS 100%, 2.12 min (3.5
minute LC-MS method), m/z=433.2. To this oil was added DCM (4 ml)
followed by TFA (1 ml) and the solution was stirred at room
temperature for 2 h after which the solvent was evaporated and
saturated aqueous NaHCO.sub.3 was added until pH 7-8. The aqueous
phase was extracted with DCM (3.times.30 ml), washed with brine (30
ml), dried with MgSO4, filtered and evaporated to afford the title
compound as a yellow oil (440 mg). LC-MS 100%, m/z=333.1.
Step 3: Synthesis of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(oxan-4-yl-
) amino]-2-methyl-5-(1-methylazetidin-3-yl)benzamide
##STR00753##
[1615] To a stirred solution of methyl
5-(azetidin-3-yl)-3-[ethyl(oxan-4-yl)amino]-2-methylbenzoate (440
mg, 1.32 mmol) in DCE (10 ml) under a nitrogen atmosphere was added
paraformaldehyde (278 mg, 9.27 mmol) followed by acetic acid (0.15
ml, 2.65 mmol) and the reaction was left to stir for 5 min before
the addition of sodium triacetoxyborohydride (1.40 g, 6.62 mmol).
The reaction was stirred for 16 h at room temperature after which
time distilled water (20 ml) was added and solid NaHCO.sub.3 was
added until the aqueous phase was pH 8 and the phases were
separated, the aqueous phase was washed with DCM (3.times.30 ml),
the combined organics were then washed with brine (50 ml), dried
using Na.sub.2SO.sub.4, filtered and evaporated. The residue was
purified by chromatography over a 10 g Isolute (elution with 0% to
5% MeOH in DCM) and evaporated followed by purification over
alumina (elution with 0% to 1% MeOH in DCM) to afford the title
compound as a yellow oil which was used directly in the next stage
of the synthesis. LC-MS 77%, 1 m/z=347.1.
[1616] Methyl
3-[ethyl(oxan-4-yl)amino]-2-methyl-5-(1-methylazetidin-3-yl)benzoate
(103 mg, 0.30 mmol) was dissolved in THF (20 ml) and MeOH (5 ml)
and 2M NaOH (1.49 ml, 2.97 mmol) was added. The reaction was
stirred at 50.degree. C. for 16 h, cooled, acidified to pH 4 using
6M HCl and the solvent evaporated under reduced pressure, and the
residue dried in a vacuum oven at 40.degree. C. for 5 h to afford
the crude acid. To the acid in DMF (10 ml) were added HATU (170 mg,
0.45 mmol) and DIPEA (1300, 0.74 mmol). The mixture was stirred at
room temperature for 5 min and then
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 66 mg,
0.39 mmol) was added. The reaction was stirred for 16 h after which
additional HATU (80 mg) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (30 mg) and
the mixture stirred for a further 5 h. The reaction mixture was
poured onto distilled after (50 ml), extracted with EtOAc
(3.times.50 ml), washed with brine (2.times.20 ml), dried with
Na.sub.2SO.sub.4, filtered and evaporated. The crude residue was
purified using preparative HPLC and evaporated to afford the title
compound as an off-white solid (28 mg, 5% over three steps). LC-MS
95%, m/z=467.3, 1H NMR (500 MHz, Acetone-d.sub.6) .delta. 10.73 (s,
1H), 7.61 (s, 1H), 7.30 (s, 1H), 7.26 (s, 1H), 5.97 (s, 1H), 4.74
(t, J=9.9 Hz, 2H), 4.60 (t, J=9.6 Hz, 2H), 4.38 (q, J=6.2 Hz, 3H),
3.83 (d, J=11.6 Hz, 2H), 3.27 (s, 3H), 3.23 (t, J=11.6 Hz, 2H),
3.10 (dd, J=14.1, 7.0 Hz, 2H), 3.04-2.97 (m, 1H), 2.34 (s, 3H),
2.31 (s, 3H), 2.25 (s, 3H), 1.67 (d, J=11.1 Hz, 2H), 1.56 (td,
J=11.8, 4.1 Hz, 2H), 0.83 (t, J=7.0 Hz, 3H).
Compound 332:
3-[benzyl(methyl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl]-2-(trifluoromethyl)benzamide
##STR00754##
[1617] Step 1: Synthesis of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-fluoro-2-(triflu-
oromethyl)benzamide
##STR00755##
[1619] To a stirred solution of 3-fluoro-2-(trifluoromethyl)benzoic
acid (1.00 g, 4.81 mmol) in DMF (25 ml) cooled in an ice bath was
added HATU (2.19 g, 5.77 mmol) and DIPEA (1.67 ml, 9.61 mmol). The
reaction was stirred at 0.degree. C. for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 904 mg,
5.29 mmol) was added and the reaction was warmed room temperature
for 4 h. The reaction mixture was then poured onto distilled water
(300 ml) and stirred for 1 h after which time the precipitate was
collected via vacuum filtration using microfibre paper. The
resulting solid was triturated using diethyl ether and filtered to
afford the title compound as a pale brown solid (1.19 g, 72%).
LC-MS 100%, m/z=343.0, 1H NMR (500 MHz, DMSO) .delta. 11.48 (s,
1H), 8.54 (t, J=4.8 Hz, 1H), 7.71 (dd, J=13.3, 8.0 Hz, 1H),
7.56-7.45 (m, 1H), 7.20 (d, J=7.6 Hz, 1H), 5.86 (s, 1H), 4.25 (d,
J=4.9 Hz, 2H), 2.18 (s, 3H), 2.11 (s, 3H).
Step 2: Synthesis of
3-(benzylamino)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2--
(trifluoromethyl)benzamide
##STR00756##
[1621] A suspension of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-fluoro-2-(triflu-
oro methyl)benzamide (0.80 g, 2.34 mmol) in benzylamine (2.6 ml,
23.4 mmol) was heated in a microwave reactor at 210.degree. C. for
4 h. After cooling distilled water (20 ml) was added to the slurry
and 2M HCl was added until the suspension was at pH 5. and the
mixture was extracted using EtOAc (3.times.50 ml). The combined
organics were washed with pH 5 HCl solution (2.times.50 ml), brine
(30 ml), dried using Na.sub.2SO.sub.4 filtered and evaporated. The
resultant oil was purified using a 50 g Isolute silica column
eluting with a gradient of 0% to 5% MeOH in DCM to afford the title
compound as an off-white solid (313 mg, 29%). LC-MS 94%, m/z=430.2,
1H NMR (500 MHz, Acetone-d6) .delta. 7.41-7.16 (m, 8H), 6.69 (d,
J=8.5 Hz, 1H), 6.50 (d, J=7.4 Hz, 1H), 5.87 (s, 1H), 5.84 (s, 1H),
4.53 (d, J=5.7 Hz, 2H), 4.35 (d, J=5.6 Hz, 2H), 2.28, (d, J=4.0 Hz,
3H), 2.21 (s, 3H).
Step 3: Synthesis of
3-[benzyl(methyl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)me-
thyl]-2-(trifluoromethyl)benzamide
##STR00757##
[1623] To a solution of
3-(benzylamino)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2--
(trifluoromethyl)benzamide (100 mg, 0.23 mmol) in DCE (5 ml) was
added acetic acid (27 .mu.l, 0.47 mmol) followed by
paraformaldehyde (7 mg, 0.23 mmol) after which the reaction was
stirred at room temperature for 10 min. Then sodium
triacetoxyborohydride (74 mg, 0.35 mmol) was added in one portion
and the reaction was stirred at room temperature for 16 h. Further
treatment with acetaldehyde (35 mg), sodium triacetoxyborohydride
(250 mg) and AcOH (100 ml) was performed. After a further 24 h
further acetaldehyde (445 mg) and sodium triacetoxyborohydride (250
mg) were added and the reaction was stirred for a further 3 h.
Water (20 ml) was then added to the reaction mixture and the
aqueous phase was neutralised by the portionwise addition of
NaHCO.sub.3. The aqueous phase was extracted with DCM (2.times.50
ml), then the combined organic phases were washed with brine, dried
(Na.sub.2SO.sub.4) and evaporated. The product was purified using
FCC over a 5 g Isolute, eluting with 0-5% MeOH in DCM to afford the
title compound as an off-white powder (24 mg, 22%). LC-MS 95%, 4.12
min (7 minute LC-MS method), m/z=444.2, 1H NMR (500 MHz,
Chloroform-d) .delta. 11.10 (br. s, 1H), 7.44 (t, J=7.8 Hz, 1H),
7.38-7.29 (m, 5H), 7.27-7.23 (m, 1H), 7.12 (d, J=7.4 Hz, 1H), 7.04
(t, J=5.6 Hz, 1H), 5.92 (s, 1H), 4.52 (d, J=5.9 Hz, 2H), 4.09 (s,
2H), 2.60 (s, 3H), 2.38 (s, 3H), 2.19 (s, 3H).
Compound 333:
3-[benzyl(ethyl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl]-2-(trifluoromethyl)benzamide
##STR00758##
[1625] To a solution of
3-(benzylamino)-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2--
(trifluoromethyl)benzamide (198 mg, 0.46 mmol) in DCE (5 ml) was
added acetic acid (80 .mu.l, 1.38 mmol) followed by acetaldehyde
(129 .mu.l, 2.31 mmol) after which the reaction was stirred at room
temperature for 10 min. Then sodium triacetoxyborohydride (490 mg,
2.31 mmol) was added in one portion and the reaction was stirred at
room temperature for 16 h. Further 4.times.130 uL treatments with
acetaldehyde and 4.times.0.5 g sodium triacetoxyborohydride were
performed over 3 days to completed the reaction. Water (100 ml) was
then added to the reaction mixture and the aqueous phase was
neutralized by the portionwise addition of NaHCO.sub.3. The aqueous
phase was extracted with DCM (2.times.50 ml), then the combined
organic phases washed with brine, dried (Na.sub.2SO.sub.4) and
evaporated. The product was purified using FCC over a 5 g Isolute,
eluting with 0-5% MeOH in DCM followed by purification via RP-HPLC
to give the title compound as an off-white powder (76 mg, 34%).
LC-MS 100%, m/z=458.3, 1H NMR (500 MHz, Chloroform-d) .delta. 11.70
(s, 1H), 7.42 (t, J=7.8 Hz, 1H), 7.36-7.27 (m, 5H), 7.24 (d, J=7.1
Hz, 1H), 7.12 (d, J=7.4 Hz, 1H), 7.03 (d, J=5.4 Hz, 1H), 5.90 (s,
1H), 4.50 (d, J=5.7 Hz, 2H), 4.05 (s, 2H), 2.96 (q, J=7.1 Hz, 2H),
2.36 (s, 3H), 2.15 (s, 3H), 0.89 (t, J=7.1 Hz, 3H).
Compound 334:
3-[benzyl(ethyl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl]-2-methylbenzamide
##STR00759##
[1626] Step 1: Synthesis of ethyl
3-(benzylamino)-2-methylbenzoate
##STR00760##
[1628] To a stirred solution of ethyl 3-amino-2-methylbenzoate
(1.00 g, 5.58 mmol) in DCE (20 ml) was added benzaldehyde (569
.mu.l, 5.58 mmol) followed by acetic acid (0.64 ml, 11.2 mmol). The
reaction was stirred at room temperature for 1 h and then sodium
triacetoxyborohydride (1.77 g, 8.37 mmol) was added and the
reaction stirred for 2 h. Distilled water (20 ml) was added and
solid NaHCO.sub.3 was added until the aqueous phase was pH 7 and
the phases were separated. The aqueous phase was washed with DCM
(2.times.5 0 ml), the combined organics were then washed with brine
(50 ml), dried using Na.sub.2SO.sub.4, filtered and evaporated to
afford the title compound as a yellow oil (1.52 g, 96%) which was
used in the next stage without any further purification. LC-MS 95%,
m/z=270.0, 1H NMR (500 MHz, Chloroform-d) .delta. 7.42-7.32 (m,
4H), 7.32-7.28 (m, 1H), 7.17-7.06 (m, 2H), 6.73 (dd, J=7.8, 1.1 Hz,
1H), 4.40 (s, 2H), 4.36 (q, J=14.3, 5.3 Hz, 2H), 4.08 (br. s, 1H),
2.35, (s, 3H), 1.39 (t, 3H).
Step 2: Synthesis of ethyl
3-[benzyl(ethyl)amino]-2-methylbenzoate
##STR00761##
[1630] To a stirred solution of ethyl
3-(benzylamino)-2-methylbenzoate (396 mg, 1.47 mmol) in DCE (10 ml)
was added acetaldehyde (164 .mu.l, 2.94 mmol) followed by acetic
acid (0.17 ml, 2.94 mmol) The reaction was stirred at room
temperature for 10 min and then sodium triacetoxyborohydride (0.62
g, 2.94 mmol) was added and the reaction stirred for 16 h.
Distilled water (30 ml) was added and solid NaHCO.sub.3 was then
added until the aqueous phase was pH 7, the phases were separated,
the aqueous phase was washed with DCM (2.times.50 ml), the combined
organics were then washed with brine (50 ml), dried using
Na.sub.2SO.sub.4, filtered and evaporated. The resultant residue
was then purified using a 25 g silica Isolute column eluting using
a gradient of 0% to 5% EtOAc in heptane to afford the title
compound as a colourless oil (372 mg, 81%). LC-MS 94%, m/z=298.1,
1H NMR (500 MHz, Chloroform-d) .delta. 7.48 (dd, J=7.5, 1.3 Hz,
1H), 7.29-7.08 (m, 8H), 4.32 (q, J=7.1 Hz, 2H), 4.05 (s, 2H), 2.92
(q, J=7.1 Hz, 2H), 2.56 (s, 3H), 1.35 (t, J=7.1 Hz, 3H), 0.92 (t,
J=7.1 Hz, 3H).
Step 3: Synthesis of 3-[benzyl(ethyl)amino]-2-methylbenzoic
acid
##STR00762##
[1632] To a stirred solution of ethyl
3-[benzyl(ethyl)amino]-2-methylbenzoate (370 mg, 1.24 mmol) in THF
(20 ml) and MeOH (5 ml) was added 2M NaOH (3.11 ml, 6.22 mmol) and
the reaction was stirred at 50.degree. C. for 16 h. The reaction
was cooled to room temperature and the solvent removed in vacuo
after which the aqueous solution was adjusted to pH 1 using 1M HCl.
The product was extracted into DCM (3.times.50 ml), washed with
brine (2 0 ml), dried with Na.sub.2SO.sub.4 concentrated and
filtered to afford the title compound as an orange oil (296 mg,
80%) which was used in the next stage without any further
purification. LC-MS 91%, m/z=270.0, 1H NMR (500 MHz, Chloroform-d)
.delta. 7.73 (d, J=7.5 Hz, 1H), 7.34-7.30 (m, 4H), 7.23 (dt,
J=15.4, 6.9 Hz, 3H), 4.12 (d, J=13.5 Hz, 2H), 3.04-2.94 (m, 2H),
2.69 (s, 3H), 0.99 (t, J=7.0 Hz, 3H).
Step 4: Synthesis of
3-[benzyl(ethyl)amino]-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl]-2-methylbenzamide
##STR00763##
[1634] To a stirred solution of
(3-[benzyl(ethyl)amino]-2-methylbenzoic acid (91% purity, 296 mg,
1.0 mmol) in DMF (10 ml) was added DIPEA (440 .mu.l, 2.5 mmol) and
HATU (570 mg, 1.5 mmol). The reaction was stirred at room
temperature for 5 min after which time
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 222 mg,
1.3 mmol) was added and the reaction was stirred at room
temperature for 16 h. The reaction mixture was then poured onto 150
ml of water and stirred for 20 min., The product was filtered, air
dried under vacuum and then purified using RP-HPLC. The resulting
solid was triturated with diethyl ether and isolated by filtration
to afford the title compound as an off white solid (86 mg, 21%).
LC-MS 99%, m/z=404.3, .sup.1H NMR (500 MHz, Chloroform-d) .delta.
12.04 (s, 1H), 7.33-7.23 (m, 4H), 7.21 (t, J=6.8 Hz, 1H), 7.14-7.05
(m, 3H), 7.05-6.99 (m, 1H), 5.92 (s, 1H), 4.53 (d, J=5.9 Hz, 2H),
4.06 (s, 2H), 2.92 (q, J=6.9 Hz, 2H), 2.41 (s, 3H), 2.39 (s, 3H),
2.16 (s, 3H), 0.92 (t, J=7.0 Hz, 3H).
Compound 191:
2-bromo-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-
-(ethyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00764##
[1635] Step 1: Synthesis of methyl
2-bromo-5-chloro-3-nitrobenzoate
##STR00765##
[1637] A solution of methyl 2-bromo-5-chlorobenzoate (10 g, 40
mmol) in concentrated H.sub.2SO.sub.4 (48 ml, 910 mmol) was cooled
to -5.degree. C. in an acetone/ice bath in air. A mixture of
concentrated nitric acid (3.35 ml, 52 mmol) and concentrated
H.sub.2SO.sub.4 (3.4 ml, 64 mmol) was added dropwise to the
reaction mixture at 0.degree. C. over 30 minutes. The yellow
reaction mixture was stirred at 0.degree. C. for 1.5 hours before
being poured onto ice. EtOAc (150 ml) was added and the phases were
separated. The organic phase was washed with deionized water
(3.times.50 ml) followed by brine (50 ml). The organic phase was
dried over MgSO4, filtered and concentrated under reduced pressure
to give 12.0 g (60%) of the title compound material as a pale
yellow oil which solidified upon standing. This material contained
40% of the 6-nitro isomer. This material was used without further
purification. LC-MS 91%, m/z=no ionisation, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 7.86 (1H, d, J=2.52 Hz), 7.77 (1H, d, J=2.52
Hz), 3.99 (3H, s).
Step 2: Synthesis of methyl 3-amino-2-bromo-5-chlorobenzoate
##STR00766##
[1639] To a solution of methyl 2-bromo-5-chloro-3-nitrobenzoate
(4.0 g, 14 mmol) in methanol (130 ml) at room temperature was added
ammonium chloride (7.13 g, 136 mmol) followed by deionized water
(65 ml). The mixture was heated to 70.degree. C. in air before the
addition of iron (4.55 g, 81.5 mmol). The reaction was stirred at
70.degree. C. for 3 hours, allowed to cool to room temperature and
filtered through Kieselgel. The filter pad was washed with MeOH (65
ml) and the filtrate concentrated under reduced pressure. The
residue was dissolved in saturated NaHCO.sub.3 (50 ml) and EtOAc
(100 ml). The phases were separated and the organic phase was
washed with saturated NaHCO.sub.3(aq) (3.times.50 ml) before being
dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by FCC (50 g silica, Isolute
cartridge, gradient of eluents; 95:5 Heptane:EtOAc to 7:3
Heptane:EtOAc) to give 1.92 g (48%) of the title compound as a
colourless oil. LC-MS 92%, m/z=268.8/265.8/267.6, .sup.1H NMR (500
MHz, Chloroform-d) .delta. 7.08 (1H, d, J=2.36 Hz), 6.86 (1H, d,
J=2.36 Hz), 4.34-4.49 (2 H, m), 3.93 (3H, s).
Step 3: Synthesis of methyl
2-bromo-5-chloro-3-(oxan-4-ylamino)benzoate
##STR00767##
[1641] To a solution of methyl 3-amino-2-bromo-5-chlorobenzoate
(1.0 g, 3.8 mmol) in 1,2-dichloroethane (15 ml) at room temperature
under nitrogen was added oxan-4-one (0.7 ml, 7.6 mmol) followed by
acetic acid (1.3 ml, 23 mmol). This solution was stirred for 5
minutes before the addition of sodium triacetoxyborohydride (2.4 g,
11 mmol) at room temperature. After 21 h, additional oxan-4-one
(0.7 ml, 7.6 mmol) was added to the reaction mixture, with stirring
for 10 minutes before adding sodium triacetoxyborohydride (2.4 g,
11 mmol). After stirring for 20 h, deionized water (30 ml) was
added and the mixture was neutralised with solid NaHCO.sub.3. The
phases were separated and the aqueous layer was extracted with
EtOAc (2.times.30 ml). The combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by FCC (25 g silica, Isolute cartridge,
gradient of eluents; 9:1 Heptane:EtOAc to 8:2 Heptane:EtOAc) to
give 903 mg (68%) of the title compound as a white solid. LC-MS
92%, 2.11 min (3 minute LC-MS method), m/z=348.2/350.2/351.9,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. 6.94-7.07 (1H, m), 6.70
(1H, d, J=2.36 Hz), 4.72 (1H, d, J=7.57 Hz), 3.96-4.15 (2H, m),
3.87-3.96 (3H, m), 3.40-3.67 (3H, m), 2.05 (2H, d, J=13.08 Hz),
1.49-1.69 (2H, m).
Step 4: Synthesis of methyl
2-bromo-5-chloro-3-[ethyl(oxan-4-yl)amino]benzoate
##STR00768##
[1643] To a solution of methyl
2-bromo-5-chloro-3-(oxan-4-ylamino)benzoate (300 mg, 0.86 mmol) in
1,2-dichloroethane (3 ml) at room temperature under nitrogen was
added acetaldehyde (96 .mu.l, 1.7 mmol) followed by acetic acid
(0.3 ml, 5.16 mmol). This solution was stirred for 5 minutes before
the addition of sodium triacetoxyborohydride (0.55 g, 2.6 mmol) at
room temperature with reaction monitoring by LCMS. After 22 h,
further acetaldehyde (96 .mu.l, 1.7 mmol) was added to the reaction
mixture and this was stirred for 10 minutes before the addition of
sodium triacetoxyborohydride (0.55 g, 2.6 mmol). After 3 days, the
reaction mixture was diluted with 1,2-dichloroethane (5 ml).
Acetaldehyde (962 .mu.l, 17.2 mmol) was added to the reaction
mixture and this was stirred for 10 minutes before the addition of
sodium triacetoxyborohydride (1.82 g, 8.6 mmol). After 15 h,
further acetaldehyde (960 .mu.l, 17.2 mmol) was added to the
reaction mixture and this was stirred for 10 minutes before the
addition of sodium triacetoxyborohydride (1.82 g, 8.6 mmol). The
reaction mixture was then stirred at room temperature for a further
22 hours before deionized water (25 ml) was added and the mixture
was neutralised with solid NaHCO.sub.3. EtOAc (25 ml) was added and
the phases were separated. The aqueous layer was then extracted
with EtOAc (2.times.25 ml) and the combined organic extracts were
dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by FCC (10 g silica, Isolute
cartridge, gradient of eluents; 99:1 Heptane:EtOAc to 8:2
Heptane:EtOAc) to give 122 mg (38%) of the title compound as a
colourless oil. LC-MS 85%, m/z=376.3/378.2/379.9, .sup.1H NMR (500
MHz, Chloroform-d) .delta. 7.31 (d, J=2.5 Hz, 1H), 7.18 (d, J=2.4
Hz, 1H), 3.98 (dd, J=8.7, 5.5 Hz, 2H), 3.94 (s, 3H), 3.35 (td,
J=11.6, 5.5 Hz, 2H), 3.19 (t, J=7.1 Hz, 1H), 3.12 (q, J=7.1 Hz,
2H), 1.75 (td, J=10.1, 8.8, 3.9 Hz, 4H), 0.92 (t, J=7.1 Hz,
3H).
Step 5: Synthesis of
2-bromo-5-chloro-3-[ethyl(oxan-4-yl)amino]benzoic acid
##STR00769##
[1645] To a solution of methyl
2-bromo-5-chloro-3-[ethyl(oxan-4-yl)amino]benzoate (122 mg, 0.32
mmol) in tetrahydrofuran (3 ml) was added 4M NaOH (3.2 ml). The
reaction mixture was stirred at 50.degree. C. for 20 hours. The
reaction mixture was acidified to pH 2-3 with 6M HCl and extracted
with DCM (3.times.100 ml). The combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated under reduced pressure
to give 109 mg (93%, 83% corrected yields) of
2-bromo-5-chloro-3-[ethyl(oxan-4-yl)amino]benzoic acid as a pale
yellow foam. LC-MS 89%, m/z=361.9/364.2/365.8, .sup.1H NMR (500
MHz, Chloroform-d) .delta. 7.50 (d, J=2.4 Hz, 1H), 7.23 (d, J=2.4
Hz, 1H), 4.00 (d, J=11.5 Hz, 2H), 3.49-3.27 (m, 2H), 3.21 (t, J=7.4
Hz, 1H), 3.14 (q, J=7.0 Hz, 2H), 1.78 (dd, J=7.4, 3.4 Hz, 4H), 0.93
(t, J=7.0 Hz, 3H).
Step 6: Synthesis of
2-bromo-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-
-(ethyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00770##
[1647] To a solution of
2-bromo-5-chloro-3-[ethyl(oxan-4-yl)amino]benzoic acid (109 mg, 0.3
mmol) in DMF (3 ml) at room temperature under nitrogen was added
PyBOP (187 mg, 0.36 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (78 .mu.l, 0.45 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 56 mg,
0.33 mmol). After stirring for 1 hour at room EtOAc (30 ml) was
added to the reaction mixture and this was then washed with
deionized water (2.times.10 ml) followed by saturated
NaHCO.sub.3(aq) (2.times.10 ml). The organic phase was dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified twice by FCC (5 g silica, Isolute cartridge,
gradient of eluents; 100% DCM to 98:2 DCM:MeOH). The sample was
dried in a vacuum oven for 18 hours to give 103 mg (65%) of the
title compound as a pale yellow solid. LC-MS 94%,
m/z=496.0/498.0/500.0, .sup.1H NMR (500 MHz, Chloroform-d) .delta.
11.00 (s, 1H), 7.20-7.10 (m, 1H), 7.10-7.05 (m, 2H), 5.94 (s, 1H),
4.54 (d, J=5.9 Hz, 2H), 3.96 (d, J=11.4 Hz, 2H), 3.42-3.26 (m, 2H),
3.28-3.12 (m, 1H), 3.09 (q, J=7.0 Hz, 2H), 2.39 (s, 3H), 2.23 (s,
3H), 1.72 (td, J=8.5, 3.8 Hz, 4H), 0.90 (t, J=7.0 Hz, 3H).
Compound 102:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(-
piperidin-4-yl)amino]-2-methylbenzamide
##STR00771##
[1648] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00772##
[1650] A solution of 5-chloro-2-methylbenzoic acid (9.9 g, 58 mmol)
in concentrated H.sub.2SO.sub.4 (70 ml, 1310 mmol) was cooled to
-5.degree. C. in an acetone/ice bath. A mixture of concentrated
nitric acid (4.9 ml, 75 mmol) and concentrated H.sub.2SO.sub.4 (5.0
ml, 94 mmol) was added dropwise to the reaction mixture at
-5.degree. C. over 30 minutes. The reaction mixture was stirred at
-5.degree. C. for 2 hours before being poured onto ice (150 g) and
the precipitate collected by filtration. The precipitate was
dissolved in EtOAc (100 ml) and washed with brine (100 ml) before
being dried over MgSO.sub.4, filtered and concentrated under
reduced pressure to give 11.9 g of the title compound as an
off-white solid. This material was used without further
purification. LC-MS 57%, m/z=214.0/216.0, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 10.37 (1H, br. s.), 8.19 (1H, d, J=2.21 Hz),
7.91 (1H, d, J=2.21 Hz), 2.67 (2H, s).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00773##
[1652] To a solution of 5-chloro-2-methyl-3-nitrobenzoic acid (1.17
g, 5.43 mmol) in DMF (11 ml) at room temperature and nitrogen was
added Na.sub.2CO.sub.3 (862 mg, 8.14 mmol) followed by iodomethane
(0.51 ml, 8.14 mmol). The reaction mixture was stirred at room
temperature for 19 hours before being diluted with deionized water
(30 ml) and extracted with EtOAc (3.times.30 ml). The combined
organic phases were washed with saturated NaHCO.sub.3(aq) (30 ml),
dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by FCC (10 g silica, Isolute
cartridge, gradient of eluents; 99:1 Heptane:EtOAc to 95:5
Heptane:EtOAc) to give 900 of the title compound as a pale yellow
oil. LC-MS 89%, m/z=no ionisation, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. 8.00 (1H, d, J=2.21 Hz), 7.86 (1H, d, J=2.21
Hz), 3.96 (3H, s), 2.60 (3H, s).
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methylbenzoate
##STR00774##
[1654] To a solution of methyl 5-chloro-2-methyl-3-nitrobenzoate
(9.19 g, 40 mmol) in methanol (200 ml) at room temperature was
added ammonium chloride (21 g, 400 mmol) followed by deionized
water (100 ml). The mixture was heated to 70.degree. C. before the
addition of iron (13.4 g, 240 mmol). The reaction turned to a dark
colour over the 2.5 hours it was stirred at 70.degree. C. This
mixture was allowed to cool to room temperature and was filtered
through Kieselgel. The filter pad was washed with MeOH (100 ml) and
the filtrate concentrated under reduced pressure. The residue was
dissolved in deionized water (100 ml) and EtOAc (100 ml). The
phases were separated and the aqueous phase was extracted with
EtOAc (2.times.100 ml). The combined organic extracts were washed
with brine (100 ml), dried over MgSO.sub.4, filtered and
concentrated under reduced pressure to give 7.6 g of the title
compound as a viscous orange oil that solidified upon standing.
This material was used without further purification. LC-MS 63%,
m/z=200.3/202.0, .sup.1H NMR (500 MHz, Chloroform-d) .delta. 7.20
(1H, d, J=2.21 Hz), 6.80 (1H, d, J=2.05 Hz), 3.89 (3H, s),
3.74-3.87 (2H, m), 2.30 (3H, s).
Step 4: Synthesis of tert-butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl]amino}piperidine-1-carbox-
ylate
##STR00775##
[1656] To a solution of methyl 3-amino-5-chloro-2-methylbenzoate
(100 mg, 0.50 mmol) in 1,2-dichloroethane (2.5 ml) at room
temperature and under nitrogen was added tert-butyl
4-oxopiperidine-1-carboxylate (200 mg, 1.0 mmol) followed by acetic
acid (0.17 ml, 3.0 mmol). This solution was stirred for 5 minutes
before the addition of sodium triacetoxyborohydride (0.32 g, 1.5
mmol) at room temperature. The resulting mixture was stirred for 20
hours under nitrogen. Deionized water (5 ml) was added and the
mixture was neutralised with solid NaHCO.sub.3. The phases were
separated and the aqueous layer was extracted with EtOAc (2.times.5
ml). The combined organic extracts were dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by FCC (10 g silica, Isolute cartridge, gradient of
eluents; 99:1 Heptane:EtOAc to 7:3 Heptane:EtOAc) to give 131 mg
(68%) of the title compound as a white solid. LC-MS 98%,
m/z=405.4/406.9 (M+Na), .sup.1H NMR (500 MHz, Chloroform-d) .delta.
ppm 7.10 (1H, d, J=2.05 Hz), 6.70 (1H, d, J=1.89 Hz), 3.98-4.16
(2H, m), 3.88 (3H, s), 3.58-3.72 (1H, m), 3.37-3.52 (1H, m), 2.97
(2H, br. t, J=11.50, 11.50 Hz), 2.24 (3H, s), 2.06 (2H, br. d,
J=10.60 Hz), 1.44-1.51 (9H, m), 1.32-1.44 (2H, m).
Step 5: Synthesis of tert-butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl](ethyl)amino}piperidine-1-
-carboxylate
##STR00776##
[1658] To a solution of tert-butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl]amino}piperidine-1-carbox-
ylate (125 mg, 0.33 mmol) in 1,2-dichloroethane (1.6 ml) at room
temperature under nitrogen was added acetaldehyde (37 .mu.l, 0.65
mmol) followed by acetic acid (0.11 ml, 1.96 mmol). This solution
was stirred for 5 minutes before adding sodium
triacetoxyborohydride (0.21 g, 0.98 mmol) at room temperature. The
reaction was monitored by LCMS and after 4.5 h a further
acetaldehyde (73 .mu.l, 1.31 mmol) and sodium triacetoxyborohydride
(0.28 g, 1.31 mmol) were added to the reaction mixture. The mixture
was stirred for a further 18 hours after which deionized water (10
ml) was added and the mixture was neutralised with solid
NaHCO.sub.3. The phases were separated and the aqueous layer was
extracted with EtOAc (2.times.10 ml). The combined organic extracts
were dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by FCC (10 g silica, Isolute
cartridge, gradient of eluents; 99:1 Heptane:EtOAc to 8:2
Heptane:EtOAc) to give 131 mg (98%) of the title compound as a
thick colourless oil. LC-MS 98%, m/z=433.5/435.1 (M+Na), .sup.1H
NMR (500 MHz, Chloroform-d) .delta. ppm 7.57 (1H, d, J=2.21 Hz),
7.22 (1H, d, J=2.05 Hz), 3.97-4.14 (2H, m), 3.90 (3H, s), 3.04 (2H,
q, J=6.99 Hz), 2.81-2.91 (1H, m), 2.70 (2H, br. t, J=11.50, 11.50
Hz), 2.45 (3H, s), 1.69-1.80 (2H, m), 1.47-1.56 (2H, m), 1.46 (9H,
s), 0.87 (3H, t, J=7.09 Hz).
Step 6: Synthesis of
3-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(ethyl)amino)-5-chloro-2-meth-
ylbenzoic acid
##STR00777##
[1660] To a solution of tert-butyl
4-{[5-chloro-3-(methoxycarbonyl)-2-methylphenyl](ethyl)amino}piperidine-1-
-carboxylate (131 mg, 0.32 mmol) in tetrahydrofuran (2 ml) was
added LiOH (8.4 mg, 0.35 mmol) in deionized water (2 ml). The
reaction mixture was stirred in air at room temperature for 24
hours. 1M NaOH (3.19 ml) was added to the reaction mixture and this
was stirred for a further 24 hours. The reaction mixture was then
warmed to 40.degree. C. for 25 h. The reaction mixture was
acidified to pH 2-3 with 1M HCl and extracted with EtOAc
(3.times.20 ml). The combined organic extracts were dried over
MgSO.sub.4, filtered and concentrated under reduced pressure to
give 122 mg of the title compound as a thick orange oil. LC-MS 76%,
m/z=397.5/399.4, .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm
7.74 (1H, s), 7.20-7.35 (2H, m), 3.92-4.11 (2H, m), 2.97-3.13 (2H,
m), 2.81-2.95 (1H, m), 2.62-2.79 (2H, m), 2.53 (3H, s), 1.75 (2H,
br. s.), 1.48-1.61 (2H, m), 1.46 (9H, s), 0.89 (3H, t, J=6.86
Hz).
Step 7: Synthesis of tert-butyl
4-[(5-chloro-3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carba-
moyl}-2-methylphenyl)(ethyl)amino]piperidine-1-carboxylate
##STR00778##
[1662] To a solution of
3-({1-[(tert-butoxy)carbonyl]piperidin-4-yl}(ethyl)amino)-5-chloro-2-meth-
ylbenzoic acid (60 mg, 0.15 mmol) in DMF (2 ml) at room temperature
under nitrogen was added PyBOP (94 mg, 0.18 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (40 .mu.l, 0.23 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 28 mg,
0.17 mmol). After stirring for 1 hour at room temperature EtOAc (30
ml) was added to the reaction mixture and this was then washed with
deionized water (5 ml) followed by saturated NaHCO.sub.3(aq)
(3.times.5 ml). The organic phase was dried over MgSO4, filtered
and concentrated under reduced pressure. The residue was purified
by FCC (2 g silica, Isolute cartridge, gradient of eluents; 100%
EtOAc to 96:4 EtOAc:MeOH) to give 71 mg (88%) of the title compound
as a pale yellow solid. LC-MS 97%, m/z=531.1/533.1, .sup.1H NMR
(250 MHz, Chloroform-d) .delta. ppm 11.17 (1H, br. s.), 7.11-7.18
(1H, m), 7.06 (2H, dd, J=7.46, 2.28 Hz), 5.96 (1H, s), 4.53 (2H, d,
J=5.94 Hz), 3.93-4.14 (2H, m), 2.97-3.08 (2H, m), 2.78-2.88 (1H,
m), 2.68 (2H, t, J=11.57 Hz), 2.39 (3H, s), 2.26 (3H, s), 2.24 (3H,
s), 1.71 (2H, d, J=11.27 Hz), 1.52 (2H, br. s.), 1.45 (9H, s), 0.86
(3H, t, J=6.93 Hz).
Step 8: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(-
piperidin-4-yl)amino]-2-methylbenzamide
##STR00779##
[1664] To a solution of tert-butyl
4-[(5-chloro-3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]carba-
moyl}-2-methylphenyl)(ethyl)amino]piperidine-1-carboxylate (140 mg,
0.26 mmol) in 1,4-dioxane (5 ml) at room temperature was added HCl
(4M in dioxane) (1.32 ml). A gum formed after stirring for a short
time which then dissolved upon the addition of deionized water (2
ml). The reaction was stirred for 21 h before being concentrated
under reduced pressure to give a thick oil. DCM (3 ml) was added
and a solid formed. This mixture was concentrated to give 105 mg of
a yellow solid which was stirred in DCM (10 ml) for 1 hour before
being filtered and dried under high vacuum in a vacuum oven for 24
hours. The sample was then dissolved in deionized water (0.2 ml)
and lyophilized to give 86 mg (70%) of the title compound as a
beige solid. LC-MS 100%, m/z=431.2/433.2, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.44 (bs, 1H), 7.26 (bs, 1H), 6.69 (bs,
1H), 4.55 (s, 2H), 3.53-3.34 (m, 3H), 3.28-3.08 (m, 2H), 3.01 (t,
J=11.8 Hz, 2H), 2.51 (s, 3H), 2.43 (s, 3H), 2.33 (s, 3H), 2.16-1.96
(m, 2H), 1.91-1.76 (m, 2H), 0.92 (t, J=6.9 Hz, 3H).
Compound 192:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(-
oxan-4-yl)amino]-2-methoxybenzamide
##STR00780##
[1665] Step 1: Synthesis of methyl
5-chloro-2-methoxy-3-nitrobenzoate
##STR00781##
[1667] To concentrated H.sub.2SO.sub.4 (90 mL) was added
portionwise 5-chloro-2-methylbenzoic acid (13.2 g, 77.6 mmol) at
0.degree. C. Then a mixture of conc. HNO.sub.3 (10.5 g, 1740 mmol)
in conc. H.sub.2SO.sub.4 (15 mL) was added dropwise at 0.degree. C.
over a period of about 1.5 hr. After the addition, the mixture was
stirred at this temperature for 2 hr. The mixture was poured into
ice water with vigorous stirring and the precipitate was collected
by filtration. The precipitate was dissolved in EtOAc, washed with
brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated to
give a crude title compound (13.2 g), which was used in the next
step without further purification. LC-MS 95%, m/z=245.9/247.9,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 8.01 (1H, d, J=2.68
Hz), 7.91 (1H, d, J=2.68 Hz), 4.01 (3H, s), 3.97-3.99 (3H, s).
Step 2: Synthesis of methyl 3-amino-5-chloro-2-methoxybenzoate
##STR00782##
[1669] To a solution of methyl 5-chloro-2-methoxy-3-nitrobenzoate
(1.21 g, 4.93 mmol) in methanol (50 ml) at room temperature was
added ammonium chloride (2.59 g, 49.2 mmol) followed by deionized
water (25 ml). The mixture was heated to 70.degree. C. before the
addition of iron (1.65 g, 29.6 mmol). The reaction turned to a dark
color over the 2 hours it was stirred at 70.degree. C. This mixture
was allowed to cool to room temperature and was filtered through
Kieselgel. The filter pad was washed with MeOH (50 ml) and the
filtrate concentrated under reduced pressure. The residue was
dissolved in deionized water (25 ml) and EtOAc (25 ml). The phases
were separated and the aqueous phase was extracted with EtOAc
(2.times.25 ml). The combined organic extracts were washed with
brine (25 ml), dried over MgSO.sub.4, filtered and concentrated
under reduced pressure to give 892 mg (67%) of the title compound
as a viscous brown oil. This material was used without further
purification. LC-MS 89%, 1.75 min (3 minute LC-MS method),
m/z=216.3/218.0, .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm
7.16 (1H, d, J=2.52 Hz), 6.87 (1H, d, J=2.52 Hz), 4.03 (2H, br.
s.), 3.89-3.94 (3H, m), 3.84 (3H, s).
Step 3: Synthesis of methyl
5-chloro-2-methoxy-3-[(oxan-4-yl)amino]benzoate
##STR00783##
[1671] To a solution of methyl 3-amino-5-chloro-2-methoxybenzoate
(440 mg, 2 mmol) in 1,2-dichloroethane (8 ml) at room temperature
under nitrogen was added oxan-4-one (0.38 ml, 4.0 mmol) followed by
acetic acid (0.7 ml, 12 mmol). This solution was stirred for 5
minutes before the addition of sodium triacetoxyborohydride (1.3 g,
6.1 mmol) at room temperature. After stirring for 23 hours
deionized water (16 ml) was added and the mixture was neutralized
with solid NaHCO.sub.3. The phases were separated and the aqueous
layer was extracted with EtOAc (2.times.16 ml). The combined
organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
FCC (10 g silica, Isolute cartridge, gradient of eluents; 9:1
Heptane:EtOAc to 7:3 Heptane:EtOAc) to give 439 mg (72%) of the
title compound as a white solid. LC-MS 97%, m/z=300.3/302.0,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 7.06 (1H, d, J=2.52
Hz), 6.73 (1H, d, J=2.52 Hz), 4.38-4.45 (1H, m), 4.02 (2H, dt,
J=11.78, 3.49 Hz), 3.91 (3H, s), 3.81 (3H, s), 3.54 (2H, td,
J=11.47, 2.13 Hz), 3.40-3.50 (1H, m), 2.03 (2H, br. d, J=13.70 Hz),
1.48-1.55 (2H, m).
Step 4: Synthesis of methyl
5-chloro-3-[ethyl(oxan-4-yl)amino]-2-methoxybenzoate
##STR00784##
[1673] To a solution of methyl
5-chloro-2-methoxy-3-[(oxan-4-yl)amino]benzoate (220 mg, 0.73 mmol)
in 1,2-dichloroethane (2 ml) at room temperature under nitrogen was
added acetaldehyde (82 .mu.l, 1.47 mmol) followed by acetic acid
(0.25 ml, 4.4 mmol). This solution was stirred for 5 minutes before
the addition of sodium triacetoxyborohydride (0.47 g, 2.2 mmol) at
room temperature. After stirring for 16 hours acetaldehyde (82
.mu.l, 1.47 mmol) was added to the reaction mixture and stirred for
5 minutes before the addition of sodium triacetoxyborohydride (0.47
g, 2.2 mmol). After stirring for 24 hours, a further acetaldehyde
(82 .mu.l, 1.47 mmol) was added to the reaction mixture and stirred
for 5 minutes before the addition of sodium triacetoxyborohydride
(0.47 g, 2.2 mmol). The reaction was stirred for 24 h after which
deionized water (10 ml) was added and the mixture was neutralized
with solid NaHCO.sub.3. The phases were separated and the aqueous
layer was extracted with EtOAc (2.times.10 ml). The combined
organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
FCC (10 g silica, Isolute cartridge, gradient of eluents; 95:5
Heptane:EtOAc to 7:3 Heptane:EtOAc) to give 127 mg (53%) of the
title compound as a yellow oil. LC-MS 93%, m/z=328.4/330.0, .sup.1H
NMR (500 MHz, Chloroform-d) .delta. ppm 7.35 (d, J=2.6 Hz, 1H),
7.09 (d, J=2.6 Hz, 1H), 3.98 (dd, J=11.3, 4.2 Hz, 2H), 3.91 (s,
3H), 3.85 (s, 3H), 3.48 (tt, J=11.6, 4.0 Hz, 1H), 3.36 (td, J=11.8,
1.9 Hz, 2H), 3.15 (q, J=7.0 Hz, 2H), 1.78 (qd, J=12.2, 4.6 Hz, 2H),
1.72-1.59 (m, 2H), 0.99 (t, J=7.0 Hz, 3H).
Step 5: Synthesis of
5-chloro-3-[ethyl(oxan-4-yl)amino]-2-methoxybenzoic acid
##STR00785##
[1675] To a solution of methyl
5-chloro-3-[ethyl(oxan-4-yl)amino]-2-methoxybenzoate (127 mg, 0.39
mmol) in THF (4 ml) was added 4M NaOH (3.87 ml). The reaction
mixture was stirred at 50.degree. C. for 24 hours. The reaction
mixture was acidified to pH 2-3 with 6M HCl and extracted with DCM
(5.times.15 ml). The combined organic extracts were dried over
MgSO4, filtered and concentrated under reduced pressure to give 111
mg (91%) of the title compound as a yellow oil. LC-MS 80%, 1.25 min
(3 minute LC-MS method), m/z=314.4/315.9, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 10.02-9.55 (m, 1H), 8.01-7.47 (m, 1H),
7.36-6.96 (m, 1H), 4.15-3.86 (m, 5H), 3.76 (t, J=6.6 Hz, 2H),
3.55-3.12 (m, 3H), 1.95-1.59 (m, 4H), 1.15-0.88 (m, 3H).
Step 6: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(-
oxan-4-yl)amino]-2-methoxybenzamide
##STR00786##
[1677] To a solution of
5-chloro-3-[ethyl(oxan-4-yl)amino]-2-methoxybenzoic acid (111 mg,
0.35 mmol) in DMF (3 ml) at room temperature under nitrogen was
added PyBOP (220 mg, 0.42 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (92 .mu.l, 0.53 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 66 mg,
0.39 mmol). After stirring for 1.5 hours at room temperature no
starting material was observed by LCMS. Deionized water (20 ml) was
added to the reaction mixture and the resultant precipitate was
collected by filtration. The solid was washed with deionized water
(3.times.5 ml) before being air dried. The solid was further
purified by FCC (5 g silica, Isolute cartridge, gradient of
eluents; 100% DCM to 97:3 DCM:MeOH). The sample was dried in a
vacuum oven for 18 hours to give 105 mg (64%) the title compound as
an off-white solid. LC-MS 98%, m/z=449.5/452.1, .sup.1H NMR (500
MHz, Chloroform-d) .delta. ppm 11.47 (s, 1H), 8.79 (t, J=5.8 Hz,
1H), 7.73 (d, J=2.6 Hz, 1H), 7.05 (d, J=2.6 Hz, 1H), 5.92 (s, 1H),
4.55 (d, J=6.0 Hz, 2H), 3.98 (dd, J=11.3, 3.8 Hz, 2H), 3.78 (s,
3H), 3.35 (t, J=11.6 Hz, 3H), 3.12 (q, J=7.0 Hz, 2H), 2.42 (s, 3H),
2.28 (s, 3H), 1.92-1.61 (m, 4H), 0.94 (t, J=7.0 Hz, 3H).
Compound 212:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methoxy-
-3-[methyl(oxan-4-yl)amino]benzamide
##STR00787##
[1678] Step 1: Synthesis of methyl
5-chloro-2-methoxy-3-[methyl(oxan-4-yl)amino]benzoate
##STR00788##
[1680] To a solution of methyl
5-chloro-2-methoxy-3-[(oxan-4-yl)amino]benzoate (500 mg, 1.67 mmol)
in 1,2-dichloroethane (16 ml) at room temperature under nitrogen
was added paraformaldehyde (300 mg, 10 mmol) followed by acetic
acid (0.57 ml, 10 mmol). This mixture was stirred for 5 minutes
before the addition of sodium triacetoxyborohydride (2.12 g, 10
mmol). After stirring for 64 hours deionized water (30 ml) was
added and the mixture was neutralized with solid NaHCO.sub.3 and
the phases were separated. The aqueous layer was extracted with
EtOAc (3.times.15 ml) and the combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
The residue was purified by FCC (10 g silica, Isolute cartridge,
gradient of eluents; 9:1 Heptane:EtOAc to 7:3 Heptane:EtOAc) to
give 431 mg (82%) of the title compound as a colorless oil. LC-MS
98%, m/z=314.0/316.0, .sup.1H NMR (500 MHz, Chloroform-d) .delta.
ppm 7.30 (d, J=2.5 Hz, 1H), 7.04 (d, J=2.5 Hz, 1H), 4.00 (dd,
J=11.3, 4.3 Hz, 2H), 3.90 (s, 3H), 3.82 (s, 3H), 3.70 (ddt, J=11.7,
7.7, 3.8 Hz, 1H), 3.39 (t, J=11.1 Hz, 2H), 2.71 (s, 3H), 1.84 (qd,
J=12.3, 4.6 Hz, 2H), 1.66-1.55 (m, 2H).
Step 2: Synthesis of
5-chloro-2-methoxy-3-[methyl(oxan-4-yl)amino]benzoic acid
##STR00789##
[1682] To a solution of methyl
5-chloro-2-methoxy-3-[methyl(oxan-4-yl)amino]benzoate (413 mg, 1.32
mmol) in THF (13 ml) and MeOH (2 ml) was added 4M NaOH (13.16 ml).
The reaction mixture was stirred at 50.degree. C. for 16 hours. The
reaction mixture was acidified to pH 2-3 with 6M HCl and extracted
with DCM (5.times.10 ml). The combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated under reduced pressure
to give 385 mg (98%) of the title compound as a thick yellow oil.
LC-MS 94%, m/z=300.0/302.0, .sup.1H NMR (500 MHz, Chloroform-d)
.delta. ppm 7.74 (d, J=2.0 Hz, 1H), 7.30-7.12 (m, 1H), 4.11-4.02
(m, 2H), 4.01 (s, 3H), 3.54 (t, J=11.6 Hz, 1H), 3.40 (t, J=11.3 Hz,
2H), 2.76 (s, 3H), 1.97-1.78 (m, 2H), 1.63 (d, J=11.8 Hz, 2H).
Step 3: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methoxy-
-3-[methyl(oxan-4-yl)amino]benzamide
##STR00790##
[1684] To a solution of
5-chloro-2-methoxy-3-[methyl(oxan-4-yl)amino]benzoic acid (200 mg,
0.67 mmol) in DMF (2 ml) at room temperature under nitrogen was
added HBTU (304 mg, 0.80 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (174 .mu.l, 1.0 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 125 mg,
0.73 mmol). After stirring for 2 hours at room temperature EtOAc
(30 ml) was added to the reaction mixture and this was then washed
with deionized water (5 ml) followed by saturated NaHCO.sub.3(aq)
(3.times.5 ml). The organic phase was dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by FCC (10 g silica, Isolute cartridge, gradient of
eluents; 100% DCM to 96:4 DCM:MeOH) to give 182 mg (63%) of the
title compound as an off-white solid. LC-MS 98%, m/z=434.1/436.1,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 12.52 (br s, 1H),
8.76 (t, J=5.8 Hz, 1H), 7.69 (d, J=2.6 Hz, 1H), 7.01 (d, J=2.6 Hz,
1H), 5.94 (s, 1H), 4.56 (d, J=6.0 Hz, 2H), 3.99 (dd, J=11.2, 3.9
Hz, 2H), 3.72 (s, 3H), 3.49 (tt, J=11.5, 3.7 Hz, 1H), 3.36 (t,
J=11.2 Hz, 2H), 2.67 (s, 3H), 2.42 (s, 3H), 2.31 (s, 3H), 1.93-1.73
(m, 2H), 1.64-1.52 (m, 2H).
Compound 205:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-[methy(oxan-4-yl)amino]benzamide
##STR00791##
[1685] Step 1: Synthesis of methyl
5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate
##STR00792##
[1687] To a solution of methyl 2-bromo-5-chlorobenzoate (14.8 g, 59
mmol) in TEA (124 ml, 890 mmol) was added copper(I) iodide (338 mg,
1.78 mmol) and triphenylphosphine (778 mg, 2.97 mmol) at room
temperature under nitrogen. This mixture had nitrogen bubbled
through it for 10 minutes before the addition of
ethynyl(trimethyl)silane (12.45 ml, 89 mmol) and Pd(OAc).sub.2 (266
mg, 1.19 mmol). The reaction mixture was stirred at 50.degree. C.
for 20 hours before being concentrated under reduced pressure. The
residue was dissolved in deionized water (50 ml) and EtOAc (50 ml)
and filtered through celite. The filter cake was washed with EtOAc
(50 ml) before the phases were separated and the aqueous layer was
extracted with EtOAc (2.times.50 ml). The combined organic extracts
were dried over MgSO.sub.4, filtered and concentrated under reduced
pressure. The residue was purified by FCC (10 g silica, Isolute
cartridge, gradient of eluents; 99:1 Heptane:EtOAc to 85:15
Heptane:EtOAc) to give 16.2 g (102.4%) of the title compound as an
orange oil that solidified upon standing and contained residual
heptane. LC-MS 91%, m/z=267.4/268.9, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.89 (d, J=2.2 Hz, 1H), 7.51 (d, J=8.3
Hz, 1H), 7.41 (dd, J=8.3, 2.3 Hz, 1H), 3.92 (s, 3H), 0.27 (s,
9H).
Step 2: Synthesis of methyl 5-chloro-2-ethynylbenzoate
##STR00793##
[1689] To a solution of methyl
5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate (10 g, 37.5 mmol) in
MeOH (150 ml) was added K.sub.2CO.sub.3 (10.4 g, 75 mmol) at room
temperature. The reaction mixture was stirred for 1 hour before
being concentrated under reduced pressure. The residue was
dissolved in deionized water (50 ml) and EtOAc (50 ml). The phases
were separated and the aqueous layer was extracted with EtOAc
(2.times.50 ml). The combined organic extracts were dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified by FCC (50 g silica, Isolute cartridge,
gradient of eluents; 95:5 Heptane:EtOAc to 9:1 Heptane:EtOAc) to
give 5.75 g (55%) of the title compound as an orange oil that
solidified upon standing. This material contained 30% of ethyl
ester and was suitable for use without any further purification.
LC-MS 38%, m/z=195.0/196.9, .sup.1H NMR (500 MHz, Chloroform-d)
.delta. ppm 7.93 (d, J=2.2 Hz, 1H), 7.55 (d, J=8.3 Hz, 1H), 7.45
(dd, J=8.3, 2.3 Hz, 1H), 3.94 (s, 3H), 3.43 (s, 1H).
Step 3: Synthesis of methyl 5-chloro-2-ethylbenzoate
##STR00794##
[1691] To a solution of methyl 5-chloro-2-ethynylbenzoate (5.34 g,
27 mmol) in ethyl acetate (135 ml) was added Pd/C (10%) (50% water,
2.92 g, 1.37 mmol). The reaction mixture was stirred under a
hydrogen atmosphere at room temperature for 3 hours. The mixture
was filtered through Celite. The filter cake washed with EtOAc (50
ml) and the filtrate was concentrated under reduced pressure to
give 5.12 g (94%) of the title compound as a brown oil which was
suitable for use without any further purification. LC-MS 56%,
m/z=198.9/200.9, .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm
7.84 (d, J=2.3 Hz, 1H), 7.39 (dd, J=8.3, 2.3 Hz, 1H), 7.21 (d,
J=8.3 Hz, 1H), 3.90 (s, 3H), 2.94 (q, J=7.5 Hz, 2H), 1.21 (t, J=7.5
Hz, 3H).
Step 4: Synthesis of methyl 5-chloro-2-ethyl-3-nitrobenzoate
##STR00795##
[1693] A solution of methyl 5-chloro-2-ethylbenzoate (5.12 g, 25.8
mmol) in concentrated H.sub.2SO.sub.4 (31 ml, 587 mmol) was cooled
to -5.degree. C. in an acetone/ice bath. A mixture of concentrated
nitric acid (2.15 ml, 33.5 mmol) and concentrated H.sub.2SO.sub.4
(2.0 ml, 37.5 mmol) was added dropwise to the reaction mixture at
-5.degree. C. over 15 minutes. The pale yellow reaction mixture was
stirred at -5.degree. C. for 1 hour before being poured onto ice
(500 ml) and this was extracted with EtOAc (3.times.100 ml). The
combined organic phases were washed with deionized water (100 ml)
and then brine (100 ml). The organic phase was dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. LCMS
and NMR showed .about.30% hydrolysis of the ester. The crude
material was dissolved in methanol (30 ml) and cooled to 0.degree.
C. under nitrogen where SOCl.sub.2 (2.25 ml, 30.93 mmol) was added
slowly. The reaction mixture was then heated to reflux for 6 hours
before being concentrated under reduced pressure to give 6.18 g
(98%) of the title compound containing material as an orange oil
which was a 1:1 mixture of 3-nitro:6-nitro isomers along with some
ethyl ester. The material was suitable for use in the next step
without any further purification.
Step 5: Synthesis of methyl 3-amino-5-chloro-2-ethylbenzoate
##STR00796##
[1695] To a solution of methyl 5-chloro-2-ethyl-3-nitrobenzoate
(6.18 g, 25.4 mmol) in methanol (250 ml) at room temperature was
added ammonium chloride (13.3 g, 253 mmol) followed by deionized
water (125 ml). The mixture was heated to 70.degree. C. before the
addition of iron (8.50 g, 152 mmol). The reaction turned to a dark
color over the 2.5 hours it was stirred at 70.degree. C. This
mixture was allowed to cool to room temperature and was filtered
through Kieselgel. The filter pad was washed with MeOH (250 ml) and
the filtrate concentrated under reduced pressure. The residue was
dissolved in saturated NaHCO.sub.3(aq) (50 ml) and EtOAc (150 ml).
The phases were separated and the organic phase was washed with
saturated NaHCO.sub.3(aq) (2.times.50 ml) before being dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified by FCC (50 g silica, Isolute cartridge,
gradient of eluents; 95:5 Heptane:EtOAc to 75:25 Heptane:EtOAc) to
give 2.42 g of crude methyl 3-amino-5-chloro-2-ethylbenzoate as a
yellow oil. The material was taken through the next step without
further purification. LC-MS 31%, m/z=295.0, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.17 (d, J=2.1 Hz, 1H), 6.79 (d, J=2.1
Hz, 1H), 3.87 (s, 3H), 3.86-3.81 (m, 2H), 2.74 (q, J=7.5 Hz, 2H),
1.20 (t, J=7.5 Hz, 3H).
Step 6: Synthesis of methyl
5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate
##STR00797##
[1697] To a solution of methyl 3-amino-5-chloro-2-ethylbenzoate
(1.5 g, 7.02 mmol) in 1,2-dichloroethane (28 ml) at room
temperature under nitrogen was added oxan-4-one (1.3 ml, 14. mmol)
followed by acetic acid (2.41 ml, 42 mmol). This solution was
stirred for 5 minutes before the addition of sodium
triacetoxyborohydride (4.46 g, 21 mmol) at room temperature. After
stirring for 20 hours, deionized water (28 ml) was added and the
mixture was neutralized with solid NaHCO.sub.3. The phases were
separated and the aqueous layer was extracted with EtOAc
(2.times.28 ml). The combined organic extracts were dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified by FCC (50 g silica, Isolute cartridge,
gradient of eluents; 95:5 Heptane:EtOAc to 8:2 Heptane:EtOAc) to
give 1.76 g of crude methyl
5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate as a white solid. The
material was taken through the next step without further
purification. LC-MS 60%, m/z=298.0/300.0, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.07 (d, J=2.0 Hz, 1H), 6.71 (d, J=1.9
Hz, 1H), 4.01 (dt, J=11.8, 3.4 Hz, 2H), 3.87 (s, 3H), 3.82-3.76 (m,
1H), 3.64-3.47 (m, 3H), 2.79-2.63 (m, 2H), 2.06 (d, J=13.2 Hz, 2H),
1.55-1.46 (m, 2H), 1.18 (t, J=7.5 Hz, 3H).
Step 7: Synthesis of methyl
5-chloro-2-ethyl-3-[methyl(oxan-4-yl)amino]benzoate
##STR00798##
[1699] To a solution of methyl
5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate (350 mg, 1.18 mmol)
in 1,2-dichloroethane (10 ml) at room temperature under nitrogen
was added paraformaldehyde (212 mg, 7.05 mmol) followed by acetic
acid (0.4 ml, 7.05 mmol). This mixture was stirred for 5 minutes
before the addition of sodium triacetoxyborohydride (1.49 g, 7.05
mmol). After stirring for 23 hours, further paraformaldehyde (212
mg, 7.05 mmol) was added to the reaction mixture and this was
stirred for 10 minutes before the addition of sodium
triacetoxyborohydride (1.49 g, 7.05 mmol). After stirring for 3
hours, deionized water (15 ml) was added and the mixture was
neutralized with solid NaHCO.sub.3 and the phases were separated.
The aqueous layer was extracted with EtOAc (2.times.15 ml) and the
combined organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
FCC (10 g silica, Isolute cartridge, gradient of eluents; 99:1
Heptane:EtOAc to 85:15 Heptane:EtOAc) to give 334 mg of the title
compound as a colorless oil. The product contains .about.22% ethyl
ester. The material was taken through the next step without further
purification. LC-MS 56%, m/z=312.0/314.0, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.52 (d, J=2.2 Hz, 1H), 7.24 (d, J=2.2
Hz, 1H), 4.01-3.93 (m, 2H), 3.89 (s, 3H), 3.34 (td, J=11.7, 2.3 Hz,
2H), 3.06-2.96 (m, 2H), 2.94-2.85 (m, 1H), 2.61 (s, 3H), 1.76-1.59
(m, 4H), 1.10 (t, J=7.4 Hz, 3H).
Step 8: Synthesis of
5-chloro-2-ethyl-3-[methyl(oxan-4-yl)amino]benzoic acid
##STR00799##
[1701] To a solution of methyl
5-chloro-2-ethyl-3-[methyl(oxan-4-yl)amino]benzoate (334 mg, 1.07
mmol) in THF (11 ml) was added 4M NaOH (10.7 ml). The reaction
mixture was stirred at 50.degree. C. for 27 hours. MeOH (5 ml) was
added to the reaction mixture and this was stirred for a further 21
hours at 50.degree. C. The reaction mixture was acidified to pH 2-3
with 6M HCl and extracted with DCM (5.times.10 ml). The combined
organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure to give 311 mg of the title
compound as an orange oil that solidified upon standing. The
material was taken through the next step without further
purification. LC-MS 79%, 2.01 min (3 minute LC-MS method),
m/z=298.0/300.0, .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm
7.72 (d, J=2.1 Hz, 1H), 7.37-7.27 (m, 1H), 4.00 (brd, J=11.1 Hz,
2H), 3.36 (td, J=11.6, 2.0 Hz, 2H), 3.11 (q, J=7.4 Hz, 2H), 2.94
(t, J=10.6 Hz, 1H), 2.64 (s, 3H), 1.78-1.56 (m, 4H), 1.15 (t, J=7.4
Hz, 3H).
Step 9: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-[methyl(oxan-4-yl)amino]benzamide
##STR00800##
[1703] To a solution of
5-chloro-2-ethyl-3-[methyl(oxan-4-yl)amino]benzoic acid (100 mg,
0.34 mmol) in DMF (2 ml) at room temperature under nitrogen was
added HBTU (153 mg, 0.4 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (88 .mu.l, 0.5 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 63 mg,
0.37 mmol). After stirring for 3 hours at room temperature EtOAc
(30 ml) was added to the reaction mixture and this was then washed
with deionized water (5 ml) followed by saturated NaHCO.sub.3(aq)
(3.times.5 ml). The organic phase was dried over MgSO4, filtered
and concentrated under reduced pressure. The residue was purified
by FCC (5 g silica, Isolute cartridge, gradient of eluents; 100%
DCM to 96:4 DCM:MeOH) to give 94 mg (65%) of the title compound as
an off-white solid. LC-MS 95%, m/z=432.1/434.1, .sup.1H NMR (500
MHz, Chloroform-d) .delta. ppm 11.34 (s, 1H), 7.14-7.08 (m, 2H),
7.03 (d, J=2.0 Hz, 1H), 5.95 (s, 1H), 4.52 (d, J=5.8 Hz, 2H), 3.96
(d, J=11.2 Hz, 2H), 3.33 (td, J=11.4, 2.3 Hz, 2H), 2.96-2.85 (m,
1H), 2.79 (q, J=7.4 Hz, 2H), 2.58 (s, 3H), 2.39 (s, 3H), 2.23 (s,
3H), 1.73-1.62 (m, 4H), 1.05 (t, J=7.4 Hz, 3H).
Compound 206:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-[ethyl(oxan-4-yl)amino]benzamide
##STR00801##
[1704] Step 1: Synthesis of methyl
5-chloro-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzoate
##STR00802##
[1706] To a solution of methyl
5-chloro-2-ethyl-3-[(oxan-4-yl)amino]benzoate (350 mg, 1.18 mmol)
in 1,2-dichloroethane (10 ml) at room temperature under nitrogen
was added acetaldehyde (0.66 ml, 11.8 mmol) followed by acetic acid
(0.4 ml, 7.05 mmol). This solution was stirred for 5 minutes before
the addition of sodium triacetoxyborohydride (2.49 g, 11.8 mmol) at
room temperature. After stirring for 23 hours acetaldehyde (0.66
ml, 11.8 mmol) was added to the dark reaction mixture and this was
stirred for 10 minutes before the addition of sodium
triacetoxyborohydride (2.49 g, 11.8 mmol). After stirring for 3
hours deionized water (15 ml) was added and the mixture was
neutralized with solid NaHCO.sub.3. The phases were separated and
the aqueous layer was extracted with EtOAc (2.times.15 ml). The
combined organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure. The residue was purified by
FCC (10 g silica, Isolute cartridge, gradient of eluents; 99:1
Heptane:EtOAc to 85:15 Heptane:EtOAc) to give 317 mg of the title
compound as a colorless oil. The product contains .about.25% ether
ester. The material was taken through the next step without further
purification. LC-MS 55%, 2 m/z=326.0/328.0, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.52 (d, J=2.2 Hz, 1H), 7.23 (d, J=2.2
Hz, 1H), 3.97 (d, J=11.1 Hz, 2H), 3.90 (s, 3H), 3.32 (td, J=11.6,
2.5 Hz, 2H), 3.04 (q, J=7.1 Hz, 4H), 2.93 (ddd, J=15.1, 10.8, 4.3
Hz, 1H), 1.76-1.62 (m, 4H), 1.08 (t, J=7.4 Hz, 3H), 0.88 (t, J=7.1
Hz, 3H).
Step 2: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-[ethyl(oxan-4-yl)amino]benzamide
##STR00803##
[1708] To a solution of methyl
5-chloro-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzoate (317 mg, 0.97
mmol) in THF (10 ml) was added 4M NaOH (9.73 ml). The reaction
mixture was stirred at 50.degree. C. for 27 hours. MeOH (5 ml) was
added to the reaction mixture and this was stirred for a further 21
hours at 50.degree. C. The reaction mixture was acidified to pH 2-3
with 6M HCl and extracted with DCM (5.times.10 ml). The combined
organic extracts were dried over MgSO.sub.4, filtered and
concentrated under reduced pressure to give 289 mg of the title
compound as an orange oil that solidified upon standing. The
material was taken through the next step without further
purification. LC-MS 79%, m/z=311.95/313.95, .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.73 (d, J=1.7 Hz, 1H), 7.29 (d, J=1.8
Hz, 1H), 3.99 (d, J=11.0 Hz, 2H), 3.38-3.29 (m, 2H), 3.20-3.03 (m,
4H), 3.02-2.91 (m, 1H), 1.78-1.61 (m, 4H), 1.13 (t, J=7.4 Hz, 3H),
0.91 (t, J=7.0 Hz, 3H).
Step 3: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-[ethyl(oxan-4-yl)amino]benzamide
##STR00804##
[1710] To a solution of
5-chloro-2-ethyl-3-[ethyl(oxan-4-yl)amino]benzoic acid (100 mg,
0.32 mmol) in DMF (2 ml) at room temperature under nitrogen was
added HBTU (146 mg, 0.38 mmol) followed by
N-ethyl-N-(propan-2-yl)propan-2-amine (84 .mu.l, 0.48 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 60.3
mg, 0.35 mmol). After stirring for 3 hours at room EtOAc (30 ml)
was added to the reaction mixture and this was then washed with
deionized water (5 ml) followed by saturated NaHCO.sub.3(aq)
(3.times.5 ml). The organic phase was dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. The residue was
purified by FCC (5 g silica, Isolute cartridge, gradient of
eluents; 100% EtOAc to 97:3 EtOAc:MeOH) to give 90 mg (63%) of the
title compound as a white solid. LC-MS 97%, m/z=446.1/448.2,
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 11.93 (s, 1H), 7.14
(t, J=5.7 Hz, 1H), 7.10 (d, J=2.0 Hz, 1H), 7.04 (d, J=2.0 Hz, 1H),
5.95 (s, 1H), 4.52 (d, J=5.8 Hz, 2H), 3.95 (d, J=11.2 Hz, 2H), 3.30
(td, J=11.3, 3.6 Hz, 2H), 3.01 (q, J=7.0 Hz, 2H), 2.93 (ddd,
J=15.3, 10.0, 5.3 Hz, 1H), 2.83 (q, J=7.4 Hz, 2H), 2.38 (s, 3H),
2.22 (s, 3H), 1.71-1.63 (m, 4H), 1.02 (t, J=7.5 Hz, 3H), 0.86 (t,
J=7.0 Hz, 3H).
Compound 243:
5-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-
-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzamide
##STR00805##
[1711] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00806##
[1713] A solution of 5-bromo-2-methylbenzoic acid (5.0 g, 23 mmol)
in concentrated H.sub.2SO.sub.4 (27 ml, 512 mmol) was cooled to
5.degree. C. in an acetone/ice bath. A mixture of concentrated
nitric acid (1.9 ml, 30 mmol) and concentrated H.sub.2SO.sub.4 (2.8
ml, 52 mmol) was added dropwise to the reaction mixture at -5 to
.degree. C. over 15 minutes. The yellow reaction mixture was
stirred at -5.degree. C. for 2 hours during which time a yellow
precipitate formed. The reaction mixture was poured onto ice (150
g) and the precipitate was then collected by filtration. The
precipitate was air dried to give the title compound (5.5 g, 52%)
as a pale yellow solid.; .sup.1H NMR (500 MHz, DMSO-d.sub.6)
.delta. ppm 8.29 (s, 1H) 8.13 (d, J=1.58 Hz, 1H) 2.43 (s, 3H).
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate
##STR00807##
[1715] To a solution of 5-bromo-2-methyl-3-nitrobenzoic acid (5.5
g, 21 mmol) in DMF (42 ml) under nitrogen, was added
Na.sub.2CO.sub.3 (3.4 g, 32 mmol) followed by iodomethane (2.0 ml,
32 mmol). The reaction mixture was stirred at room temperature for
2 hours. The mixture was diluted with deionized water (150 ml) and
extracted with EtOAc (4.times.50 ml). The combined organic phases
were washed with saturated NaHCO.sub.3 (aq) (2.times.50 ml), dried
over MgSO.sub.4, filtered and concentrated in-vacuo to give the
title compound (6.3 g, 61%) as a yellow oil. .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.38 (d, J=2.05 Hz, 1H) 7.23 (d, J=2.05
Hz, 1H) 3.20 (s, 3H) 1.82 (s, 3H).
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00808##
[1717] To a solution of methyl 5-bromo-2-methyl-3-nitrobenzoate
(6.3 g, 21 mmol) in methanol (150 ml) was added ammonium chloride
(11.0 g, 210 mmol) followed by deionized water (75 ml). The mixture
was heated to 70.degree. C. before the addition of iron (7.0 g, 125
mmol). The reaction mixture was stirred at 70.degree. C. for 2
hours, before being allowed to cool to room temperature and
filtered through Kieselgel. The filter pad was washed with MeOH
(150 ml) and the filtrate concentrated in-vacuo. The residue was
dissolved in saturated NaHCO.sub.3 (aq) (50 ml) and EtOAc (150 ml).
The phases were separated and the organic phase was washed with
saturated NaHCO.sub.3 (aq) (3.times.50 ml), dried over MgSO.sub.4,
filtered and concentrated in-vacuo. The residue was purified by
flash column chromatography (50 g silica Isolute cartridge, 5-20%
EtOAc:Heptanes) to give the title compound (3.0 g, 51%) as a thick
pale yellow oil. LC-MS 87%, m/z=243.9, 244.9, 245.9, 246.9; .sup.1H
NMR (500 MHz, Chloroform-d) .delta. ppm 7.34 (d, J=1.89 Hz, 1H)
6.95 (d, J=1.89 Hz, 1H) 3.88 (s, 3H) 3.80 (br. s., 2H) 2.29 (s,
3H).
Step 4: Synthesis of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate
##STR00809##
[1719] To a solution of methyl 3-amino-5-bromo-2-methylbenzoate
(3.0 g, 12 mmol) in 1,2-dichloroethane (48 ml) under nitrogen, was
added oxan-4-one (2.3 ml, 25 mmol) followed by acetic acid (4.2 ml,
74 mmol). The reaction mixture was stirred for 5 minutes before the
addition of sodium triacetoxyborohydride (7.8 g, 37 mmol). After
stirring for 64 hours, deionized water (100 ml) was added and the
mixture was neutralized with solid NaHCO.sub.3. The phases were
separated and the aqueous layer was extracted with EtOAc
(4.times.50 ml). The combined organic extracts were dried over
MgSO.sub.4, filtered and concentrated in-vacuo. The residue was
purified by flash column chromatography (50 g silica, Isolute
cartridge, 10-30% EtOAc:Heptanes) to give the title compound (3.5 g
85%) as a white solid. LC-MS 99.8 m/z=327.9, 328.9, 329.9, 330.9;
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 7.24 (d, J=1.73 Hz,
1H) 6.85 (d, J=1.58 Hz, 1H) 4.03 (dt, J=11.82, 3.31 Hz, 2H) 3.88
(s, 3H) 3.66 (br.s., 1H) 3.56 (td, J=11.55, 1.97 Hz, 2H) 3.47-3.55
(m, 1H) 2.24 (s, 3H) 2.06 (d, J=13.56 Hz, 2H) 1.47-1.60 (m,
2H).
Step 5: Synthesis of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate
##STR00810##
[1721] To a solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (500 mg, 1.5 mmol)
and TFA (15 ml), was added NaBH.sub.4 (1.0 g, 26 mmol) portionwise
over 5 minutes. The reaction mixture was stirred at room
temperature for 2 hours and then heated to 50.degree. C. for 3
hours and treated with a further aliquot of NaBH.sub.4 (300 mg)
over 25 minutes. The reaction mixture was then heated to 60.degree.
C. for 2 hours and left to stir at room temperature for 17 hours.
The reaction mixture was treated with TFA (5 ml) and NaBH.sub.4
(200 mg) and heated back up to 60.degree. C. for 3.5 hours. A
further aliquot of NaBH.sub.4 (200 mg) was added over 15 minutes,
along with TFA (5 ml) and heating continued for a further 3 hours,
before being left to stand at room temperature overnight. The
reaction mixture was poured over ice (75 ml) and stirred until the
ice had melted. The reaction mixture was then basified by the
addition of 6M NaOH (aq) (40 ml) and re-adjusted to pH 7 using 1M
HCl (aq) (40 ml). The resulting white suspension was collected by
filtration, the solid washed with water (20 ml) and dried in-vacuo
at 40.degree. C. for 3 hours to give the title compound (577 mg,
91%) as a white solid. LC-MS 98%), m/z=409.90, 410.9, 411.90,
412.9; .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 7.80 (d,
J=1.73 Hz, 1H) 7.41 (d, J=1.73 Hz, 1H) 4.01 (dd, J=11.51, 4.10 Hz,
2H) 3.91 (s, 3H) 3.64 (d, J=5.20 Hz, 2H) 3.32 (t, J=11.82 Hz, 2H)
2.99 (tt, J=11.43, 3.63 Hz, 1H) 2.48 (s, 3H) 1.80 (dd, J=12.53,
1.50 Hz, 2H) 1.54-1.62 (m, 2H).
Step 6: Synthesis of
5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoic
acid
##STR00811##
[1723] To a stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate
(572 mg, 1.4 mmol) in a mixture of THF (14 ml) and MeOH (2.1 ml),
was added 4M NaOH (aq) (13.9 ml). The reaction mixture was stirred
at 50.degree. C. for 5.5 hours and then stirred at room temperature
for 17 hours. The THF was removed by concentrating in-vacuo and the
aqueous residue was acidified to pH 4 with 6M HCl (9.5 ml). The
resulting suspension was allowed to stand at room temperature for
20 minutes before collecting the solid by filtration. The solid
cake was washed with water (20 ml) and dried under high vacuum for
2 hours to give the title compound (507 mg, 90%) as a white solid.
LC-MS 98%, m/z=395.9, 396.9, 397.9, 398.9; .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.97 (d, J=1.73 Hz, 1H) 7.48 (d, J=1.73
Hz, 1H) 4.02 (dd, J=11.35, 3.94 Hz, 2H) 3.65 (br. s, 2H) 3.33 (t,
J=11.59 Hz, 2H) 3.00 (tt, J=11.49, 3.80 Hz, 1H) 2.55 (s, 3H) 1.82
(d, J=11.98 Hz, 2H) 1.55-1.69 (m, 2H). OH not visible.
Step 7: Synthesis of
5-bromo-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-3-
-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzamide
##STR00812##
[1725] A stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)(2,2,2-trifluoroethyl)amino]benzoate
(250 mg, 0.63 mmol) in dry DMF (3.0 ml) at 0.degree. C. under
nitrogen, was treated with HATU (288 mg, 0.76 mmol) and DIPEA (220
.mu.l, 1.3 mmol) dropwise. The resulting solution was stirred for 5
minutes and then treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 119 mg,
0.69 mmol). The resulting suspension was stirred at 0.degree. C.
for 20 minutes and then stirred at room temperature for 16.5 hours.
The reaction mixture was treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (30 mg).
Stirring was continued for 23 hours and the reaction mixture was
then partitioned between water (30 ml) and CH.sub.2Cl.sub.2 (20
ml). The layers were separated and the aqueous phase was extracted
with CH.sub.2Cl.sub.2 (3.times.20 ml). The combined organics were
washed with a saturated solution of NaHCO.sub.3) (50 ml), water (60
ml), brine (2.times.40 ml), dried over MgSO.sub.4, filtered and
concentrated in-vacuo. The crude residue was purified by flash
column chromatography (10 g SNAP cartridge, Isolera, 0-10%
MeOH/CH.sub.2Cl.sub.2) and triturated from ether (10 ml) with
sonication. The resulting precipitate was collected by filtration
and dried in-vacuo to give the title compound (249 mg, 74%) as a
white solid. LC-MS 100%, m/z=530.0, 531.0, 532.0, 533.0; .sup.1H
NMR (500 MHz, Acetone) .delta. 10.67 (s, 1H), 7.55 (d, J=1.8 Hz,
2H), 7.27 (d, J=1.9 Hz, 1H), 5.90 (s, 1H), 4.40 (d, J=5.5 Hz, 2H),
3.90 (dd, J=11.2, 4.6 Hz, 4H), 3.28 (t, J=11.6 Hz, 2H), 3.07-2.97
(m, 1H), 2.32 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H), 1.76 (dd,
J=12.3, 1.6 Hz, 2H), 1.61 (qd, J=12.0, 4.5 Hz, 2H).
Compound 244:
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-N-[(4,6-dimethyl-2-oxo-1,-
2-dihydropyridin-3-yl)methyl]-2-methylbenzamide
##STR00813##
[1726] Step 1: Synthesis of methyl
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoate
##STR00814##
[1728] A stirred solution of methyl
5-bromo-2-methyl-3-[(oxan-4-yl)amino]benzoate (500 mg, 1.5 mmol) in
difluoroacetic acid (15 ml), was treated with sodium
tetrahydroborate (1000 mg, 26 mmol) portionwise over 12 minutes.
The reaction mixture was warmed to 50.degree. C. and stirred for 4
hours. The reaction mixture was allowed to reach room temperature
and then poured over ice (130 ml) and left for 5 minutes. The
mixture was basified by the addition of 6M NaOH (aq) (35 ml) and
the pH adjusted to 7 using 1M HCl (aq) (20 ml). The resulting
suspension was allowed to stand until the solution was clear and
the resulting solid collected by filtration and dried in-vacuo at
40.degree. C. to give the title compound (572 mg, 96%) as a white
solid. LC-MS 100%, m/z=391.9, 392.9, 393.9, 394.9; .sup.1H NMR (500
MHz, Chloroform-d) .delta. ppm 7.79 (d, J=1.89 Hz, 1H) 7.44 (d,
J=1.89 Hz, 1H) 5.44-5.71 (m, 1H) 4.00 (dd, J=11.51, 4.10 Hz, 2H)
3.91 (s, 3H) 3.41 (td, J=13.99, 4.18 Hz, 2H) 3.32 (t, J=11.27 Hz,
2H) 2.97 (tt, J=11.37, 3.84 Hz, 1H) 2.47 (s, 3H) 1.72-1.81 (m, 2H)
1.59-1.67 (m, 2H).
Step 2: Synthesis of
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoic
acid
##STR00815##
[1730] To a stirred solution of methyl
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoate
(571 mg, 1.5 mmol) in a mixture of THF (14.6 ml) and MeOH (2.2 ml),
was added 4M NaOH (14.6 ml). The reaction mixture was stirred at
50.degree. C. for 7 hours. The heat was switched off and the
reaction mixture was stirred at room temperature for 16.5 hours.
THF was removed in-vacuo and the aqueous residue was acidified to
pH 4 by the addition of 6M HCl (aq) (10 ml) with ice cooling. The
resulting solid was collected by filtration, washed with water (20
ml), and dried in-vacuo to give the title compound (526 mg, 96%) as
a light beige solid. LC-MS 100%, m/z=377.9, 378.9, 379.9, 380.9;
.sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 7.91 (d, J=1.58 Hz,
1H) 7.49 (d, J=1.58 Hz, 1H) 5.43-5.75 (m, 1H) 4.01 (dd, J=11.43,
3.55 Hz, 2H) 3.42 (td, J=13.95, 3.78 Hz, 2H) 3.32 (t, J=11.35 Hz,
2H) 2.98 (tt, J=11.37, 3.53 Hz, 1H) 2.52 (s, 3H) 1.77 (d, J=10.88
Hz, 2H) 1.56-1.69 (m, 2H). OH not visible.
Step 3: Synthesis of
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-N-[(4,6-dimethyl-2-oxo-1,-
2-dihydropyridin-3-yl)methyl]-2-methylbenzamide
##STR00816##
[1732] A stirred solution of
5-bromo-3-[(2,2-difluoroethyl)(oxan-4-yl)amino]-2-methylbenzoic
acid (250 mg, 0.66 mmol) in dry DMF (3.0 ml) at 0.degree. C. under
nitrogen, was treated with HATU (327 mg, 0.86 mmol) and DIPEA (230
.mu.l, 1.3 mmol) dropwise. The resulting solution was stirred for 5
minutes and then treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 136 mg,
0.79 mmol). The resulting suspension was stirred at 0.degree. C.
for 20 minutes and then stirred at room temperature overnight.
After 18 hours,
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (25 mg) was
added and stirring continued for a further 25 hours. The reaction
mixture was diluted with water (30 ml) and CH.sub.2Cl.sub.2 (30
ml). The layers were separated and the aqueous phase was extracted
with CH.sub.2Cl.sub.2 (3.times.15 ml). The combined organic phases
were washed with a saturated solution of NaHCO.sub.3 (aq) (45 ml),
water (2.times.50 ml), brine (2.times.50 ml), dried (MgSO.sub.4),
filtered and concentrated in-vacuo. The residue was purified by
column chromatography (10 g SNAP cartridge, Isolera, 0-3%
MeOH:CH.sub.2Cl.sub.2) and then triturated with ether. The
resulting solid was collected by filtration and dried in-vacuo at
40.degree. C. to give the title compound (259 mg, 77%) as an off
white solid. LC-MS 100%, m/z=512.0, 513.0, 514.0, 515.0; .sup.1H
NMR (500 MHz, Acetone) .delta. 10.71 (s, 1H), 7.57-7.49 (m, 2H),
7.25 (d, J=1.9 Hz, 1H), 5.91 (s, 1H), 5.76 (tt, J=56.2, 4.3 Hz,
1H), 4.40 (d, J=5.5 Hz, 2H), 3.88 (dd, J=11.3, 4.2 Hz, 2H), 3.52
(td, J=14.6, 4.2 Hz, 2H), 3.33-3.23 (m, 2H), 3.02 (tt, J=11.6, 3.9
Hz, 1H), 2.32 (s, 3H), 2.28 (s, 3H), 2.24 (s, 3H), 1.73 (dd,
J=12.4, 1.9 Hz, 2H), 1.59 (qd, J=12.2, 4.5 Hz, 2H).
Compound 328:
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-[ethyl(oxan-4-yl-
)amino]-3-methylpyridine-4-carboxamide
##STR00817##
[1733] Step 1: Synthesis of
2-chloro-N-phenylpyridine-4-carboxamide
##STR00818##
[1735] To a stirred suspension of 2-chloroisonicotinic acid (5.0 g,
0.03 mol) in dry toluene (100 ml) and 4 drops of DMF, was added
dropwise thionyl chloride (7.4 ml, 0.10 mol) under nitrogen. The
reaction mixture was stirred at 60.degree. C. for 2.5 hours and the
temperature was gradually increased to 100.degree. C. over 1.5
hours and held at this temperature for 2 hours. The reaction
mixture was stirred at room temperature for 65 hours, before being
treated with another equivalent of thionyl chloride (2.3 ml) and
heated to 95.degree. C. for 40 minutes. The reaction was allowed to
cool to room temperature and was then concentrated to dryness
in-vacuo and azeotroped with dry toluene (2.times.10 ml) to give a
yellow mobile oil which was used directly in the next step without
further purification.
[1736] To a stirred solution of aniline (2.9 ml, 0.03 mol) and
N-ethyl-N-(propan-2-yl)propan-2-amine (13.6 ml, 83 mmol) in dry THF
(50 ml) was added the acid chloride prepared above dropwise. The
reaction mixture was stirred under nitrogen for 19 hours. The
reaction mixture was then quenched by the addition of a saturated
solution of sodium hydrogen carbonate (aq) (100 ml) and stirred for
35 minutes. The layers were separated and the aqueous phase
extracted with ethyl acetate (3.times.55 ml). The combined organic
extracts were washed with water (3.times.66 ml), brine (80 ml),
dried (MgSO.sub.4), filtered and concentrated in-vacuo. The residue
was suspended in diethyl ether (100 ml), briefly sonicated and the
product collected by filtration. It was dried under high vacuum for
3 hours at room temperature, then left under vacuum overnight to
give the title compound (6.1 g, 83%) as a white solid. LC-MS 100%,
m/z=233.0, 234.9; .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm
8.58 (d, J=5.04 Hz, 1H) 7.81 (br. s., 1H) 7.77 (s, 1H) 7.63 (d,
J=7.41 Hz, 3H) 7.42 (t, J=7.96 Hz, 2H) 7.19-7.26 (m, 1H).
Step 2: Synthesis of
2-chloro-N,3-dimethyl-N-phenylpyridine-4-carboxamide
##STR00819##
[1738] To a stirred solution of
2-chloro-N-phenylpyridine-4-carboxamide (2.0 g, 8.6 mmol) in dry
THF (40 ml) at -78.degree. C. under nitrogen, was added dropwise
1.6M butyllithium in n-hexanes (11.8 ml, 18.9 mmol) over 24
minutes. The reaction mixture was stirred at -78.degree. C. for 50
minutes and then treated with iodomethane (1.8 ml, 28 mmol)
dropwise over 8 minutes. The reaction mixture was stirred at
-78.degree. C. for 1 hour and then left to slowly reach room
temperature and stirred for 20 hours. The reaction mixture was
quenched with water (20 ml), and the layers were separated. The
aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.20 ml).
The combined organic phases were washed with brine (60 ml), dried
(MgSO.sub.4), filtered, concentrated in-vacuo and to give the title
compound (2.2 g, 93%) as a thick brown oil. LC-MS 95%, m/z=261.0,
263.0; .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 8.03 (d,
J=4.89 Hz, 1H) 7.23 (d, J=7.88 Hz, 2H) 7.16-7.21 (m, 1H) 7.02 (d,
J=7.25 Hz, 2H) 6.89 (d, J=4.89 Hz, 1H) 3.51 (s, 3H) 2.34 (s,
3H).
Step 3: Synthesis of 2-chloro-3-methylpyridine-4-carboxylic
acid
##STR00820##
[1740] 2-chloro-N,3-dimethyl-N-phenylpyridine-4-carboxamide (2.2 g,
8.5 mmol) was treated with a mixture of concentrated
H.sub.2SO.sub.4 (23.2 ml) and water (16.8 ml) and then heated to
130.degree. C. for 46 hours. After which time, the reaction mixture
was allowed to reach room temperature and poured onto ice (200 ml)
and stirred until the ice had melted. The mixture was made alkaline
(pH 8) by the addition of solid Na.sub.2CO.sub.3 (50.3 g)
portionwise. The suspension was filtered and the filtrate acidified
to pH 3 by the addition of 6M HCl (aq) (.about.1.5 ml). The
resulting solid was collected by filtration, washed with water (20
ml) and dried in-vacuo at 40.degree. C. for 10 hours to give the
title compound (779 mg, 54%) as a pink solid. LC-MS 100%,
m/z=171.9, 173.9; .sup.1H NMR (500 MHz, MeOD) .delta. ppm 8.30 (d,
J=4.89 Hz, 1H) 7.64 (d, J=5.04 Hz, 1H) 2.58 (s, 3H).
Step 4: Synthesis of methyl
2-chloro-3-methylpyridine-4-carboxylate
##STR00821##
[1742] To a stirred solution of
2-chloro-3-methylpyridine-4-carboxylic acid (780 mg, 4.5 mmol) in
anhydrous DMF (5.0 ml), was added potassium carbonate (1.25 g, 9.1
mmol), followed by methyl iodide (0.42 ml, 6.8 mmol) dropwise. A
further 4 ml of DMF was added and the reaction mixture was stirred
at room temperature under nitrogen for 18.5 hours. The solvent was
removed in-vacuo and the residue was then partitioned between
CH.sub.2Cl.sub.2 (20 ml) and water (20 ml). The layers were
separated and the aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.15 ml). The combined organic layers were washed with water
(2.times.30 ml), brine (40 ml), dried (MgSO.sub.4), filtered and
concentrated in-vacuo. The residue was purified by column
chromatography (10 g SNAP cartridge, Isolera, 0-6% ethyl
acetate:heptanes) to give the title compound (591 mg, 59%) as a
colourless oil. LC-MS 84%, m/z=185.9, 187.9; .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 8.33 (d, J=5.04 Hz, 1H) 7.54 (d, J=5.04
Hz, 1H) 3.95 (s, 3H) 2.60 (s, 3H).
Step 5: Synthesis of methyl
3-methyl-2-[(oxan-4-yl)amino]pyridine-4-carboxylate
##STR00822##
[1744] A nitrogen purged suspension of Pd(OAc).sub.2 (77 mg, 0.34
mmol) and 1,1'-binaphthalene-2,2'-diylbis(diphenylphosphane) (430
mg, 0.69 mmol) in 1,4-dioxane (11 ml) was heated at 40.degree. C.
for 1 hour. Oxan-4-amine (237 .mu.l, 2.3 mmol), a solution of
methyl 2-chloro-3-methylpyridine-4-carboxylate (213 mg, 1.2 mmol)
in degassed dioxane (2 ml) and Cs.sub.2CO.sub.3 (562 mg, 1.7 mmol)
were then added and the red suspension was heated at 100.degree. C.
for 6 hours. After cooling to room temperature, the mixture was
diluted with CH.sub.2Cl.sub.2 (30 ml) and water (20 ml). The layers
were separated and the aqueous phase was extracted with
CH.sub.2Cl.sub.2 (3.times.15 ml). The combined organic phases were
washed with brine (25 ml), dried (MgSO.sub.4), filtered and
concentrated in-vacuo. The crude residue was purified by column
chromatography (25 g SNAP cartridge, Isolera, 0-28% ethyl
acetate:heptanes) followed by prep-HPLC (MeCN/Water) to give the
title compound (112 mg, 39%) as a white crystalline solid. LC-MS
100%, m/z=251.0; .sup.1H NMR (500 MHz, MeOD) .delta. ppm 7.91 (d,
J=5.20 Hz, 1H) 6.77 (d, J=5.36 Hz, 1H) 4.07-4.17 (m, 1H) 3.98 (dd,
J=12.06, 1.81 Hz, 2H) 3.88 (s, 3H) 3.55 (td, J=11.78, 1.66 Hz, 2H)
2.22 (s, 3H) 1.97 (dd, J=12.69, 1.97 Hz, 2H) 1.62 (qd, J=12.03,
4.41 Hz, 2H). NH not observed.
Step 6: Synthesis of
2-[ethyl(oxan-4-yl)amino]-3-methylpyridine-4-carboxylic acid
##STR00823##
[1746] A solution of methyl
3-methyl-2-[(oxan-4-yl)amino]pyridine-4-carboxylate (67 mg, 0.27
mmol) in dry DMF (1.0 ml), was added to a stirred suspension of
sodium hydride (60% dispersion in mineral oil, 16 mg, 0.4 mmol) in
DMF (0.3 ml) at 0.degree. C. under nitrogen. The reaction mixture
was allowed to warm to room temperature and stirred for 30 minutes.
The reaction was then treated with iodoethane (16 .mu.l, 0.20 mmol)
and stirred for 30 minutes, and then treated again with iodoethane
(16 .mu.l, 0.20 mmol) and stirred for a further 30 minutes. The
reaction mixture was added to a suspension of sodium hydride (16
mg, 0.4 mmol) in DMF (0.3 ml) at 0.degree. C. Once addition was
complete, the reaction mixture was allowed to reach room
temperature and stirred for 1 hour before treating with iodoethane
(2.times.16 .mu.l, 0.40 mmol). The reaction mixture was stirred for
a further 30 minutes. Sodium hydride (16 mg, 0.4 mmol) was added to
the reaction mixture and the mixture stirred for 45 minutes before
treating with iodoethane (2.times.16 .mu.l, 0.40 mmol) and stirred
for 15 minutes before adding under nitrogen to a solution of 4M
HCl/dioxane (5.0 ml) with cooling.
[1747] The reaction mixture was then concentrated in-vacuo at
40.degree. C. was treated with 1M NaOH (aq) (5 ml) and basified by
the addition of 10 NaOH pellets to pH 12 with cooling. The pH was
adjusted back to pH 10 by the addition of 6M HCl. The resulting
solution was stirred at room temperature for 18 hours. The reaction
mixture was then acidified to pH 5 by the addition of 1M HCl (80
drops). The resulting solution was extracted with CHCl.sub.3/IPA
(1:1) (6.times.10 ml). The combined extracts were concentrated
in-vacuo to give the crude title compound (207 mg) as a yellow oil.
LC-MS 100%, m/z=265.1. This material was used directly in the
coupling reaction as described below.
Step 7: Synthesis of
N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-[ethyl(oxan-4-yl-
)amino]-3-methylpyridine-4-carboxamide
##STR00824##
[1749] A stirred solution of
2-[ethyl(oxan-4-yl)amino]-3-methylpyridine-4-carboxylic acid (20
mg, 0.08 mmol) in dry DMF (1.0 ml) at 0.degree. C. under nitrogen,
was treated with HATU (37 mg, 0.1 mmol) and DIPEA (26 .mu.l, 0.15
mmol) dropwise. The resulting solution was stirred for 10 minutes
and then treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 15 mg,
0.09 mmol). The resulting suspension was stirred at 0.degree. C.
for 20 minutes and then stirred at room temperature for 18.5 hours.
The reaction mixture was cooled to 0.degree. C. and retreated with
HATU (120 mg, 0.31 mmol) and DIPEA (84 .mu.l, 0.48 mmol). After
stirring for 10 minutes,
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 49 mg,
0.29 mmol) was added. The reaction mixture was stirred for 1 hour
at 0.degree. C. and was then lifted out of the ice bath stirred at
room temperature for 48 hours. The reaction mixture was diluted
with water (20 ml) and CH.sub.2Cl.sub.2 (20 ml). The layers were
separated and the aqueous phase was extracted with CH.sub.2Cl.sub.2
(3.times.15 ml). The combined organic phases were washed with water
(2.times.25 ml), brine (2.times.25 ml), dried (MgSO.sub.4),
filtered and concentrated in-vacuo. The crude residue was purified
by column chromatography (10 g SNAP cartridge, Isolera, 0-40%
MeOH:CH.sub.2Cl.sub.2 and 0-25% MeOH (containing 10%
NH.sub.4OH):CH.sub.2Cl.sub.2) to give 6.7 mg (22% Yield) of the
title compound as an off white solid. LC-MS 100%, m/z=399.1;
.sup.1H NMR (250 MHz, DMSO) .delta. 11.49 (s, 1H), 8.32 (t, J=4.9
Hz, 1H), 8.14 (d, J=4.9 Hz, 1H), 6.84 (d, J=4.9 Hz, 1H), 5.86 (s,
1H), 4.26 (d, J=4.7 Hz, 2H), 3.84 (dd, J=10.3, 3.5 Hz, 2H), 3.21
(dt, J=13.6, 4.6 Hz, 5H), 2.19 (s, 3H), 2.15 (s, 3H), 2.11 (s, 3H),
1.62 (dt, J=21.1, 6.6 Hz, 4H), 0.80 (t, J=6.9 Hz, 3H).
Compound 356:
5-chloro-3-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}-N-[(5-fluoro-
-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methylbenzamide
##STR00825##
[1750] Step 1: Synthesis of
5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile
##STR00826##
[1752] To a stirred solution of 2-cyanoacetamide (689 mg, 8.2 mmol)
in anhydrous EtOH (7.0 ml) at 75.degree. C., was added
3-fluoropentane-2,4-dione (880 mg, 7.5 mmol), followed by
piperidine (96 .mu.l, 0.97 mmol). The reaction mixture was stirred
at this temperature for 3 hours and the reaction mixture left to
reach room temperature before being stored in the refrigerator for
4 days. The beige solid was collected by filtration and rinsed with
cold EtOH (4.times.0.4 ml) until the filtrate ran clear. The
resulting beige solid was dried in-vacuo at 40.degree. C. for 5
hours to give the title compound (733 mg, 58%) as a beige solid.
LC-MS 97%, m/z=166.9, .sup.1H NMR (500 MHz, Chloroform-d) .delta.
ppm 13.67 (br. s., 1H) 2.46 (d, J=2.05 Hz, 3H) 2.45 (d, J=2.84 Hz,
3H).
Step 2: Synthesis of
3-(aminomethyl)-5-fluoro-4,6-dimethyl-1,2-dihydropyridin-2-one
##STR00827##
[1754] A solution of 0.05M
5-fluoro-4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (731
mg, 4.4 mmol) in 1.75M NH.sub.3/MeOH (87 ml) was passed through the
H-Cube flow hydrogenator at 80.degree. C. and 50 bar at a flow rate
of 1 ml/min. The resulting solution was concentrated in-vacuo. The
resulting solid was split into 2 batches and 350 mg of the crude
product was purified by column chromatography (25 g SNAP cartridge,
Isolera, 0-25% MeOH (containing 10% NH.sub.4OH):CH.sub.2Cl.sub.2)
to give the title compound (307 mg, 20%) as an off white solid and
a 1:1 mixture of product:starting material. LC-MS (ELS) 100%,
m/z=170.9, .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 3.79 (s,
2H) 2.31 (d, J=2.84 Hz, 3H) 2.25 (d, J=2.05 Hz, 3H).
Step 3: Synthesis of
5-chloro-3-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}-2-methylbenz-
oic acid
##STR00828##
[1756] To a stirred solution of methyl
5-chloro-3-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}-2-methylbenz-
oate (1.0 g, 2.8 mmol) in THF (21 ml), was added a solution of 4M
NaOH (aq) (21.3 ml, 85.0 mmol), followed by MeOH (8.0 ml). The
resulting solution was stirred at 50.degree. C. for 16.5 hours and
cooled to room temperature. The organic solvents were removed
in-vacuo and the remaining aqueous phase was treated with a
solution of 6M HCl (aq) (14 ml) to adjust the pH to 7 and then 0.1
M HCl (aq) (24 ml) to adjust the pH to 4. The mixture was extracted
with ethyl acetate (3.times.35 ml). The aqueous extract was further
extracted with a mixture of CHCl.sub.3:IPA (1:1) (3.times.30 ml).
The combined CHCl.sub.3:IPA extracts were dried (MgSO.sub.4),
filtered and concentrated in-vacuo and then thoroughly dried on the
high vac line. The resulting solid was azeotroped with toluene
(3.times.20 ml). CH.sub.2Cl.sub.2 (10 ml) was then added to the
sample, concentrated in-vacuo and further dried on the high vac
line for several hours. This gave the title compound (831 mg, 88%)
as a white solid. LC-MS 92 m/z=339.0, 341.0; .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 12.21 (s, 1H) 7.69 (d, J=2.05 Hz, 1H)
7.22-7.26 (m, 1H) 2.99-3.11 (m, 3H) 2.69-2.79 (m, 7H) 2.47 (s, 3H)
2.28 (d, J=11.98 Hz, 2H) 2.05 (d, J=13.08 Hz, 2H) 1.35-1.65 (m, 4H)
0.88 (t, J=7.01 Hz, 3H).
Step 4: Synthesis of
5-chloro-3-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}-N-[(5-fluoro-
-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-methylbenzamide
##STR00829##
[1758] To a stirred solution of
5-chloro-3-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}-2-methylbenz-
oic acid (100 mg, 0.27 mmol) in anhydrous DMF (2.0 ml) at 0.degree.
C. under a f nitrogen, was treated with HATU (121 mg, 0.32 mmol)
and DIPEA (93 .mu.l, 0.53 mmol) dropwise. The resulting solution
was stirred for 10 minutes and then treated with
3-(aminomethyl)-5-fluoro-4,6-dimethyl-1,2-dihydropyridin-2-one
(50%, 99 mg, 0.29 mmol). The resulting suspension was stirred at
0.degree. C. for 1 hour and then stirred for 17 hours at room
temperature. The reaction mixture was partitioned between water (15
ml) and CH.sub.2Cl.sub.2 (20 ml). The layers were separated and the
aqueous phase was extracted with CH.sub.2Cl.sub.2 (2.times.15 ml).
The combined organics were washed with a saturated solution of
NaHCO.sub.3 (aq) (30 ml), water (2.times.25 ml), brine (20 ml),
dried (MgSO.sub.4), filtered and concentrated in-vacuo. The crude
residue was purified by flash column chromatography (10 g SNAP
cartridge, Isolera, 0-29% MeOH:CH.sub.2Cl.sub.2) to give the title
compound (68 mg, 52%) as a beige solid. LC-MS 99%, m/z=491.1,
493.1; .sup.1H NMR (500 MHz, Methanol-d.sub.4) .delta. 7.17 (d,
J=1.9 Hz, 1H), 7.02 (d, J=2.0 Hz, 1H), 4.59 (s, 1H), 4.44 (s, 2H),
3.06 (q, J=7.0 Hz, 2H), 2.69 (tt, J=12.1, 3.2 Hz, 1H), 2.41-2.29
(m, 10H), 2.24 (d, J=2.8 Hz, 3H), 2.20 (s, 3H), 1.91 (dd, J=27.7,
12.2 Hz, 4H), 1.44 (q, J=12.3, 11.1 Hz, 2H), 1.30-1.20 (m, 2H),
0.83 (t, J=7.0 Hz, 3H). One proton assumed to be coincident with a
solvent peak.
Compound 347:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}benzamide and
Compound 348:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-et-
hyl-3-{ethyl[(1s,4s)-4-(dimethylamino)cyclohexyl]amino}benzamide
##STR00830##
[1759] Step 1: Synthesis of methyl
5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate
##STR00831##
[1761] To a stirred solution of methyl 2-bromo-5-chlorobenzoate
(14.8 g, 59 mmol) in triethylamine (124 ml, 890 mmol) was added
copper iodide (339 mg, 1.8 mmol) and triphenylphosphine (778 mg,
3.0 mmol) at room temperature, under nitrogen. This mixture was
purged with nitrogen before the addition of
ethynyl(trimethyl)silane (12.5 ml, 89 mmol) and Pd(OAc).sub.2 (266
mg, 1.2 mmol). The reaction mixture was stirred at 50.degree. C.
for 20 hours and concentrated in-vacuo. The residue was dissolved
in deionized water (50 ml) and EtOAc (50 ml) and filtered through
Celite. The filter cake was washed with EtOAc (50 ml) before the
phases were separated and the aqueous layer was extracted with
EtOAc (2.times.50 ml). The combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography (10 g Silica Isolute
cartridge, 1-15% EtOAc:Heptanes) to give the title compound (16.2
g, 93%) as an orange solid. LC-MS 91%, m/z=267.4, 268.9; .sup.1H
NMR (500 MHz, Chloroform-d) .delta. ppm 7.90 (d, J=2.21 Hz, 1H)
7.52 (d, J=8.35 Hz, 1H) 7.42 (dd, J=8.28, 2.29 Hz, 1H) 3.93 (s, 3H)
0.28 (s, 9H).
Step 2: Synthesis of methyl 5-chloro-2-ethynylbenzoate
##STR00832##
[1763] A suspension of potassium carbonate (5.0 g, 36 mmol) in
methanol (20 ml) was stirred under nitrogen whilst adding a
solution of methyl 5-chloro-2-[2-(trimethylsilyl)ethynyl]benzoate
(4.8 g, 18.0 mmol) in methanol (50 ml). The resulting mixture was
stirred at 20.degree. C. for 2 hours. The reaction mixture was
filtered, and the solid washed with methanol. The filtrate was
concentrated in-vacuo and then re-dissolved in methanol (50 ml) and
carefully treated with acetyl chloride (4.0 ml). The reaction
mixture was stirred overnight and concentrated in-vacuo. TBME (50
ml) was added to the residue and the resulting suspension filtered.
The filtrate was washed with a saturated solution of NaHCO.sub.3
(40 ml). The layers were separated and the aqueous phase was
extracted with TBME (3.times.40 ml). The combined organics were
dried over MgSO.sub.4, filtered and concentrated in-vacuo. The
crude residue was purified by flash column chromatography (50 g
Isolute cartridge, 0-10% TBME:Heptanes) to give the title compound
(1.2 g, 33%) as a white solid. LC-MS 93%, m/z=194.9, 196.9; .sup.1H
NMR (500 MHz, Chloroform-d) .delta. ppm 7.94 (s, 1H), 7.56 (d,
J=8.4 Hz, 1H), 7.46 (d, J=8.4 Hz, 1H), 3.95 (s, 3H), 3.44 (s,
1H).
Step 3: Synthesis of methyl 5-chloro-2-ethylbenzoate
##STR00833##
[1765] To a solution of methyl 5-chloro-2-ethynylbenzoate (1.13 g,
5.8 mmol) in TBME (50 ml) was added Pd/C (10%) (50% water, 0.61 g,
0.29 mmol). The reaction mixture was stirred under a hydrogen
atmosphere for 130 minutes. The mixture was filtered through Celite
and the filter cake was washed with TBME. The filtrate was
concentrated in-vacuo to give the title compound (1.08 g, 74%) as a
pale brown oil. LC-MS 79%, m/z=198.9/200.9; .sup.1H NMR (500 MHz,
Chloroform-d) .delta. ppm 7.85 (s, 1H) 7.40 (d, J=8.20 Hz, 1H) 7.22
(d, J=8.20 Hz, 1H) 3.91 (s, 3H) 2.95 (q, J=7.46 Hz, 2H) 1.22 (t,
J=7.49 Hz, 3H).
Step 4: Synthesis of methyl 5-chloro-2-ethyl-3-nitrobenzoate
##STR00834##
[1767] A solution of methyl 5-chloro-2-ethylbenzoate (1.08 g, 5.4
mmol) in concentrated sulphuric acid (7.0 ml) was cooled to
-5.degree. C. in an acetone/ice bath. A mixture of 70% nitric acid
(0.45 ml, 7.1 mmol) and concentrated sulphuric acid (0.5 ml) was
added dropwise to the reaction mixture at -5.degree. C. over 15
minutes. The resulting pale yellow reaction mixture was stirred at
-5.degree. C. for 1 hour before being poured onto ice (100 ml) and
extracted with CH.sub.2Cl.sub.2 (3.times.20 ml). The combined
organic phases were washed with deionized water (20 ml), brine (20
ml), dried over Na.sub.2SO.sub.4, filtered and concentrated
in-vacuo. The resulting oil was added to a solution containing
methanol (25 ml) and thionyl chloride (0.75 ml) at 0.degree. C.
Following addition, the mixture was heated under gentle reflux for
6 hours, before cooling to room temperature and concentrating
in-vacuo to give the title compound (1.2 g) as an orange oil which
was used without further purification in the next step.
Step 5: Synthesis of methyl 3-amino-5-chloro-2-ethylbenzoate
##STR00835##
[1769] To a solution of methyl 5-chloro-2-ethyl-3-nitrobenzoate
(1.2 g, 5.0 mmol) in methanol (50 ml) and deionised water (25 ml),
was added ammonium chloride (2.6 g, 50 mmol). The mixture was
heated to 70.degree. C. before the addition of iron (1.7 g, 30
mmol). The reaction mixture was stirred at 70.degree. C. for 3.5
hours, hot filtered through Celite and the filter pad was washed
with MeOH (20 ml). The filtrate was concentrated in-vacuo and the
aqueous residue was dissolved in a saturated solution of
NaHCO.sub.3(aq) (50 ml) and EtOAc (100 ml). The phases were
separated and the organic phase was washed with saturated
NaHCO.sub.3(aq) (2.times.65 ml), brine (30 ml), dried over
MgSO.sub.4, filtered and concentrated in-vacuo. The residue was
purified by column chromatography (25 g SNAP cartridge, Isolera,
0-11% ethyl acetate/heptanes) to give the title compound (372 mg,
33%) as a yellow oil. LC-MS 94%, m/z=213.9, 215.9; .sup.1H NMR (500
MHz, Chloroform-d) d ppm 7.18 (d, J=2.21 Hz, 1H) 6.80 (d, J=2.05
Hz, 1H) 3.88 (s, 3H) 3.85 (d, J=2.52 Hz, 2H) 2.75 (q, J=7.57 Hz,
2H) 1.21 (t, J=7.49 Hz, 3H).
Step 6: Synthesis of methyl
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)amino]-5-chloro-2-ethylben-
zoate
##STR00836##
[1771] To a stirred solution of methyl
3-amino-5-chloro-2-ethylbenzoate (365 mg, 1.7 mmol) in
1,2-dichloroethane (20 ml) under nitrogen, was added tert-butyl
(4-oxocyclohexyl)carbamate (364 mg, 1.7 mmol) followed by acetic
acid (587 .mu.l, 10.3 mmol). The solution was stirred for 10
minutes before the portionwise addition of sodium
triacetoxyborohydride (1.09 g, 5.1 mmol) over 1 hour. The resulting
solution was stirred at room temperature for 17 hours, before
treating with tert-butyl (4-oxocyclohexyl)carbamate (364 mg, 1.7
mmol). The reaction mixture was stirred for 30 minutes and then
treated with sodium triacetoxyborohydride (1.09 g, 5.1 mmol) over
2.5 hours. The reaction mixture was stirred at room temperature for
a further 26 hours before addition of deionized water (40 ml). The
mixture was neutralised with solid NaHCO.sub.3 (2.95 g) and was
extracted with ethyl acetate (3.times.50 ml). The combined organic
extracts were washed with brine (35 ml), dried (MgSO.sub.4),
filtered and concentrated in-vacuo. The crude residue was purified
by column chromatography (25 g SNAP cartridge, Isolera, 0-18%
EtOAc/Heptanes) to give as an oil the title compound (623 mg, 85%)
as a mixture of cis/trans isomers. LC-MS 46.1%, 2.61 min (3.5
minute LC-MS method), m/z=411.1, 413.1 and 50.2%, 2.66 min (3.5
minute LC-MS method), m/z=411.1, 413.1; 1H NMR (500 MHz,
Chloroform-d) d ppm 7.05 (dd, J=7.09, 2.05 Hz, 1H) 6.68 (t, J=2.36
Hz, 1H) 4.36-4.63 (m, 1H) 3.88 (d, J=3.31 Hz, 3H) 3.62-3.78 (m, 1H)
3.49 (br. s., 1H) 3.23 (br. s., 1H) 2.63-2.76 (m, 2H) 2.07-2.18 (m,
2H) 1.77-1.89 (m, 2H) 1.69 (br. s., 1H) 1.46 (s, 9H) 1.27 (m, 3H)
1.13-1.24 (m, 3H).
Step 7: Synthesis of methyl
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)(ethyl)amino]-5-chloro-2-e-
thylbenzoate
##STR00837##
[1773] To a stirred solution of methyl
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)amino]-5-chloro-2-ethylben-
zoate (200 mg, 0.49 mmol) in dry 1,2-dichloroethane (5.0 ml), was
added acetaldehyde (54 .mu.l, 1.0 mmol), followed by acetic acid
(167 .mu.l, 2.9 mmol). The reaction mixture was stirred for 50
minutes, sodium triacetoxyborohydride (309 mg, 1.5 mmol) was added
over 2 hours and the reaction was stirred for 20 hours. The
reaction mixture was treated with acetaldehyde (54 .mu.l, 1.0 mmol)
stirred for 1 hour and sodium triacetoxyborohydride (309 mg, 1.5
mmol) was added portionwise over 2 hours. The reaction mixture was
stirred for a further 22 hours and then treated with acetaldehyde
(136 .mu.l, 2.4 mmol) and stirred for 1 hour, followed by sodium
triacetoxyborohydride (516 mg, 2.4 mmol) over 1.5 hours and stirred
for 22 hours. The reaction mixture was treated with acetaldehyde
(109 .mu.l, 2.0 mmol) and stirred for 1.5 hours before sodium
triacetoxyborohydride (413 mg, 2.0 mmol) was added over 2 hours,
along with dichloroethane (5.0 ml) to aid stirring. The reaction
mixture was stirred for 19 hours before the addition of
acetaldehyde (82 .mu.l, 1.5 mmol), stirred for 1.5 hours, followed
by sodium triacetoxyborohydride (309 mg, 1.5 mmol) over 45 minutes.
The reaction mixture was stirred for a further 70 hours before
being diluted with water (20 ml) and the pH was adjusted to 8 by
the gradual addition of solid NaHCO.sub.3 (2.53 g). The layers were
separated and the aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.20 ml). The combined organic phases were washed with brine
(60 ml), dried (MgSO.sub.4), filtered and concentrated in-vacuo.
The crude residue was purified by flash column chromatography (10 g
SNAP cartridge, Isolera, 0-30% EtOAc/Heptanes) to give the title
compound (163 mg, 73%) as a yellow oil and a mixture of cis/trans
isomers. LC-MS 100%, 5.83 min (7 minute LC-MS method), m/z=439.2,
441.2; .sup.1H NMR (500 MHz, Chloroform-d) .delta. ppm 7.45-7.55
(m, 1H) 7.22 (dd, J=4.33, 2.13 Hz, 1H) 4.25-4.67 (m, 1H) 3.90 (d,
J=2.21 Hz, 3H) 3.29-3.76 (m, 1H) 2.95-3.11 (m, 4H) 2.58-2.96 (m,
1H) 1.62-2.03 (m, 4H) 1.38-1.53 (m, 11H) 1.01-1.14 (m, 4H)
0.81-0.94 (m, 4H).
Step 8: Synthesis of
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)(ethyl)amino]-5-chloro-2-e-
thylbenzoic acid
##STR00838##
[1775] To a stirred solution of methyl
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)(ethyl)amino]-5-chloro-2-e-
thylbenzoate (156 mg, 0.36 mmol) in THF (4.0 ml) and MeOH (0.29
ml), was added a solution of 4M NaOH (aq) (2.7 ml, 10.7 mmol). The
resulting solution was stirred at 50.degree. C. for 17 hours. The
reaction mixture was treated with 4M NaOH (aq) (0.5 ml, 2.0 mmol)
and stirring was continued at 50.degree. C. for 32 hours. The heat
was removed and the reaction mixture was left to stir at room
temperature for 65 hours. The reaction mixture was treated with 4M
NaOH (aq) (1.0 ml, 4.0 mmol) and heated back up to 50.degree. C.
for 24 hours. The organic solvents were removed in-vacuo and the
aqueous phase was treated with a solution of 0.5M citric acid (20
ml) to adjust the pH to 4/5. The product was extracted with ethyl
acetate (3.times.25 ml). The combined organic phases were washed
with brine (30 ml), dried (MgSO.sub.4), filtered and concentrated
in-vacuo and then further dried on the high vac line for 4 hours to
give the title compound (142 mg, 92%) as a white solid and a
mixture of cis/trans isomers. LC-MS 98%, 5.49 min (7 minute LC-MS
method), m/z=425.1, 427.1. .sup.1H NMR shows a mixture of cis/trans
isomers.
Step 9: Synthesis of tert-butyl
N-{4-[(5-chloro-3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]ca-
rbamoyl}-2-ethylphenyl)(ethyl)amino]cyclohexyl} carbamate
##STR00839##
[1777] A stirred solution of
3-[(4-{[(tert-butoxy)carbonyl]amino}cyclohexyl)(ethyl)amino]-5-chloro-2-e-
thylbenzoic acid (139 mg, 0.33 mmol) in anhydrous DIPEA (2.0 ml) at
0.degree. C. under nitrogen, was treated with HATU (149 mg, 0.39
mmol) and DIPEA (114 .mu.l, 0.65 mmol). The resulting solution was
stirred for 10 minutes and then treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 61 mg,
0.36 mmol). The resulting suspension was stirred at 0.degree. C.
for 1 hour and then stirred at room temperature for 98.5 hours. The
reaction mixture was treated with HATU (37 mg, 0.1 mmol), stirred
for 1 hour and then
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 11 mg,
0.07 mmol). The reaction mixture was stirred at room temperature
for a further 23 hours and then partitioned between water (20 ml)
and CH.sub.2Cl.sub.2 (20 ml). The layers were separated and the
aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.10 ml).
The combined organics were washed with a saturated solution of
NaHCO.sub.3 (aq) (40 ml), water (20 ml), brine (2.times.20 ml),
dried (MgSO.sub.4), filtered and concentrated in-vacuo. The crude
residue was purified by flash column chromatography (10 g SNAP
cartridge, Isolera, 0-13% MeOH/CH.sub.2Cl.sub.2) and triturated
from ether/heptane (2.0 ml) with sonication and the liquor
decanted. The remaining solid was dried under high vacuum to give
the title compound (101 mg, 52%) as a beige solid. LC-MS 93%, 4.54
min (7 minute LC-MS method), m/z=559.2, 561.2. .sup.1H NMR shows a
mixture of cis/trans isomers.
Step 10: Synthesis of
3-[(4-aminocyclohexyl)(ethyl)amino]-5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-d-
ihydropyridin-3-yl)methyl]-2-ethylbenzamide
##STR00840##
[1779] To a stirred solution of tert-butyl
N-{4-[(5-chloro-3-{[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]ca-
rbamoyl}-2-ethylphenyl)(ethyl)amino]cyclohexyl} carbamate (100 mg,
0.18 mmol) in CH.sub.2Cl.sub.2 (4.0 ml) at 0.degree. C., was added
trifluoroacetic acid (1.0 ml). The reaction mixture was stirred at
0.degree. C. for 40 minutes and then stirred for 2 hours at room
temperature. The reaction mixture was concentrated in-vacuo and the
residue was basified with a saturated solution of NaHCO.sub.3 (aq)
(10 ml) until the pH was 8. The aqueous residue was extracted with
20% MeOH/CH.sub.2Cl.sub.2 (4.times.15 ml). The combined organic
layers were dried (MgSO.sub.4), filtered and concentrated in-vacuo
to give the title compound (108 mg, 113%) as a yellow solid. LC-MS
48%, 2.88 min (7 minute LC-MS method), m/z=459.1 and 39%, 2.96 min
(7 minute LC-MS method), m/z=459.1. .sup.1H NMR shows a mixture of
cis/trans isomers.
Step 11: Synthesis of
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}benzamide and
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-{ethyl [(1s,4s)-4-(dimethylamino)cyclohexyl]amino}benzamide
##STR00841##
[1781] A stirred solution of
3-[(4-aminocyclohexyl)(ethyl)amino]-5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-d-
ihydropyridin-3-yl)methyl]-2-ethylbenzamide (80 mg, 0.18 mmol) in
1,2-dichloroethane (3.0 ml), was treated with acetic acid (60
.mu.l, 1.0 mmol) and paraformaldehyde (31 mg, 1.0 mmol). The
reaction mixture was stirred for 35 minutes under nitrogen before
the addition of sodium triacetoxyborohydride (222 mg, 1.0 mmol).
The reaction mixture was stirred for 16.5 hours before
paraformaldehyde (16 mg, 0.52 mmol) was added and stirring was
continued for 1 hour prior to the addition of sodium
triacetoxyborohydride (111 mg, 0.52 mmol). The reaction mixture was
stirred for a further 6 hours and then treated with
paraformaldehyde (10 mg, 0.35 mmol), stirred for 10 minutes before
the addition of sodium triacetoxyborohydride (74 mg, 0.35 mmol) and
1,2-dichloroethane (1.0 ml) and stirred for a further 89 hours. The
reaction mixture was diluted with water (20 ml) and basified to pH
8 by the addition of solid NaHCO.sub.3 (1.45 g). CH.sub.2Cl.sub.2
(20 ml) was added and the layers were separated. The aqueous phase
was extracted with CH.sub.2Cl.sub.2 (3.times.10 ml). The combined
organic phases were washed with brine (30 ml), dried (MgSO.sub.4),
filtered and concentrated in-vacuo. The crude residue was purified
by preparative-HPLC (MeCN/water+0.2% ammonium hydroxide) to give
pure trans isomer title compound 347:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}benzamide (22 mg,
26%) as a white solid and pure cis isomer title compound 348:
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3-
-{ethyl[(1s,4s)-4-(dimethylamino)cyclohexyl]amino}benzamide-(18 mg,
21%) as a white solid.
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-ethyl-3--
{ethyl[(1r,4r)-4-(dimethylamino)cyclohexyl]amino}benzamide
[1782] LC-MS 100%, 3.07 min (7 minute LC-MS method), m/z=487.2,
489.2; .sup.1H NMR (500 MHz, Chloroform-d) .delta. 11.09 (s, 1H),
7.11-7.04 (m, 2H), 7.00 (d, J=2.1 Hz, 1H), 5.94 (s, 1H), 4.51 (d,
J=5.9 Hz, 2H), 3.00 (q, J=7.0 Hz, 2H), 2.79 (q, J=7.4 Hz, 2H), 2.65
(t, J=11.5 Hz, 1H), 2.38 (s, 3H), 2.24 (s, 6H), 2.22 (s, 3H), 2.14
(t, J=11.4 Hz, 1H), 1.87 (d, J=10.3 Hz, 4H), 1.37 (q, J=12.1 Hz,
2H), 1.17 (q, J=11.5 Hz, 2H), 1.02 (t, J=7.4 Hz, 3H), 0.85 (t,
J=7.0 Hz, 3H).
[1783]
5-chloro-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-e-
thyl-3-{ethyl[(1s,4s)-4-(dimethylamino)cyclohexyl]amino}benzamide
LC-MS 100%, 3.13 min (7 minute LC-MS method), m/z=487.2, 489.2;
.sup.1H NMR (500 MHz, chloroform-d) .delta. 10.72 (s, 1H), 7.09 (d,
J=2.0 Hz, 2H), 7.01 (d, J=2.1 Hz, 1H), 5.93 (s, 1H), 4.51 (d, J=5.9
Hz, 2H), 3.01 (q, J=6.7 Hz, 3H), 2.89 (q, J=7.4 Hz, 2H), 2.39 (s,
3H), 2.23 (s, 9H), 2.06 (s, 1H), 1.87-1.72 (m, 4H), 1.43 (td,
J=8.3, 3.8 Hz, 2H), 1.33 (dd, J=12.6, 9.2 Hz, 2H), 1.01 (t, J=7.5
Hz, 3H), 0.84 (t, J=7.0 Hz, 3H).
Compound 92: 3-(allyl (tetrahydro-2H-pyran-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methylbenzamide
##STR00842##
[1784] Step 1: Synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR00843##
[1786] 5-chloro-2-methylbenzoic acid (4.0 g, 23 mmol) was added
portionwise to cooled conc. H.sub.2SO.sub.4 (27 mL) at -10.degree.
C. After 10 minutes a nitrating mixture {prepared by mixing conc.
HNO3 (3.3 g, 52.68 mmol) with conc. H.sub.2SO.sub.4 (4.4 mL)} was
added dropwise at -10.degree. C. The resulting reaction mass was
stirred at -10.degree. C. for 30 minutes. On completion, the
reaction mixture was poured on ice cold water, the solid
precipitate was filtered, washed with water and dried under vacuum
giving the title compound (4.95 g, 99%).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR00844##
[1788] To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic
acid (6.75 g, 31.3 mmol) in DMF (33 mL), sodium carbonate (13.2 g,
125 mmol) and methyl iodide (17.8 g, 125 mmol) were added. The
mixture was heated at 60.degree. C. for 4 h. On completion, water
was added to the reaction mass and extraction was carried out using
DCM. The combined organic layers were dried, concentrated under
reduced pressure and purified by column chromatography over silica
gel giving the title compound (6.0 g, 83%).
Step 3: Synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR00845##
[1790] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6.0 g, 26 mmol) in ethanol (60
mL), ammonium chloride (6.0 g, 110 mmol) dissolved in water (60 mL)
and iron powder (11.9 g, 208 mmol) were added under stirring. The
resulting reaction mass was heated at 80.degree. C. for 1 h. On
completion, water was added to reaction mass and reaction mixture
was filtered through celite, the filtrate was extracted with ethyl
acetate. The combined organic layers were washed with water, dried,
and concentrated under reduced pressure giving the title compound
which was used as is.
Step 4: Synthesis of methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate
##STR00846##
[1792] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (5.0 g, 19 mmol) and dihydro-2H-pyran-4(3H)-one (2.86 g,
28.6 mmol) in methanol (50 mL), acetic acid (2.3 g, 38 mmol) was
added and reaction stirred at room temperature for 8 h. Then sodium
cyanoborohydride (3.0 g, 48 mmol) was added at 0.degree. C. and
reaction stirred overnight at room temperature. On completion, the
solvent was removed under reduced pressure and crude material was
purified by column chromatography to afford the title compound (3.0
g, 42%).
Step 5: Synthesis of methyl 3-(allyl (tetrahydro-2H-pyran-4-yl)
amino)-5-chloro-2-methylbenzoate
##STR00847##
[1794] To a stirred solution of methyl
5-chloro-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate
(1.0 g, 3.3 mmol) in DMF (10 mL), NaH (0.25 g, 10 mmol) was added
at 0.degree. C. and the mixture stirred for 20 minutes.
3-bromoprop-1-ene (1.7 g, 14.13 mmol) was added and the mixture was
heated at 80.degree. C. for 15 h. On completion, the reaction was
quenched with ice cold water and extraction was carried out using
DCM. The combined organic layers were dried, concentrated and
resulting crude was purified by column chromatography giving the
title compound (0.50 g, 44%).
Step 6: Synthesis of 3-(allyl (tetrahydro-2H-pyran-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methylbenzamide
##STR00848##
[1796] Aqueous NaOH (0.092 g, 2.32 mmol) was added to a solution of
methyl 3-(allyl (tetrahydro-2H-pyran-4-yl)
amino)-5-chloro-2-methylbenzoate (0.50 g, 1.54 mmol) in ethanol (15
mL) and the mixture stirred at 60.degree. C. for 1 h. After
completion of the reaction, ethanol was removed under reduced
pressure and acidified using dilute HCl up to pH 6 and pH 4 was
adjusted using citric acid. Extraction was carried out using DCM.
The combined organic layers were dried and concentrated giving
respective acid (0.47 g, 98%).
[1797] The above acid (0.47 g, 1.52 mmol) was then dissolved in
DMSO (5 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one
(0.462 g, 3.04 mmol) was added. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (1.18 g, 2.28 mmol) was
added to it and stirring was continued overnight. After completion
of the reaction, reaction mass was poured into ice to obtain a
solid which was filtered and dried to afford the title compound
(0.40 g, 59%). LCMS: 444.25 (M+1).sup.+; HPLC: 96.85% (@254 nm)
(R.sub.t; 6.310; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.47 (s, 1H), 8.22 (t, 1H), 7.16 (s, 1H), 6.93 (s, 1H), 5.85 (s,
1H), 5.56-5.62 (m, 1H), 5.02-5.07 (m, 1H), 4.94-4.96 (m, 1H), 4.23
(d, 2H, J=3.6 Hz), 3.83 (d, 2H, J=9.6 Hz), 3.64 (d, 2H, J=4 Hz),
3.23 (t, 2H, J=10 Hz), 2.97 (m, 1H), 2.18 (s, 3H), 2.17 (s, 3H),
2.10 (s, 3H), 1.54-1.60 (m, 4H).
Compound 98:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(propyl(tetrahydro-2H-pyran-4-yl)amino)benzamide
##STR00849##
[1799] To a stirred solution of 3-(allyl (tetrahydro-2H-pyran-4-yl)
amino)-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2--
methylbenzamide (0.10 g) in MeOH (10 mL) was added 10% Pd/C (0.03
g) and the reaction stirred at room temperature under hydrogen
(balloon pressure) for 2 h. On completion, the reaction mixture was
filtered through celite and the filtrate was concentrated under
reduced pressure to obtain crude solid which was purified by column
chromatography to afford the title compound (0.02 g, 20%). LCMS:
445.25 (M+1).sup.+; HPLC: 90.40% (@254 nm) (R.sub.t; 5.609; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.23 (t,
1H), 7.19 (s, 1H), 6.93 (d, 1H, J=1.6 Hz), 5.88 (s, 1H), 4.24 (d,
2H, J=4.4 Hz), 3.83 (d, 2H, J=10.4 Hz), 3.22-3.25 (m, 2H), 2.94 (t,
2H, J=7.2 Hz), 2.88-2.90 (m, 1H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10
(s, 3H), 1.56 (m, 4H), 1.14-1.23 (m, 2H), 0.75 (t, 3H, J=8 Hz).
Compound 104:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isobut-
yl(methyl)amino)-2-methylbenzamide
##STR00850##
[1800] Step 1: Synthesis of methyl 5-chloro-3-(isobutyl
amino)-2-methylbenzoate
##STR00851##
[1802] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (5.0 g, 25 mmol) and isobutyraldehyde (4.5 g, 62 mmol) in
methanol (50 mL), acetic acid (3.0 g, 50 mmol) was added and
reaction stirred at room temperature for 2 h. Then sodium
cyanoborohydride (3.94 g, 62 mmol) was added at 0.degree. C. and
reaction stirred overnight at room temperature. On completion, the
solvent was removed under reduced pressure and the crude material
was purified by column chromatography to afford the title compound
(6.0 g, 94%).
Step 2: Synthesis of methyl 5-chloro-3-(isobutyl (methyl)
amino)-2-methylbenzoate
##STR00852##
[1804] To a stirred solution of methyl 5-chloro-3-(isobutyl
amino)-2-methylbenzoate (1.0 g, 3.9 mmol) in dry acetonitrile (10
mL), cesium carbonate (2.55 g, 7.8 mmol) and methyl iodide (5.55 g,
39 mmol) were added. The mixture was heated at 80.degree. C. for 12
h. On completion, the mixture was cooled to room temperature and
filtered. The residue was washed with ethyl acetate and the
filtrate was concentrated to the title compound (1.0 g, 95%).
Step 3: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(isobut-
yl(methyl)amino)-2-methylbenzamide
##STR00853##
[1806] Aqueous NaOH (0.22 g, 5.5 mmol) was added to a solution of
methyl 5-chloro-3-(isobutyl (methyl) amino)-2-methylbenzoate (1.0
g, 3.7 mmol) in EtOH (5 mL) and the mixture stirred at 60.degree.
C. for 1 h. After completion of the reaction, ethanol was removed
under reduced pressure. The residue was acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. The combined organic layers were
dried and concentrated giving the respective acid (0.900 g, 95%).
The acid (0.30 g, 1.17 mmol) was then dissolved in DMSO (1.5 mL)
and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.357 g, 2.34
mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.915 g, 1.76 mmol) was added
to it and stirring was continued overnight. After completion of the
reaction, the reaction mass was poured into ice to obtain a solid,
this was filtered and purified by column chromatography to afford
the title compound (0.245 g, 53%). LCMS: 390.25 (M+1).sup.+; HPLC:
97.59% (@254 nm) (R.sub.t; 6.063; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.20 (t, 1H), 7.07
(s, 1H), 6.90 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4.4 Hz), 2.63
(d, 2H, J=7.2 Hz), 2.54 (s, 3H), 2.18 (s, 3H), 2.16 (s, 3H), 2.10
(s, 3H), 1.75-1.78 (m, 1H), 0.85 (d, 6H, J=6 Hz).
Compound 105:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-3-(ethyl(isobutyl)amino)-2-methylbenzamide
##STR00854##
[1807] Step 1: Synthesis of methyl 5-chloro-3-(ethyl (isobutyl)
amino)-2-methylbenzoate
##STR00855##
[1809] To a stirred solution of methyl 5-chloro-3-(isobutyl
amino)-2-methylbenzoate (1.0 g, 3.9 mmol) in dry DMF (10 mL),
cesium carbonate (2.55 g, 7.82 mmol) and ethyl iodide (6.09 g, 39.1
mmol) were added. The resulting reaction mass was heated at
80.degree. C. for 12 h. On completion, the reaction mass was cooled
to room temperature and filtered. The residue was washed with ethyl
acetate and the filtrate was concentrated to give crude material
which then used for next step as is (1.0 g, 91%).
Step 2: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-3-(ethyl (isobutyl) amino)-2-methylbenzamide
##STR00856##
[1811] Aqueous NaOH (0.212 g, 5.28 mmol) was added to a solution of
compound 7 (1.0 g, 3.50 mmol) in EtOH (5 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. The combined organic layers were
dried concentrated giving the respective acid (0.900 g).
[1812] The acid (0.90 g, 3.33 mmol) was then dissolved in DMSO (4.5
mL) and 3-(amino methyl)-4,6-dimethylpyridin-2(1H)-one (1.01 g,
6.67 mmol) was added to it. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (2.60 g, 4.9 mmol) was
added and stirring was continued for overnight. The mixture was
poured into ice to give a solid, this was filtered and purified by
acetonitrile and diethyl ether washings to afford the title
compound (1.0 g, 75%). LCMS: 404.25 (M+1).sup.+; HPLC: 90.28% (@254
nm) (R.sub.t; 6.063; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (s, 1H), 8.19 (t, 1H), 7.10 (s, 1H), 6.90 (s, 1H), 5.85 (s,
1H), 4.24 (d, 2H, J=4.8 Hz), 2.86 (q, 2H, J=7.2 Hz), 2.73 (d, 2H,
J=7.6 Hz), 2.18 (s, 3H), 2.16 (s, 3H), 2.10 (s, 3H), 1.57-1.61 (m,
1H), 0.91 (t, 3H, J=6.8 Hz), 0.82 (d, 6H, J=6.4 Hz).
Compound 117: 5-chloro-3-((cyclopentylmethyl)(methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00857##
[1813] Step 1: Synthesis of methyl 5-chloro-3-((cyclopentylmethyl)
amino)-2-methylbenzoate
##STR00858##
[1815] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (3.0 g, 15 mmol) and cyclopentanecarbaldehyde (2.2 g, 22
mmol) in methanol (30 mL), acetic acid (1.8 g, 30 mmol) was added
and reaction stirred at room temperature for 8 h. Then sodium
cyanoborohydride (2.4 g, 37 mmol) was added at 0.degree. C. and the
reaction stirred overnight at room temperature. On completion, the
solvent was removed under reduced pressure and the crude material
was purified by column chromatography to afford the title compound
(4.2 g, 99%).
Step 2: Synthesis of methyl 5-chloro-3-((cyclopentylmethyl)
(methyl) amino)-2-methylbenzoate
##STR00859##
[1817] To a stirred solution of methyl
5-chloro-3-((cyclopentylmethyl) amino)-2-methylbenzoate (1.0 g, 3.5
mmol) in acetonitrile (10 mL), cesium carbonate (2.3 g, 7.1 mmol)
and methyl iodide (5.0 g, 35 mmol) were added. The mixture was
heated at 80.degree. C. for 12 h. cooled to room temperature and
filtered. The residue was washed with ethyl acetate and the
filtrate was concentrated and then purified by column
chromatography to afford the title compound (1.0 g, 95%).
Step 3: Synthesis of 5-chloro-3-((cyclopentylmethyl) (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00860##
[1819] Aqueous NaOH (0.206 g, 15.1 mmol) was added to a solution of
methyl 5-chloro-3-((cyclopentylmethyl) (methyl)
amino)-2-methylbenzoate (1.0 g, 3.38 mmol) in ethanol (10 mL) and
the mixture stirred at 60.degree. C. for 1 h. The ethanol was
removed under reduced pressure and the residue acidified using
dilute HCl up to pH 6 and pH 4 was adjusted using citric acid.
Extraction was carried out using DCM. The combined organic layers
were dried and concentrated giving the respective acid (0.9 g) The
above acid (0.3 g, 1.06 mmol) was then dissolved in DMSO (1.5 mL)
and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.324 g, 2.12
mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.831 g, 1.57 mmol) was added
and stirring was continued overnight. The mixture was poured into
ice to obtain a solid, this was filtered and washed with
acetonitrile to provide the title compound (0.25 g, 56%). LCMS:
416.15 (M+1).sup.+; HPLC: 92.58% (@254 nm) (R.sub.t; 6.169; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.19 (t,
1H), 7.08 (s, 1H), 6.89 (s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4.4
Hz), 2.77 (d, 2H, J=7.2 Hz), 2.57 (s, 3H), 2.18 (s, 3H), 2.15 (s,
3H), 2.10 (s, 3H), 2.04-2.06 (m, 1H), 1.63-1.65 (m, 2H), 1.49-1.52
(m, 2H), 1.45-1.46 (m, 2H), 1.15-1.18 (m, 2H).
Compound 118: 5-chloro-3-((cyclopentylmethyl) (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00861##
[1820] Step 1: Synthesis of methyl 5-chloro-3-((cyclopentylmethyl)
(ethyl) amino)-2-methylbenzoate
##STR00862##
[1822] To a stirred solution of methyl
5-chloro-3-((cyclopentylmethyl) amino)-2-methylbenzoate (1.0 g,
3.54 mmol) in DMF (10 mL), cesium carbonate (2.31 g, 7.09 mmol) and
ethyl iodide (5.53 g, 35.4 mmol) were added. The resulting reaction
mass was heated at 80.degree. C. for 12 h. The mixture was cooled
to room temperature and filtered. The residue was washed with ethyl
acetate and the filtrate was concentrated to a residue which was
purified by column chromatography to afford the title compound (1.0
g, 91%).
Step 2: Synthesis of 5-chloro-3-((cyclopentylmethyl) (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00863##
[1824] Aqueous NaOH (0.193 g, 15.1 mmol) was added to a solution of
methyl 5-chloro-3-((cyclopentylmethyl) (ethyl)
amino)-2-methylbenzoate (1.0 g, 3.22 mmol) in ethanol (10 mL) and
the mixture stirred at 60.degree. C. for 1 h. After completion of
the reaction, ethanol was removed under reduced pressure and the
residue acidified using dilute HCl up to pH 6 and pH 4 was adjusted
using citric acid. Extraction was carried out using DCM. The
combined organic layers were dried concentrated giving respective
acid (0.9 g).
[1825] The above acid (0.3 g, 1.01 mmol) was then dissolved in DMSO
(1.5 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.308
g, 2.12 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.791 g, 1.52 mmol) was added
and stirring was continued overnight. After completion of the
reaction, the reaction mass was poured into ice to obtain a solid,
this was filtered and washed with acetonitrile to provide the title
compound (0.25 g, 57%). LCMS: 430.2 (M+1).sup.+; HPLC: 91.54% (@210
nm) (R.sub.t; 9.378; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (s, 1H), 8.20 (t, 1H), 7.12 (s, 1H), 6.91 (s, 1H), 5.85 (s,
1H), 4.23 (d, 2H, J=3.6 Hz), 2.83-2.89 (m, 4H), 2.18 (s, 3H), 2.14
(s, 3H), 2.10 (s, 3H), 1.85-1.89 (m, 1H), 1.51-1.58 (m, 4H),
1.41-1.46 (m, 2H), 1.14-1.15 (m 2H), 0.91 (t, 3H, J=8 Hz).
Compound 123: 5-chloro-3-(cyclobutyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00864##
[1826] Step 1: Synthesis of methyl
5-chloro-3-(cyclobutylamino)-2-methylbenzoate
##STR00865##
[1828] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (2.0 g, 10 mmol) and
cyclobutanone (1.4 g, 20 mmol) in dichloroethane (20 mL), acetic
acid (3.6 g, 60 mmol) was added and reaction stirred at room
temperature for 10 minutes. Then sodium triacetoxyborohydride (6.3
g, 30 mmol) was added at 0.degree. C. and the reaction stirred for
3 h at room temperature. On completion, the solvent was removed
under reduced pressure and the crude material was purified by
column chromatography to afford the title compound (1.4 g,
56%).
Step 2: Synthesis of methyl 5-chloro-3-(cyclobutyl (methyl)
amino)-2-methylbenzoate
##STR00866##
[1830] To a stirred solution of methyl
5-chloro-3-(cyclobutylamino)-2-methylbenzoate (0.7 g, 2.8 mmol) in
acetonitrile (10 mL), cesium carbonate (2.2 g, 6.8 mmol) and methyl
iodide (3.9 g, 28 mmol) were added. The resulting reaction mass was
heated at 80.degree. C. for 8 h. On completion, the reaction mass
was cooled to room temperature and filtered. The residue was washed
with ethyl acetate and the filtrate was concentrated to give the
title compound (0.62 g, 84%).
Step 3: Synthesis of 5-chloro-3-(cyclobutyl (methyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide
##STR00867##
[1832] Aqueous NaOH (0.134 g, 3.34 mmol) was added to a solution of
methyl 5-chloro-3-(cyclobutyl (methyl) amino)-2-methylbenzoate
(0.60 g, 2.23 mmol) in ethanol (10 mL) and the mixture stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using DCM. The combined organic layers were dried
concentrated giving respective acid (0.5 g).
[1833] The above acid (0.25 g, 0.98 mmol) was then dissolved in
DMSO (5 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one
(0.22 g, 1.41 mmol) was added. The reaction mixture was stirred at
room temperature for 15 min before PYBOP (0.767 g, 1.47 mmol) was
added and stirring was continued overnight. After completion of the
reaction, the reaction mass was poured into ice and extraction was
carried out using 10% MeOH/DCM. The combined organic layers were
dried over sodium sulphate and concentrated under reduced pressure
to obtain crude material which was then purified by column
chromatography giving the title compound (0.36 g, 95%). LCMS:
388.09 (M+1).sup.+; HPLC: 96.62% (@254 nm) (R.sub.t; 5.461; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.44 (s, 1H), 8.18 (t,
1H), 6.90 (d, 2H, J=7.2 Hz), 5.85 (s, 1H), 4.24 (d, 2H, J=4.4 Hz),
3.65 (m, 1H), 2.45 (s, 3H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10 (s,
3H), 2.03 (bs, 2H), 1.76-1.81 (m, 2H), 1.61-1.62 (m, 2H).
Compound 137: 5-chloro-3-(cyclobutyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00868##
[1834] Step 1: Synthesis of methyl 5-chloro-3-(cyclobutyl (ethyl)
amino)-2-methylbenzoate
##STR00869##
[1836] To a stirred solution of methyl
5-chloro-3-(cyclobutylamino)-2-methylbenzoate (0.70 g, 2.73 mmol)
and acetaldehyde (0.36 g, 8.20 mmol) in dichloroethane (15 mL),
acetic acid (0.98 g, 16.4 mmol) was added and the reaction stirred
at room temperature for 10 minutes. Then sodium
triacetoxyborohydride (1.7 g, 8.2 mmol) was added at 0.degree. C.
and the reaction stirred for 3 h at room temperature. On
completion, the solvent was removed under reduced pressure to give
title compound (0.71 g, 91%).
Step 2: Synthesis of 5-chloro-3-(cyclobutyl (ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1, 2-dihydropyridin-3-yl)
methyl)-2-methylbenzamide
##STR00870##
[1838] Aqueous NaOH (0.149 g, 3.7 mmol) was added to a solution of
methyl 5-chloro-3-(cyclobutyl (ethyl) amino)-2-methylbenzoate (0.70
g, 2.48 mmol) in ethanol (10 mL) and the mixture stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and the residue acidified using
dilute HCl up to pH 6 and pH 4 was adjusted using citric acid.
Extraction was carried out using DCM. The combined organic layers
were dried and concentrated giving respective acid (0.6 g,).
[1839] The above acid (0.6 g, 2.2 mmol) was then dissolved in DMSO
(5 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.67 g,
4.44 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (1.7 g, 3.33 mmol) was added
and stirring was continued overnight. After completion of the
reaction, the reaction mass was poured into ice; extraction was
carried out using 10% MeOH/DCM. The combined organic layers were
dried over sodium sulphate and concentrated under reduced pressure
to obtain crude material which was then purified by solvent
washings giving the title compound (0.45 g, 50%). LCMS: 402.20
(M+1).sup.+; HPLC: 98.73% (@254 nm) (R.sub.t; 4.096; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 8.22 (t,
1H, J=4.4 Hz), 6.97 (s, 1H), 6.92 (s, 1H), 5.85 (s, 1H), 4.23 (d,
2H, J=4.4 Hz), 3.75-3.79 (m, 1H), 2.87-2.93 (m, 2H), 2.18 (s, 3H),
2.15 (s, 3H), 2.10 (s, 3H), 1.99 (m, 2H), 1.58-1.69 (m, 4H), 0.77
(t, 3H, J=6.8 Hz).
Compound 126: 5-chloro-N-((4,
6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-3-(methyl ((tetrahydro-2H-pyran-4-yl) methyl)
amino) benzamide
##STR00871##
[1840] Step 1: Synthesis of methyl
5-chloro-2-methyl-3-(((tetrahydro-2H-pyran-4-yl) methyl) amino)
benzoate
##STR00872##
[1842] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (2.0 g, 10 mmol) and tetrahydro-2H-pyran-4-carbaldehyde
(1.71 g, 15 mmol) in methanol (20 mL), acetic acid (1.2 g, 20 mmol)
was added and the reaction stirred at room temperature for 8 h.
Then sodium cyanoborohydride (2.11 g, 25.1 mmol) was added at
0.degree. C. and the reaction stirred overnight at room
temperature. On completion, the solvent was removed under reduced
pressure and the crude material was purified by column
chromatography to afford the title compound (1.8 g, 60%).
Step 2: Synthesis of methyl 5-chloro-2-methyl-3-(methyl
((tetrahydro-2H-pyran-4-yl) methyl) amino) benzoate
##STR00873##
[1844] To a stirred solution of methyl
5-chloro-2-methyl-3-(((tetrahydro-2H-pyran-4-yl) methyl) amino)
benzoate (0.80 g, 2.71 mmol) in acetonitrile (20 mL), cesium
carbonate (1.75 g, 5.42 mmol) and methyl iodide (1.93 g, 13.6 mmol)
were added. The resulting reaction mass was heated at 80.degree. C.
for 8 h. On completion, the mixture was cooled to room temperature
and filtered. The residue was washed with ethyl acetate and the
filtrate was concentrated to give crude material which was purified
by column chromatography to give the title compound (0.70 g,
84%).
Step 3: Synthesis of 5-chloro-N-((4,
6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-2-methyl-3-(methyl ((tetrahydro-2H-pyran-4-yl) methyl)
amino) benzamide
##STR00874##
[1846] Aqueous NaOH (0.18 g, 4.5 mmol) was added to a solution of
methyl 5-chloro-2-methyl-3-(methyl ((tetrahydro-2H-pyran-4-yl)
methyl) amino) benzoate (0.70 g, 2.25 mmol) in ethanol (10 mL) and
the mixture stirred at 60.degree. C. for 1 h. The ethanol was
removed under reduced pressure and the residue acidified using
dilute HCl up to pH 6 and pH 4 was adjusted using citric acid.
Extraction was carried out using DCM. The combined organic layers
were dried and concentrated giving the respective acid (0.2 g).
[1847] The above acid (0.2 g, 0.68 mmol) was then dissolved in DMSO
(0.5 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.206
g, 1.35 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.528 g, 1.01 mmol) was added
t and stirring was continued overnight. The mixture was poured into
ice and extraction was carried out using 10% MeOH/DCM. The combined
organic layers were dried over sodium sulphate and concentrated
under reduced pressure to obtain crude material which was purified
by column chromatography giving the title compound (0.2 g, 69%).
LCMS: 432.25 (M+1).sup.+; HPLC: 99.95% (@254 nm) (R.sub.t; 5.750;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.45 (s, 1H), 8.19
(t, 1H), 7.10 (d, 1H, J=1.6 Hz), 6.90 (d, 1H, J=2 Hz), 5.85 (s,
1H), 4.23 (d, 2H, J=4.8 Hz), 3.80 (d, 2H, J=8.4 Hz), 3.21-3.27 (m,
2H), 2.73 (d, 2H, J=7.2 Hz), 2.55 (s, 3H), 2.18 (s, 3H), 2.15 (s,
3H), 2.10 (s, 3H), 1.74-1.75 (m, 1H), 1.58-1.61 (m, 2H), 1.07-1.16
(m, 2H).
Compound 127:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(tetrahydro-2H-pyran-4-yl)methyl)amino)-2-methylbenzamide
##STR00875##
[1848] Step 1: Synthesis of methyl 5-chloro-3-(ethyl
((tetrahydro-2H-pyran-4-yl) methyl) amino)-2-methylbenzoate
##STR00876##
[1850] To a stirred solution of methyl
5-chloro-2-methyl-3-(((tetrahydro-2H-pyran-4-yl) methyl) amino)
benzoate (0.40 g, 1.35 mmol) and acetaldehyde (0.120 g, 2.71 mmol)
in dichloroethane (5 mL), acetic acid (0.48 g, 8.1 mmol) was added
and the reaction stirred at room temperature for 10 minutes. Then
sodium triacetoxyborohydride (0.663 g, 4.05 mmol) was added at
0.degree. C. and the reaction stirred for 3 h at room temperature.
On completion, the solvent was removed under reduced pressure and
the crude material purified by column chromatography to the title
compound (0.38 g, 86%).
Step 2: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(tetrahydro-2H-pyran-4-yl)methyl)amino)-2-methylbenzamide
##STR00877##
[1852] Aqueous NaOH (0.099 g, 2.46 mmol) was added to a solution of
methyl 5-chloro-3-(ethyl ((tetrahydro-2H-pyran-4-yl) methyl)
amino)-2-methylbenzoate (0.38 g, 1.2 mmol) in ethanol (20 mL) and
the mixture stirred at 60.degree. C. for 1 h. After completion of
the reaction, ethanol was removed under reduced pressure and the
residue acidified using dilute HCl up to pH 6 and pH 4 was adjusted
using citric acid. Extraction was carried out using DCM. The
combined organic layers were dried concentrated giving the
respective acid (0.2 g).
[1853] The above acid (0.2 g, 0.64 mmol) was then dissolved in DMSO
(2 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.196
g, 1.29 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min before PYBOP (0.504 g, 0.97 mmol) was added
and stirring was continued overnight. The mixture was poured into
ice and extraction was carried out using 10% MeOH/DCM. The combined
organic layers were dried over sodium sulphate and concentrated
under reduced pressure to obtain crude material which was purified
by column chromatography to afford the title compound (0.15 g,
52%). LCMS: 446.20 (M+1).sup.+; HPLC: 99.89% (@254 nm) (R.sub.t;
5.381; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.47 (s, 1H), 8.21 (t, 1H), 7.14 (s, 1H), 6.91 (s, 1H), 5.85 (s,
1H), 4.23 (d, 2H, J=4.4 Hz), 3.78 (d, 2H, J=8.8 Hz), 3.18 (t, 2H,
J=11.6 Hz), 2.82-2.89 (m, 4H), 2.18 (s, 3H), 2.13 (s, 3H), 2.10 (s,
3H), 1.54-1.57 (m, 3H), 1.09-1.15 (m, 2H), 0.91 (t, 3H, J=6.8
Hz).
Compound 157:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydr-
o-2H-pyran-4-yl)amino)-2,5-dimethylbenzamide
##STR00878##
[1854] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00879##
[1856] To stirred solution of 2-methyl-3-nitrobenzoic acid (50 g,
276 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 152 mmol)
was added portionwise at room temperature. The mixture was stirred
at room temperature for 5 h. and poured on ice cold water. The
precipitate was filtered, washed with water and dried under vacuum
giving the title compound (71 g, 99%).
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzene
##STR00880##
[1858] To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic
acid (287 g, 1100 mmol) in DMF (150 mL), were added sodium
carbonate (468 g, 4415 mmol) and methyl iodide (627 g, 4415 mmol).
The mixture was heated at 60.degree. C. for 8 h. The precipitate
was filtered and washed with diethyl ether (5 times). The combined
organic layers were dried, concentrated under reduced pressure
giving the title compound (302 g, 99%) which was used directly
without further purification.
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00881##
[1860] To a stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzene (150 g, 544 mmol) in ethanol (750
mL), ammonium chloride (150 g, 2777 mmol) dissolved in water (750
mL) and iron powder (93.3 g, 1636 mmol) were added under stirring.
The mixture was heated at 80.degree. C. for 7 h. and filtered
through celite.; The solids were washed with water and ethyl
acetate and the combined filtrates were extracted with ethyl
acetate. The combined organic layers were dried and concentrated
under reduced pressure to give the title compound (175 g) which was
used without further purification.
Step 4: Synthesis of methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate
##STR00882##
[1862] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (15 g, 61 mmol) and
dihydro-2H-pyran-4(3H)-one (9.2 g, 92 mmol) in dichloroethane (300
mL), acetic acid (22 g, 368.4 mmol) was added and the reaction
stirred at room temperature for 2 h. Then sodium
triacetoxyborohydride (39 g, 184 mmol) was added at 0.degree. C.
and the reaction stirred overnight at room temperature. The solvent
was removed under reduced pressure and the crude material was
purified by column chromatography to afford the title compound (14
g, 69%).
Step 5: Synthesis of methyl 5-bromo-3-(ethyl
(tetrahydro-2H-pyran-4-yl) amino)-2-methylbenzoate
##STR00883##
[1864] To a stirred solution of methyl
5-bromo-2-methyl-3-((tetrahydro-2H-pyran-4-yl) amino) benzoate (14
g, 43 mmol) and acetaldehyde (3.75 g, 85 mmol) in dichloroethane
(150 mL), was added acetic acid (15.4 g, 256 mmol) and the reaction
stirred at room temperature for 20 minutes. Then sodium
triacetoxyborohydride (27.0 g, 128 mmol) was added at 0.degree. C.
and the reaction stirred at room temperature for 2 h. The solvent
was removed under reduced pressure and water added to the residue.
Extraction was carried out using DCM and the combined organic
layers were dried over sodium sulphate and concentrated under
reduced pressure to give crude material was purified by column
chromatography to afford the title compound (14 g, 93%).
Step 6: Synthesis of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)
amino)-2-methylbenzamide
##STR00884##
[1866] Aqueous NaOH (2.36 g, 59 mmol) was added to a solution of
methyl 5-bromo-3-(ethyl (tetrahydro-2H-pyran-4-yl)
amino)-2-methylbenzoate (14 g, 39 mmol) in ethanol (100 mL) and the
mixture stirred at 60.degree. C. for 1 h. The ethanol was removed
under reduced pressure and the residue acidified using dilute HCl
up to pH 6 and pH 4 was adjusted using citric acid. Extraction was
carried out using ethyl acetate. The combined organic layers were
dried and concentrated giving the respective acid (13.9 g).
[1867] The above acid (10 g, 29 mmol) was then dissolved in DMSO
(25 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (8.8 g,
58 mmol) and triethyl amine (5.6 g, 58 mmol) was added. The
reaction mixture was stirred at room temperature for 15 min before
PYBOP (22 g, 43.8 mmol) was added and stirring was continued
overnight. The mixture was poured into ice and extracted with 10%
MeOH/DCM. The combined organic layers were dried and concentrated
to obtain crude material which was purified by solvent washings to
afford the title compound (14 g, 74%).
Step 7: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydr-
o-2H-pyran-4-yl)amino)-2,5-dimethylbenzamide
##STR00885##
[1869] To stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)
methyl)-3-(ethyl (tetrahydro-2H-pyran-4-yl)
amino)-2-methylbenzamide (0.30 g, 0.63 mmol) in DMF (2 mL), was
added dichlorobis (triphenylphosphine) palladium (II) (0.028 g,
0.034 mmol) followed by tetramethyl tin (0.124 g, 0.69 mmol). The
mixture was heated at 160.degree. C. for 30 minutes in a microwave
reactor. The reaction mass was quenched with water and extraction
was carried out using 10% MeOH/DCM. The combined organic layers
were dried and concentrated giving crude material. Purification by
column chromatography and then by preparative HPLC gave the title
compound as the corresponding TFA salt (0.15 g, 57.7%). LCMS:
412.15 (M+1).sup.+; HPLC: 94.29% (@254 nm) (R.sub.t; 4.245; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (CD3OD, 400 MHz) .delta. 7.54 (s, 1H), 7.33 (s, 1H),
6.13 (s, 1H), 4.47 (s, 2H), 3.95-3.98 (m, 4H), 3.74 (bs, 2H), 3.37
(m, 1H), 2.42 (s, 3H), 2.38 (s, 3H), 2.37 (s, 3H), 2.24 (s, 3H),
1.61-1.91 (bs, 4H), 1.01 (t, 3H, J=3.2 Hz).
Compound 222:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4-
s)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00886##
[1870] Step 1: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate
##STR00887##
[1872] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (5.0 g, 25 mmol) and tert-butyl (4-oxocyclohexyl)carbamate
(6.95 g, 32.6 mmol) in 25 ml of dichloroethane, was added acetic
acid (9.0 mL, 450.75 mmol) at room temperature. The reaction
mixture was cooled and sodium triacetoxyborohydride (22.8 g, 108
mmol) was added and the mixture stirred at room temperature
overnight. The mixture was neutralized with sat. NaHCO.sub.3 and
extracted with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure and the
product isomers separated by silica gel (100-200) column
chromatography to give the less polar as cis-isomer, methyl
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate (5.2 g, 52%) and the more polar title compound
trans-isomer, methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chlo-
ro-2-methylbenzoate (3.5 g, 35%).
Step 2: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-chloro-2-methylbenzoate
##STR00888##
[1874] To a stirred solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate (3.0 g, 7.6 mmol) and acetaldehyde (0.66 g, 15 mmol)
in 15 ml of dichloroethane, was added acetic acid (2.72 mL, 45.45
mmol) and the mixture stirred at room temperature for 20 minutes.
The reaction mixture was cooled to 0.degree. C. and sodium
triacetoxyborohydride (4.81 g, 22.72 mmol) was added and the
mixture stirred at room temperature overnight. The reaction mixture
was neutralized with sat. NaHCO.sub.3 extracted with DCM. The
combined organic phases were dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure and the crude material purified
by silica gel (100-200) column chromatography to yield the title
compound (3.5 g, 96%).
Step 3: Synthesis of
tert-butyl((1r,rs)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)
carbamate
##STR00889##
[1876] A mixture of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-chloro-2-methylbenzoate (3.5 g, 8.2 mmol) and NaOH (0.49
g, 12 mmol) in 20 ml of ethanol:water (4:1) was heated at
70.degree. C. for 2 h. The reaction mixture was cooled to 0.degree.
C. and acidified to pH 6 by using 1N HCl. The mixture was
concentrated and the residue partitioned between water and ethyl
acetate. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to afford 3.2 g of crude acid.
A mixture of the crude acid (3.2 g, 7.8 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2.36 mg, 15.5 mmol)
and PyBOP (6.07 mg, 11.7 mmol) was stirred in 15 ml of DMSO at room
temperature overnight. The reaction mixture was diluted with water
and extracted with 10% MeOH in DCM. The organic layer was dried
over Na.sub.2SO.sub.4, concentrated under reduced pressure and the
residue purified by silica gel (100-200) column chromatography to
give the title compound (3.0 g, 71%).
Step 4: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00890##
[1878] To a cooled solution of
tert-butyl((1r,4r)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)
carbamate (400 mg, 0.73 mmol) in 5 ml of DCM, 2 ml of TFA was added
and the reaction mixture stirred at room temperature for 2 h. The
reaction mixture was concentrated to dryness under reduced
pressure. The crude product was dissolved in 10% MeOH in DCM and
washed with sat NaHCO.sub.3, water and brine. The organic phase was
dried over Na.sub.2SO.sub.4, concentrated under reduced pressure to
afford the title compound (320 mg, 98%).
Step 5: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4-
r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00891##
[1880] To a stirred solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (300 mg, 0.67
mmol) and formaldehyde (0.5 ml of 38% solution, 6.75 mmol) in 3 ml
of methanol, sodium triacetoxyborohydride (83.0 mg, 1.35 mmol) was
added at 0.degree. C. and the mixture stirred at room temperature
for 3 h. The reaction mixture was concentrated to dryness and
partitioned between water and 10% MeOH in DCM. The organic layer
was dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the crude material was purified over basic alumina to
yield the title compound (120 mg, 32%).
Compound 268:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00892##
[1882] To a solution of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4-
r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (80
mg, 0.16 mmol) in 5 mL of methanol, was added a catalytic amount of
10% palladium on carbon. The mixture was stirred under hydrogen
atmosphere at balloon pressure for 13 h. The reaction mixture was
filtered through a bed of celite, concentrated under reduced
pressure to a viscous oil which on ether washing afforded the title
compound (40 mg, 54%) as an off-white solid.
[1883] LCMS: 439.25 (M+1).sup.+; HPLC: 88.61% (@254 nm)
(R.sub.t4.084; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (bs, 1H), 8.03 (t, 1H), 7.15-7.13 m, 2H), 6.94 (d, 1H), 4.26
(d, 2H, J=4.4 Hz), 3.02 (d, 2H, J=6.8 Hz), 2.61 (s, 6H), 2.18 (s
6H), 2.10 (s, 3H), 1.97-1.83 (m, 4H), 1.39 (m, 4H), 0.8 (t, 3H,
J=6.8 Hz), (3H merged in solvent peak).
Compound 273:
5-chloro-3-(((1s,4s)-4-(diethylamino)cyclohexyl)(ethyl)amino)-N-((4,6-dim-
ethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00893##
[1885] To a stirred solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (150 mg, 0.33
mmol) and acetaldehyde (0.15 ml, 2.70 mmol) in 5 ml of
dichloroethane, acetic acid (0.14 mL, 2.36 mmol) was added and the
mixture stirred at room temperature for 20 minutes. The reaction
mixture was cooled to 0.degree. C. and sodium triacetoxyborohydride
(286 mg, 1.35 mmol) was added and the mixture stirred at room
temperature overnight. The reaction mixture was neutralized with
sat. NaHCO.sub.3 and extracted with DCM, dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to a
residue. Purification by prep. HPLC gave the title compound (100
mg, 59%). LCMS: 501.50 (M+1).sup.+; HPLC: 93.90% (@210-370 nm)
(R.sub.t; 4.543; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (bs, 1H), 8.21 (t, 1H), 7.13 (s, 1H), 6.92 (s, 1H), 5.85 (s,
1H), 4.25 (d, 2H, J=3.6 Hz), 3.03-3.01 (m, 2H), 2.67-2.61 (m, 2H),
2.42-2.33 (m, 4H), 2.19 (s, 3H), 2.14 (s, 3H), 2.11 (s, 3H), 1.73
(m, 4H), 1.39-1.19 (m, 4H), 0.92 (m, 6H), 0.78 (t, 3H).
Compound 277:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-ethynyl-2-methylbenzamide
##STR00894##
[1886] Step 1: Synthesis of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-2-methylbenzoate
##STR00895##
[1888] To a stirred solution of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)
cyclohexyl)amino)-2-methylbenzoate 6.0 g, 13.0 mmol) and
acetaldehyde (1.5 ml, 27.0 mmol) in dichloroethane (60 mL), was
added acetic acid (5.4 ml, 82 mmol) and the mixture was stirred at
room temperature for 20 minutes. Sodium triacetoxyborohydride (8.6
g, 41 mmol) was added at 0.degree. C. and the mixture stirred at
room temperature for 2 h. The reaction mixture was neutralized with
sat. NaHCO.sub.3 and was extracted with DCM. The combined organic
phases were dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure. The crude material was purified by silica gel
(100-200) column chromatography to afford the title compound (5.0
g, 79%).
Steps 2 and 3: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl) (ethyl)
amino)-5-ethynyl-2-methylbenzoate
##STR00896##
[1890] A solution of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)
cyclohexyl)(ethyl)amino)-2-methylbenzoate (1.5 g, 3.20 mmol),
trimethylsilyl acetylene (0.52 ml, 3.84 mmol), copper iodide (12
mg, 0.06 mmol) and N,N-diisopropyl amine (1.0 ml, 8.38 mmol) in 20
ml of toluene was purged with argon. Dichloro-bis(triphenyl
phosphine) palladium (II) (44 mg, 0.06 mmol) was added the reaction
mixture heated at 80.degree. C. for 16 h. The reaction mixture was
cooled to room temperature, diluted with ethyl acetate and washed
with water. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to obtain 1.8 g of crude silyl
protected compound. This was dissolved in 15 ml of THF and
tetra-butylammonium fluoride (7.5 ml of 1M solution in THF, 7.40
mmol) was added at room temperature. The mixture stirred for 2 h,
diluted with ethyl acetate and washed with water. The organic layer
was dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure, and the residue purified by silica gel (100-200) column
chromatography to give the title compound (800 mg, 52%).
Steps 4 and 5: Synthesis of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-5-ethynyl-2-methylphenyl)(ethyl)amino)cyclohexyl)
carbamate
##STR00897##
[1892] A mixture of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl) (ethyl)
amino)-5-ethynyl-2-methylbenzoate (800 mg, 1.92 mmol) and NaOH (115
mg, 2.89 mmol) in 12 ml of ethanol:water (3:1) was heated at
70.degree. C. for 2 h. The reaction mixture was concentrated to
dryness and the residue was partitioned between water and DCM. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford 700 mg of the crude acid. The
crude acid (700 mg, 1.75 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (532 mg, 3.5 mmol)
and PyBOP (1.36 g, 2.62 mmol) were stirred in 7 ml of DMSO at room
temperature overnight. The reaction mixture was diluted with water,
and the precipitated product was filtered and purified by silica
gel column chromatography to give the title compound (750 mg,
80%).
Step 6: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-5-ethynyl-2-methylbenzamide
##STR00898##
[1894] To a cooled solution of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-5-ethynyl-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate
(750 mg, 1.40 mmol) in 10 ml of DCM, was added 3 ml of TFA The
mixture was stirred at rt for 2 h and was concentrated to dryness
under reduced pressure. The residue was dissolved in 10% MeOH in
DCM and washed with sat NaHCO.sub.3, water and brine. The organic
phase was dried over Na.sub.2SO.sub.4 and concentrated under
reduced pressure to give the crude title compound (600 mg,
98%).
Step 7: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-ethynyl-2-methylbenzamide
##STR00899##
[1896] To a stirred solution of crude
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-5-ethynyl-2-methylbenzamide (600 mg, 1.38
mmol) and formaldehyde (0.4 ml of 38% solution, 6.75 mmol) in 10 ml
of dichloroethane was added sodium triacetoxyborohydride (731 mg,
3.45 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 3 h and partitioned between water and 10% MeOH in
DCM The organic layer was separated, dried over Na.sub.2SO.sub.4
and concentrated under reduced pressure. The residue was purified
over basic alumina to give the title compound (450 mg, 70%).
[1897] Analytical Data for
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-ethynyl-2-methylbenzamide-TFA
salt: LCMS: 463.65 (M+1).sup.+; HPLC: 92.65% (@254 nm) (R.sub.t;
4.748; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (bs, 1H), 9.43 (bs, 1H), 8.23 (t, 1H), 7.36 (s, 1H), 7.12 (s,
1H), 5.87 (s, 1H), 4.26 (d, 2H, J=4.0 Hz), 3.17-3.03 (m, 4H),
2.69-2.68 (m, 6H), 2.50 (1H merged in solvent peak), 2.21 (s, 3H),
2.19 (s, 3H), 2.11 (s, 3H), 1.96-1.84 (m, 4H), 1.43 (m, 4H), 0.79
(t, 3H).
Compound 278:
5-cyano-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00900##
[1898] Step 1: Synthesis of methyl
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-cyano-2-methylbenzoate
##STR00901##
[1900] A solution of compound methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-2-methylbenzoate (1.0 gm, 2.13 mmol) and Zn(CN).sub.2
(370 mg, 3.19 mmol) in 6 ml of DMF was degassed with argon for 20
min followed by addition of Pd(PPh.sub.3).sub.4 (240 mg, 0.21
mmol). The mixture was heated at 100.degree. C. overnight, diluted
with water and was extracted with ethyl acetate. The combined
organic layers were dried over Na.sub.2SO.sub.4, concentrated and
the residue purified by silica gel column chromatography to give
the title compound (280 mg, 82%).
Steps 2 and 3: Synthesis of tert-butyl
((1r,4r)-4-((5-cyano-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl) carbamate
##STR00902##
[1902] A mixture of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-cyano-2-methylbenzoate (735 mg, 1.78 mmol) and NaOH
(85 mg, 2.13 mmol) in 12 ml of ethanol:water (3:1) was heated at
70.degree. C. for 2 h. The mixture was concentrated to dryness and
the crude product was partitioned between water and DCM. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford 380 mg of crude acid. A mixture of
the crude acid (380 mg, 0.94 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (287 mg, 1.89 mmol)
and PyBOP (737 mg, 1.41 mmol) in 4 ml of DMSO was stirred at room
temperature overnight. The reaction mixture was diluted with water
and was extracted with 10% MeOH in DCM. The combined organic phases
were dried over Na2SO4, concentrated under reduced pressure and the
residue purified by silica column chromatography to afford the
title compound (350 mg, 75%).
Step 4: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-cyano-N-((4,6-dimethyl-2-ox-
o-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00903##
[1904] To a cooled solution of compound tert-butyl
((1r,4r)-4-((5-cyano-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate (350
mg, 0.80 mmol) in 5 ml of DCM, was added 1 ml of TFA. The mixture
was stirred at rt for 2 h. and concentrated to dryness under
reduced pressure. The residue was dissolved in 10% MeOH in DCM and
washed with sat NaHCO.sub.3, water and brine. The organic phase was
dried over Na.sub.2SO.sub.4, and concentrated under reduced
pressure to obtain 284 mg of the crude title compound.
Step 5: Synthesis of
5-cyano-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00904##
[1906] To a stirred solution of crude
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-cyano-N-((4,6-dimethyl-2-ox-
o-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (325 mg, 0.74
mmol) and formaldehyde (0.3 ml of 38% solution, 3.72 mmol) in 10 ml
of methanol, sodium cyanoborohydride (100 mg, 1.49 mmol) was added
at 0.degree. C. The mixture was stirred at room temperature for 4 h
and partitioned between water and 10% MeOH in DCM. The organic
layer was dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the residue was purified by basic alumina column
purification to give the title compound (75 mg, 22%).
[1907] Analytical Data of
5-cyano-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide:
LCMS: 464.30 (M+1).sup.+; HPLC: 87.24% (@254 nm) (R.sub.t; 4.540;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (bs, 1H),
8.31 (t, 1H), 7.61 (s, 1H), 7.35 (s, 1H), 5.87 (s, 1H), 4.26 (d,
2H, J=4.4 Hz), 3.07-3.05 (m, 3H), 2.73-2.63 (m, 7H), 2.25 (s, 3H),
2.20 (s, 3H), 2.11 (s, 3H), 1.96-1.82 (m, 4H), 1.41-1.37 (m, 4H),
0.78 (t, 3H).
Compound 279:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methyl(tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)amino)-2-methy-
lbenzamide
##STR00905##
[1908] Step 1: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methyl(tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)amino)-2-methy-
lbenzamide
##STR00906##
[1910] To a stirred solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (300 mg, 0.67
mmol) and dihydro-2H-pyran-4(3H)-one (99 mg, 1.01 mmol) in 5 ml of
dichloroethane, was added acetic acid (0.24 mL, 4.05 mmol) and the
mixture stirred at room temperature for 20 minutes. The mixture was
cooled to 0.degree. C. and sodium triacetoxyborohydride (429 mg,
2.02 mmol) was added and the reaction stirred at room temperature
overnight. The reaction mixture was neutralized with sat.
NaHCO.sub.3 and extracted with DCM. The combined organic phases
were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the residue purified by silica gel column
chromatography to give the title compound (175 mg, 57%).
Step 2: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methyl(tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)amino)-2-methy-
lbenzamide
##STR00907##
[1912] To a stirred solution of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-((tetrahydro-2H-pyran-4-yl)amino)cyclohexyl)amino)-2-methylbenza-
mide (175 mg, 0.33 mmol) and formalin (99 mg, 3.31 mmol) in 2 ml of
methanol was added sodium cyanoborohydride (41 mg, 0.66 mmol) at
0.degree. C. The mixture was stirred at room temperature overnight,
neutralized with sat. NaHCO.sub.3 and extracted with DCM. The
combined organic phases were dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure and the residue purified by
preparative HPLC to afford the title compound (70 mg, 39%).
[1913] LCMS: 543.65 (M+1).sup.+; HPLC: 98.19% (@210-370 nm)
(R.sub.t; 4.515; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (bs, 1H), 8.21 (t, 1H), 7.13 (s, 1H), 6.92 (s, 1H), 5.85 (s,
1H), 4.25 (d, 2H, J=4 Hz), 3.84-3.82 (m, 2H), 3.26-3.23 (m, 3H),
3.03-3.01 (m, 2H), 2.67-2.53 (m, 2H), 2.19 (s, 3H), 2.14 (s, 3H),
2.11 (s, 6H), 1.75-1.57 (m, 6H), 1.41-1.24 (m, 6H), 0.78 (t,
3H).
Compound 276: Synthesis of compound
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00908##
[1914] Step 1: Synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00909##
[1916] To stirred solution of compound 2-methyl-3-nitrobenzoic acid
(50.0 g, 276 mmol) in conc. H.sub.2SO.sub.4 (200 mL),
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 152 mmol)
was added portionwise at room temperature and stirring was
continued for 5 h. On completion, the reaction mass was poured on
ice cold water, the solid precipitate was filtered, washed with
water and dried under vacuum giving the crude title compound (71.7
g, 99.9%) which was used without further purification.
Step 2: Synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate
##STR00910##
[1918] To a stirred solution of crude
5-bromo-2-methyl-3-nitrobenzoic acid (287 g, 1103 mmol) in DMF (150
mL), sodium carbonate (468 g, 4415 mmol) and methyl iodide (626 g,
4415 mmol) were added. The mixture was heated at 60.degree. C. for
8 h, cooled to room temperature. The solid precipitate was filtered
and washed with diethyl ether (5 times). The combined organic
layers were dried and concentrated under reduced pressure to give
the title compound (302 g, 99%) which was used directly.
Step 3: Synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00911##
[1920] To a stirred solution of above crude compound methyl
5-bromo-2-methyl-3-nitrobenzoate (150 g, 0.54 mol) in ethanol (750
mL), ammonium chloride (150 g, 2.78 mol) dissolved in 750 ml of
water was added followed by iron powder (93.3 g, 1.64 mol) with
vigorous stirring. The mixture was heated at 80.degree. C. for 7
hand filtered through celite. The filter pad was washed with water
and ethyl acetate and the filtrates were extracted with ethyl
acetate. The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
the title compound (175 g crude).
Step 4: Synthesis of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
amino)-2-methylbenzoate
##STR00912##
[1922] To a stirred solution of crude compound methyl
3-amino-5-bromo-2-methylbenzoate (15 g, 62 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (17 g, 8 mmol) in dichloroethane (200
mL), was added acetic acid (23 ml, 368 mmol) and the reaction
stirred at room temperature for 10 min. Sodium
triacetoxyborohydride (40 g, 185 mmol) was added at 0.degree. C.
and the mixture stirred at room temperature for overnight. The
reaction mixture was neutralized with sat. NaHCO.sub.3 and
extracted with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure.
Separation by column chromatography of the crude isomeric mixture
gave desired the title compound (6.0 g, 22%).
Step 5: Synthesis of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-2-methylbenzoate
##STR00913##
[1924] To a stirred solution of compound methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl) amino)cyclohexyl)
amino)-2-methylbenzoate (6.0 g, 13.0 mmol) and acetaldehyde (1.5
ml, 27.0 mmol) in dichloroethane (60 mL) was added acetic acid (5.4
ml, 82 mmol) and the mixture was stirred at room temperature for 20
minutes. Sodium triacetoxyborohydride (8.6 g, 40.9 mmol) was added
at 0.degree. C. and the mixture stirred at room temperature for 2
h. The reaction mixture was neutralized with sat. NaHCO.sub.3 and
extracted with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude
material was purified by silica gel column chromatography to afford
the title compound (5.0 g, 79%).
Steps 6 and 7: Synthesis of tert-butyl
((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)(ethyl)amino) cyclohexyl)carbamate
##STR00914##
[1926] A mixture of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-2-methylbenzoate (1.00 g, 2.13 mmol) and NaOH (102
mg, 2.56 mmol) in 6 ml of ethanol:water (2:1) was heated at
70.degree. C. for 2 h. The reaction mixture was concentrated to
dryness and the residue was partitioned between water and DCM. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford 1.0 g of crude acid. The crude
acid (1.0 g, 2.20 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (669 mg, 4.40 mmol),
PyBOP (1.71 g, 3.30 mmol) and 1 ml of triethyl amine were stirred
in 2 ml of DMSO at room temperature overnight. The reaction mixture
was diluted with water and extracted with DCM. The combined organic
layers were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the residue purified by silica gel column
chromatography to give the title compound (1.0 g, 83%).
Step 8: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-ox-
o-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00915##
[1928] To a cooled solution of compound tert-butyl
((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)(ethyl)amino) cyclohexyl)carbamate
(1.0 g, 1.60 mmol) in 10 ml of DCM, was added 2 ml of TFA. The
reaction mixture was stirred at rt for 2 h, and concentrated to
dryness under reduced pressure. The residue was dissolved in 10%
MeOH in DCM and washed with sat NaHCO.sub.3, water and brine. The
organic phase was dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to give the crude title compound (650 mg,
81%).
Step 9: Synthesis of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00916##
[1930] To a stirred solution of crude
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-bromo-N-((4,6-dimethyl-2-ox-
o-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (650 mg, 1.32
mmol) and formaldehyde (0.5 ml of 38% solution, 13.3 mmol) in 10 ml
of methanol, was added sodium cyanoborohydride (82 mg, 1.32 mmol at
0.degree. C. and the mixture stirred at room temperature overnight.
The reaction mixture was partitioned between water and 10% MeOH in
DCM. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. The residue was purified by
basic alumina column purification to give the title compound (450
mg, 65%). Analytical data of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide:
LCMS: 519.30 (M+1).sup.+; HPLC: 98.35% (@254 nm) (R.sub.t; 4.392);
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (bs, 1H),
9.39 (bs, 1H), 8.23 (t, 1H), 7.30 (s, 1H), 7.09 (s, 1H), 5.86 (s,
1H), 4.25 (d, 2H, J=4.0 Hz), 3.03-3.01 (m, 3H), 2.69-2.69 (m, 6H),
2.50 (1H merged in solvent peak), 2.18 (s, 3H), 2.14 (s, 3H), 2.11
(s, 3H), 1.94-1.84 (m, 4H), 1.42 (m, 4H), 0.79 (t, 3H, J=6.8
Hz).
Compound 282:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-ethyl-2-methylbenzamide
##STR00917##
[1931] Step 1: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-vinylbenzamide
##STR00918##
[1933] A mixture of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (200
mg, 0.38 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (71
mg, 0.46 mmol) and sodium carbonate (147 mg, 1.39 mmol) in 10 ml of
dioxane was degassed with argon for 20 min, Pd(PPh.sub.3).sub.4 (44
mg, 0.04 mmol) was added and the mixture heated at 100.degree. C.
overnight. The reaction was cooled to room temperature and diluted
with water. The mixture was extracted with 10% MeOH in DCM, the
combined organic phases dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by silica gel chromatography
to obtain the title compound (150 mg, 83%).
Step 2: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-ethyl-2-methylbenzamide
##STR00919##
[1935] To a solution of compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-vinylbenzamide (150
mg, 0.32 mmol) in 5 mL of methanol, was added a catalytic amount of
10% palladium on carbon The mixture was stirred under hydrogen
atmosphere at balloon pressure for 12 h, filtered through celite,
and concentrated under reduced pressure. The residue was purified
by prep. HPLC to give the title compound as a TFA salt (65 mg,
43%).
[1936] LCMS: 467.35 (M+1).sup.+; HPLC: 93.41% (@254 nm) (R.sub.t;
10.946; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 10 m M ammonium formate in water+0.1% NH.sub.3, B;
Acetonitrile+5% solvent A+0.1% NH.sub.3; Inj. Vol: 10 .mu.L, Col.
Temp.: 30.degree. C.; Flow rate: 1.0 mL/min.; Gradient: 15% B to
95% B in 6.0 min, Hold till 8.0 min, 15% B at 8.5 min hold up to 15
min); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.47 (bs, 1H),
9.45 (s, 1H), 8.38 (s, 1H), 8.03 (s, 2H), 7.26-6.84 (m, 2H), 5.87
(s, 1H), 4.26 (d, J=4.0 Hz, 2H), 3.17-2.89 (m, 3H), 2.70, 2.68 (2s,
6H), 2.19, 2.11 (3s, 9H), 1.96-1.87 (m, 4H), 1.42 (m, 4H), 1.15 (t,
J=7.2 Hz, 3H), 0.80 (s, 3H).
Compound 283:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(3-(4-methylpiperazin-1-yl)-
prop-1-yn-1-yl)benzamide
##STR00920##
[1937] Step 1 Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-2-methyl-5-(3-(4-methyl
piperazin-1-yl)prop-1-yn-1-yl)benzoate
##STR00921##
[1939] A solution of methyl
5-(3-bromoprop-1-yn-1-yl)-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclo-
hexyl)(ethyl)amino)-2-methylbenzoate (500 mg, 1.0 mmol) and
1-methylpiperazine (0.49 ml, 4.9 mmol) in 5 ml of DMF was stirred
at room temperature overnight. The reaction mixture was diluted
with water and extracted with DCM. The combined organic phases were
dried over Na.sub.2SO.sub.4, and concentrated under vacuum to give
the title compound (550 mg).
Steps 2 and 3: Synthesis of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methyl-5-(3-(4-methylpiperazin-1-yl)prop-1-yn-1-yl)phenyl)
(ethyl)amino)cyclohexyl)carbamate
##STR00922##
[1941] A mixture of compound methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-2-methyl-5-(3-(4-methylpiperazin-1-yl)prop-1-yn-1-yl)benzoate
(550 mg, 1.0 mmol) and NaOH (83 mg, 2.1 mmol) in 8 ml of
ethanol:water (3:1) was heated at 70.degree. C. for 2 h. The
reaction mixture was concentrated to dryness and the residue was
partitioned between water and DCM. The organic layer was dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure to afford 470
mg of acid. The crude acid (470 mg, 0.92 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (279 mg, 1.83 mmol),
PyBOP (716 mg, 1.37 mmol) and triethyl amine (0.38 ml, 2.75 mmol)
mixture were stirred in 5 ml of DMSO at room temperature overnight.
The reaction mixture was diluted with water and the compound was
extracted into 10% MeOH in DCM. The organic layer was dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure and the
residue purified by silica gel column chromatography to give the
title compound (350 mg, 50%).
Step 4: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methyl-5-(3-(4-methylpiperazin-1-yl)prop-1-yn--
1-yl)benzamide
##STR00923##
[1943] To a cooled solution of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methyl-5-(3-(4-methylpiperazin-1-yl)prop-1-yn-1-yl)phenyl)(ethyl)a-
mino)cyclohexyl)carbamate (350 mg, 0.54 mmol) in 5 ml of DCM, were
added 2 ml of TFA. The reaction mixture was stirred at rt for 2 h,
concentrated to dryness under reduced pressure. The residue was
dissolved in 10% MeOH in DCM and washed with sat NaHCO.sub.3, water
and brine. The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to obtain the crude title
compound (250 mg, 84%).
Step 5: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(3-(4-methylpiperazin-1-yl)-
prop-1-yn-1-yl)benzamide
##STR00924##
[1945] To a stirred solution of crude
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methyl-5-(3-(4-methylpiperazin-1-yl)prop-1-yn--
1-yl)benzamide (250 mg, 0.45 mmol) and formaldehyde (0.35 ml of 38%
solution, 4.56 mmol) in 3 ml of methanol was added sodium
cyanoborohydride (85 mg, 1.37 mmol) at 0.degree. C. The mixture was
stirred at room temperature for 4 hand partitioned between water
and 10% MeOH in DCM. The organic layer was separated, dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure. The
residue was purified by prep. HPLC to afford the title compound as
a TFA salt (40 mg, 15%).
[1946] LCMS: 575.45 (M+1).sup.+; HPLC: 99.30% (@254 nm) (R.sub.t;
3.849; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (D.sub.2O, 400 MHz) .delta. 7.75 (s,
1H), 7.60 (s, 1H), 6.28 (s, 1H), 4.45 (d, J=4.0 Hz, 2H), 3.80-3.63
(m, 5H), 3.40-3.21 (m, 9H), 2.92 (s, 3H), 2.79 (2s, 6H), 2.35 (m,
3H), 2.32 (s, 3H), 2.26 (s, 3H), 2.19-2.05 (m, 4H), 1.65-1.57 (m,
4H), 0.97 (t, J=7.2 Hz, 3H).
Compound 284:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methylamino)cyclohexyl)amino)-2-methylbenzamide
##STR00925##
[1947] Step 1: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate
##STR00926##
[1949] To a stirred solution of compound methyl
3-amino-5-chloro-2-methylbenzoate (5.0 g, 25 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (6.95 g, 32.7 mmol) in 25 ml of
dichloroethane was added acetic acid (9.0 mL, 450 mmol) and the
mixture stirred at room temperature for 10 min. The mixture was
cooled and sodium triacetoxyborohydride (22.8 g, 108 mmol) was
added and the mixture stirred at room temperature overnight. The
mixture was neutralized with sat. NaHCO.sub.3 and was extracted
with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure and the
residue was purified by silica gel column chromatography to give
(3.5 g, 35%) of the more polar trans-isomer title compound
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate, along with (5.2 g, 52%) of the less polar cis-isomer
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate.
Step 2: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-chloro-2-methylbenzoate
##STR00927##
[1951] To a stirred solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)
cyclohexyl)amino)-5-chloro-2-methylbenzoate (3.0 g, 7.6 mmol) and
acetaldehyde (0.66 g, 15 mmol) in 15 ml of dichloroethane, was
added acetic acid (2.7 mL, 45 mmol) and the mixture was stirred at
room temperature for 20 minutes. The mixture was cooled to
0.degree. C., sodium triacetoxyborohydride (4.8 g, 22 mmol) was
added and the mixture stirred at room temperature overnight. The
mixture was neutralized with sat. NaHCO.sub.3 and was extracted
with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure and the
residue purified by silica gel column chromatography to give the
title compound (3.5 g, 96%).
Step 3: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)(methyl)amino)cyclohexyl)
(ethyl)amino)-5-chloro-2-methylbenzoate
##STR00928##
[1953] To a cooled solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)
cyclohexyl)(ethyl)amino)-5-chloro-2-methylbenzoate (1.1 g, 3.3
mmol) in 12 ml of THF, was added sodium hydride (410 mg, 10 mmol)
followed by methyl iodide (2.1 ml, 34 mmol), The mixture was
stirred at room temperature overnight, quenched by slow addition of
ice water and acidified with citric acid solution. Extraction with
10% MeOH in DCM followed by drying and concentration of the
combined organic phases under reduced pressure gave the title
compound (1.0 g, 88%).
Steps 4 and 5: Synthesis of tert-butyl
((1r,4r)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)(methyl)
carbamate
##STR00929##
[1955] A mixture of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)(methyl)amino)
cyclohexyl)(ethyl)amino)-5-chloro-2-methylbenzoate (1.0 g, 2.4
mmol) and NaOH (140 mg, 3.5 mmol) in 10 ml of ethanol:water (4:1)
was heated at 70.degree. C. for 2 h. The reaction mixture was
concentrated to dryness and the residue was partitioned between
water and DCM. The organic layer was dried over Na.sub.2SO.sub.4,
and concentrated under reduced pressure to afford 1.0 g of crude
acid. A mixture of the crude acid (1.0 g, 2.4 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (716 mg, 4.71 mmol),
PyBOP (1.83 g, 3.53 mmol) and 2 ml of triethyl amine was stirred in
5 ml of DMSO at room temperature overnight. The mixture was diluted
with water and extracted with 10% MeOH in DCM. The combined organic
layers were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the residue purified by silica gel column
chromatography to give the title compound (700 mg, 53%).
Step 6: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methylamino)cyclohexyl)amino)-2-methylbenzamide
##STR00930##
[1957] To a cooled solution of tert-butyl
((1s,4s)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)met-
hyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)(methyl)
carbamate (700 mg, 1.25 mmol) in 20 ml of DCM, were added 3 ml of
TFA. The reaction mixture was stirred at room temperature for 2 h,
and concentrated to dryness under reduced pressure. The residue was
dissolved in 10% MeOH in DCM and washed with sat NaHCO.sub.3, water
and brine. The organic phase was dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure and the residue purified by
preparative HPLC to obtain the title compound as its TFA salt (500
mg, 87%).
[1958] LCMS: 459.35 (M+1).sup.+; HPLC: 97.63% (@254 nm) (R.sub.t;
4.565; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-D.sub.6, 400 MHz) .delta.
11.46 (bs, 1H), 8.32 (s, 2H), 8.22 (m, 1H), 7.20 (s, 1H), 7.00 (s,
1H), 5.86 (s, 1H), 4.25 (d, J=4.0 Hz, 2H), 3.74 (m, 3H), 3.03-3.01
(m, 3H), 2.67 (m, 1H), 2.19 (s, 3H), 2.14 (s, 3H), 2.11 (s, 3H),
1.33 (m, 2H), 1.82-1.80 (m, 2H), 1.42-1.23 (m, 2H+2H), 0.78 (t,
J=7.2 Hz, 3H).
Compound 285:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(ethyl(methyl)amino)cyclohexyl)amino)-2-methylbenzamide
##STR00931##
[1959] Step 1: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(ethyl(methyl)amino)cyclohexyl)amino)-2-methylbenzamide
##STR00932##
[1961] To a stirred solution of compound
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methylamino)cyclohexyl)amino)-2-methylbenzamide (200 mg,
0.43 mmol) and acetaldehyde (0.03 ml, 0.43 mmol) in 5 ml of
dichloroethane, was added acetic acid (0.15 mL, 2.62 mmol) and the
mixture stirred at room temperature for 20 minutes. The mixture was
cooled to 0.degree. C. and sodium triacetoxyborohydride (0.28 g,
1.31 mmol) was added and the mixture stirred at room temperature
overnight. The reaction mixture was neutralized with sat.
NaHCO.sub.3 and was extracted with DCM. The combined organic layers
were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure, and the residue purified by prep. HPLC to give the title
compound as a TFA salt (80 mg, 37%). LCMS: 487.35 (M+1).sup.+;
HPLC: 99.87% (@254 nm) (R.sub.t; 4.711; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-D.sub.6, 400 MHz) .delta. 11.47 (bs, 1H), 9.15 (s, 1H), 8.22
(s, 1H), 7.19 (s, 1H), 6.97 (s, 1H), 5.86 (s, 1H), 4.24 (d, J=4.0
Hz, 2H), 3.16-3.01 (m, 5H), 2.64-2.63 (m, 4H), 2.19 (s, 3H), 2.15
(s, 3H), 2.10 (s, 3H), 1.91-1.83 (m, 4H), 1.47 (m, 4H), 1.19 (m,
3H), 0.79 (t, J=7.2 Hz, 3H).
Compound 286:
5-chloro-3-(((1r,4r)-4-((cyclopropylmethyl)(methyl)amino)cyclohexyl)
(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-me-
thylbenzamide
##STR00933##
[1962] Step 1: Synthesis of
5-chloro-3-(((1r,r)-4-((cyclopropylmethyl)(methyl)amino)cyclohexyl)
(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-me-
thylbenzamide
##STR00934##
[1964] To a stirred solution of d
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methylamino)cyclohexyl)amino)-2-methylbenzamide (200 mg,
0.43 mmol) and cyclopropanecarbaldehyde (0.03 gm, 0.43 mmol) in 5
ml of dichloroethane, was added acetic acid (0.15 mL, 2.62 mmol)
and the mixture stirred at room temperature for 20 minutes. The
mixture was cooled to 0.degree. C. and sodium triacetoxyborohydride
(0.277 g, 1.31 mmol) was added and the mixture stirred at room
temperature overnight. The mixture was neutralized with sat.
NaHCO.sub.3 and was extracted with DCM. The combined organic phases
were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure, and the residue purified by prep. HPLC to give the title
compound as a TFA salt (100 mg, 44%). LCMS: 513.40 (M+1).sup.+;
HPLC: 94.81% (@254 nm) (R.sub.t; 4.924; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-D.sub.6, 400 MHz) .delta. 11.47 (bs, 1H), 9.09 (s, 1H), 8.21
(s, 1H), 7.18 (s, 1H), 6.97 (s, 1H), 5.86 (s, 1H), 4.25 (d, J=4.0
Hz, 2H), 3.24 (m, 1H), 3.03-3.01 (m, 3H), 2.89-2.87 (m, 1H),
2.71-2.70 (m, 4H), 2.18 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H),
1.93-1.83 (m, 4H), 1.46-1.44 (m, 4H), 1.05 (m, 1H), 0.79 (t, J=7.2
Hz, 3H), 0.63 (m, 2H), 0.38-0.31 (m, 2H).
Compound 287: Synthesis of compound
5-chloro-3-(((1r,4r)-4-(cyclobutyl(methyl)amino)cyclohexyl)(ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide
##STR00935##
[1965] Steps 1 and 2: Synthesis of
tert-butyl((1r,4r)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)
carbamate
##STR00936##
[1967] A mixture of compound methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-chloro-2-methylbenzoate (3.5 g, 8.2 mmol) and NaOH
(0.5 g, 12 mmol) in 20 ml of ethanol:water (4:1) was heated at
70.degree. C. for 2 h. The reaction mixture was cooled to 0.degree.
C. and acidified to pH 6 by using 1N HCl and concentrated. The
residue was partitioned between water and ethyl acetate, the
organic layer was dried over Na.sub.2SO.sub.4 and concentrated
under reduced pressure to afford 3.2 g of crude acid. The crude
acid (3.2 g, 7.8 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2.36 mg, 15.6 mmol)
and PyBOP (6.1 mg, 11 mmol) were stirred in 15 ml of DMSO at room
temperature overnight. The reaction mixture was diluted with water
and extracted with 10% MeOH in DCM. The combined organic layers
were dried over Na.sub.2SO.sub.4, concentrated under reduced
pressure and the residue purified by silica gel column
chromatography to give the title compound (3.0 g, 71%).
Step 3: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00937##
[1969] To a cooled solution of
tert-butyl((1s,4s)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridi-
n-3-yl)methyl)carbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl)
carbamate (400 mg, 0.73 mmol) in 5 ml of DCM, were added 2 ml of
TFA. The reaction mixture was stirred at room temperature for 2 h
and concentrated to dryness under reduced pressure. The residue was
dissolved in 10% MeOH in DCM and washed with sat NaHCO.sub.3, water
and brine. The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure to give the title compound (320
mg, 98%).
Step 4: Synthesis of
5-chloro-3-(((1r,4r)-4-(cyclobutylamino)cyclohexyl)(ethyl)amino)-N-((4,6--
dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00938##
[1971] To a stirred solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (300 mg, 0.67
mmol) and cyclobutanone (141 mg, 2.02 mmol) in 20 ml of methanol,
was added sodium cyanoborohydride (127 mg, 2.02 mmol) at 0.degree.
C. The mixture was stirred at rt overnight, neutralized with sat.
NaHCO.sub.3 and extracted with DCM. The combined organic phases
were dried over Na.sub.2SO.sub.4 and concentrated under reduced
pressure to obtain crude title compound (260 mg, 77%).
Step 5: Synthesis of
5-chloro-3-(((1r,4r)-4-(cyclobutyl(methyl)amino)cyclohexyl)(ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide
##STR00939##
[1973] To a stirred solution of
5-chloro-3-(((1r,4r)-4-(cyclobutylamino)cyclohexyl)
(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-me-
thylbenzamide (230 mg, 0.46 mmol) and formalin (138 mg, 4.61 mmol)
in 10 ml of methanol, was added sodium cyanoborohydride (43 mg,
0.69 mmol) at 0.degree. C. The mixture was stirred at rt for
overnight neutralized with sat. NaHCO.sub.3 and extracted with DCM.
The combined organic phases were dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure and the residue purified by
silica gel column chromatography to give the title compound (150
mg, 63%).
[1974] Analytical Data of
5-chloro-3-(((1r,4r)-4-(cyclobutyl(methyl)amino)cyclohexyl)(ethyl)
amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylben-
zamide-TFA salt: LCMS: 513.25 (M+1).sup.+; HPLC: 96.89% (@254 nm)
(R.sub.t; 4.886; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-D.sub.6, 400 MHz) .delta.
11.45 (bs, 1H), 9.40 (s, 1H), 8.21 (s, 1H), 7.18 (s, 1H), 6.97 (s,
1H), 5.86 (s, 1H), 4.25 (d, J=4.0 Hz, 2H), 3.77-3.75 (m, 1H),
3.12-3.02 (m, 3H), 2.69 (m, 1H), 2.50 (4H merged in solvent peak),
2.18, 2.15, 2.11 (3s, 6H+3H+3H), 1.83-1.44 (m, 10H), 0.78 (t,
3H).
Compound 290:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2--
methylbenzamide
##STR00940##
[1975] Steps 1 and 2: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-(3-hydroxyprop-1-yn-1-yl)-2-methylbenzoate
##STR00941##
[1977] A solution of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-2-methylbenzoate (1.5 g, 3.2 mmol),
tert-butyldimethyl(prop-2-yn-1-yloxy)silane (1.6 g, 9.6 mmol),
copper iodide (183 mg, 0.96 mmol) and triethyl amine (1.30 ml, 9.60
mmol) in 10 ml of DMF was purged with argon gas for 20 min.
Tetrakis-(triphenylphosphine)palladium (369 mg, 0.32 mmol) was
added to the reaction mixture and the mixture heated at 100.degree.
C. for 5 h and cooled to room temperature. The mixture was diluted
with ethyl acetate, washed with water, and the organic layer was
dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure
to obtain crude 1.9 g of the silyl protected intermediate product.
The product was dissolved in 15 ml of THF and tetra-butyl ammonium
fluoride (6.81 ml of 1M solution in THF, 6.81 mmol) was added at
room temperature and the mixture stirred for 2 h. The reaction
mixture was diluted with ethyl acetate and washed with water. The
organic layer was dried over Na.sub.2SO.sub.4, concentrated under
reduced pressure, and the residue purified by silica gel column
chromatography to give the title compound (950 mg, 630%).
Step 3: Synthesis of methyl
5-(3-bromoprop-1-yn-1-yl)-3-(((1r,4r)-4-((tert-butoxycarbonyl)
amino)cyclohexyl)(ethyl)amino)-2-methylbenzoate
##STR00942##
[1979] To a solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-(3-hydroxyprop-1-yn-1-yl)-2-methylbenzoate (950 mg,
2.14 mmol) in 13 ml of DCM, triphenylphosphine (840 mg, 3.21 mmol)
and carbon tetrabromide (1.06 gm, 3.21 mmol) were added at room
temperature. The mixture was stirred for 3 h, diluted with DCM and
washed with water. The organic layer was dried over
Na.sub.2SO.sub.4, concentrated under reduce pressure and the
residue column purified over silica gel to give the title compound
(900 mg, 82%).
Step 4: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylbenzoate
##STR00943##
[1981] A solution of methyl
5-(3-bromoprop-1-yn-1-yl)-3-(((1r,4r)-4-((tert-butoxycarbonyl)
amino)cyclohexyl)(ethyl)amino)-2-methylbenzoate (500 mg, 0.98 mmol)
and a 2M solution in THF of N,N-dimethylamine (2.5 ml, 4.9 mmol) in
5 ml of DMF was stirred at room temperature overnight. The mixture
was concentrated under reduced pressure, diluted with water and
extracted with DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4 and concentrated under vacuum to afford the title
compound (450 mg, 96%).
Steps 5 and 6: Synthesis of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylphenyl)(ethyl)
amino)cyclohexyl)carbamate
##STR00944##
[1983] A mixture of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)
amino)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylbenzoate (450
mg, 0.95 mmol) and NaOH (57 mg, 1.40 mmol) in 10 ml of
ethanol:water (4:1) was heated at 70.degree. C. for 2 h. The
reaction mixture was concentrated to dryness and the residue
partitioned between water and DCM. The organic layer was dried over
Na.sub.2SO.sub.4c and concentrated under reduced pressure to afford
430 mg of acid. The crude acid (430 mg, 0.94 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (280 mg, 1.88 mmol),
PyBOP (733 mg, 1.40 mmol) and triethyl amine (0.1 ml, 0.94 mmol)
were stirred in 7 ml of DMSO at rt overnight. The reaction mixture
was diluted with water, the precipitated compound was filtered and
dried under vacuum to obtain the crude title compound (450 mg,
81%).
Step 7: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylbenz-
amide
##STR00945##
[1985] To a cooled solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)
cyclohexyl)(ethyl)amino)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylbenz-
oate (450 mg, 0.76 mmol) in 6 ml of DCM, was added 1 ml of TFA. The
reaction mixture was stirred at rt for 2 h, and concentrated to
dryness under reduced pressure. The residue was dissolved in 10%
MeOH in DCM and washed with sat NaHCO.sub.3, water and brine. The
organic phase was dried over Na.sub.2SO.sub.4, concentrated under
reduced pressure to obtain the crude title compound (350 mg,
93%).
Step 8: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2--
methylbenzamide
##STR00946##
[1987] To a stirred solution of crude compound
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-5-(3-(dimethylamino)prop-1-yn-1-yl)-2-methylbenz-
amide (350 mg, 0.71 mmol) and formaldehyde (0.56 ml of 38%
solution, 7.10 mmol) in 8 ml of methanol, was added sodium
cyanoborohydride (132 mg, 2.10 mmol) at 0.degree. C. The mixture
was stirred at room temperature for 4 h, and partitioned between
water and 10% MeOH in DCM. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The
residue was purified by basic alumina column chromatography to
afford the title compound (120 mg, 32%).
[1988] LCMS: 520.6 (M+1).sup.+; HPLC: 92.27% (@254 nm) (R.sub.t;
3.886; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (bs, 1H), 8.18 (t, 1H), 7.15 (s, 1H), 6.96 (s, 1H), 5.86 (s,
1H), 4.25 (d, 2H), 3.02-3.00 (m, 2H), 2.50 (13 protons merged in
solvent peak), 2.33 (s, 3H), 2.23 (s, 3H), 2.18 (s, 3H), 2.11 (s,
3H), 1.82 (m, 4H), 1.39-1.28 (m, 4H), 0.77 (t, 3H).
Compound 301:
N-((1-benzyl-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-chloro--
3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide
##STR00947##
[1990]
5-Chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-me-
thylbenzoic acid (1 eq) was dissolved in DMSO and
3-(aminomethyl)-1-(2-hydroxyethyl)-4,6-dimethylpyridin-2(1H)-one (2
eq) and triethylamine (1 eq) was added to it. The reaction mixture
was stirred at room temperature for 15 min, PYBOP (1.5 eq) was
added and stirring was continued overnight. The reaction mixture
was poured into ice and extracted with 10% MeOH/DCM. The combined
organic layers were dried and concentrated to obtain crude product
which was purified by prep. HPLC to afford the title compound
(0.011 g, 16%) as the TFA salt. LCMS: 563.40 (M+1).sup.+; HPLC:
90.27% (@210 nm-370 nm) (R.sub.t; 6.122; Method: Column: YMC ODS-A
150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in
water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR (MeOD, 400
MHz) .delta., 7.31-7.26 (m, 2H), 7.24-7.04 (m, 5H), 6.23 (s, 1H),
5.40 (d, 2H), 4.50 (s, 2H), 3.34 (1 Protons merged in solvent
peak), 3.11-3.08 (m, 2H+1H), 2.81 (s, 3H+3H), 2.42 (s, 3H), 2.29
(s, 3H), 2.21 (s, 3H), 2.08-1.99 (m, 2H+2H), 1.52-1.49 (m, 4H),
0.86 (t, 3H J=7.2 Hz).
Compound 302:
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-N-((1-(2-h-
ydroxyethyl)-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbe-
nzamide
##STR00948##
[1991] Step 1: Synthesis of methyl
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbe-
nzoate
##STR00949##
[1993] To a stirred solution of ethyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-5-chlo-
ro-2-methylbenzoate (3.4 g, 8.0 mmol) in DCM (35 mL) was added TFA
(10 mL). The mixture was stirred at room temperature for 1 hour,
the solvent was removed under reduced pressure and saturated
NaHCO.sub.3 solution was added. Extraction was carried out using
10% MeOH/DCM and the combined organic layers were washed with water
and brine and dried over anhydrous Na.sub.2SO.sub.4. Evaporation of
the solvent under reduced pressure gave methyl
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-2-methylbenzo-
ate (2.5 g, 96%).
[1994] The above compound (2.4 g, 7.4 mmol) was dissolved in
methanol (25 mL) and the mixture cooled to 0.degree. C. Formalin
(2.21 g, 73.95 mmol) was added and the resulting reaction mixture
was stirred at same temperature for 20 minutes. Sodium
cyanoborohydride (0.92 g, 14.8 mmol) was added and the mixture
stirred at room temperature for 1 hour. The solvent was removed
under reduced pressure, water added to the residue, and extraction
was carried out using 10% MeOH/DCM. The combined organic layers
were dried and concentrated under reduced pressure giving the title
compound (2.3 g, 88) which was used further without further
purification.
Step 2: Synthesis of
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbe-
nzoic acid
##STR00950##
[1996] Aqueous NaOH (0.73 g, 18.35 mmol) was added to a solution of
ethyl
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbe-
nzoate (2.3 g, 9.1 mmol) in ethanol (25 mL) and the mixture stirred
at 60.degree. C. for 1 hour. The ethanol was removed under reduced
pressure and the residue acidified using dilute HCl up to pH 6 and
adjusted to pH 4 using citric acid. Extraction was carried out
using ethyl acetate. The combined organic layers were dried and
concentrated giving the title compound (2.0 g, 92%).
Step 3: Synthesis of
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-N-((1-(2-h-
ydroxyethyl)-4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbe-
nzamide
##STR00951##
[1998]
5-Chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-me-
thylbenzoic acid (1 eq) was dissolved in DMSO and
3-(aminomethyl)-1-(2-hydroxyethyl)-4,6-dimethylpyridin-2(1H)-one (2
eq) and triethylamine (1 eq) was added. The reaction mixture was
stirred at room temperature for 15 min, PYBOP (1.5 eq) was added
and stirring was continued overnight. After completion, the
reaction mixture was poured into ice and extracted with 10%
MeOH/DCM. The combined organic layers were dried and concentrated
to obtain a crude product which was purified by prep. HPLC to
afford the title compound (0.07 g, 20%) as the TFA salt. LCMS:
517.35 (M+1).sup.+; HPLC: 91.15% (@254 nm) (R.sub.t; 5.134; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-D.sub.2O Exchange, 400 MHz) .delta. 7.17 (s, 1H),
6.94 (s, 1H), 6.01 (s, 1H), 4.26 (m, 2H), 3.99 (s, 2H), 3.01-2.99
(m, 3H), 2.67 (s, 3H+3H), 2.35 (1 Protons merged in solvent peak),
2.33 (m, 2H+2H), 2.19 (s, 3H), 2.12 (s, 3H), 1.94-1.81 (m, 5H) 1.40
(m, 4H), 0.77 (t, 3H).
Compound 306:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(trifluoromethyl)benzamide
##STR00952##
[1999] Step 1: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-methyl-5-(t-
rifluoromethyl)benzoate
##STR00953##
[2001] Acetic acid (3.28 g, 54.8 mmol) was added to a stirred
solution of methyl 3-amino-2-methyl-5-(trifluoromethyl)benzoate
(2.0 g, 9.1 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (2.52
g, 11.87 mmol) in dichloroethane (20 mL) and the reaction mixture
was stirred at room temperature for 10 minutes. Then sodium
triacetoxyborohydride (5.80 g, 27.4 mmol) was added at 0.degree. C.
and the reaction mixture was stirred overnight at room temperature.
On completion the reaction was quenched with aqueous sodium
bicarbonate. The organic phase was separated and the aqueous phase
extracted with dichloromethane. The combined organic layers were
washed with water, dried, and concentrated under reduced pressure.
The crude material was purified by column chromatography to afford
the title compound (2.8 g, 82%).
Step 2: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)(ethyl)amino)-2-met-
hyl-5-(trifluoromethyl)benzoate
##STR00954##
[2003] Acetic acid (0.85 g, 14.2 mmol) was added to a stirred
solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-meth-
yl-5-(trifluoromethyl)benzoate (0.9 g, 2.40 mmol) and acetaldehyde
(0.21 g, 4.80 mmol) in dichloroethane (10 mL) and the reaction
mixture stirred at room temperature for 20 minutes. Then sodium
triacetoxyborohydride (1.53 g, 7.21 mmol) was added at 0.degree. C.
and the reaction mixture stirred overnight at room temperature. The
reaction was quenched with aqueous sodium bicarbonate, the organic
phase separated and the aqueous phase extracted with
dichloromethane. The combined organic layers were washed with
water, dried, concentrated under reduced pressure, and the crude
material was purified by column chromatography to afford the title
compound (0.8 g, 83%).
Step 3: Synthesis of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methyl-5-(trifluoromethyl)phenyl)(ethyl)amino)cyclohexyl)carbamate
##STR00955##
[2005] Aqueous sodium hydroxide (0.12 g, 2.97 mmol) was added to a
solution of compound methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-2-meth-
yl-5-(trifluoromethyl)benzoate (0.80 g, 1.98 mmol) in ethanol (8
mL) and the mixture was stirred at 60.degree. C. for 1 hour. After
completion of the reaction, ethanol was removed under reduced
pressure and the residue acidified using dilute HCl up to pH 6 and
then adjusted to pH 4 using citric acid. Extraction was carried out
using 10% MeOH/DCM. The combined organic layers were dried and
concentrated giving the respective acid (0.7 g,) The acid (0.7 g,
1.8 mmol) was then dissolved in DMSO (10 mL) and 3-(amino
methyl)-4, 6-dimethylpyridin-2(1H)-one (0.54 g, 3.59 mmol) was
added. The reaction mixture was stirred at room temperature for 15
min, PYBOP (1.40 g, 2.69 mmol) was added and stirring was continued
overnight. After completion of the reaction, the reaction mixture
was poured into ice and extracted with 10% MeOH/DCM. The combined
organic layers were dried, concentrated to obtain crude product
which was then purified by washing with acetonitrile to afford the
title compound (0.7 g, 74%).
Step 4: Synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamide
##STR00956##
[2007] To a solution of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-2-methyl-5-(trifluoromethyl)phenyl)(ethyl)amino)cyclohexyl)carbamate
(0.7 g, 1.3 mmol) in DCM (10 mL) was added TFA (3 mL) and the
reaction mixture stirred at room temperature for 1 h. After
completion of reaction, the solvent was removed under reduced
pressure and saturated NaHCO.sub.3 solution was added. Extraction
was carried out using 10% MeOH/DCM; the combined organic layers
were washed with water and brine; dried over anhydrous
Na.sub.2SO.sub.4; filtered and concentrated under reduced pressure
to give the title compound (0.56 g, 98%) which was used directly in
the next step.
Step 5: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(trifluoromethyl)benzamide
##STR00957##
[2009] To a solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-di-
hydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamide
(0.5 g, 1.2 mmol) in DCM (5 mL) at 0.degree. C. was added formalin
(0.124 g, 4.13 mmol). The reaction mixture was stirred at 0.degree.
C. for 20 minutes. Sodium triacetoxyborohydride (0.63 g, 2.95 mmol)
was added and the reaction mixture was stirred at room temperature
for 1 h. After completion, water was added and extraction was
carried out using 10% MeOH/DCM. The combined organic layers were
dried and concentrated under reduced pressure. The crude solid
obtained was washed with acetonitrile and ether to afford the title
compound (0.4 g, 67%). LCMS: 507.45 (M+1).sup.+; HPLC: 96.81%
(@210-370 nm) (R.sub.t; 4.857; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 11.46 (bs, 1H), 8.34 (s, 1H), 7.38 (s, 1H), 7.20 (s,
1H), 5.86 (s, 1H), 4.27 (d, 2H), 3.08 (m, 2H), 2.67 (m, 1H), 2.40
(s, 6H), 2.24 (s, 3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.85 (m, 2H),
1.80 (m, 2H), 1.42 (m, 2H), 1.25 (m, 2H), 0.78 (t, 3H).
Compound 390:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-4-methylnicotinamide
##STR00958##
[2010] Step 1: Synthesis of methyl 5-amino-4-methylnicotinate
##STR00959##
[2012] To a stirred solution of 5-amino-4-methylnicotinic acid (5.0
g, 32 mmol) in methanol (75 mL), was added conc. H.sub.2SO.sub.4 (5
mL) slowly at room temperature. The mixture was stirred at
70.degree. C. for 12 h. On completion, the solvent was removed
under reduced pressure and the residue was neutralized with
saturated NaHCO.sub.3 solution. Extraction was carried out using
EtOAc; the combined organic layers were washed with water, brine
and dried over anhydrous Na.sub.2SO.sub.4 and concentrated under
reduced pressure to give the title compound (4.1 g, 75%) which used
without further purification.
Step 2: Synthesis of methyl
5-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-4-methylnicot-
inate
##STR00960##
[2014] To a stirred solution of methyl 5-amino-4-methylnicotinate
(1.20 g, 7.22 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (2.30
g, 10.8 mmol) in dichloroethane (20 mL), was added acetic acid
(2.59 g, 43 mmol) and the mixture stirred at room temperature for
10 minutes. Then sodium triacetoxyborohydride (4.59 g, 21.56 mmol)
was added at 0.degree. C. and the reaction stirred overnight at
room temperature. On completion, the reaction was quenched with
aqueous sodium bicarbonate, the organic phase separated and the
aqueous phase extracted with dichloromethane. The combined organic
layers were dried over anhydrous sodium sulphate and concentrated
under reduced pressure. The crude compound was purified by silica
gel column chromatography to afford the title compound (0.65 g,
25%).
Step 3: Synthesis of methyl
5-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-4--
methylnicotinate
##STR00961##
[2016] To a stirred solution of methyl
5-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-4-methylnicot-
inate (0.65 g, 1.79 mmol) and acetaldehyde (0.39 g, 8.95 mmol) in
dichloroethane (20 mL) was added acetic acid (0.64 g, 10.7 mmol)
and the mixture stirred at room temperature for 10 minutes. Then
sodium triacetoxyborohydride (1.13 g, 5.33 mmol) was added at
0.degree. C. and the reaction stirred overnight at room
temperature. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were dried over anhydrous sodium sulphate and concentrated under
reduced pressure to afford the title compound (0.55 g, 79%).
Step 4: Synthesis of tert-butyl
((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-4-methylpyridin-3-yl)(ethyl)amino)cyclohexyl)carbamate
##STR00962##
[2018] Aqueous NaOH (0.11 g, 2.81 mmol) was added to a solution of
methyl
5-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-amino)-4--
methylnicotinate (0.55 g, 1.40 mmol) in ethanol (7 mL) and the
mixture stirred at 60.degree. C. for 1 h. After completion of the
reaction, ethanol was removed under reduced pressure and the
mixture acidified using dilute HCl up to pH 6 and pH 4 was adjusted
using citric acid. Extraction was carried out using ethyl acetate.
The combined organic layers were dried concentrated giving
respective acid (0.5 g). The above acid (0.5 g, 1.3 mmol) was then
dissolved in DMSO (5 mL) and 3-(amino methyl)-4,
6-dimethylpyridin-2(1H)-one (0.48 g, 2.65 mmol) was added. The
reaction mixture was stirred at room temperature for 15 min, PYBOP
(1.02 g, 1.58 mmol) was added and stirring was continued overnight.
After completion of the reaction, the mixture was poured into ice
and extracted with 10% MeOH/DCM. The combined organic layers were
dried and concentrated to a residue which was purified by silica
gel chromatography to afford the title compound (0.5 g, 74%).
Step 5: Synthesis of
5-(((1r,r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dih-
ydropyridin-3-yl)methyl)-4-methylnicotinamide
##STR00963##
[2020] To a stirred solution of tert-butyl
((1r,4r)-4-((5-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-4-methylpyridin-3-yl)(ethyl)amino)cyclohexyl)carbamate (0.5
g, 0.97 mmol) in DCM (10 mL) was added TFA (2 mL). The mixture was
stirred at rt for 1 the solvent was removed under reduced pressure.
Saturated NaHCO.sub.3 solution was added to The residue and
extraction was carried out using 10% MeOH/DCM. The combined organic
layers were washed with water and brine; dried over anhydrous
Na.sub.2SO.sub.4; filtered and concentrated under reduced pressure
to give the title compound (0.35 g, 88%).
Step 6: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-4-methylnicotinamide
##STR00964##
[2022]
5-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo--
1,2-dihydropyridin-3-yl)methyl)-4-methylnicotinamide (0.35 g, 0.85
mmol) was dissolved in DCM (5 mL) and formalin (0.089 g, 2.96 mmol)
was added at 0.degree. C. The mixture was stirred for 20 minutes,
sodium triacetoxyborohydride (0.45 g, 2.12 mmol) was added and the
mixture stirred at room temperature for 1 h. After completion, the
mixture was diluted with water and extraction was carried out using
10% MeOH/DCM. The combined organic layers were dried and
concentrated under reduced pressure. The solid obtained was further
purified by washings with acetonitrile and ether to afford the
title compound (0.3 g, 80%). LCMS: 440.45 (M+1).sup.+; HPLC: 73.44%
& 21.34% (@210-370 nm) (R.sub.t; 3.769 & 3.965); Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.45 (bs, 1H), 8.34 (s,
1H), 8.29 (s, 1H), 8.07 (s, 1H), 5.86 (s, 1H), 4.27 (d, 2H), 3.18
(m, 1H), 3.01 (m, 2H), 2.22 (s, 3H), 2.19 (s, 3H), 2.12 (s, 6H),
2.10 (s, 3H), 1.74 (m, 2H), 1.64 (m, 2H), 1.40-1.20 (m, 4H), 0.81
(t, 3H).
Compound 308:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-methoxy-2-methylbenzamide
##STR00965##
[2023] Step 1: Synthesis of methyl
5-methoxy-2-methyl-3-nitrobenzoate
##STR00966##
[2025] To a stirred solution of 5-hydroxy-2-methyl-3-nitrobenzoic
acid (1.50 g, 7.61 mmol) in DMF (15 mL) was added sodium carbonate
(3.23 g, 30.5 mmol) and methyl iodide (1.88 mL, 30.5 mmol). The
resulting reaction mixture was heated at 60.degree. C. for 8 h. On
completion, the reaction mixture was diluted with water and
extraction was carried out using ethyl acetate. The combined
organic layers were dried and concentrated under reduced pressure
to give the crude title compound (1.7 g) which was used directly in
the next step.
Step 2: Synthesis of methyl 3-amino-5-methoxy-2-methylbenzoate
##STR00967##
[2027] To a stirred solution of methyl
5-methoxy-2-methyl-3-nitrobenzoate (1.7 g, 7.6 mmol) in ethanol (20
mL) was added ammonium chloride (1.7 g, 32 mmol) dissolved in water
(20 mL) and iron powder (3.38 g, 60.44 mmol). The mixture was
heated at 80.degree. C. for 3 h, filtered and the residue washed
well with hot ethanol. The filtrate was concentrated and the
residue was basified with aqueous sodium bicarbonate. The aqueous
phase was extracted with ethyl acetate. The combined organic layers
were washed with water, dried, and concentrated under reduced
pressure to give the crude title compound (1.2 g) which was used
directly in the next step.
Step 3: Synthesis methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-methoxy-2-m-
ethylbenzoate
##STR00968##
[2029] To a stirred solution of methyl
3-amino-5-methoxy-2-methylbenzoate (1.2 g, 6.15 mmol) and
tert-butyl (4-oxocyclohexyl)carbamate (1.70 g, 7.99 mmol) in
dichloroethane (20 mL) was added acetic acid (2.21 g, 36.8 mmol).
The reaction was stirred at room temperature for 20 minutes. Sodium
triacetoxyborohydride (3.91 g, 18.4 mmol) was added at 0.degree. C.
and the reaction mixture was stirred at room temperature overnight.
On completion, the reaction was quenched with aqueous sodium
bicarbonate, the organic phase separated and the aqueous phase
extracted with dichloromethane. The combined organic layers were
washed with water, dried, and concentrated under reduced pressure.
The crude material obtained was purified by column chromatography
to afford the title compound (0.35 g, 14%).
Step 4: Synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-5-met-
hoxy-2-methylbenzoate
##STR00969##
[2031] To a stirred solution of
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-methoxy-2-m-
ethylbenzoate (0.35 g, 0.89 mmol) and acetaldehyde (0.2 g, 4.46
mmol) in dichloroethane (5 mL) was added acetic acid (0.32 g, 5.35
mmol) and the reaction mixture stirred at room temperature for 20
minutes. Then sodium triacetoxyborohydride (0.57 g, 2.67 mmol) was
added at 0.degree. C. and the reaction mixture was stirred at room
temperature for 2 h. On completion, the reaction was quenched with
aqueous sodium bicarbonate, the organic phase separated and the
aqueous phase extracted with dichloromethane. The combined organic
layers were washed with water, dried, and concentrated under
reduced pressure. The crude material was purified by column
chromatography to afford the title compound (0.3 g, 80%).
Step 5: Synthesis of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-5-methoxy-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate
##STR00970##
[2033] Aqueous NaOH (0.06 g, 1.42 mmol) was added to a solution of
methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-5-met-
hoxy-2-methylbenzoate (0.3 g, 0.71 mmol) in ethanol (4 mL) and the
mixture stirred at 60.degree. C. for 1 h. After completion of the
reaction, ethanol was removed under reduced pressure and the
residue acidified using dilute HCl up to pH 6 and pH 4 was adjusted
using citric acid. Extraction was carried out using ethyl acetate.
The combined organic layers were dried concentrated to give the
crude acid (0.25 g).
[2034] The acid (0.25 g, 0.62 mmol) was dissolved in DMSO (3 mL)
and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.19 g, 1.23
mmol) was added. The reaction mixture was stirred at room
temperature for 15 min, PYBOP (0.48 g, 0.92 mmol) was added and the
mixture was stirred overnight. The reaction was quenched by pouring
into ice, and the mixture was extracted with 10% MeOH/DCM. The
combined organic layers were washed with water, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to afford crude material which was purified by column
chromatography to give the title compound (0.2 g, 60%).
Step 6: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-methoxy-2-methylbenzamide
##STR00971##
[2036] To a solution of tert-butyl
((1r,4r)-4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carba-
moyl)-5-methoxy-2-methylphenyl)(ethyl)amino)cyclohexyl)carbamate
(0.2 g, 0.37 mmol) in DCM (4 mL) was added TFA (2 mL) and the
reaction mixture was stirred at rt for 1 h. After completion of
reaction the solvent was removed under reduced pressure and
saturated NaHCO.sub.3 solution was added. Extraction was carried
out using 10% MeOH/DCM; the combined organic layers were washed
with water and brine; dried over anhydrous Na.sub.2SO.sub.4;
filtered and concentrated under reduced pressure to give
Boc-deprotected compound (0.15 g,).
[2037] The Boc deprotected compound (0.15 g, 0.34 mmol) was
dissolved in DCM (2 mL) and formalin (0.035 g, 1.19 mmol) was added
to it at 0.degree. C. The resulting reaction mixture was stirred at
0.degree. C. for 10 minutes. Sodium triacetoxyborohydride (0.18 g,
0.85 mmol) was the added and the mixture stirred at room
temperature carried for 1 h. After reaction completion, water was
added and extraction was carried out using 10% MeOH/DCM. The
combined organic layers were dried and concentrated under reduced
pressure to afford crude material which was purified by prep. HPLC
to give the title compound as a TFA salt (0.02 g, 12%). LCMS:
469.80 (M+1).sup.+; HPLC: 96.80% (@210-370 nm) (R.sub.t; 3.802);
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.46 (s, 1H), 9.42 (s,
1H), 8.06 (s, 1H), 6.75 (s, 1H), 6.58 (s, 1H), 5.86 (s, 1H), 4.26
(d, 2H, J=4 Hz), 3.72 (s, 3H), 3.09 (m, 3H), 2.69 (s, 3H), 2.68 (s,
3H), 2.19 (s, 3H), 2.10 (s, 6H), 1.95 (m, 2H), 1.87 (m, 2H), 1.41
(m, 4H), 0.801 (t, 3H).
Compound 309:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)(ethyl)amino)-5-(2-hydroxyethoxy)-2-methylbenzamide
##STR00972##
[2038] Step 1: synthesis of methyl
5-hydroxy-2-methyl-3-nitrobenzoate
##STR00973##
[2040] To a stirred solution of 5-hydroxy-2-methyl-3-nitrobenzoic
acid (3.50 g, 17.8 mmol) in methanol (40 mL) was added thionyl
chloride (3.9 mL, 53 mmol) at 0.degree. C. The reaction mixture was
heated at reflux for 3 h. On completion, the solvent was removed
under reduced pressure. Aqueous sodium bicarbonate was added,
followed by extraction with ethyl acetate. The combined organic
layers were washed with water, dried and concentrated under reduced
pressure. The crude material was purified by column chromatography
to afford the title compound (3.0 g, 80%).
Step 2: synthesis of methyl
5-(2-hydroxyethoxy)-2-methyl-3-nitrobenzoate
##STR00974##
[2042] To a stirred solution of methyl
5-hydroxy-2-methyl-3-nitrobenzoate (1.5 g, 7.1 mmol) in ACN (15
mL), were added cesium carbonate (4.6 g, 14 mmol) and
2-bromoethanol (2.5 mL, 35 mmol). The resulting reaction mixture
was heated at 80.degree. C. overnight. On completion, the reaction
mixture was diluted with water and extraction was carried out using
ethyl acetate. The combined organic layers were washed with water,
dried and concentrated under reduced pressure. The crude material
was purified by silica gel column chromatography to afford the
title compound (1.35 g, 74%).
Step 3: synthesis of methyl
3-amino-5-(2-hydroxyethoxy)-2-methylbenzoate
##STR00975##
[2044] To a stirred solution of methyl
5-(2-hydroxyethoxy)-2-methyl-3-nitrobenzoate (1.5 g, 5.88 mmol) in
ethanol (20 mL), were added ammonium chloride (1.5 g, 28 mmol)
dissolved in water (20 mL) and iron powder (1.3 g, 23 mmol). The
resulting reaction mixture was heated at 80.degree. C. for 3 h. On
completion, the reaction mixture was filtered and the residue
washed well with hot ethanol. The filtrate was concentrated and the
residue was basified by aqueous sodium bicarbonate. The aqueous
phase was extracted with ethyl acetate. The combined organic layers
were washed with water, dried, and concentrated under reduced
pressure giving the crude title compound (1.3 g) which was used
directly.
Step 4: synthesis of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(2-hydroxyethoxy)-2-
-methylbenzoate
##STR00976##
[2046] To a stirred solution of methyl
3-amino-5-(2-hydroxyethoxy)-2-methylbenzoate (1.3 g, 5.77 mmol) and
tert-butyl (4-oxocyclohexyl)carbamate (1.6 g, 7.5 mmol) in
dichloroethane (20 mL) was added acetic acid (2.07 g, 34.66 mmol)
and the reaction mixture stirred at room temperature for 20
minutes. Then sodium triacetoxyborohydride (3.7 g, 17 mmol) was
added at 0.degree. C. and the reaction mixture was stirred at room
temperature overnight. On completion, the reaction was quenched
with aqueous sodium bicarbonate, the organic phase separated and
the aqueous phase extracted with dichloromethane. The combined
organic layers were washed with water, dried, and concentrated
under reduced pressure. The crude material was purified by silica
gel column chromatography to afford the title compound (2.0 g, 82%)
as a mixture of cis/trans-isomers which was carried forward as a
mixture through to the final compound 309 which was also isolate as
a mixture of cis/trans-isomers.
Step 5: synthesis of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-5-(2-hydroxye-
thoxy)-2-methylbenzoate
##STR00977##
[2048] To a stirred solution of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(2-hydroxyethoxy)-2-
-methylbenzoate (2.0 g, 4.7 mmol) and acetaldehyde (0.63 g, 14.2
mmol) in dichloroethane (20 mL) was added acetic acid (1.7 g, 28
mmol) and the reaction mixture stirred at room temperature for 20
minutes. Then sodium triacetoxyborohydride (3.0 g, 14.2 mmol) was
added at 0.degree. C. and reaction was stirred at room temperature
for 2 h. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase was separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were washed with water, dried, concentrated under reduced pressure;
Crude material was purified by column chromatography to afford the
title compound (1.9 g, 89%).
Step 6: synthesis of tert-butyl
(4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5--
(2-hydroxyethoxy)-2-methylphenyl)-(ethyl)-amino)cyclohexyl)carbamate
##STR00978##
[2050] Aqueous NaOH (0.25 g, 6.33 mmol) was added to a solution of
methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)amino)-5-(2-hydroxye-
thoxy)-2-methylbenzoate (1.9 g, 4.2 mmol) in ethanol (20 mL) and
the mixture stirred at 60.degree. C. for 1 h. After completion of
the reaction, ethanol was removed under reduced pressure and
acidified using dilute HCl up to pH 6 and pH 4 was adjusted using
citric acid. Extraction was carried out using ethyl acetate. The
combined organic layers were dried and concentrated giving the
corresponding crude acid (1.8 g).
[2051] The acid (1.8 g, 4.1 mmol) was then dissolved in DMSO (15
mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (1.25 g,
8.25 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min, PYBOP (3.22 g, 6.19 mmol) was added and
stirring was continued overnight. After completion of the reaction,
the reaction mixture was poured into ice and extracted with 10%
MeOH/DCM. The combined organic layers were washed with water, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford crude material which was purified by
column chromatography to give the title compound (1.4 g, 59%).
Step 7: synthesis of
3-((4-aminocyclohexyl)-(ethyl)-amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydrop-
yridin-3-yl)methyl)-5-(2-hydroxyethoxy)-2-methylbenzamide
##STR00979##
[2053] To a solution of tert-butyl
(4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5--
(2-hydroxyethoxy)-2-methylphenyl)-(ethyl)-amino)cyclohexyl)carbamate
(0.80 g, 1.40 mmol) in DCM (5 mL) was added TFA (2 mL) and the
reaction mixture was stirred at rt for 1 h. After completion of
reaction, the solvent was removed under reduced pressure and
saturated NaHCO.sub.3 solution was added. Extraction was carried
out using 10% MeOH/DCM; the combined organic layers were washed
with water and brine; dried over anhydrous Na.sub.2SO.sub.4;
filtered and concentrated under reduced pressure to give the title
compound (0.5 g, 76%).
Step 8: synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)(ethyl)amino)-5-(2-hydroxyethoxy)-2-methylbenzamide
##STR00980##
[2055] To a solution of
3-((4-aminocyclohexyl)-(ethyl)-amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydrop-
yridin-3-yl)methyl)-5-(2-hydroxyethoxy)-2-methylbenzamide (0.30 g,
0.63 mmol) in DCM (3 mL) was added formalin (0.07 g, 2.23 mmol) at
0.degree. C. The resulting reaction mixture was stirred at
0.degree. C. for 20 minutes. Sodium triacetoxyborohydride (0.34 g,
1.59 mmol) was added and the reaction mixture stirred at room
temperature for 1 h. After completion, solvent was removed under
reduced pressure and water was added to the residue and extraction
was carried out using 10% MeOH/DCM. The combined organic layers
were dried and concentrated under reduced pressure. The solid
obtained was further purified by washings with acetonitrile and
ether to afford the title compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)(ethyl)amino)-5-(2-hydroxyethoxy)-2-methylbenzamide
as a mixture of cis/trans-isomers (0.25 g, 78%). LCMS: 499.55
(M+1).sup.+; HPLC: 51.61 & 47.78% (@210-370 nm) (R.sub.t; 3.539
& 3.751); Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d6, 400 MHz) .delta.
11.45 (bs, 1H), 8.04 (s, 1H), 6.68 (s, 1H), 6.51 (s, 1H), 5.85 (s,
1H), 4.25 (d, 2H), 3.92 (t, 2H), 3.67 (t, 2H), 2.97 (m, 2H), 2.18
(s, 3H), 2.16 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H), 2.08 (s, 3H),
1.89 (s, 3H), 1.76 (m, 3H), 1.65 (bs, 1H), 1.33 (m, 3H), 1.10 (m,
1H), 0.788 (bs, 3H).
Compound 310:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)-(ethyl)-amino)-5-(2-methoxyethoxy)-2-methylbenzamide
##STR00981##
[2056] Step 1: synthesis of methyl
5-(2-methoxyethoxy)-2-methyl-3-nitrobenzoate
##STR00982##
[2058] To stirred solution of methyl
5-hydroxy-2-methyl-3-nitrobenzoate (1.50 g, 7.61 mmol) in ACN (15
mL) was added cesium carbonate (4.96 g, 15.2 mmol) and
1-bromo-2-methoxyethane (1.57 g, 11.4 mmol). The resulting reaction
mixture was heated at 80.degree. C. overnight. On completion, the
reaction mixture was diluted with water and extraction was carried
out using ethyl acetate. The combined organic layers were washed
with water, dried, and concentrated under reduced pressure. The
crude material was purified by column chromatography to afford the
title compound (1.3 g, 68).
Step 2: synthesis of methyl
3-amino-5-(2-methoxyethoxy)-2-methylbenzoate
##STR00983##
[2060] To stirred solution of methyl
5-(2-methoxyethoxy)-2-methyl-3-nitrobenzoate (1.35 g, 4.83 mmol) in
ethanol (20 mL) were added ammonium chloride (1.35 g, 25.0 mmol)
dissolved in water (20 mL) and iron powder (1.07 g, 19.3 mmol). The
resulting reaction mixture was heated at 80.degree. C. for 3 h. On
completion, the reaction mixture was filtered and the residue
washed well with hot ethanol. The filtrate was concentrated and the
residue was basified by aqueous sodium bicarbonate. The aqueous
phase was extracted with ethyl acetate. The combined organic layers
were washed with water, dried, concentrated under reduced pressure
to give the crude title compound (1.23 g) which was used directly
in the next step.
Step 3: synthesis of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(2-methoxyethoxy)-2-
-methylbenzoate
##STR00984##
[2062] To a stirred solution of methyl
3-amino-5-(2-methoxyethoxy)-2-methylbenzoate (1.23 g, 5.14 mmol)
and tert-butyl (4-oxocyclohexyl)carbamate (1.42 g, 6.69 mmol) in
dichloroethane (15 mL) was added acetic acid (1.8 g, 31 mmol) and
the reaction mixture stirred at room temperature for 20 minutes.
Then sodium triacetoxyborohydride (3.2 g, 15 mmol) was added at
0.degree. C. and the reaction was stirred at room temperature
overnight. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were washed with water, dried and concentrated under reduced
pressure. The crude material was purified by column chromatography
to afford methyl the title compound (1.6 g, 72%) as a mixture of
cis/trans-isomers which was carried forward through to the final
compound as a cis/trans-isomer mixture.
Step 4: synthesis of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)-amino)-5-(2-methoxy-
ethoxy)-2-methylbenzoate
##STR00985##
[2064] To a stirred solution of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(2-methoxyethoxy)-2-
-methylbenzoate (1.60 g, 3.67 mmol) and acetaldehyde (0.48 g, 10.9
mmol) in dichloroethane (20 mL) was added acetic acid (1.3 g, 22
mmol) and the reaction mixture stirred at room temperature for 20
minutes. Then sodium triacetoxyborohydride (2.33 g, 11.0 mmol) was
added at 0.degree. C. and the reaction was stirred at room
temperature for 2 h. The reaction was quenched with aqueous sodium
bicarbonate, the organic phase separated and the aqueous phase
extracted with dichloromethane. The combined organic layers were
washed with water, dried, concentrated under reduced pressure, the
crude material was purified by column chromatography to afford the
title compound (1.6 g, 94%).
Step 5: synthesis of tert-butyl
(4-((3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-5--
(2-methoxyethoxy)-2-methylphenyl)-(ethyl)-amino)-cyclohexyl)carbamate
##STR00986##
[2066] Aqueous NaOH (0.2 g, 5.3 mmol) was added to a solution of
methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)-amino)-5-(2-methoxy-
ethoxy)-2-methylbenzoate (1.6 g, 3.5 mmol) in ethanol (20 mL) and
stirred at 60.degree. C. for 1 h. After completion of the reaction,
ethanol was removed under reduced pressure and acidified using
dilute HCl up to pH 6 and pH 4 was adjusted using citric acid.
Extraction was carried out using ethyl acetate. The combined
organic layers were concentrated to give the corresponding crude
acid (1.4 g).
[2067] The acid (1.4 g, 3.1 mmol) was then dissolved in DMSO (15
mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.95 g,
6.2 mmol) was added. The reaction mixture was stirred at room
temperature for 15 min, PYBOP (2.4 g, 4.6 mmol) was added and
stirring was continued overnight. The reaction mixture was poured
into ice and extracted with 10% MeOH/DCM. The combined organic
layers were washed with water, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure
to afford crude material which was purified by column
chromatography to afford the title compound (1.9 g, 95%).
Step 5: synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)-(ethyl)-amino)-5-(2-methoxyethoxy)-2-methylbenzamide
##STR00987##
[2069] To a solution of methyl
3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)-(ethyl)-amino)-5-(2-methoxy-
ethoxy)-2-methylbenzoate (1.2 g, 2.67 mmol) in DCM (10 mL) was
added TFA (2 mL) and the reaction mixture stirred at rt for 1 h.
After completion of reaction, the solvent was removed under reduced
pressure and saturated NaHCO.sub.3 solution was added to the
residue. Extraction was carried out using 10% MeOH/DCM; the
combined organic layers were washed with water and brine; dried
over anhydrous Na.sub.2SO.sub.4; filtered and concentrated under
reduced pressure to give the Boc deprotected-compound (0.78
g,).
[2070] The Boc deprotected compound (0.30 g, 0.62 mmol) was
dissolved in DCM (3 mL) and formalin (0.07 g, 2.33 mmol) was added
at 0.degree. C. The resulting reaction mixture was stirred at
0.degree. C. for 20 minutes. Sodium triacetoxyborohydride (0.33 g,
1.56 mmol) was added and the reaction mixture was stirred at room
temperature for 1 h. After completion, the solvent was removed
under reduced pressure and water was added to the residue,
extraction was carried out using 10% MeOH/DCM. The combined organic
layers were dried and concentrated under reduced pressure to afford
crude material which was purified by prep. HPLC to give the title
compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)-(ethyl)-amino)-5-(2-methoxyethoxy)-2-methylbenzamide
as a mixture of cis/trans-isomers (0.02 g, 6.3%). LCMS: 513.60
(M+1).sup.+; HPLC: 44.48% & 50.77% (@210-370 nm) (R.sub.t;
3.879 & 4.316); Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d6, 400 MHz) .delta.
11.45 (s, 1H), 8.04 (s, 1H), 6.68 (s, 1H), 6.52 (s, 1H), 5.85 (s,
1H), 4.25 (d, 2H, J=4.4 Hz), 4.03 (bs, 2H), 3.61 (bs, 2H), 3.28 (s,
3H), 3.10 (m, 1H), 2.95 (m, 2H), 2.18 (s, 6H), 2.12 (s, 3H), 2.10
(s, 3H), 2.08 (s, 3H), 1.77 (m, 2H), 1.65 (bs, 2H), 1.33 (m, 3H),
1.15 (m, 1H), 0.78 (t, 3H, J=4 Hz).
Compound 311:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)-(ethyl)-amino)-5-(ethylthio)-2-methylbenzamide
##STR00988##
[2071] Step 1: synthesis of 5-bromo-2-methyl-3-nitrobenzoic
acid
##STR00989##
[2073] To stirred solution of 2-methyl-3-nitrobenzoic acid (50.0 g,
276 mmol) in conc. H.sub.2SO.sub.4 (200 mL) was added
1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (43.4 g, 151.8
mmol) portion-wise at room temperature and the reaction mixture was
stirred at room temperature for 5 h. On completion, the reaction
mixture was poured on ice cold water, the resulting precipitate was
filtered, washed with water, and dried under vacuum giving desired
crude title compound (71.7 g, 99.9%) which was used directly in the
next step.
Step 2: synthesis of methyl 5-bromo-2-methyl-3-nitrobenzoate
##STR00990##
[2075] To stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid
(285 g, 1104 mmol) in DMF (2.8 L) was added sodium carbonate (468
g, 4415 mmol) followed by addition of methyl iodide (626 g, 4415
mmol) at room temperature. The resulting reaction mixture was
heated at 60.degree. C. for 8 h. After completion, the reaction
mixture was filtered and washed with ethyl acetate. The combined
filtrates were washed with water and the aqueous phase was back
extracted with ethyl acetate. The combined organic layers were
dried over anhydrous sodium sulphate, filtered and concentrated
under reduced pressure to afford the title compound as an off-white
solid (290 g, 97% yield). The isolated compound was taken directly
into the next step.
Step 3: synthesis of methyl 3-amino-5-bromo-2-methylbenzoate
##STR00991##
[2077] To stirred solution of methyl
5-bromo-2-methyl-3-nitrobenzoate (290 g, 1060 mmol) in ethanol (1.5
L) was added aqueous ammonium chloride (283 g, 5290 mmol dissolved
in 1.5 L water). The resulting mixture was stirred and heated at
80.degree. C. followed by addition of iron powder (472 g, 8450
mmol) in portions at 80.degree. C. The resulting reaction mixture
was heated at 80.degree. C. for 12 h. After completion, the
reaction mixture was hot filtered over celite and the celite bed
was washed with methanol (5 L) followed by washing with 30% MeOH in
DCM (5 L). The combined filtrate was concentrated in-vacuo, the
residue obtained was diluted with aqueous bicarbonate and extracted
with ethyl acetate. The combined organic layers were dried over
anhydrous sodium sulphate, filtered and concentrated under reduced
pressure to afford the title compound as brown solid (220 g, 89%
yield).
Step 4: synthesis of methyl
5-bromo-3-(((1R,4R)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-m-
ethylbenzoate
##STR00992##
[2079] To a stirred solution of methyl
3-amino-5-bromo-2-methylbenzoate (15.0 g, 617 mmol) and tert-butyl
(4-oxocyclohexyl)carbamate (17.1 g, 80.2 mmol) in dichloroethane
(150 mL) was added acetic acid (22.2 g, 370 mmol) and the reaction
mixture stirred at room temperature for 10 minutes. Then sodium
triacetoxyborohydride (39.3 g, 185 mmol) was added at 0.degree. C.
and the reaction stirred overnight at room temperature. On
completion, the reaction was quenched with aqueous sodium
bicarbonate, the organic phase separated and the aqueous phase
extracted with dichloromethane. The combined organic layers were
washed with water, dried, and concentrated under reduced pressure.
The crude material was purified by column chromatography to afford
the title compound methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-m-
ethylbenzoate (10.5 g, 39%).
Step 5: synthesis of methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-a-
mino)-2-methylbenzoate
##STR00993##
[2081] To a stirred solution of
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)amino)-2-m-
ethylbenzoate (10.0 g, 22.7 mmol) and acetaldehyde (2.99 g, 68
mmol) in dichloroethane (100 mL) was added acetic acid (8.18 g, 136
mmol) and the reaction mixture was stirred at room temperature for
20 minutes. Then sodium triacetoxyborohydride (14.5 g, 68.1 mmol)
was added at 0.degree. C. and the reaction stirred overnight at
room temperature. On completion, the reaction was quenched with
aqueous sodium bicarbonate, the organic phase separated and the
aqueous phase extracted with dichloromethane. The combined organic
layers were washed with water, dried, concentrated under reduced
pressure, crude material was purified by column chromatography to
afford the title compound (9.0 g, 84%).
Step 6: synthesis of tert-butyl
((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)-(ethyl)-amino)-cyclohexyl)carbamate
##STR00994##
[2083] Aqueous NaOH (1.15 g, 28.8 mmol) was added to a solution of
methyl
5-bromo-3-(((1r,4r)-4-((tert-butoxycarbonyl)-amino)-cyclohexyl)-(ethyl)-a-
mino)-2-methylbenzoate (9.0 g, 19 mmol) in ethanol (100 mL) and the
mixture stirred at 60.degree. C. for 1 h. After completion of the
reaction, ethanol was removed under reduced pressure and the
residue was acidified using dilute HCl up to pH 6 and adjusted to
pH 4 with citric acid. Extraction was carried out using 10%
MeOH/DCM. The combined organic layers were dried and concentrated
to give the crude acid (8.6 g,).
[2084] The acid (8.6 g, 19 mmol) was then dissolved in DMSO (10 mL)
and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (5.74 g, 37.8
mmol) was added. The reaction mixture was stirred at room
temperature for 15 min, PYBOP (14.7 g, 2845 mmol) was added and
stirring was continued overnight. The reaction mixture was poured
into ice and extracted with 10% MeOH/DCM. The combined organic
layers were dried, concentrated to obtain crude material which was
purified by washing with acetonitrile to afford the title compound
(10.2 g, 92%).
Step 7: synthesis of
3-(((1r,4r)-4-aminocyclohexyl)-(ethyl)-amino)-5-bromo-N-((4,6-dimethyl-2--
oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR00995##
[2086] To a solution of tert-butyl
((1r,4r)-4-((5-bromo-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)meth-
yl)carbamoyl)-2-methylphenyl)-(ethyl)-amino)-cyclohexyl)carbamate
(3.0 g, 5.10 mmol) in DCM (20 mL) was added TFA (5 mL) and the
reaction mixture was stirred at rt for 1 h. After completion of
reaction, the solvent was removed under reduced pressure and
saturated NaHCO.sub.3 solution was added to the residue. Extraction
was carried out using 10% MeOH/DCM; the combined organic layers
were washed with water and brine; dried over anhydrous
Na.sub.2SO.sub.4; filtered and concentrated under reduced pressure
to give the title compound (2.2 g, 88%) which was used directly in
the next step.
Step 8: synthesis of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-2-methylbenzamide
##STR00996##
[2088] To a solution of
3-(((1r,4r)-4-aminocyclohexyl)-(ethyl)-amino)-5-bromo-N-((4,6-dimethyl-2--
oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (2.20 g, 4.50
mmol) in DCM (25 mL) was added formalin (0.49 g, 16.3 mmol) at
0.degree. C. The resulting reaction mixture was stirred at
0.degree. C. for 20 minutes. Sodium triacetoxyborohydride (2.39 g,
11.2 mmol) was added and the reaction mixture was stirred at room
temperature for 1 h. After completion, water was added and
extraction was carried out using 10% MeOH/DCM. The combined organic
layers were dried and concentrated under reduced pressure to give
the title compound (2.3 g, 98%) which was used directly in the next
step.
Step 9: synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)-(ethyl)-amino)-5-(ethylthio)-2-methylbenzamide
##STR00997##
[2090] To a stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-2-methylbenzamide
(0.40 g, 0.77 mmol) in dioxane was added ethanethiol (0.048 g, 0.77
mmol) and DIPEA (2.70 mL, 1.55 mmol) and the reaction mixture then
purged with argon for 10 min. Then, Pd (OAc).sub.2 (0.009 g, 0.038
mmol) and Xantphos (0.045 g, 0.077 mmol) were added and the
reaction mixture was stirred at 100.degree. C. overnight. After
completion of the reaction, water was added and extraction was
carried out using ethyl acetate. The combined organic layers were
washed with water, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure to afford crude material
which was purified by column chromatography to give the title
compound (0.17 g, 44%). LCMS: 499.55 (M+1).sup.+; HPLC: 99.30% (at
210-370 nm) (R.sub.t; 4.264); Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 11.46 (bs, 1H), 9.37 (s, 1H), 8.14 (s, 1H), 7.08 (s,
1H), 6.89 (s, 1H), 5.86 (s, 1H), 4.25 (d, 2H, J=4 Hz), 3.15-2.90
(m, 6H), 2.69 (s, 3H), 2.68 (s, 3H), 2.18 (s, 3H), 2.14 (s, 3H),
2.10 (s, 3H), 1.94 (m, 2H), 1.85 (m, 2H), 1.41 (m, 4H), 1.20 (t,
3H, J=7.2), 0.78 (t, 3H, J=6 Hz).
Compound 313:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
4-hydroxycyclohexyl)-amino)-2-methylbenzamide
##STR00998##
[2091] Step 1: synthesis of methyl
3-(1,4-dioxaspiro[4.5]decan-8-ylamino)-5-chloro-2-methylbenzoate
##STR00999##
[2093] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (5.0 g, 25 mmol) and
1,4-dioxaspiro[4.5]decan-8-one (5.86 g, 37.8 mmol) in
dichloroethane (50 mL) was added acetic acid (9.0 g, 150 mmol) and
the reaction mixture stirred at room temperature for 10 minutes.
Then sodium triacetoxyborohydride (15.9 g, 752 mmol) was added at
0.degree. C. and the reaction stirred overnight at room
temperature. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were dried over anhydrous sodium sulphate and concentrated under
reduced pressure. The crude material was purified by silica gel
column chromatography to afford the title compound (6.6 g,
76%).
Step 2: synthesis of methyl
5-chloro-3-(ethyl(1,4-dioxaspiro[4.5]decan-8-yl)amino)-2-methylbenzoate
##STR01000##
[2095] To a stirred solution of methyl
3-(1,4-dioxaspiro[4.5]decan-8-ylamino)-5-chloro-2-methylbenzoate
(6.6 g, 19 mmol) and acetaldehyde (2.56 g, 58.4 mmol) in
dichloroethane (70 mL) was added acetic acid (7.0 g, 120 mmol) and
the reaction mixture stirred at room temperature for 10 minutes.
Then sodium triacetoxyborohydride (12 g, 57 mmol) was added at
0.degree. C. and the reaction stirred overnight at room
temperature. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were dried over anhydrous sodium sulphate and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to afford the title compound (4.0 g, 56%).
Step 3: synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
1,4-dioxaspiro[4.5]decan-8-yl)amino)-2-methylbenzamide
##STR01001##
[2097] Aqueous NaOH (1.5 g, 38 mmol) was added to a solution of
methyl
5-chloro-3-(ethyl(1,4-dioxaspiro[4.5]decan-8-yl)amino)-2-methylbenzoate
(6.9 g, 19 mmol) in ethanol (70 mL) and the mixture stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and adjusted to pH 4 with citric acid. Extraction was
carried out using ethyl acetate. The combined organic layers were
dried concentrated to give the crude acid (6.5 g).
[2098] The crude acid (6.5 g, 18 mmol) was dissolved in DMSO (50
mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (5.6 g, 37
mmol) was added. The reaction mixture was stirred at room
temperature for 15 min, PYBOP (14.3 g, 27.5 mmol) was added and
stirring was continued overnight. The reaction mixture was poured
into ice and extracted with 10% MeOH/DCM. The combined organic
layers were washed with water, dried, concentrated under reduced
pressure to give the title compound (5.9 g 66%) which was used
directly in the next step.
Step 4: synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
4-oxocyclohexyl)amino)-2-methylbenzamide
##STR01002##
[2100] To a stirred solution of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
1,4-dioxaspiro[4.5]decan-8-yl)amino)-2-methylbenzamide (2.0 g, 4.1
mmol) in acetone: water (14 mL+6 mL) was added PTSA (3.1 g, 16
mmol). The resulting reaction mixture was stirred at 70.degree. C.
overnight. On completion, the solvent was removed under reduced
pressure and the residue was basified with aqueous sodium
bicarbonate. The aqueous phase was extracted with ethyl acetate.
The combined organic layers were washed with water, dried,
concentrated under reduced pressure to give the title compound (1.5
g, 83%) which was used directly in the next step.
Step 5: synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
4-hydroxycyclohexyl)-amino)-2-methylbenzamide
##STR01003##
[2102] To a stirred solution of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
4-oxocyclohexyl)amino)-2-methylbenzamide (0.30 g, 0.68 mmol) in
MeOH was added NaBH.sub.4 (0.038 g, 1.01 mmol) slowly at 0.degree.
C. The resulting reaction mixture was stirred at 0.degree. C. for 2
h. On completion, the reaction was quenched with water and
extracted with ethyl acetate. The combined organic layers were
washed with water, dried, concentrated under reduced pressure and
the crude material obtained was purified by acetonitrile and ether
washings to afford the title compound as a mixture of isomers (175
mg, 58%). LCMS: 446.35 (M+1).sup.+; HPLC: 74.32% & 24.18%
(@210-370 nm) (R.sub.t; 5.157 & 5.276); Method: Column: YMC
ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA
in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col.
Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95%
B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d6, 400 MHz) .delta. 11.49 (s, 1H), 8.25 (s, 1H), 7.17 (s,
1H), 6.96 (s, 1H), 5.86 (s, 1H), 4.25 (d, 2H, J=4 Hz), 3.33 (m,
1H), 3.04 (m, 2H), 2.19 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 1.78
(m, 2H), 1.66 (m, 2H), 1.38 (m, 3H), 1.10 (m, 2H), 0.78 (t,
3H).
Compound 315:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(tetrahydro-2H-pyran-4-yl)b-
enzamide
##STR01004##
[2103] Step 1: Synthesis of
5-(3,6-dihydro-2H-pyran-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-
-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-
benzamide
##STR01005##
[2105] To a stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (0.30
g, 0.58 mmol) and
2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(0.18 g, 0.88 mmol) in dioxane-water mixture was added
Na.sub.2CO.sub.3 (0.22 g, 2.07 mmol). The solution was purged with
argon for 15 min, Pd (PPh.sub.3).sub.4 (0.067 g, 0.05 mmol) was
added and the solution was purged again with argon for 10 min. The
reaction mixture was heated at 100.degree. C. for 3 hours. On
completion, the reaction mixture was diluted with water and
extracted with 10% MeOH/DCM. The combined organic layers were dried
over Na.sub.2SO.sub.4 and the solvent removed under reduced
pressure to afford the title compound (0.30 g, 98%).
Step 2: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(tetrahydro-2H-pyran-4-yl)b-
enzamide
##STR01006##
[2107] To a stirred solution of
5-(3,6-dihydro-2H-pyran-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-
-yl)methyl)-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methyl-
benzamide (0.3 g, 0.576 mmol) in methanol, was added a catalytic
amount of 10% Pd/C. The reaction mixture was stirred at room
temperature under hydrogen balloon pressure for 1 hour. On
completion, the reaction mixture was filtered through a celite bed,
the celite washed with methanol and the filtrate concentrated under
reduced pressure, to yield a crude compound which was purified by
prep. HPLC to afford the target compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1s,4s)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(tetrahydro-2H-pyran-4-yl)b-
enzamide as the TFA salt (0.08 g, 26%).
[2108] LCMS: 523.60 (M+1).sup.+; HPLC: 48.41+51.05% (@210 nm-370
nm) (R.sub.t; 3.93& 3.99); Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.47 (bs, 1H), 8.39 (t, 1H), 8.08
(s, 1H), 7.07 (s, 1H), 6.86 (s, 1H), 5.87 (s, 1H), 4.26 (d, 2H,
J=2.8 Hz), 3.94-3.92 (m, 2H), 3.41 (m, 2H), 3.09-2.89 (m, 3H), 2.69
(s, 3H+3H), 2.19 (s, 3H+3H), 2.11 (s, 3H), 1.99-1.86 (m, 5H), 1.64
(m, 4H), 1.42 (m, 3H), 1.25 (m, 2H), 0.79 (t, 3H).
Compound 319:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(ethylsulfonyl)-2-methylbenzamide
##STR01007##
[2109] Step 1: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(ethylthio)-2-methylbenzamide
[2110] To a stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (0.4
g, 0.77 mmol.) in dioxane was added ethanethiol (0.048 g, 0.77
mmol) and DIPEA (2.7 mL, 1.55 mmol). The solution was purged with
argon for 10 min, Pd (OAc).sub.2 (0.009 g, 0.038 mmol) and Xantphos
(0.045 g, 0.077 mmol) were added to it and argon was purged again
for 10 min. The reaction mixture was stirred at 100.degree. C. for
overnight, water was and extraction was carried out using ethyl
acetate. The combined organic layers were washed with water, dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure to afford crude material which was purified by
column chromatography to afford compound the title compound (0.17
g, 44%). LCMS: 499.55 (M+1).sup.+; HPLC: 99.30% (@210-370 nm)
(R.sub.t; 4.264); Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d6, 400 MHz) .delta.
11.46 (bs, 1H), 9.37 (s, 1H), 8.14 (s, 1H), 7.08 (s, 1H), 6.89 (s,
1H), 5.86 (s, 1H), 4.25 (d, 2H, J=4 Hz), 3.15-2.90 (m, 6H), 2.69
(s, 3H), 2.68 (s, 3H), 2.18 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H),
1.94 (m, 2H), 1.85 (m, 2H), 1.41 (m, 4H), 1.20 (t, 3H, J=7.2), 0.78
(t, 3H, J=6 Hz).
Step 2: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(ethylsulfonyl)-2-methylbenzamide
##STR01008##
[2112] To a stirred solution of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-(ethylthio)-2-methylbenzamide
(0.085 g, 0.17 mmol.) in methanol was added oxone (0.105 g, 0.34
mmol) at room temperature. The reaction mixture was stirred at room
temperature for 4 h, water was added and extraction was carried out
using 10% MeOH/DCM. The combined organic layers were washed with
water, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to afford crude material which
was purified by prep. HPLC to afford the title compound as the TFA
salt (0.07 g, 77%). LCMS: 531.60 (M+1).sup.+; HPLC: 89.24% (@210
nm-370 nm) (R.sub.t; 4.082; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.47 (s, 1H), 9.36 (s, 1H), 8.36
(s, 1H), 7.53 (s, 1H), 7.38 (s, 1H), 7.22 (m, 1H), 7.09 (s, 1H),
6.96 (s, 1H), 5.87 (s, 1H), 4.29-4.28 (d, 2H, J=4.8 Hz), 3.0-3.20
(m, 4H), 2.60-2.80 (m, 8H), 2.27 (s, 9H), 2.20 (s, 3H), 2.11 (s,
3H), 1.90-2.0 (m, 2H), 1.80-1.90 (m, 2H), 1.40-1.50 (m, 4H),
1.05-1.15 (t, 3H, J=7.6 Hz), 0.75-0.85 (t, 3H, J=6.8 Hz).
Compound 320:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-5-isopropyl-2-methylbenzamide
##STR01009##
[2113] Step 1: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(prop-1-en-2-yl)benzamide
##STR01010##
[2115] To a stirred solution of
5-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r-
)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-2-methylbenzamide (0.30
g, 0.58 mmol) and 4, 4,
5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (0.147 g,
0.875 mmol) in dioxane/water mixture was added Na.sub.2CO.sub.3
(0.22 g, 2.07 mmol) and the solution purged with argon for 15 min.
Then Pd (PPh.sub.3).sub.4 (0.067 g, 0.058 mmol) was added and argon
was purged again for 10 min. The reaction mixture was heated at
100.degree. C. for 2 hours diluted with water and extracted with
10% MeOH/DCM. The combined organic layers were dried over
Na.sub.2SO.sub.4 and the solvent removed under reduced pressure to
afford crude material which was purified by column chromatography
over silica gel to afford the title compound (0.20 g, 71%).
Step 2:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-
-4-(dimethylamino)cyclohexyl)(ethyl)amino)-5-isopropyl-2-methylbenzamide
##STR01011##
[2117] To a stirred solution of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(((1r,4r)-4-(dim-
ethylamino)cyclohexyl)(ethyl)amino)-2-methyl-5-(prop-1-en-2-yl)benzamide
(0.20 g, 0.41 mmol) in methanol was added a catalytic amount of 10%
Pd/C. The reaction mixture was stirred at room temperature under
hydrogen pressure (balloon pressure) for 2 h. The mixture was
filtered through a celite bed, washed with methanol and the
filtrate was concentrated under reduced pressure. The resulting
crude compound was purified by prep. HPLC to afford the title
compound as the TFA salt (0.07 g, 35%). LCMS: 481.40 (M+1).sup.+;
HPLC: 97.01% (@210 nm-370 nm) (R.sub.t; 3.996; Method: Column: YMC
ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA
in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col.
Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95%
B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.46 (bs, 1H), 9.37 (bs, 1H), 8.00
(s, 1H), 7.01 (s, 1H), 6.83 (s, 1H), 5.86 (s, 1H), 4.26-4.25 (d,
2H, J=4.0 Hz), 3.0-3.25 (m, 3H), 2.70-2.8 (m, 1H), 2.60-2.75 (m,
6H), 2.19 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 1.90-2.0 (m, 2H),
1.80-1.90 (m, 2H), 1.35-1.50 (m, 4H), 1.16-1.20 (d, 6H, J=6.8 Hz),
0.75-0.85 (t, 3H).
Compound 321:
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-N-((4-et-
hyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01012##
[2118] Step 1: synthesis of methyl
5-chloro-2-methyl-3-nitrobenzoate
##STR01013##
[2120] To stirred solution of 5-chloro-2-methyl-3-nitrobenzoic acid
(126 g, 0.59 mol) in DMF (1.0 L) was added sodium carbonate (249 g,
2.34 mol) and methyl iodide (145 mL, 2.34 mol). The reaction
mixture was heated at 60.degree. C. for 4 h. On completion, the
reaction mixture was filtered and the residue washed with DCM. The
aqueous phase was separated and extracted with DCM. The combined
organic layers were dried, concentrated under reduced pressure and
purified by column chromatography over silica eluting with ethyl
acetate: hexane to afford the title compound (85 g, 63%). .sup.1H
NMR (DMSO-d.sub.6, 400 MHz): .delta. 8.25 (d, 1H), 8.05 (s, 1H),
3.9 (s, 1H), 2.4 (s, 1H).
Step 2: synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR01014##
[2122] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (85 g, 0.37 mol) in ethanol (425
mL) were added ammonium chloride (85 g, 112 mmol) dissolved in
water (425 mL) and iron powder (169 g, 2.96 mol) with stirring. The
resulting reaction mixture was heated at 80.degree. C. for 16 h
filtered and the residue washed well with hot ethanol. The filtrate
was concentrated and the residue was basified with aqueous sodium
bicarbonate. The aqueous phase was extracted with ethyl acetate.
The combined organic layers were washed with water, dried,
concentrated under reduced pressure to give the crude title
compound (70 g) which was used directly. .sup.1H NMR (DMSO-d.sub.6,
400 MHz): .delta. 6.8 (s, 2H), 5.45 (s, 2H), 3.75 (s, 3H), 2.15 (s,
3H)
Step 3: synthesis of methyl 3-(((1r,
4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-methylbenz-
oate
##STR01015##
[2124] To a stirred solution of methyl 3-amino-5-chloro-2-methyl
benzoate (12 g, 60 mmol) and tert-butyl (4-oxocyclohexyl)carbamate
(15.4 g, 72 mmol) in dichloroethane (120 mL) was added acetic acid
(21 g, 360 mmol) and the reaction mixture was stirred at room
temperature for 10 minutes. Then sodium triacetoxyborohydride (38.2
g, 180 mmol) was added and the reaction stirred for 2 h at room
temperature. On completion, water was added to the reaction mixture
and extraction was carried out using DCM. The combined organic
layers were washed with bicarbonate solution, dried over sodium
sulphate and concentrated under reduced pressure to give crude
material which was then column purified to give trans-isomer title
compound methyl 3-(((1r,
4rs)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-methylben-
zoate (8.0 g, 33%).
Step 4: synthesis of methyl 3-(((1r,
4r)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)-(ethyl)amino)-5-chloro-2-m-
ethylbenzoate
##STR01016##
[2126] To a stirred solution of methyl 3-(((1r,
4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-methylbenz-
oate (11.4 g, 28.8 mmol) and acetaldehyde (2.5 g, 56 mmol) in
dichloroethane (120 mL) was added acetic acid (10.7 g, 169 mmol)
and the reaction mixture was stirred at room temperature for 10
minutes. Then sodium triacetoxyborohydride (17.95 g, 84.6 mmol) was
added and reaction stirred for 2 h at room temperature. On
completion, water was added to the reaction mixture and extraction
was carried out using DCM. Combined organic layers were washed with
bicarbonate solution, dried over sodium sulphate and concentrated
under reduced pressure to give crude material which was then column
purified to give the title compound (9.8 g, 80%).
Step 5: synthesis of methyl 5-chloro-3-(((1r,
4r)-4-(dimethylamino)-cyclohexyl)-(ethyl)amino)-2-methylbenzoate
##STR01017##
[2128] To a stirred solution of methyl 3-(((1r,
4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-methylbenz-
oate (9.8 g, 23 mmol) in DCM (50 mL) at 0.degree. C. was added TFA
(10 mL) and the reaction mixture was stirred for 2 h at room
temperature. After completion, the reaction was concentrated to
dryness. The residue was then basified with aqueous sat.
bicarbonate solution (100 mL) to pH 8 and the aqueous layer
extracted with 20% methanol in DCM (100 mL.times.4). The combined
organic layers were dried over Na.sub.2SO.sub.4 and the solvent
removed under reduced pressure to afford crude amine (7.4 g) which
was used as such for next reaction.
[2129] To a stirred solution of the amine (7.4 g, 23 mmol) in DCM
(70 mL) at 0.degree. C., was added 37-41% aq. formalin solution
(2.4 g, 81 mmol) and the reaction mixture stirred at room
temperature for 10 min. NaBH(OAc).sub.3 (12.1 g, 57 mmol) was then
added and the reaction stirred for 2 h. On completion, the reaction
mixture was quenched with water. MeOH (8 mL) was added and the
layers were separated. Extraction was performed with 10% MeOH in
DCM and the combined organic phases were dried and concentrated.
The residue was column purified over basic alumina to afford the
title compound (8.0 g, 99%).
Step 6: synthesis of 5-chloro-3-(((1r,
4r)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-2-methyl-benzoic
acid
##STR01018##
[2131] Aqueous NaOH (1.8 g, 45 mmol in 7 mL H.sub.2O) was added to
a solution of compound methyl 3-(((1r,
4r)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)-(ethyl)amino)-5-chloro-2-m-
ethylbenzoate (8.0 g, 23 mmol) in ethanol (70 mL) and stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and the residue acidified using
dilute HCl up to pH 6 and adjusted to pH 4 with citric acid.
Extraction was carried out using 10% MeOH in DCM. The combined
organic layers were dried over Na.sub.2SO.sub.4 and concentrated to
afford the title compound (7.6 g, 99%).
Step 7: synthesis of
3-(aminomethyl)-4-ethyl-6-methylpyridin-2(1H)-one
##STR01019##
[2133] To a solution of
4-ethyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (1 eq) in
methanol and aq. ammonia solution (9:1) was added a catalytic
amount of Raney Nickel. The reaction mixture was stirred at room
temperature under hydrogen pressure (balloon pressure) for 2-5 h.
On completion of reaction, it was filtered through a celite bed and
the filtrate was concentrated under reduced pressure to afford the
title compound.
Step 8: synthesis of
5-chloro-3-(((1r,4r)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-N-((4-et-
hyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01020##
[2135] The above acid,
5-chloro-3-(((trans)-4-(dimethylamino)cyclohexyl)-(ethyl)-amino)-2-methyl-
-benzoic acid, (1 eq) was dissolved in DMSO and
3-(aminomethyl)-4-ethyl-6-methylpyridin-2(1H)-one (2 eq.) was
added. The reaction mixture was stirred at room temperature for 15
min, PYBOP (1.5 eq.) and triethylamine (1 eq.) were added. The
reaction mixture was stirred overnight. The reaction mixture was
then poured into ice, and extracted with 10% MeOH/DCM. The combined
organic layers were dried, and concentrated to obtain which was
purified by column chromatography followed by prep. HPLC to afford
the title compound as a TFA salt (0.1 g, 70%). LCMS: 487.45
(M+1).sup.+; HPLC: 96.17% (@254 nm) (R.sub.t; 6.026; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.46 (bs, 1H), 9.33
(s, 1H), 8.22 (s, 1H), 7.19 (s, 1H), 6.97 (s, 1H), 5.88 (s, 1H),
4.20-4.25 (d, 2H), 3.0-3.15 (m, 1H+2H), 2.60-2.75 (m, 1H+3H+3H),
2.35-2.45 (m, 2H), 2.20 (s, 3H), 2.15 (s, 3H), 1.90-2.0 (m, 2H),
1.80-1.90 (m, 2H), 1.35-1.50 (m, 4H), 1.10-1.20 (t, 3H, J=7.2 Hz),
0.75-0.85 (t, 3H).
Compound 335:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(di-
methylamino)cyclohexyl)thio)-2-methylbenzamide
##STR01021##
[2136] Step 1: Synthesis of methyl
3-bromo-5-chloro-2-methylbenzoate
##STR01022##
[2138] To stirred solution of CuBr.sub.2 (12.3 g, 55.3 mmol) in
acetonitrile (150 mL), tert-butyl nitrite (7.77 g, 75.4 mmol) was
added at 0.degree. C. and then methyl
3-amino-5-chloro-2-methylbenzoate (10.0 g, 50.3 mmol) dissolved in
acetonitrile was added to it. The resulting reaction mixture was
stirred at 0.degree. C. for 2 hours and then at room temperature
for 18 h. On completion, the reaction mixture was diluted with
water and extracted with ethyl acetate. The combined organic layers
were dried, concentrated under reduced pressure and purified by
silica gel column chromatography giving the title compound (6.6 g,
50%).
Step 2: Synthesis of
3-bromo-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)--
2-methylbenzamide
##STR01023##
[2140] Aqueous NaOH (0.45 g, 11.4 mmol) was added to a solution of
methyl 3-bromo-5-chloro-2-methylbenzoate (2.0 g, 7.6 mmol) in EtOH
(20 mL) and the mixture stirred at 60.degree. C. for 1 hour. After
completion of the reaction, ethanol was removed under reduced
pressure and the residue acidified using dilute HCl up to pH 8 and
then with citric acid till pH 6. The aqueous phase was extracted
with 10% methanol in DCM. The combined organic layers were dried
and concentrated to give respective crude acid intermediate (1.5
g).
[2141] The acid (1.5 g, 6.2 mmol) was dissolved in DMSO (15 mL) and
3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (1.67 g, 12.43
mmol) and triethylamine (0.61 g, 6.01 mmol) was added. The reaction
mixture was stirred at room temperature for 15 min, PyBop (4.81 g,
9.26 mmol) was added and stirring was continued overnight at room
temperature. After completion, the reaction mixture was poured into
ice water. The suspended solid was collected by filtration, washed
well with water and dried. The solid obtained was further purified
by washings with acetonitrile to afford the title compound (2.18 g,
92%).
Step 3: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-(4-oxocyclohexyl)thio)benzamide
##STR01024##
[2143] A solution of
3-bromo-5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)--
2-methylbenzamide (0.35 g, 0.91 mmol.), 4-mercaptocyclohexanone
(0.142 g, 1.09 mmol) and DIPEA (0.235 g, 1.82 mmol) in dioxane was
purged with argon for 10 min. Then, Pd(OAc).sub.2 (0.01 g, 0.044
mmol) and Xantphos (0.052 g, 0.089 mmol) were added and argon was
purged again for 10 min. The reaction mixture was stirred at
100.degree. C. overnight. After completion of the reaction, water
was added and extraction was carried out using ethyl acetate. The
combined organic layers were washed with water, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure to afford crude material which was purified by column
chromatography to afford the title compound (0.50 g, 88%).
Step 4: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(di-
methylamino)cyclohexyl)thio)-2-methylbenzamide
##STR01025##
[2145] To a stirred solution of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl--
3-((4-oxocyclohexyl)thio)benzamide (0.45 g, 1.03 mmol) and
dimethylamine (0.14 g, 3.11 mmol) in dichloroethane (5 mL) was
added acetic acid (0.37 g, 6.16 mmol) and the mixture stirred at
room temperature for 20 minutes. Then sodium triacetoxyborohydride
(0.66 g, 3.11 mmol) was added at 0.degree. C. and the mixture
stirred overnight at room temperature. On completion of the
reaction, the solvent was removed under reduced pressure. Water was
added to the residue and extraction was performed using 5%
MeOH/DCM. The combined organic layers were dried and concentrated
to afford crude material which was purified by column
chromatography to afford the title compound (0.016 g, 3.3%).
Analytical Data: LCMS: 462.35 (M+1).sup.+; HPLC: 89.00% (@210-370
nm) (R.sub.t; 4.949; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
7.44 (s, 1H), 7.16 (s, 1H), 6.10 (s, 1H), 4.44 (s, 2H), 3.63 (m,
1H), 3.13 (m, 1H), 2.38 (s, 3H), 2.36 (s, 3H), 2.35 (s, 3H), 2.31
(s, 3H), 2.24 (s, 3H), 1.75-2.08 (m, 8H).
Compound 336:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-hyd-
roxycyclohexyl)thio)-2-methylbenzamide
##STR01026##
[2147] This compound was also isolated as an additional product
from the purification of compound 335 (0.235 g, 50%) Analytical
Data of: LCMS: 435.30 (M+1).sup.+; HPLC: 99.37% (@210-370 nm)
(R.sub.t; 5.900; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (bs, 1H), 8.31 (t, 1H), 7.72-7.68 (m, 1H), 7.41 (s, 1H), 7.07
(s, 1H), 5.85 (s, 1H), 4.58 (d, 1H, J=4 Hz), 4.24 (d, 2H, J=4 Hz),
4.13 (m, 1H), 3.43 (m, 1H), 2.20 (s, 3H), 2.18 (s, 3H), 2.10 (s,
3H), 1.95-1.80 (m, 1H), 1.75-1.55 (m, 1H), 1.40-1.20 (m, 4H), 0.87
(m, 2H).
Compound 339:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-hydroxy-
-4-methylcyclohexyl)amino)-2-methylbenzamide
##STR01027##
[2148] Step 1: synthesis of methyl 2-methyl-3-nitrobenzoate
##STR01028##
[2150] To a solution of 2-methyl-3-nitrobenzoic acid (10.0 g, 55.0
mmol) in 120 ml of methanol, was added conc. H.sub.2SO.sub.4 (15
ml) at room temperature and the mixture stirred at 60.degree. C.
overnight. The mixture was concentrated under reduced pressure, the
crude material was neutralized with sat. NaHCO.sub.3, and extracted
with ethyl acetate. The organic phase was dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure to afford
the title compound (10 g).
Step 2: synthesis of methyl 3-amino-2-methylbenzoate
##STR01029##
[2152] To a stirred solution of methyl 2-methyl-3-nitrobenzoate
(10.0 g, 56.4 mmol) in 60 ml of ethanol were added ammonium
chloride (18.0 g, 338 mmol) dissolved in 60 mL of water and iron
powder (18.8 g, 338.2 mmol). The resulting reaction mixture was
heated at 80.degree. C. for 3 h. Water was added to the reaction
mixture and then it was filtered through celite. The filtrate was
extracted with ethyl acetate. The combined organic layers were
washed with water, dried over Na.sub.2SO.sub.4, concentrated under
reduced pressure to give the title compound (7.0 g, 83%).
Step 3: synthesis of methyl
3-((4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzoate
##STR01030##
[2154] To a stirred solution of methyl 3-amino-2-methylbenzoate
(1.5 g, 9.1 mmol) and 4-hydroxy-4-methylcyclohexanone (1.51 ml,
11.8 mmol) in 10 ml of dichloroethane was added acetic acid (3.2
mL, 55 mmol) and the reaction mixture stirred at room temperature
for 20 minutes. The reaction mixture was cooled to 0.degree. C. and
sodium triacetoxyborohydride (5.78 g, 27.3 mmol) was added and the
mixture stirred at room temperature overnight. The reaction mixture
was neutralized with sat. NaHCO.sub.3, extracted with DCM, dried
over Na.sub.2SO.sub.4, concentrated under reduced pressure, and
purified by silica gel column chromatography to afford the title
compound which was carried forward as a mixture of cis and trans
isomers (1.4 g, 56%).
Step 4: synthesis of methyl
3-(ethyl(4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzoate
##STR01031##
[2156] To a stirred solution of methyl
3-((4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzoate (1.4 g,
5.05 mmol) and acetaldehyde (0.7 ml, 13 mmol) in 20 ml of
dichloroethane was added acetic acid (1.7 mL, 30 mmol) and the
reaction mixture was stirred at room temperature for 20 minutes.
The reaction mixture was cooled to 0.degree. C. and sodium
triacetoxyborohydride (3.2 g, 15 mmol) was added and the mixture
was stirred at room temperature for overnight. The reaction mixture
was neutralized with sat. NaHCO.sub.3 and extracted with DCM. The
combined organic layers were dried over Na.sub.2SO.sub.4,
concentrated under reduced pressure, and the residue purified by
silica gel column chromatography to afford the title compound (1.42
g, 94%).
Step 5: synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-hydroxy-
-4-methylcyclohexyl)amino)-2-methylbenzamide
##STR01032##
[2158] A mixture of methyl
3-(ethyl(4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzoate (300
mg, 0.98 mmol) and NaOH (58 mg, 1.47 mmol) in 5 ml of ethanol and
water (3:2) was heated at 70.degree. C. for 2 h. The reaction
mixture was concentrated to dryness and the residue was partitioned
between water and ethyl acetate. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to afford
270 mg of the crude acid. The crude acid (270 g, 0.92 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (282 mg, 1.85 mmol),
PyBOP (717 mg, 1.38 mmol) and triethylamine (0.12 ml, 0.92 mmol)
were stirred in 3 ml of DMSO at room temperature overnight. The
reaction mixture was diluted with water and extracted with 10% MeOH
in DCM. The combined organic phases were dried over
Na.sub.2SO.sub.4, concentrated and the residue purified by silica
gel column chromatography to give the title compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzamide as a mixture
of isomers (120 mg, 30%). LCMS: 426.45 (M+1).sup.+; HPLC: 62.09
& 33.24% (@210-370 nm) (R.sub.t; 3.982 & 4.164; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6 400 MHz) .delta. 11.44 (s, 1H), 8.04 (m,
1H), 7.12 (s, 2H), 6.91 (m, 1H), 5.85 (s, 1H), 4.25 (d, 2H, J=4
Hz), 4.15 (m, 1H), 3.00 (m, 2H), 2.84 (m, 1H), 2.18 (s, 6H), 2.10
(s, 3H), 1.75-1.60 (m, 2H), 1.55-1.35 (m, 4H), 1.35-1.15 (m, 4H),
1.08 & 1.03 (s, 3H), 0.87 & 0.77 (t, 3H).
Compound 340:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-methoxy-
-4-methylcyclohexyl)amino)-2-methylbenzamide
##STR01033##
[2159] Step 1: synthesis of methyl
3-(ethyl(4-methoxy-4-methylcyclohexyl)amino)-2-methylbenzoate
##STR01034##
[2161] To a cooled solution of methyl
3-(ethyl(4-hydroxy-4-methylcyclohexyl)amino)-2-methylbenzoate (300
mg, 0.98 mmol) in THF (5 ml), was added sodium hydride (235 mg,
5.89 mmol) portion-wise followed by methyl iodide (0.61 ml, 9.8
mmol). The mixture was stirred at room temperature for 1 h, cooled
to 0.degree. C. quenched with ice cold water and extracted with
ethyl acetate. The combined organic layers were dried over
Na.sub.2SO.sub.4 and concentrated under reduced pressure to give
300 mg of the crude title compound.
Step 2: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-methoxy-
-4-methylcyclohexyl)amino)-2-methylbenzamide
##STR01035##
[2163] A mixture of
3-(ethyl(4-methoxy-4-methylcyclohexyl)amino)-2-methylbenzoate (300
mg, 0.94 mmol) and NaOH (56 mg, 1.41 mmol) in 5 ml of ethanol and
water (4:1) was heated at 70.degree. C. for 2 h. The reaction
mixture was concentrated to dryness and the crude material was
partitioned between water and ethyl acetate. The organic layer was
dried over Na.sub.2SO.sub.4, and concentrated under reduced
pressure to afford 280 mg of the crude acid. A mixture of the crude
acid (280 mg, 0.91 mmol),
3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (279 mg, 1.83 mmol),
PyBOP (716 mg, 1.37 mmol) and triethylamine (0.12 ml, 0.91 mmol) in
DMSO (3 ml) was stirred at room temperature overnight. The reaction
mixture was diluted with water, extracted with 10% MeOH in DCM,
dried over Na.sub.2SO.sub.4, and concentrated to a residue which
was purified by silica gel column chromatography to afford the
title compound
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(4-methoxy-
-4-methylcyclohexyl)amino)-2-methylbenzamide as a mixture of
isomers (70 mg, 17%). LCMS: 440.45 (M+1).sup.+; HPLC: 45.77 &
48.47% (@210-370 nm) (R.sub.t; 4.297 & 4.430; Method: Column:
YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05%
TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col.
Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95%
B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6 400 MHz) .delta. 11.45 (s, 1H), 8.03 (bs, 1H), 7.15
(bs, 2H), 6.91 (s, 1H), 5.85 (s, 1H), 4.25 (d, 2H, J=2.8 Hz),
3.10-2.90 (m, 7H), 2.19 (s, 6H), 2.10 (s, 3H), 1.72-1.10 (m, 8H),
1.08 & 0.99 (s, 3H), 0.77 (t, 3H).
Compound 344:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
1s,4s)-4-methoxycyclohexyl)amino)-2-methylbenzamide
##STR01036##
[2164] Step 1: Synthesis of methyl
5-chloro-3-(((1s,4s)-4-methoxycyclohexyl)amino)-2-methylbenzoate
##STR01037##
[2166] To a stirred solution of methyl
3-amino-5-chloro-2-methylbenzoate (1.0 g, 5.0 mmol) and
4-methoxycyclohexanone (1.28 g, 10.0 mmol) in dichloroethane (10
mL) was added acetic acid (1.8 g, 30 mmol) and the mixture stirred
at room temperature for 10 minutes. Then sodium
triacetoxyborohydride (3.2 g, 15 mmol) was added at 0.degree. C.
and the reaction stirred overnight at room temperature. On
completion, the reaction was quenched with aqueous sodium
bicarbonate, the organic phase separated and the aqueous phase
extracted with dichloromethane. The combined organic layers were
dried over anhydrous sodium sulphate and concentrated under reduced
pressure. The crude material obtained was purified by silica gel
column chromatography and the cis-isomer title compound was
isolated as the less polar product isomer (0.60 g, 38%). The trans
isomer can also be carried forward to produce the corresponding
trans final product, compound 343.
Step 2: Synthesis of methyl
5-chloro-3-(ethyl((1s,4s)-4-methoxycyclohexyl)amino)-2-methylbenzoate
##STR01038##
[2168] To a stirred solution of methyl
5-chloro-3-(((1s,4s)-4-methoxycyclohexyl)amino)-2-methylbenzoate
(0.60 g, 1.92 mmol) and acetaldehyde (0.25 g, 5.78 mmol) in
dichloroethane (10 mL), was added acetic acid (0.69 g, 11.6 mmol)
and the mixture stirred at room temperature for 10 minutes. Then
sodium triacetoxyborohydride (1.22 g, 5.78 mmol) was added at
0.degree. C. and the reaction stirred overnight at room
temperature. On completion, the reaction was quenched with aqueous
sodium bicarbonate, the organic phase separated and the aqueous
phase extracted with dichloromethane. The combined organic layers
were dried over anhydrous sodium sulphate and concentrated under
reduced pressure and the crude product purified by silica gel
column chromatography to afford the title compound (0.60 g,
92%).
Step 3: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1s,4s)-4-methoxycyclohexyl)amino)-2-methylbenzamide
##STR01039##
[2170] Aqueous NaOH (0.07 g, 1.76 mmol) was added to a solution of
methyl
5-chloro-3-(ethyl((1r,4r)-4-methoxycyclohexyl)amino)-2-methylbenzoate
(0.4 g, 1.17 mmol) in ethanol (5 mL) and the mixture was stirred at
60.degree. C. for 1 h. After completion of the reaction, ethanol
was removed under reduced pressure and acidified using dilute HCl
up to pH 6 and adjusted using to pH 4 citric acid. Extraction was
carried out using ethyl acetate and the combined organic layers
were dried concentrated giving the respective crude acid (0.35
g).
[2171] The above crude acid (0.35 g, 1.1 mmol) was dissolved in
DMSO (4 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one
(0.33 g, 2.14 mmol) was added. The reaction mixture was stirred at
room temperature for 15 min, PYBOP (0.84 g, 1.61 mmol) was added
and stirring was continued overnight. After completion of the
reaction, the mixture was poured into ice and extracted with 10%
MeOH/DCM. The combined organic layers were dried over anhydrous
sodium sulphate and concentrated under reduced pressure to yield a
crude product which was purified by silica gel column
chromatography to afford the title compound (0.55 g, 79%). LCMS:
460.30 (M+1).sup.+; HPLC: 97.37% (@210-370 nm) (R.sub.t; 5.026;
Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile
Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj.
Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.;
Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5%
B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 11.44 (bs, 1H),
8.21 (t, 1H), 7.13 (s, 1H), 6.93 (s, 1H), 5.85 (s, 1H), 4.25 (d,
2H, J=3.2 Hz), 3.27 (m, 1H), 3.18 (s, 3H), 2.99 (q, 2H), 2.79 (m,
1H), 2.18 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 1.77 (m, 2H), 1.60
(m, 2H), 1.45 (m, 2H), 1.33 (m, 2H), 0.78 (t, 3H, J=6 Hz).
Compound 349:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)(ethyl)amino)-2-methyl-5-((1-methylazetidin-3-yl)oxy)benzami-
de
##STR01040##
[2172] Step 1: Synthesis of methyl
5-hydroxy-2-methyl-3-nitrobenzoate
##STR01041##
[2174] To a stirred solution of 5-hydroxy-2-methyl-3-nitrobenzoic
acid (3.50 g, 17.8 mmol) in methanol (40 mL), was added thionyl
chloride (3.9 mL, 53 mmol) at 0.degree. C. The mixture was heated
under reflux for 3 hours and concentrated under reduced pressure.
Aqueous sodium bicarbonate was added to the residue which was
extracted with ethyl acetate. The combined organic layers were
washed with water, dried and concentrated under reduced pressure.
The crude material was purified by column chromatography to afford
the title compound (3.0 g, 80%).
Step 2: Synthesis of tert-butyl
3-(3-(methoxycarbonyl)-4-methyl-5-nitrophenoxy)azetidine-1-carboxylate
##STR01042##
[2176] To stirred solution of 5-hydroxy-2-methyl-3-nitrobenzoate
(1.5 g, 7.1 mmol) in ACN (15 mL) were added cesium carbonate (4.64
g, 14.2 mmol) and tert-butyl 3-iodoazetidine-1-carboxylate (3.0 g,
11 mmol). The resulting mixture was heated at 80.degree. C.
overnight. On completion, the reaction mixture was diluted with
water and extraction was carried out using ethyl acetate. The
combined organic layers were washed with water, dried and
concentrated under reduced pressure. The crude material was
purified by column chromatography to afford the title compound (1.2
g, 46%).
Step 3: Synthesis of tert-butyl
3-(3-amino-5-(methoxycarbonyl)-4-methylphenoxy)azetidine-1-carboxylate
##STR01043##
[2178] To stirred solution of tert-butyl
3-(3-(methoxycarbonyl)-4-methyl-5-nitrophenoxy)azetidine-1-carboxylate
(1.0 g, 2.7 mmol) in methanol (10 mL) was added a catalytic amount
of 10% Pd/C. The mixture was stirred at room temperature under a
hydrogen atmosphere (balloon pressure) for 3 hours. On completion,
the reaction mixture was filtered through a celite bed which was
further washed with methanol. The combined filtrates were
concentrated under reduced pressure to afford the title compound
(0.90 g, 98%).
Step 4: Synthesis of tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(methoxycarbonyl-
)-4-methylphenoxy)azetidine-1-carboxylate
##STR01044##
[2180] To a stirred solution of tert-butyl
3-(3-amino-5-(methoxycarbonyl)-4-methylphenoxy)azetidine-1-carboxylate
(0.90 g, 2.67 mmol) and tert-butyl (4-oxocyclohexyl)carbamate
(0.741 g, 3.47 mmol) in dichloroethane (10 mL) was added acetic
acid (0.96 g, 16.1 mmol) and the mixture stirred at room
temperature for 20 minutes. Then sodium triacetoxyborohydride (1.7
g, 8.0 mmol) was added at 0.degree. C. and the reaction stirred at
room temperature for 2 h. On completion, the reaction was quenched
with aqueous sodium bicarbonate, the organic phase separated and
the aqueous phase extracted with dichloromethane. The combined
organic layers were washed with water, dried, concentrated under
reduced pressure and the crude material purified by column
chromatography to afford the title compound (0.9 g, 63%) as a
mixture of cis/trans isomers.
Step 5: Synthesis of tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-5-(methoxyc-
arbonyl)-4-methylphenoxy)azetidine-1-carboxylate
##STR01045##
[2182] To a stirred solution of tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(methoxycarbonyl-
)-4-methylphenoxy)azetidine-1-carboxylate (0.90 g, 1.69 mmol) and
acetaldehyde (0.22 g, 5.06 mmol) in dichloroethane (10 mL), was
added acetic acid (0.61 g, 10 mmol) and the reaction stirred at
room temperature for 20 minutes. Then sodium triacetoxyborohydride
(1.07 g, 5.05 mmol) was added at 0.degree. C. and the reaction was
stirred at room temperature for 2 h. On completion, the reaction
was quenched with aqueous sodium bicarbonate, the organic phase
separated and the aqueous phase extracted with dichloromethane. The
combined organic layers were washed with water, dried, concentrated
under reduced pressure giving the title compound which was used
without further purification (0.85 g).
Step 6: Synthesis of tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-5-(((4,6-di-
methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenoxy)aze-
tidine-1-carboxylate
##STR01046##
[2184] Aqueous NaOH (0.12 g, 3.03 mmol) was added to a solution of
tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-5-(methoxyc-
arbonyl)-4-methylphenoxy)azetidine-1-carboxylate (0.85 g, 1.52
mmol) in ethanol (10 mL) and the mixture stirred at 60.degree. C.
for 1 h. After completion of the reaction, ethanol was removed
under reduced pressure and acidified using dilute HCl up to pH 6
and adjusted using citric acid to pH 4. Extraction was carried out
using ethyl acetate. The combined organic layers were dried and
concentrated giving the respective acid (0.75 g).
[2185] The above acid (0.75 g, 1.37 mmol) was then dissolved in
DMSO (7 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one
(0.42 g, 2.74 mmol) was added to it. The reaction mixture was
stirred at room temperature for 15 min, PYBOP (1.06 g, 2.05 mmol)
was added to it and stirring was continued overnight. After
completion, the reaction mixture was poured into ice, extracted
with 10% MeOH/DCM. The combined organic layers were washed with
water, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure to afford the title compound
(0.80 g, 85%).
Step 7: Synthesis of
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-((4-(dimethylami-
no)cyclohexyl)(ethyl)amino)-2-methyl-5-((1-methylazetidin-3-yl)oxy)benzami-
de
##STR01047##
[2187] To a cooled stirred solution of tert-butyl
3-(3-((4-((tert-butoxycarbonyl)amino)cyclohexyl)(ethyl)amino)-5-(((4,6-di-
methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-4-methylphenoxy)aze-
tidine-1-carboxylate (0.80 g, 1.17 mmol) in DCM (10 mL) was added
TFA (2 mL). The mixture was stirred at rt for 1 h, concentrated
under reduced pressure and saturated NaHCO.sub.3 solution was added
to the residue. Extraction was carried out using 10% MeOH/DCM and
the combined organic layers were washed with water and brine and
dried over anhydrous Na.sub.2SO.sub.4. The solvent was removed
under reduced pressure to give the Boc deprotected intermediate
compound (0.5 g).
[2188] The above intermediate (0.5 g, 1.0 mmol) was dissolved in
MeOH (3 mL) and formalin (0.16 g, 5.33 mmol) was added at 0.degree.
C. The mixture was stirred at same temperature for 20 minutes.
Sodium triacetoxyborohydride (0.55 g, 2.59 mmol) was added and the
mixture stirred at room temperature for 1 h. After completion, the
solvent was removed under reduced pressure and water added to the
residue. Extraction was carried out using 10% MeOH/DCM. The
combined organic layers were dried and concentrated under reduced
pressure to afford crude material which was purified by column
chromatography to afford the title compound (0.03 g, 5.5%).
[2189] LCMS: 524.55 (M+1).sup.+; HPLC: 37.93+61.46% (@210-370 nm)
(R.sub.t; 3.462 and 3.810; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 11.45 (bs, 1H), 8.05 (bs, 1H), 6.56
(s, 1H), 6.36 (s, 1H), 5.85 (s, 1H), 4.69 (bs, 1H), 4.24 (bs, 2H),
3.67 (m, 2H), 3.08-2.92 (m, 4H), 2.33-2.11 (m, 20H), 1.73-1.65 (m,
4H), 1.35-1.29 (m, 4H), 0.77 (t, 3H).
Compound 119:
3-(((1s,4s)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01048##
[2190] Step 1: synthesis of 5-chloro-2-methyl-3-nitrobenzoic
acid
##STR01049##
[2192] To a stirred solution of 5-chloro-2-methylbenzoic acid (4.0
g, 24 mmol) in concentrated H.sub.2SO.sub.4 (27 mL) at -10.degree.
C. was added a nitrating solution comprised of concentrated
HNO.sub.3 (3.3 g, 53 mmol) dissolved in concentrated
H.sub.2SO.sub.4 (4.4 mL). The resulting mixture was stirred at
-10.degree. C. for 3 h and poured onto ice water. The resulting
precipitate was filtered, washed with water and dried under vacuum
to give the title compound (4.95 g, 99%).
Step 2: synthesis of methyl 5-chloro-2-methyl-3-nitrobenzoate
##STR01050##
[2194] To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic
acid (6.75 g, 31.3 mmol) in DMF (33 mL), were added sodium
carbonate (13.2 g, 125 mmol) and methyl iodide (17.8 g, 125 mmol).
The mixture was heated at 60.degree. C. for 4 h. Water was added to
the reaction mixture followed by extraction with dichloromethane.
The combined organic extracts were dried over sodium sulfate,
filtered and concentrated under reduced pressure. The crude product
was purified by column chromatography over silica gel to afford the
title compound (6.0 g, 84%).
Step 3: synthesis of methyl 3-amino-5-chloro-2-methyl benzoate
##STR01051##
[2196] To stirred solution of methyl
5-chloro-2-methyl-3-nitrobenzoate (6.0 g, 26 mmol) in ethanol (60
mL), were added a solution of ammonium chloride (6.0 g, 112 mmol)
in water (60 mL) and iron powder (11.9 g, 208 mmol). The mixture
was heated at 80.degree. C. for 1 h. Water was added and the
resulting mixture was filtered through celite followed by
extraction with ethyl acetate. The combined organic layers were
washed with water, dried over sodium sulfate and concentrated under
reduced pressure to afford the crude title compound (5.0 g).
Step 4: synthesis of methyl
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
amino)-5-chloro-2-methylbenzoate
##STR01052##
[2198] To a stirred solution of 3-amino-5-chloro-2-methyl benzoate
(5.0 g, 25 mmol) and tert-butyl (4-oxocyclohexyl)carbamate (6.95 g,
32.7 mmol) in dichloroethane (25 mL), was added acetic acid (9.0 g,
150 mmol) at room temperature. Sodium triacetoxyborohydride (15.97
g, 75.4 mmol) was added and reaction stirred 1 h at room
temperature. The solvent was removed under reduced pressure and the
crude material was purified by column chromatography over silica
gel to afford to afford 5.2 g (52%) of the first eluting title
compound cis-isomer product methyl
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
amino)-5-chloro-2-methylbenzoate and 3.5 g (35%) the more polar
isomer trans-isomer product methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-chloro-2-me-
thylbenzoate.
Step 5: synthesis of methyl
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-chloro-2-methylbenzoate
##STR01053##
[2200] To a stirred solution of
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
amino)-5-chloro-2-methylbenzoate (5.20 g, 13.1 mmol) and
acetaldehyde (1.15 g, 26.3 mmol) in dichloroethane (15 mL), was
added acetic acid (4.72 g, 78.8 mmol) at room temperature. Sodium
triacetoxyborohydride (8.35 g, 39.4 mmol) was added and the mixture
was stirred for 1 h. Water was added the mixture was extracted with
dichloromethane. The combined organic layers were dried over sodium
sulfate and concentrated under reduced pressure. The crude product
was purified by column chromatography over silica gel to afford the
title compound (5.3 g, 99%).
Step 6: synthesis of tert-butyl ((1s,
4s)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)c-
arbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl) carbamate
##STR01054##
[2202] To a stirred solution of
3-(((1s,4s)-4-((tert-butoxycarbonyl)amino)-cyclohexyl)(ethyl)amino)-5-chl-
oro-2-methylbenzoate (5.20 g, 12.2 mmol) in ethanol (15 mL) was
added an aqueous solution of NaOH (0.73 g, 18.4 mmol). The mixture
was stirred at 60.degree. C. for 1 h, concentrated under reduced
pressure and then acidified to pH 4 by addition of dilute HCl
followed by citric acid. The resulting mixture was extracted with
ethyl acetate and the combined extracts were dried and concentrated
under reduced pressure to afford the corresponding crude acid (4.4
g). To a stirred solution of the acid (4.4 g, 10.7 mmol),
3-(aminomethyl)-4, 6-dimethylpyridin-2(1H)-one (3.25 g, 21.4 mmol)
and triethylamine (1.08 g, 10.70 mmol) in DMSO (15 mL) was added
PYBOP (8.34 g, 16.05 mmol) at 0.degree. C. After stirring overnight
at room temperature, the mixture was poured onto ice and extracted
with 10% MeOH/CH.sub.2Cl.sub.2. The combined organic layers were
dried over sodium sulfate and concentrated under reduced pressure
to obtain crude material which was purified by column
chromatography over silica gel to afford the title compound (4.20
g, 72%). LCMS: 548.40 (M+1).sup.+; HPLC: 98.62% (@254 nm) (R.sub.t;
5.736; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.45 (s, 1H), 8.20 (t, 1H), 7.13 (s, 1H), 6.91 (m, 2H), 5.85 (s,
1H), 4.22 (d, 2H), 3.47 (bs, 1H), 3.00-3.02 (m, 3H), 2.18 (s, 3H),
2.16 (s, 3H), 2.10 (s, 3H), 1.71 (m, 2H), 1.53 (m, 2H), 1.39 (m,
4H), 1.37 (s, 9H), 0.78 (t, 3H, J=6.8 Hz).
Step 7: synthesis of
3-(((1s,4s)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01055##
[2204] To a stirred solution of tert-butyl ((1s,
4s)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)c-
arbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl) carbamate (0.60
g, 1.10 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added trifluoroacetic
acid (2 mL) at 0.degree. C. The mixture was stirred at room
temperature for 2 h and concentrated to dryness. The residue was
dissolved in aqueous sodium bicarbonate followed by extraction with
20% MeOH/CH.sub.2Cl.sub.2. The combined organic layers were dried
over sodium sulfate and concentrated under reduced pressure to
afford a residue which was triturated to afford the title compound
(0.5 g, 100%). LCMS: 445.30 (M+1).sup.+; HPLC: 99.3% (@254 nm)
(R.sub.t; 4.524; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
8.21 (t, 1H), 7.13 (s, 1H), 6.91 (d, 1H, J=1.6 Hz), 5.85 (s, 1H),
4.24 (d, 2H, J=4.8 Hz), 2.99-3.04 (m, 2H), 2.82-2.85 (m, 2H), 2.18
(s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 1.75-1.77 (m, 2H), 1.36-1.39
(m, 6H), 0.79 (t, 3H, J=6.8 Hz).
Compound 121:
3-(((1s,4s)-4-acetamidocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-
-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methylbenzamide
##STR01056##
[2206] To a stirred solution of
3-(((1s,4s)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (0.30 g, 0.68
mmol) and EDCI.HCl (0.194 g, 1.01 mmol) in DMF (5 mL) were added
HOBt (0.091 g, 0.675 mmol) and acetic acid (0.081 g, 1.35 mmol) at
room temperature. After stirring for 18 h water was added and the
mixture was extracted with 10% MeOH/CH.sub.2Cl.sub.2. The combined
organic layers were dried over sodium sulfate and concentrated to a
residue which was purified by column chromatography over silica gel
to afford the title compound (0.10 g, 30%). LCMS: 487.30
(M+1).sup.+; HPLC: 91.72% (@254 nm) (R.sub.t; 4.585; Method:
Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.; Mobile Phase: A;
0.05% TFA in water/B; 0.05% TFA in acetonitrile; Inj. Vol: 10
.mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4 mL/min.; Gradient:
5% B to 95% B in 8 min, Hold for 1.5 min, 9.51-12 min 5% B);
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 8.22 (t, 1H), 7.75 (d,
1H, J=7.2 Hz), 7.15 (s, 1H), 6.93 (s, 1H), 5.85 (s, 1H), 4.24 (d,
2H, J=4.4 Hz), 3.69 (bs, 1H), 3.01-3.03 (m, 2H), 2.94 (m, 1H), 2.18
(s, 6H), 2.10 (s, 3H), 1.80 (s, 3H), 1.69-1.71 (m, 2H), 1.37-1.51
(m, 6H), 0.79 (t, 3H, J=6.8. Hz).
Compound 120:
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01057##
[2207] Step 1: synthesis of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-chloro-2-methylbenzoate
##STR01058##
[2209] To a stirred solution of methyl
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
amino)-5-chloro-2-methylbenzoate (3.5 g, 8.8 mmol) prepared as
described above and acetaldehyde (0.77 g, 17.7 mmol) in
dichloroethane (15 mL), was added acetic acid (3.17 g, 53 mmol) at
room temperature. After stirring for 10 minutes, sodium
triacetoxyborohydride (5.61 g, 26.5 mmol) was added and the
reaction was stirred for 1 h at room temperature. Water was added
and the mixture was extracted with dichloromethane. The combined
organic layers were dried over sodium sulfate and concentrated
under reduced pressure to give crude material which was purified by
column chromatography over silica gel to afford the title compound
(3.5 g, 93%).
Step 2: synthesis of tert-butyl ((1r,
4r)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)c-
arbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl) carbamate
##STR01059##
[2211] To a stirred solution of
3-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)
(ethyl)amino)-5-chloro-2-methylbenzoate (3.50 g, 8.23 mmol) in
ethanol (15 mL) was added an aqueous solution of NaOH (0.49 g, 12.4
mmol). The mixture was stirred at 60.degree. C. for 1 h,
concentrated under reduced pressure and acidified to pH 4 by
addition of dilute HCl followed by citric acid solution. The
resulting mixture was extracted with ethyl acetate, and the
combined organic layers were dried under reduced pressure to give
the corresponding crude acid (3.2 g). To a stirred solution of the
acid (3.2 g, 7.8 mmol), 3-(aminomethyl)-4,
6-dimethylpyridin-2(1H)-one (2.36 g, 15.6 mmol) and triethylamine
(0.79 g, 7.8 mmol) in DMSO (15 mL) was added PYBOP (6.07 g, 11.7
mmol) at 0.degree. C. After stirring overnight at room temperature,
the mixture was poured onto ice and extracted with 10%
MeOH/CH.sub.2Cl.sub.2. The combined organic layers were dried over
sodium sulfate and concentrated under reduced pressure to obtain
crude material which was purified by column chromatography over
silica gel to afford the title compound (3.0 g, 70%). Analytical
Data: LCMS: 545.25 (M+1).sup.+; HPLC: 99.92% (@254 nm) (R.sub.t;
5.677; Method: Column: YMC ODS-A 150 mm.times.4.6 mm.times.5.mu.;
Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in acetonitrile;
Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow rate: 1.4
mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5 min,
9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.46 (s, 1H), 8.23 (t, 1H), 7.14 (s, 1H), 6.92 (s, 1H), 6.65 (d,
1H, J=7.2 Hz), 5.85 (s, 1H), 4.23 (d, 2H, J=4 Hz), 3.14 (bs, 1H),
3.00-3.02 (m, 2H), 2.18 (s, 3H), 2.11 (s, 3H), 2.10 (s, 3H),
1.67-1.76 (m, 4H), 1.40 (m, 2H), 1.35 (m, 9H), 1.04-1.13 (m, 2H),
0.77 (t, 3H, J=6 Hz).
Step 3: synthesis of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
##STR01060##
[2213] To a stirred solution of tert-butyl (1r,
4r)-4-((5-chloro-3-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)c-
arbamoyl)-2-methylphenyl)(ethyl)amino)cyclohexyl) carbamate (0.6 g,
1.10 mmol) in dichloromethane (2 mL) was added TFA (2 mL) at
0.degree. C. The mixture was stirred at room temperature for 2 h
and concentrated to dryness. The residue was dissolved in aqueous
sodium bicarbonate and extracted with 20% MeOH/CH.sub.2Cl.sub.2.
The combined organic layers were dried over sodium sulfate and
concentrated under reduced pressure to afford crude product which
was purified by column chromatography over silica get to afford the
title compound (0.40 g, 81%). LCMS: 445.18 (M+1).sup.+; HPLC:
98.13% (@254 nm) (R.sub.t; 4.096; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 8.22 (t, 1H), 7.13 (s, 1H), 6.92
(s, 1H), 5.85 (s, 1H), 4.23 (d, 2H, J=4 Hz), 3.00-3.01 (m, 2H),
2.59 (m, 1H), 2.18 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H), 1.73-1.76
(m, 2H), 1.65-1.68 (m, 2H), 1.36-1.39 (m, 2H), 0.97-1.00 (m, 2H),
0.77 (t, 3H, J=6 Hz). [1H merged in solvent peak].
Compound 122:
3-(((1r,4r)-4-acetamidocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-
-2-oxo-1,2-dihydropyridin-3-yl) methyl)-2-methylbenzamide
##STR01061##
[2215] To a stirred solution of
3-(((1r,4r)-4-aminocyclohexyl)(ethyl)amino)-5-chloro-N-((4,6-dimethyl-2-o-
xo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide (0.30 g, 0.68
mmol) and EDCI.HCl (0.19 g, 1.0 mmol) in DMF (5 mL) were added HOBt
(0.09 g, 0.68 mmol) and acetic acid (0.081 g, 1.35 mmol) at room
temperature. After stirring for 18 h, water was added and the
mixture was extracted with 10% MeOH/DCM. The combined organic
layers were dried and concentrated giving crude material which was
then purified by column chromatography over silica get to afford
the title compound (0.10 g, 30%). LCMS: 487.25 (M+1).sup.+; HPLC:
92.95% (@254 nm) (R.sub.t; 4.445; Method: Column: YMC ODS-A 150
mm.times.4.6 mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B;
0.05% TFA in acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.:
30.degree. C.; Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8
min, Hold for 1.5 min, 9.51-12 min 5% B); .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta.11.45 (s, 1H), 8.21 (t, 1H), 7.64
(d, 1H, J=7.6 Hz), 7.14 (s, 1H), 6.92 (s, 1H), 5.85 (s, 1H), 4.24
(d, 2H, J=4.8 Hz), 3.41-3.43 (m, 1H), 3.01-3.03 (m, 2H), 2.61 (m,
1H), 2.18 (s, 3H), 2.13 (s, 3H), 2.10 (s, 3H), 1.74 (t, 7H, J=18
Hz), 1.40-1.43 (m, 2H), 1.07-1.12 (m, 2H), 0.77 (t, 3H, J=6.8
Hz).
Compound 207:
5-chloro-3-(cyclohexylthio)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-2-methylbenzamide
##STR01062##
[2216] Step 1: Synthesis of methyl
3-bromo-5-chloro-2-methylbenzoate
[2217] To a stirred solution of CuBr.sub.2 (12.3 g, 55.3 mmol) in
acetonitrile (100 mL), t-butyl nitrite (7.79 g, 75.4 mmol) was
added at 0.degree. C. To this solution, methyl
3-amino-5-chloro-2-methylbenzoate (10 g, 50.3 mmol) in acetonitrile
(50 mL) was added and stirred at 0.degree. C. for 2 h and at rt for
overnight. On completion, water added and the product extracted
with ethyl acetate. The organic layer was washed with aq.
NH.sub.4Cl solution and dried over sodium sulphate, then
concentrated under reduced pressure. Silica gel column purification
afforded the title compound (6.6 g, 50%).
Step 2: Synthesis of methyl
5-chloro-3-(cyclohexylthio)-2-methylbenzoate
[2218] To a stirred solution of methyl
3-bromo-5-chloro-2-methylbenzoate (1.0 g, 3.79 mmol) in 1,4-dioxane
was added N,N-diisopropyl ethylamine (0.98 g, 7.59 mmol). The
reaction mixture was purged with argon for 20 min. then
Pd(OAc).sub.2 (0.042 g, 0.189 mmol), Xanthphos (0.22 g, 0.38 mmol)
and cyclohexylthiol (0.44 g, 3.79 mmol) was added to it
sequentially and again purged with argon for 10 min. The reaction
mixture was then refluxed for 16 h. On completion, water was added,
and the product extracted with ethyl acetate. The organic layer was
dried over sodium sulphate and the product was purified by silica
gel column to afford the title compound (1.0 g, 83%)
Step 3: Synthesis of
5-chloro-3-(cyclohexylthio)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-2-methylbenzamide
[2219] Aqueous NaOH (0.2 g in 2.5 mL H.sub.2O, 5.02 mmol) was added
to a solution of methyl
5-chloro-3-(cyclohexylthio)-2-methylbenzoate (1.0 g, 3.35 mmol) in
ethanol (10 mL) and the solution stirred at 60.degree. C. for 1 h.
After completion of the reaction, ethanol was removed under reduced
pressure and the aqueous layer was acidified using dilute HCl to pH
6 and then citric acid to pH 4. The product was extracted was using
10% methanol in DCM. The combined organic layers were dried and
concentrated to give the respective acid (0.91 g, 95%).
[2220] The above acid (0.91 g, 3.2 mmol) was then dissolved in DMSO
(2 mL) and 3-(amino methyl)-4, 6-dimethylpyridin-2(1H)-one (0.974
g, 6.41 mmol) and triethyl amine (0.65 g, 3.2 mmol) was added to
it. The reaction mixture was stirred at room temperature for 15 min
before PyBOP (2.5 g, 4.8 mmol) was added to it and stirring was
continued for overnight. After completion of the reaction, the
solution was poured into ice and the product was extracted with 10%
MeOH in DCM. The combined organic layers were dried, concentrated
to obtain crude product. Purification by solvent washings afforded
the title compound (1.3 g, 93%).
[2221] Analytical Data: LCMS: 419.2 (M+1).sup.+; HPLC: 98.38% (@254
nm) (R.sub.t; 7.989; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta.
11.47 (bs, 1H), 8.32 (t, 1H), 7.40 (s, 1H), 7.07 (s, 1H), 5.85 (s,
1H), 4.24-4.25 (d, 1H, J=4 Hz), 2.21 (s, 3H), 2.18 (s, 3H), 2.11
(s, 3H), 1.88-1.91 (m, 2H), 1.56-1.69 (m, 3H), 1.25-1.38 (m, 5H), 2
protons merged in solvent peak.
Compound 208:
5-chloro-3-(cyclohexylsulfinyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydro-pyridi-
n-3-yl)methyl)-2-methylbenzamide
##STR01063##
[2223] To a stirred solution of
5-chloro-3-(cyclohexylthio)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-2-methylbenzamide (0.2 g, 0.48 mmol) in DCM (7 mL) at
0.degree. C., m-CPBA (0.098 g, 0.57 mmol) was added and stirred at
0.degree. C. for 2 h. Reaction was monitored by TLC. After
completion, the reaction mixture was diluted with DCM and washed
with aqs. NaHCO.sub.3. The product was purified by solvent washings
to afford the title compound (0.17 g, 82.3%).
[2224] Analytical Data: LCMS: 435.1 (M+1).sup.+; HPLC: 92.68% (@220
nm) (R.sub.t; 5.996; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (CDCl3, 400 MHz) .delta. 11.44
(bs, 1H), 7.82 (s, 1H), 7.45 (t, 1H), 7.39 (s, 1H), 5.97 (s, 1H),
4.52 (bs, 2H), 2.53-2.59 (m, 1H), 2.38 (s, 3H), 2.34 (s, 3H), 2.23
(s, 3H), 1.85-1.93 (m, 3H), 1.65 (m, 2H), 1.20-1.29 (m, 3H).
Compound 209:
5-chloro-3-(cyclohexylsulfonyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydro-pyridi-
n-3-yl)methyl)-2-methylbenzamide
##STR01064##
[2226] To a stirred solution of
5-chloro-3-(cyclohexylthio)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-y-
l)methyl)-2-methylbenzamide (0.1 g, 0.238 mmol) in DCM (4 mL) at
0.degree. C., m-CPBA (0.082 g, 0.48 mmol) was added and stirred at
rt for 2 h. Reaction was monitored by TLC. After completion
reaction mixture was diluted with DCM and washed with aqs.
NaHCO.sub.3. The product was purified by prep. HPLC to afford the
title compound (0.075 g, 23%).
[2227] Analytical Data: LCMS: 451.1 (M+1).sup.+; HPLC: 83.44% (@254
nm) (R.sub.t; 6.547; Method: Column: YMC ODS-A 150 mm.times.4.6
mm.times.5.mu.; Mobile Phase: A; 0.05% TFA in water/B; 0.05% TFA in
acetonitrile; Inj. Vol: 10 .mu.L, Col. Temp.: 30.degree. C.; Flow
rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5
min, 9.51-12 min 5% B); .sup.1H NMR (CDCl3, 400 MHz) .delta. 11.44
(bs, 1H), 7.98 (s, 1H), 7.50 (s, 1H), 7.45 (t, 1H), 5.97 (s, 1H),
4.51 (d, 2H, J=4 Hz), 2.94-3.00 (m, 1H), 2.62 (s, 3H), 2.38 (s,
3H), 2.21 (s, 3H), 1.88-1.95 (m, 4H), 1.70 (m, 1H), 1.52-1.55 (m,
2H), 1.21 (m, 3H).
Compound 416:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylbenzamide
##STR01065##
[2228] Step 1: Synthesis of 4-methanesulfonylcyclohexan-1-one
##STR01066##
[2230] A solution containing
(buta-1,3-dien-2-yloxy)(trimethyl)silane (12.5 ml, 70.7 mmol) in
toluene (50 ml), was treated with (methylsulfonyl)ethene (4.1 ml,
47.1 mmol) and heated under reflux for 76 hours. After which time,
the reaction mixture was cooled to room temperature and
concentrated in-vacuo to give a yellow oil. The oil was dissolved
in CH.sub.2Cl.sub.2 (120 ml) and filtered through Celite.RTM.. The
filtrate was concentrated in-vacuo and the resulting residue was
then dissolved in MeOH (8 ml) with cooling (ice/water). It was then
treated with TFA (2.0 ml). The reaction mixture was stirred at
0.degree. C. for a further 10 minutes and then at room temperature
for 25 hours. The reaction mixture was concentrated in-vacuo and
the residue was purified by flash column chromatography (0-10%
MeOH/CH.sub.2Cl.sub.2) 100 g SNAP cartridge on the Biotage Isolera
to give the title compound 1.2 g (14%) as a thick yellow oil. LC-MS
94%, 0.39 min (3.5 minute LC-MS method), m/z=177.0 (ELS visible
only). .sup.1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm 3.24-3.32
(m, 1H) 2.94 (s, 3H) 2.62-2.70 (m, 2H) 2.50-2.56 (m, 2H) 2.38-2.45
(m, 2H) 2.07-2.18 (m, 2H).
Step 2: Synthesis of methyl
5-chloro-2-methyl-3-(((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)benzoate
##STR01067##
[2232] To a stirred solution of 4-methanesulfonylcyclohexan-1-one
(94%, 0.56 g, 3.0 mmol) in 1,2-dichloroethane (10 ml) under a
balloon of nitrogen, was added methyl
3-amino-5-chloro-2-methylbenzoate (0.60 g, 3.0 mmol), followed by
acetic acid (1.0 ml, 18.0 mmol). The solution was stirred for 50
minutes before the portionwise addition of sodium
triacetoxyborohydride (1.9 g, 9.0 mmol) over 4 hours. The resulting
suspension was stirred overnight before the addition of
4-methanesulfonylcyclohexan-1-one (0.28 g, 1.5 mmol) in
1,2-dichloroethane (1.5 ml) and sodium triacetoxyborohydride (0.96
g, 4.5 mmol) over 2.5 hours. The reaction mixture was stirred for a
further 67 hours before the addition of deionized water (40 ml) to
the reaction mixture and neutralisation (pH 8) by the gradual
addition of solid NaHCO.sub.3 (4.8 g). The mixture was extracted
with CH.sub.2Cl.sub.2 (3.times.25 ml). The combined organic
extracts were washed with brine (40 ml), dried over MgSO.sub.4,
filtered and concentrated in-vacuo. The residue was pre-absorbed
onto silica and purified by flash column chromatography (0-55%
ethyl acetate/heptanes) 100 g SNAP cartridge on the Biotage Isolera
to give the title compound (131 mg, 12%). LC-MS 96%, 2.09 min (3.5
minute LC-MS method), m/z=359.85, 360.85, 361.90, 362.85; .sup.1H
NMR (500 MHz, CHLOROFORM-d) .delta. ppm 7.12 (br. s., 1H) 6.68 (br.
s., 1H) 3.89 (s, 3H) 3.82 (br. s., 1H) 3.71 (br. s., 1H) 2.93-3.02
(m, 1H) 2.89 (s, 3H) 2.29 (s, 3H) 2.03-2.18 (m, 4H) 1.91-2.02 (m,
2H) 1.71-1.83 (m, 2H).
[2233] The other isomer methyl
5-chloro-2-methyl-3-(((1r,4r)-4-(methylsulfonyl)cyclohexyl)amino)benzoate
was also isolated
##STR01068##
[2234] (92 mg, 8%), LC-MS 96%, 2.06 min (3.5 minute LC-MS method),
m/z=359.85, 360.90, 361.90, 362.85; .sup.1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 7.11 (br. s, 1H) 6.68 (br. s., 1H) 3.89
(s, 3H) 3.62 (br. s., 1H) 3.27-3.36 (m, 1H) 2.84-2.95 (m, 4H)
2.32-2.42 (m, 4H) 2.24 (br. s., 3H) 1.72-1.85 (m, 2H) 1.27-1.35 (m,
2H).
[2235] The assignment of cis and trans from further NMR experiments
could not be made and assignment is arbitrary.
Step 3: Synthesis of methyl
5-chloro-3-(ethyl((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylben-
zoate
##STR01069##
[2237] To a stirred solution of methyl
5-chloro-2-methyl-3-(((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)benzoate
(246 mg, 0.68 mmol) in dry 1,2-dichloroethane (6.0 ml), was added
acetaldehyde (115 .mu.l, 2.1 mmol), followed by acetic acid (235
.mu.l, 4.1 mmol). The reaction was stirred for 35 minutes before
sodium triacetoxyborohydride (435 mg, 2.1 mmol) was added in
portions over 50 minutes and the reaction was stirred overnight and
then treated with acetaldehyde (115 .mu.l, 2.1 mmol). The reaction
mixture was stirred for 1 hour before addition of sodium
triacetoxyborohydride (435 mg, 2.1 mmol) in portions over 1 hour
and the reaction mixture stirred overnight. Acetaldehyde (229
.mu.l, 4.1 mmol) was added and the reaction stirred for 2.5 hours,
followed by the addition of sodium triacetoxyborohydride (869 mg,
4.1 mmol) in portions over 1.5 hours and stirring was continued
overnight. Acetaldehyde (229 .mu.l, 4.1 mmol) was added along with
1,2-dichloroethane (4.0 ml) to aid solution and the reaction
stirred for 4 hours. Sodium triacetoxyborohydride (869 mg, 4.1
mmol) was added in portions over 2 hours and stirring was continued
over the weekend. Acetaldehyde (229 .mu.l, 4.1 mmol) was added and
the reaction stirred for 1.5 hours, followed by the addition of
sodium triacetoxyborohydride (869 mg, 4.1 mmol) in equal portions
over 1 hour, along with 1,2-dichloroethane (5.0 ml) and stirring
was continued overnight. Acetaldehyde (382 .mu.l, 6.8 mmol) was
added and the reaction stirred for 2.5 hours, followed by the
addition of sodium triacetoxyborohydride (1.4 g, 6.8 mmol) in equal
portions over 4 hours, along with 1,2-dichloroethane (10 ml) and
stirring was continued overnight. The reaction mixture was diluted
with water (40 ml) and neutralised (to pH 8) by the gradual
addition of solid NaHCO.sub.3 (5.79 g). The layers were separated
and the aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.25 ml). The combined organic phases were washed with brine
(20 ml), dried (MgSO.sub.4), filtered and concentrated in-vacuo.
The crude residue was pre-absorbed onto silica and purified by
column chromatography (0-50% EtOAc/Heptanes) 10 g SNAP cartridge on
the Biotage Isolera to give the title compound (140 mg, 51%) as a
colourless glassy solid. LC-MS 97%, 2.29 min (3 minute LC-MS
method), m/z=387.95, 388.95, 389.90, 390.95. .sup.1H NMR (500 MHz,
CHLOROFORM-d) .delta. ppm 7.59 (d, J=1.89 Hz, 1H) 7.21 (d, J=1.89
Hz, 1H) 3.91 (s, 3H) 3.45 (br. s., 1H) 3.01 (m, 2H) 2.86-2.95 (m,
1H) 2.82 (s, 3H) 2.52 (s, 3H) 2.08 (br. s., 2H) 1.94 (br. s., 2H)
1.86 (br. s., 2H) 1.46-1.54 (m, 2H) 0.89 (t, J=7.01 Hz, 3H).
Step 4: Synthesis of
5-chloro-3-(ethyl((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylben-
zoic acid
##STR01070##
[2239] To a stirred solution of methyl
5-chloro-3-(ethyl((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylben-
zoate (140 mg, 0.36 mmol) in THF (5.0 ml) and MeOH (0.3 ml), was
added a solution of 4M NaOH (2.7 ml). The resulting solution was
stirred at 50.degree. C. for 21 hours and then treated with 4M NaOH
(0.9 ml, 3.6 mmol) and heating was continued for a further 4 hours.
After which time, the reaction mixture was left to reach room
temperature and then concentrated in-vacuo. The resulting aqueous
residue was extracted with CH.sub.2Cl.sub.2 (20 ml) and ether (20
ml). The pH of the aqueous phase was then adjusted to 3-4 by the
addition of 1M HCl (aq) (13 ml) and extracted with ethyl acetate
(3.times.25 ml). The combined ethyl acetate extracts were dried
over MgSO.sub.4, filtered and concentrated in-vacuo to give the
title compound (122 mg, 80% yield) as a light yellow solid. LC-MS
89%, 4.03 min (7 minute LC-MS method), m/z=373.95, 374.90, 375.95,
377.00. .sup.1H NMR (500 MHz, CHLOROFORM-d) .delta. ppm 7.77 (d,
J=1.89 Hz, 1H) 3.47 (br. s., 1H) 3.03 (br. s., 2H) 2.88-2.97 (m,
1H) 2.84 (s, 3H) 2.60 (s, 3H) 2.12 (br. s, 2H) 1.96 (br. s, 2H)
1.88 (br. s., 2H) 1.55 (br. s., 2H) 0.91 (t, J=7.01 Hz, 3H).
1.times.ArH assumed to be under the CDCl.sub.3 peak and the
CO.sub.2H peak also not visible.
Step 5: Synthesis of
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylbenzamide
##STR01071##
[2241] A stirred solution of
5-chloro-3-(ethyl((1s,4s)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylben-
zoic acid (90%, 122 mg, 0.29 mmol) in anhydrous DMF (2.0 ml) at
0.degree. C. under a balloon of N.sub.2, was treated with HATU (134
mg, 0.35 mmol) and DIPEA (102 .mu.l, 0.59 mmol) dropwise. The
resulting solution was stirred for 10 minutes and then treated with
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one (89%, 60 mg,
0.35 mmol). The resulting suspension was stirred at 0.degree. C.
for 15 minutes and then stirred at room temperature over the
weekend. The reaction mixture was partitioned between water (10 ml)
and CH.sub.2Cl.sub.2 (10 ml). The layers were separated and the
aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.15 ml).
The combined organics were washed with a saturated solution of
NaHCO.sub.3 (aq) (10 ml), water (40 ml), brine (2.times.25 ml),
dried (MgSO.sub.4), filtered and concentrated in-vacuo. The residue
was purified by flash column chromatography (0-9%
MeOH/CH.sub.2Cl.sub.2) 10 g SNAP cartridge on the Biotage Isolera
to give the title compound (120 mg, 76% yield) as an off-white
solid. LC-MS: 95%, 3.71 min (7 minute LC-MS method), m/z=508.10,
509.10, 510.10, 511.10. .sup.1H NMR (500 MHz, Chloroform-d) .delta.
10.84 (s, 1H), 7.07 (s, 2H), 7.00 (m, 1H), 6.10 (s, 1H), 4.53 (d,
J=5.8 Hz, 2H), 3.45 (s, 1H), 3.07-2.95 (m, 2H), 2.93-2.85 (m, 1H),
2.81 (s, 3H), 2.45 (s, 3H), 2.31 (s, 3H), 2.29 (s, 3H), 2.14-2.01
(m, 2H), 1.97-1.89 (m, 2H), 1.87-1.80 (m, 2H), 1.50 (br s, 2H),
0.88 (t, J=5.9 Hz, 3H).
Compound 417:
5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(-
(1r,4r)-4-(methylsulfonyl)cyclohexyl)amino)-2-methylbenzamide
[2242] This compound was prepared in an analogous manner to that of
compound 416 using the other isomer obtained in step 2
##STR01072##
Compound 392:
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(1-methylp-
iperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzamide
##STR01073##
[2244] To a stirred solution of
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoi-
c acid (crude material, 580 mg, 1.68 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt (477
mg, 2.53 mmol) in DMSO (6 mL) was added PyBOP (1.57 g, 3.02 mmol)
and hunig base (650 mg, 5.04 mmol). The reaction mixture was
stirred at 23.degree. C. for 20 hours. The reaction mixture was
quenched with water and resulting precipitate was collected. The
solid was purified by silica gel column chromatography
(NH--SiO.sub.2 ethylacetate/MeOH=25/1) to give the titled compound
as a white amorphous (386 mg, 48% yield). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.31 (s, 1H), 7.27 (s, 1H), 7.18 (t, J=5.7
Hz, 1H), 5.95 (s, 1H), 4.55 (d, J=5.9 Hz, 2H), 3.17 (dq, J=6.4, 3.3
Hz, 2H), 3.08 (q, J=7.0 Hz, 2H), 2.81 (m, 2H), 2.70 (m, 1H), 2.40
(s, 3H), 2.33 (s, 3H), 2.23 (s, 3H), 2.21 (s, 3H), 1.78-1.93 (m,
2H), 1.70 (m, 2H), 0.84 (t, J=7.0 Hz, 3H); MS(ES) [M+H] 479.2,
[M+Na] 501.3; HPLC 96.8% purity.
Compound 393:
3-[Ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-[(6-methyl-2-oxo-4-prop-
yl-1,2-dihydropyridin-3-yl)methyl]-5-(trifluoromethyl)benzamide
##STR01074##
[2246] To a stirred solution of
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoi-
c acid (crude material, 680 mg, 1.97 mmol) and
3-(aminomethyl)-6-methyl-4-propyl-1,2-dihydropyridin-2-one HCl salt
(642 mg, 2.96 mmol) in DMSO (7 mL) was added PyBOP (1.85 g, 3.55
mmol) and hunig base (764 mg, 5.91 mmol). The reaction mixture was
stirred at 23.degree. C. for 20 hours. The reaction mixture was
quenched with water and resulting precipitate was collected. The
solid was purified by silica gel column chromatography
(NH--SiO.sub.2 ethylacetate/MeOH=20/1) to give the titled compound
as a white solid (224 mg, 22% yield). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.31 (s, 1H), 7.26 (s, 1H), 7.16 (m, 1H),
5.94 (s, 1H), 4.55 (d, J=5.9 Hz, 2H), 3.08 (q, J=7.0 Hz, 2H), 2.82
(m, 2H), 2.66-2.73 (m, 3H), 2.33 (s, 3H), 2.24 (s.times.2, 6H),
1.79-1.98 (m, 2H), 1.67-1.75 (m, 4H), 1.59-1.66 (m, 2H), 1.02 (t,
J=7.2 Hz, 3H), 0.84 (t, J=7.0 Hz, 3H); MS (ES) [M-H] 505.2; HPLC
98.1% purity.
Compound 394:
3-[Ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-oxo-4-(pro-
pan-2-yl)-1,2-dihydropyridin-3-yl]methyl}-5-(trifluoromethyl)benzamide
##STR01075##
[2248] To a stirred solution of
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoi-
c acid (crude material, .about.0.313 mmol) and
3-(aminomethyl)-6-methyl-4-(propan-2-yl)-1,2-dihydropyridin-2-one
HCl salt (88.0 mg, 0.407 mmol) in DMSO (2.4 mL) was added PyBOP
(244 mg, 0.470 mmol) and hunig base (164 ul, 0.939 mmol). The
reaction mixture was stirred at rt for 16 hours. The reaction
mixture was quenched with water and diluted with ethylacetate. The
aqueous layer was extracted with dichloromethane. The combined
organic layer was washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by silica gel
column chromatography (NH--SiO.sub.2 ethylacetate/MeOH=20/1+5%
triethylamine). The crude compound was dissolved with DMSO, diluted
with water. The precipitated solid was collected and triturated
with ethylacetate/TBME/hexane to give the titled compound as a
white solid (80.0 mg, 51% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.31 (s, 1H), 7.26 (s, 1H), 7.03-7.10 (m, 1H), 6.04
(s, 1H), 4.59 (d, J=6.0 Hz, 2H), 3.49-3.55 (m, 1H), 3.09 (q, J=6.8
Hz, 2H), 2.81-2.84 (m, 2H), 2.70-2.78 (m, 1H), 2.32 (s, 3H), 2.26
(s, 3H), 2.24 (s, 3H), 1.86-1.89 (m, 2H), 1.68-1.71 (m, 4H), 1.22
(d, J=6.4 Hz, 6H), 0.84 (t, J=6.8 Hz, 3H); MS (ES) [M+H] 507.2,
[M+Na] 529.2; HPLC 97.7% purity.
Compound 395:
3-[Ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-oxo-4-(tri-
fluoromethyl)-1,2-dihydropyridin-3-yl]methyl}-5-(trifluoromethyl)benzamide
##STR01076##
[2250] To a stirred solution of
3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzoi-
c acid (crude material, .about.0.313 mmol) and
3-(aminomethyl)-6-methyl-4-(trifluoromethyl)-1,2-dihydropyridin-2-one
hydrochloride HCl salt (100 mg, 0.91 mmol) in DMSO (2.4 mL) was
added PyBOP (248 mg, 1.05 mmol) and hunig base (166 ul, 2.10 mmol).
The reaction mixture was stirred at rt for 16 hours. The reaction
mixture was quenched with water and resulting precipitate was
collected. The solid was triturated from ethylacetate/hexane. The
resulting solid was purified by silica gel column chromatography
(NH--SiO.sub.2 ethylacetate/MeOH=10/1+10% triethylamine). The crude
compound was triturated from ethylacetate/hexane, and dried to give
the titled compound as a white solid (75.0 mg, 44% yield).
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. ppm; 7.32 (s, 1H), 7.26
(s, 1H), 6.86-6.89 (m, 1H), 6.34 (s, 1H), 4.73 (d, J=6.4 Hz, 2H),
3.08 (q, J=7.2 Hz, 2H), 2.80-2.85 (m, 2H), 2.70-2.78 (m, 1H), 2.37
(s, 3H), 2.35 (s, 3H), 2.24 (s, 3H), 1.85-1.89 (m, 2H), 1.69-1.72
(m, 4H), 0.84 (t, J=7.2 Hz, 3H); MS (ES) [M+H] 533.1, [M+Na] 555.2;
HPLC 95.8% purity.
Compound 395:
3-(ethyl(1-methylpiperidin-4-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-(tri-
fluoromethyl)-1,2-dihydropyridin-3-yl)methyl)-5-(trifluoromethyl)benzamide
diformate
##STR01077##
[2252] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.39 (s, 2H),
7.48 (s, 1H), 7.36 (s, 1H), 6.36 (s, 1H), 4.52 (s, 2H), 3.38 (br,
2H), 3.18 (br, m, 1H), 3.11 (q, J=7.2 Hz, 2H), 2.96 (br, m, 2H),
2.77 (s, 3H), 2.35 (s, 3H), 2.31 (s, 3H), 1.97 (br, d, J=12.0 Hz,
2H), 1.83 (br, m, 2H), 0.84 (t, J=7.2 Hz, 3H); MS (ES)
(M+H)=533.53.
[2253] Following the same preparation method above of
3-[Ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-oxo-4-(tri-
fluoromethyl)-1,2-dihydropyridin-3-yl]methyl}-5-(trifluoromethyl)benzamide-
, the following analogs have been prepared using the corresponding
carboxylic acid and amine, and purified by reverse phase HPLC
(ACN-H.sub.2O containing 0.1% formic acid).
Compound 396:
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-6-methyl-2-oxo-4-prop-
yl-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamide
formate
##STR01078##
[2255] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.31 (s, 1H),
7.49 (s, 1H), 7.34 (s, 1H), 4.45 (s, 2H), 3.39 (br, 2H), 3.17 (br,
1H), 3.10 (q, J=7.2 Hz, 2H), 2.97 (br, m, 2H), 2.77 (s, 3H), 2.74
(br, m, 2H), 2.33 (s, 3H), 2.22 (d, J=2.8 Hz, 3H), 1.97 (br, m,
2H), 1.85 (br, m, 2H), 1.60 (m, 2H), 1.02 (t, J=7.2 Hz, 3H), 0.84
(t, J=6.8 Hz, 3H); MS (ES) (M+H)=525.58.
Compound 397:
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-4-isopropyl-6-methyl--
2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamid-
e formate
##STR01079##
[2257] LCMS ES+ (M+1)=525.58. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. ppm 8.31 (s, 1H), 7.50 (s, 1H), 7.34 (s, 1H), 4.49 (s, 2H),
3.48 (m, 1H), 3.39 (br, 2H), 3.18 (br, m, 1H), 3.09 (q, J=6.8 Hz,
2H), 2.97 (br, m, 2H), 2.78 (s, 3H), 2.34 (s, 3H), 2.21 (d, J=3.2
Hz, 3H), 1.96 (br, d, J=12.0 Hz, 2H), 1.86 (br, m, 2H), 1.32 (dd,
J=1.6 Hz, J=7.2 Hz, 6H), 0.84 (t, J=6.8 Hz, 3H); MS (ES)
(M+H)=525.58.
Compound 398:
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-4,6-dimethyl-2-oxo-1,-
2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamide
formate
##STR01080##
[2259] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.29 (s, 1H),
7.49 (s, 1H), 7.35 (s, 1H), 4.45 (s, 2H), 3.39 (br, 2H), 3.19 (br,
1H), 3.09 (q, J=7.2 Hz, 2H), 2.97 (br, m, 2H), 2.77 (s, 3H), 2.34
(d, J=2.4 Hz, 3H), 2.33 (s, 3H), 2.22 (d, J=3.2 Hz, 3H), 1.97 (br,
m, 2H), 1.86 (br, m, 2H), 0.84 (t, J=6.8 Hz, 3H); MS (ES)
(M+H)=497.51.
Compound 399:
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-6-methyl-2-oxo-4-(tri-
fluoromethyl)-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)-
benzamide diformate
##STR01081##
[2261] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.32 (s, 2H),
7.50 (s, 1H), 7.34 (s, 1H), 4.59 (s, 2H), 3.40 (br, 2H), 3.18 (br,
m, 2H), 3.09 (q, J=6.8, 2H), 2.96 (br, m, 2H), 2.78 (s, 3H), 2.35
(s, 3H), 2.29 (d, J=3.2 Hz, 3H), 1.98 (br, d, J=13.2 Hz, 2H), 1.86
(m, 1H), 1.80 (br, m, 2H), 0.84 (t, J=6.8 Hz, 3H); MS (ES)
(M+H)=551.53.
Compound 399:
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-6-methyl-2-oxo-4-(tri-
fluoromethyl)-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)-
benzamide diformate
##STR01082##
[2263]
6-Methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-3-carbonitri-
le was fluorinated as described for
5-fluoro-4-isopropyl-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile.
The product was isolated as a 3:1 mixture of
6-methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-3-carbonitrile
(starting material) and
5-fluoro-6-methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-3-carboni-
trile. For
5-fluoro-6-methyl-2-oxo-4-(trifluoromethyl)-1,2-dihydropyridine-
-3-carbonitrile, LCMS E-S (M+H)=221.2. This mixture of nitriles was
subjected to the reduction conditions as described above for
3-(Aminomethyl)-5-fluoro-4-isopropyl-6-methylpyridin-2(1H)-one. For
fluorinated compound LCMS (ES) (M+H) showed 225.2 and for
non-fluorinated analog LCMS (ES) (M+H) showed 207.2. Subsequent
coupling reaction was performed with the crude mixture of amines
without further purifications. For
3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-6-methyl-2-oxo-4--
(trifluoromethyl)-1,2-dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromet-
hyl)benzamide diformate; .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
ppm 8.32 (s, 2H), 7.50 (s, 1H), 7.34 (s, 1H), 4.59 (s, 2H), 3.40
(br, 2H), 3.18 (br, m, 1H), 3.09 (q, J=6.8, 2H), 2.96 (br, m, 2H),
2.78 (s, 3H), 2.35 (s, 3H), 2.29 (d, J=3.2 Hz, 3H), 1.98 (br, d,
J=13.2 Hz, 2H), 1.80 (br, m, 2H), 0.84 (t, J=6.8 Hz, 3H); MS (ES)
(M+H) 551.5.
Compound 400:
N-[(4,6-Eimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-{ethyl[1-(propan-
-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoromethyl)benzamide
##STR01083##
[2265] To a stirred solution of
3-{ethyl[1-(propan-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoromethyl-
)benzoic acid (crude material, 510 mg, 0.569 mmol) and
3-(aminomethyl)-4,6-dimethyl-1,2-dihydropyridin-2-one HCl salt (161
mg, 0.854 mmol) in DMSO (2 mL) was added PyBOP (534 mg, 1.02 mmol)
and hunig base (220 mg, 1.71 mmol). The reaction mixture was
stirred at 23.degree. C. for 20 hours. The reaction mixture was
quenched with water and resulting precipitate was collected. The
solid was purified by silica gel column chromatography
(NH--SiO.sub.2 ethylacetate/MeOH=20/1) to give the titled compound
as a white solid (129 mg, 44% yield). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.30 (s, 1H), 7.26 (s, 1H), 7.22 (t, J=5.9
Hz, 1H), 5.95 (s, 1H), 4.54 (d, J=5.9 Hz, 2H), 3.16 (m, 2H), 3.08
(q, J=7.0 Hz, 2H), 2.85 (m, 2H), 2.62-2.74 (m, 1H), 2.38 (s, 3H),
2.33 (s, 3H), 2.19 (s, 3H), 2.00-2.09 (m, 1H), 1.81 (m, 2H),
1.58-1.76 (m, 2H), 0.99 (s.times.2, 6H), 0.83 (t, J=7.0 Hz, 3H); MS
(ES) [M+H] 507.2, [M+Na] 529.3; HPLC 97.2% purity.
Compound 401:
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-{[(2R*,6R*)-2,6--
dimethylpiperidin-4-yl](ethyl)amino}-2-methyl-5-(trifluoromethyl)benzamide
##STR01084##
[2267] To a stirred solution of 3-{[(2R,
6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-2-me-
thyl-5-(trifluoromethyl)benzamide (267 mg, 0.459 mmol) in MeOH (5
mL) was added Pd/C (250 mg). The reaction mixture was stirred at et
for 2 hour under hydrogen atmosphere. The reaction mixture was
filtered and the filtrate was concentrated in vacuo. The residue
was purified by silica gel column chromatography (1. NAM-300H
silica gel produced by Nagara Science Co., Ltd.,
ethylacetate/MeOH=4/1 to ethylacetate/MeOH/triethylamine=1/1/0.01,
2. NH--SiO.sub.2, heptane/ethylacetate=30/1 to 8/1) to give the
titled compound as a white solid (127 mg, 56% yield). .sup.1H-NMR
(400 MHz, CDCl.sub.3) .delta. ppm; 7.30 (s, 1H), 7.26 (s, 1H), 7.14
(t, J=6.1 Hz, 1H), 5.94 (s, 1H), 4.54 (dd, J=5.9, 1.6 Hz, 2H),
3.41-3.49 (m, 1H), 3.01-3.13 (m, 3H), 2.85-2.92 (m, 1H), 2.40 (s,
3H), 2.32 (s, 3H), 2.22 (s, 3H), 1.63-1.80 (m, 4H), 1.13 (d, J=7.0
Hz, 3H), 1.02 (d, J=6.2 Hz, 3H), 0.83 (t, J=7.0 Hz, 3H); LC-MS: m/z
[M+H] 493.1, [M+Na] 515.3; HPLC; 97.2% purity.
Compound 402:
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-{[(2R*,4R*,6S*)--
2,6-dimethylpiperidin-4-yl](ethyl)amino}-2-methyl-5-(trifluoromethyl)benza-
mide
##STR01085##
[2269] To a stirred solution of 3-{[(2R, 4R,
6S)-1-benzyl-2,6-dimethylpiperidin-4-yl](ethyl)amino}-N-[(4,6-dimethyl-2--
oxo-1,2-dihydropyridin-3-yl)methyl]-2-methyl-5-(trifluoromethyl)benzamide
(35.6 mg, 0.0611 mmol) in MeOH (3.0 ml) was added Pd/C (120 mg).
The reaction mixture was stirred for 30 minutes under hydrogen
atmosphere. After purging with nitrogen, the mixture was filtered
and the filtrate was concentrated in vacuo. The residue was
purified by preparative TLC (TLC silica gel 60F254, MERCK,
MeOH/Et.sub.3N=100/1) twice to give title compound as a white solid
(4.4 mg, 15% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm;
7.33 (s, 1H), 7.30 (s, 1H), 7.18 (m, 1H), 5.96 (s, 1H), 4.54 (d,
J=6.0 Hz, 2H), 3.73 (m, 1H), 3.26-2.69 (m, 4H), 2.40 (s, 3H), 2.38
(s, 3H), 2.23 (s, 3H), 1.90-1.10 (m, 4H), 1.10-0.86 (m, 6H), 0.83
(t, J=7.2 Hz, 3H). (-2H); LC-MS: m/z [M+H] 493.2, [M+Na] 515.3;
HPLC; 88.8% purity.
Compound 403:
3-(ethyl(1-methylazetidin-3-yl)amino)-2-methyl-N-((6-methyl-2-oxo-4-propy-
l-1,2-dihydropyridin-3-yl)methyl)-5-(trifluoromethyl)benzamide
formate
##STR01086##
[2271] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.33 (s, 1H),
7.37 (s, 1H), 7.26 (s, 1H), 6.11 (s, 1H), 4.46 (s, 2H), 4.42 (br,
m, 1H), 4.21 (br, 2H), 3.82 (br, 2H), 3.00 (q, J=6.8 Hz, 2H), 2.88
(s, 3H), 2.65 (m, 2H), 2.40 (s, 3H), 2.22 (s, 3H), 1.61 (m, 2H),
0.99 (t, J=7.2 Hz, 3H), 0.86 (t, J=6.8 Hz, 3H); MS (ES) (M+1)
480.53.
Compound 404:
3-(ethyl(1-methylazetidin-3-yl)amino)-N-((4-isopropyl-6-methyl-2-oxo-1,2--
dihydropyridin-3-yl)methyl)-2-methyl-5-(trifluoromethyl)benzamide
formate
##STR01087##
[2273] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.31 (s, 1H),
7.37 (s, 1H), 7.27 (s, 1H), 6.21 (s, 1H), 4.50 (s, 2H), 4.41 (br,
1H0, 4.21 (br, 2H), 3.81 (br, 2H), 3.38 (m, 1H), 3.00 (q, J=6.8 Hz,
2H), 2.87 (s, 3H), 2.40 (s, 3H), 2.24 (s, 3H), 1.20 (d, J=6.4 Hz,
6H), 0.86 (t, J=6.8 Hz, 3H); MS (ES) (M+1) 479.54.
Compound 405:
N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(1-methyla-
zetidin-3-yl)amino)-2-methyl-5-(trifluoromethyl)benzamide
formate
##STR01088##
[2275] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. ppm 8.32 (s, 1H),
7.39 (s, 1H), 7.26 (s, 1H), 6.08 (s, 1H), 4.44 (s, 2H), 4.41 (br,
m, 1H), 4.21 (br, 2H), 3.82 (br, 2H), 3.00 (q, J=6.8 Hz, 2H), 2.88
(s, 3H), 2.39 (s, 3H), 2.34 (d, J=1.2 Hz, 3H), 2.21 (s, 3H), 0.86
(t, J=6.8 Hz, 3H); MS (ES) (M+1) 451.49.
Compound 406:
5-Chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-[(6-methyl-2-o-
xo-4-propyl-1,2-dihydropyridin-3-yl)methyl]benzamide
##STR01089##
[2277] To a stirred solution of
5-chloro-3-[ethyl(1-methylpiperidin-4-yl) amino]-2-methylbenzoic
acid (crude material, 1.80 g, 3.26 mmol) and
3-(aminomethyl)-6-methyl-4-propyl-1,2-dihydropyridin-2-one HCl salt
(916 mg, 4.24 mmol) in DMSO (10 mL) was added PyBOP (3.10 g, 5.87
mmol) and hunig base (1.20 g, 9.78 mmol). The reaction mixture was
stirred at 23.degree. C. for 16 hours. The reaction mixture was
quenched with water and resulting precipitate was collected. The
solid was purified by silica gel column chromatography
(NH--SiO.sub.2 Ethylacetate/MeOH=10/1) to give the titled compound
as a white solid (264 mg, 17% yield). .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. ppm; 7.20 (t, J=5.9 Hz, 1H), 7.06 (d, J=2.0 Hz,
1H), 7.02 (d, J=2.0 Hz, 1H), 5.96 (s, 1H), 4.54 (d, J=5.9 Hz, 2H),
3.02 (q, J=7.0 Hz, 2H), 2.81 (m, 2H), 2.64-2.71 (m, 3H), 2.21-2.27
(s.times.3, 9H), 1.84-1.94 (m, 2H), 1.55-1.65 (m, 6H), 1.00 (t,
J=7.4 Hz, 3H), 0.84 (t, J=7.0 Hz, 3H); MS (ES) [M+H] 473.1, [M+Na]
495.3; HPLC 94.7% purity.
Compound 407:
5-Chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-o-
xo-4-(propan-2-yl)-1,2-dihydropyridin-3-yl]methyl}benzamide
##STR01090##
[2279] To a stirred solution of
5-chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methylbenzoic
acid (crude material, .about.0.7 mmol) and
3-(aminomethyl)-6-methyl-4-(propan-2-yl)-1,2-dihydropyridin-2-one
hydrochloride HCl salt (200 mg, 0.91 mmol) in DMSO (5.3 mL) was
added PyBOP (550 mg, 1.05 mmol) and hunig base (366 ul, 2.10 mmol).
The reaction mixture was stirred at 23.degree. C. for 7 hours. The
reaction mixture was quenched with water and resulting precipitate
was collected. The solid was purified by silica gel column
chromatography (NH--SiO.sub.2 Ethylacetate/MeOH=10/1+10%
triethylamine). The crude compound was triturated from
ethylacetate/hexane, and dried to give the titled compound as a
white solid (166 mg, 50% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. ppm; 7.06 (d, J=2.4 Hz, 1H), 7.01-7.05 (m, 1H), 7.00 (d,
J=2.4 Hz, 1H), 6.04 (s, 1H), 4.57 (d, J=6.0 Hz, 2H), 3.49-3.57 (m,
1H), 3.02 (q, J=6.8 Hz, 2H), 2.80-2.84 (m, 2H), 2.61-2.69 (m, 1H),
2.29 (s, 3H), 2.23 (s, 3H), 2.23 (s, 3H), 1.86-1.90 (m, 2H),
1.66-1.70 (m, 4H), 1.21 (d, J=6.8 Hz, 6H), 0.84 (t, J=6.8 Hz, 3H);
MS (ES) [M+Na] 495.2; HPLC 95.7% purity.
Compound 408:
5-Chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-o-
xo-4-(trifluoromethyl)-1,2-dihydropyridin-3-yl]methyl}benzamide
##STR01091##
[2281] To a stirred solution of
5-chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methylbenzoic
acid (crude material, .about.0.7 mmol) and
3-(aminomethyl)-6-methyl-4-(trifluoromethyl)-1,2-dihydropyridin-2-one
hydrochloride HCl salt (200 mg, 0.91 mmol) in DMSO (5.3 mL) was
added PyBOP (550 mg, 1.05 mmol) and hunig base (366 ul, 2.10 mmol).
The reaction mixture was stirred at 23.degree. C. for 7 hours. The
reaction mixture was quenched with water and diluted with
ethylacetate. The aqueous layer was extracted with dichloromethane.
The combined organic layer was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel column chromatography (SiO.sub.2
Ethylacetate/MeOH=10/1+5% triethylamine). The crude compound was
triturated with ethylacetate/hexane, and dried to give the titled
compound as a white solid (145 mg, 42% yield). .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta. ppm; 7.08 (d, J=2.4 Hz, 1H), 7.03 (d,
J=2.4 Hz, 1H), 6.81-6.85 (m, 1H), 6.35 (s, 1H), 4.71 (d, J=5.2 Hz,
2H), 3.03 (q, J=6.8 Hz, 2H), 2.81-2.85 (m, 2H), 2.68-2.72 (m, 1H),
2.39 (s, 3H), 2.27 (s, 3H), 2.24 (s, 3H), 1.88-1.94 (m, 2H),
1.67-1.72 (m, 4H), 0.85 (t, J=6.8 Hz, 3H); MS (ES) [M+H] 499.1,
[M+Na] 521.1; HPLC 96.0% purity.
[2282] Following the same preparation method described above for
5-chloro-3-[ethyl(1-methylpiperidin-4-yl)amino]-2-methyl-N-{[6-methyl-2-o-
xo-4-(trifluoromethyl)-1,2-dihydropyridin-3-yl]methyl}benzamide,
the following analogs were prepared using the corresponding
pyridone fragments described earlier, and purified by reverse
HPLC/MS (ACN-H.sub.2O, containing 0.1% formic acid).
Compound 409:
5-chloro-3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-4,6-dimethyl-
-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
formate
##STR01092##
[2284] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 8.45-8.25 (br,
1H), 7.30 (d, J=2.0 Hz, 1H), 7.13 (d, J=2.0 Hz, 1H), 4.47 (s, 2H),
3.50-3.38 (br, 2H), 3.25-3.15 (br, 1H), 3.08 (q, J=7.0 Hz, 2H),
3.06-2.98 (br, 2H), 2.83 (s, 3H), 2.38 (d, J=2.0 Hz, 3H), 2.27 (s,
3H), 2.26 (d, J=2.0 Hz, 3H), 2.03 (bd, J=14.75 Hz, 2H), 1.90-1.77
(br, 2H), 0.89 (t, J=7.0 Hz, 3H); MS (ES) (M+H) 463.50.
Compound 410:
5-chloro-3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-6-methyl-2-o-
xo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
formate
##STR01093##
[2286] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 8.43-8.26 (br,
1H), 7.30 (d, J=2.0 Hz, 1H), 7.12 (d, J=2.0 Hz, 1H), 4.47 (s, 2H),
3.47-3.39 (br, 2H), 3.23-3.15 (br, 1H), 3.08 (q, J=7.0 Hz, 2H),
3.05-2.96 (br, 2H), 2.82 (s, 3H), 2.79 (bd, J=8.2 Hz, 2H),
2.28-2.26 (br, 6H), 2.06-1.99 (br, 2H), 1.93-1.82 (br, 2H),
1.68-1.59 (m, 2H), 1.06 (t, J=7.5 Hz, 3H), 0.89 (t, J=7.0 Hz, 3H);
MS (ES) (M+H) 491.52.
Compound 411:
5-chloro-3-(ethyl(1-methylpiperidin-4-yl)amino)-N-((5-fluoro-4-isopropyl--
6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide
formate
##STR01094##
[2288] .sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 8.48-8.34 (br,
1H), 7.35 (d, J=2.0 Hz, 1H), 7.16 (d, J=2.0 Hz, 1H), 4.57 (s, 2H),
3.61-3.53 (m, 1H), 3.52-3.44 (br, 2H), 3.30-3.20 (br, 1H), 3.13 (q,
J=7.0 Hz, 2H), 3.10-3.01 (br, 2H), 2.87 (s, 3H), 2.33 (s, 3H), 2.31
(d, J=2.8 Hz, 3H), 2.10-2.04 (br, 2H), 1.98-1.87 (br, 2H), 1.41 (d,
J=6.5 Hz, 6H), 0.93 (t, J=7.0 Hz, 3H); MS (ES) (M+H) 491.51.
Compound 412:
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methyl-N-((6-methyl-2-ox-
o-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide formate
##STR01095##
[2290] Methyl
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methylbenzoate was
hydrolyzed to
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methylbenzoic acid
following the same preparation method for
3-{Ethyl[1-(propan-2-yl)piperidin-4-yl]amino}-2-methyl-5-(trifluoromethyl-
)benzoic acid from its corresponding carboxylate. The crude
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methylbenzoic acid
was then coupled with
3-(Aminomethyl)-6-methyl-4-propyl-1,2-dihydropyridin-2-one HCl salt
prepared earlier following the preparation method described for
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-[ethyl(1-methylp-
iperidin-4-yl)amino]-2-methyl-5-(trifluoromethyl)benzamide. After
purification by reverse phase HPLC/MS (CAN-H.sub.2O containing 0.1%
formic acid), the titled compound was obtained. .sup.1H-NMR (500
MHz, CD.sub.3OD) .delta. ppm 8.58-8.24 (br, 1H), 7.15 (d, J=2.0 Hz,
1H), 7.08 (d, J=2.0 Hz, 1H), 6.14 (s, 1H), 4.48 (s, 2H), 4.41-4.34
(m, 1H), 4.25 (br, 2H), 3.85 (br, 2H), 3.00 (q, J=7.0 Hz, 2H), 2.92
(s, 3H), 2.72-2.66 (m, 2H), 2.34 (s, 3H), 2.26 (s, 3H), 1.70-1.61
(m, 2H), 1.04 (t, J=7.2 Hz, 3H), 0.91 (t, J=7.0 Hz, 3H); MS (ES)
(M+H) 445.48.
Compound 413:
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-N-((4-isopropyl-6-methyl-2-
-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methylbenzamide formate
##STR01096##
[2292] The same way for the previous compound
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methyl-N-((6-methyl-2-ox-
o-4-propyl-1,2-dihydropyridin-3-yl)methyl)benzamide formate
prepared, the titled compound prepared using
5-chloro-3-(ethyl(1-methylazetidin-3-yl)amino)-2-methylbenzoic acid
and
3-(Aminomethyl)-6-methyl-4-(propan-2-yl)-1,2-dihydropyridin-2-one
HCl salt. After purification by reverse phase HPLC/MS (CAN-H.sub.2O
containing 0.1% formic acid), the titled compound was obtained.
.sup.1H-NMR (500 MHz, CD.sub.3OD) .delta. ppm 8.43-8.25 (br, 1H),
7.15 (d, J=2.0 Hz, 1H), 7.08 (d, J=2.0 Hz, 1H), 6.25 (s, 1H), 4.52
(s, 2H), 4.40-4.34 (m, 1H), 4.29-4.21 (br, 2H), 3.90-3.79 (br, 2H),
3.48-3.40 (m, 1H), 3.00 (q, J=7.0 Hz, 2H), 2.92 (s, 3H), 2.34 (s,
3H), 2.28 (s, 3H), 1.24 (d, J=7.0 Hz, 6H), 0.90 (t, J=7.0 Hz, 3H).
MS (ES) (M+H) 445.47.
Compound 414:
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-{[(2R,6R)-2,6-di-
methylpiperidin-4-yl](ethyl)amino}-5-fluoro-2-methylbenzamide
##STR01097##
[2294] To a stirred solution of
3-{[(2R,6R)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-fl-
uoro-2-methylbenzamide (378 mg, 0.709 mmol) in MeOH (7 mL) was
added Pd/C (150 mg). The reaction mixture was stirred at 23.degree.
C. for 1 hour under hydrogen atmosphere. The reaction mixture was
filtered and the filtrate was concentrated in vacuo. The residue
was purified by silica gel column chromatography (1. NAM-300H
silica gel produced by Nagara Science Co., Ltd.,
ethylacetate/MeOH=4/1 to ethylacetate/MeOH/triethylamine=1/1/0.01,
2. NH--SiO.sub.2, heptane/ethylacetate=30/1 to 8/1) to give title
compound as a white solid (235 mg, 75% yield). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. ppm; 7.08 (t, J=6.2 Hz, 1H), 6.82 (dd,
J=10.7, 2.5 Hz, 1H), 6.75 (dd, J=8.2, 2.7 Hz, 1H), 5.93 (s, 1H),
4.51 (d, J=6.2 Hz, 2H), 3.40-3.48 (m, 1H), 2.95-3.09 (m, 3H),
2.83-2.92 (m, 1H), 2.39 (s, 3H), 2.22 (s.times.2, 6H), 1.63-1.78
(m, 4H), 1.13 (d, J=7.0 Hz, 3H), 1.02 (d, J=6.2 Hz, 3H), 0.84 (t,
J=7.0 Hz, 3H); MS (ES) [M+H] 443.1, [M+Na] 465.2; HPLC; 99.3%
purity.
Compound 415:
N-[(4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-3-{[(2R,4R,6S)-2,6-
-dimethylpiperidin-4-yl](ethyl)amino}-5-fluoro-2-methylbenzamide
##STR01098##
[2296] To a stirred solution of
3-{[(2R,4R,6S)-1-benzyl-2,6-dimethylpiperidin-4-yl]
(ethyl)amino}-N-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-5-fl-
uoro-2-methylbenzamide (50.1 mg, 0.0860 mmol) in MeOH (2 ml) was
added Pd/C (100 mg). The reaction mixture was stirred for 3 hours
under hydrogen atmosphere at 23.degree. C. After purging with
nitrogen, the mixture was filtered and the filtrate was
concentrated in vacuo. The residue was purified by silica gel
column chromatography (NAM-300H silica gel produced by Nagara
Science Co., Ltd., ethylacetate/MeOH=4/1 to
ethylacetate/MeOH/triethylamine=1/1/0.01). The target fractions
were collected and concentrated in vacuo. The residue was purified
by silica gel column chromatography (NH--SiO.sub.2,
ethylacetate/MeOH=30/1 to 20/1 to 8/1). The target fractions were
collected and concentrated in vacuo. The residue was purified with
preparative TLC (TLC silica gel 60F.sub.254, MERCK,
MeOH/Et.sub.3N=100/1) to give the titled compound as a white solid
(17.2 mg, 45% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm; 7.10 (m, 1H), 6.81 (m, 2H), 5.91 (s, 1H), 4.52 (d, J=6.0 Hz,
2H), 3.61 (m, 1H), 3.23-2.73 (m, 4H), 2.40 (s, 3H), 2.28 (s, 3H),
2.24 (s, 3H), 1.64-1.05 (m, 4H), 1.05-0.86 (m, 6H), 0.82 (t, J=7.2
Hz, 3H). (-2H); LC-MS: m/z [M+H] 443.2, [M+Na] 465.3; HPLC; 97.4%
purity.
Example 45
Syntheses of Compounds 6, 9, 11-12, 19, 22, 24, 27, 29, 34-35,
37-47, 65-90, 93-96, 99-101, 103, 106-116, 124, 125, 128-136,
138-143, 145-156, 158-190, 193-204, 210, 211, 214-221, 223-242,
244-267, 270-272, 274, 275, 280, 281, 288, 289, 291-300, 303-305,
307, 312, 314, 316-318, 323-327, 329-331, 337, 338, 341-343, 345,
346, 350-355, 357-385, 388, 391, and 418
[2297] Compounds 6, 9, 11-12, 19, 22, 24, 27, 29, 34-35, 37-47,
65-90, 93-96, 99-101, 103, 106-116, 124, 125, 128-136, 138-143,
145-156, 158-190, 193-204, 210, 211, 214-221, 223-242, 244-267,
270-272, 274, 275, 280, 281, 288, 289, 291-300, 303-305, 307, 312,
314, 316-318, 323-327, 329-331, 337, 338, 341-343, 345, 346,
350-355, 357-385, 388, 391, and 418 were synthesized by methods
similar to those described for Examples 1-44 or by reaction schemes
depicted in the general schemes.
Example 46
Bioassay Protocol and General Methods
Protocol for Wild-Type and Mutant PRC2 Enzyme Assays
[2298] General Materials.
[2299] S-adenosylmethionine (SAM), S-adenosylhomocyteine (SAH),
bicine, KCl, Tween20, dimethylsulfoxide (DMSO) and bovine skin
gelatin (BSG) were purchased from Sigma-Aldrich at the highest
level of purity possible. Dithiothreitol (DTT) was purchased from
EMD. .sup.3H-SAM was purchased from American Radiolabeled Chemicals
with a specific activity of 80 Ci/mmol. 384-well streptavidin
Flashplates were purchased from PerkinElmer.
[2300] Substrates.
[2301] Peptides representative of human histone H3 residues 21-44
containing either an unmodified lysine 27 (H3K27me0) or
dimethylated lysine 27 (H3K27me2) were synthesized with a
C-terminal G(K-biotin) linker-affinity tag motif and a C-terminal
amide cap by 21.sup.st Century Biochemicals. The peptides were
high-performance liquid chromatography (HPLC) purified to greater
than 95% purity and confirmed by liquid chromatography mass
spectrometry (LC-MS). The sequences are listed below.
TABLE-US-00002 H3K27me0: (SEQ ID NO: 1)
ATKAARKSAPATGGVKKPHRYRPGGK(biotin)-amide H3K27me2: (SEQ ID NO: 2)
ATKAARK(me2)SAPATGGVKKPHRYRPGGK(biotin)-amide
[2302] Chicken erythrocyte oligonucleosomes were purified from
chicken blood according to established procedures.
[2303] Recombinant PRC2 Enzymes.
[2304] Human PRC2 enzymes were purified as 4-component enzyme
complexes co-expressed in Spodoptera frugiperda (sf9) cells using a
baculovirus expression system. The subunits expressed were
wild-type EZH2 (NM_004456) or EZH2 Y641F, N, H, S or C mutants
generated from the wild-type EZH2 construct, EED (NM_003797), Suz12
(NM_015355) and RbAp48 (NM_005610). The EED subunit contained an
N-terminal FLAG tag that was used to purify the entire 4-component
complex from sf9 cell lysates. The purity of the complexes met or
exceeded 95% as determined by SDS-PAGE and Agilent Bioanalyzer
analysis. Concentrations of enzyme stock concentrations (generally
0.3-1.0 mg/mL) was determined using a Bradford assay against a
bovine serum albumin (BSA) standard.
[2305] General Procedure for PRC2 Enzyme Assays on Peptide
Substrates.
[2306] The assays were all performed in a buffer consisting of 20
mM bicine (pH=7.6), 0.5 mM DTT, 0.005% BSG and 0.002% Tween20,
prepared on the day of use. Compounds in 100% DMSO (1 .mu.L) were
spotted into polypropylene 384-well V-bottom plates (Greiner) using
a Platemate 2.times.3 outfitted with a 384-channel pipet head
(Thermo). DMSO (1 .mu.L) was added to columns 11, 12, 23, 24, rows
A-H for the maximum signal control, and SAH, a known product and
inhibitor of PRC2 (1 .mu.L) was added to columns 11, 12, 23, 24,
rows I-P for the minimum signal control. A cocktail (40 .mu.L)
containing the wild-type PRC2 enzyme and H3K27me0 peptide or any of
the Y641 mutant enzymes and H3K27me2 peptide was added by Multidrop
Combi (Thermo). The compounds were allowed to incubate with PRC2
for 30 min at 25.degree. C., then a cocktail (10 .mu.L) containing
a mixture of non-radioactive and .sup.3H-SAM was added to initiate
the reaction (final volume=51 .mu.L). In all cases, the final
concentrations were as follows: wild-type or mutant PRC2 enzyme was
4 nM, SAH in the minimum signal control wells was 1 mM and the DMSO
concentration was 1%. The final concentrations of the rest of the
components are indicated in Table 2, below. The assays were stopped
by the addition of non-radioactive SAM (10 .mu.L) to a final
concentration of 600 .mu.M, which dilutes the .sup.3H-SAM to a
level where its incorporation into the peptide substrate is no
longer detectable. 50 .mu.L of the reaction in the 384-well
polypropylene plate was then transferred to a 384-well Flashplate
and the biotinylated peptides were allowed to bind to the
streptavidin surface for at least 1 h before being washed three
times with 0.1% Tween20 in a Biotek ELx405 plate washer. The plates
were then read in a PerkinElmer TopCount platereader to measure the
quantity of .sup.3H-labeled peptide bound to the Flashplate
surface, measured as disintegrations per minute (dpm) or
alternatively, referred to as counts per minute (cpm).
TABLE-US-00003 TABLE 2 Final concentrations of components for each
assay variation based upon EZH2 identity (wild-type or Y641 mutant
EZH2) PRC2 Enzyme (denoted by EZH2 Peptide Non-radioactive SAM
identity) (nM) (nM) .sup.3H-SAM (nM) Wild-type 185 1800 150 Y641F
200 850 150 Y641N 200 850 150 Y641H 200 1750 250 Y641S 200 1300 200
Y641C 200 3750 250
[2307] General Procedure for Wild-Type PRC2 Enzyme Assay on
Oligonucleosome Substrate.
[2308] The assays was performed in a buffer consisting of 20 mM
bicine (pH=7.6), 0.5 mM DTT, 0.005% BSG, 100 mM KCl and 0.002%
Tween20, prepared on the day of use. Compounds in 100% DMSO (1
.mu.L) were spotted into polypropylene 384-well V-bottom plates
(Greiner) using a Platemate 2.times.3 outfitted with a 384-channel
pipet head (Thermo). DMSO (1 .mu.L) was added to columns 11, 12,
23, 24, rows A-H for the maximum signal control, and SAH, a known
product and inhibitor of PRC2 (1 .mu.L) was added to columns 11,
12, 23, 24, rows I-P for the minimum signal control. A cocktail (40
.mu.L) containing the wild-type PRC2 enzyme and chicken erythrocyte
oligonucleosome was added by Multidrop Combi (Thermo). The
compounds were allowed to incubate with PRC2 for 30 min at
25.degree. C., then a cocktail (10 .mu.L) containing a mixture of
non-radioactive and .sup.3H-SAM was added to initiate the reaction
(final volume=51 .mu.L). The final concentrations were as follows:
wild-type PRC2 enzyme was 4 nM, non-radioactive SAM was 430 nM,
.sup.3H-SAM was 120 nM, chicken erythrocyte oligonucleosome was 120
nM, SAH in the minimum signal control wells was 1 mM and the DMSO
concentration was 1%. The assay was stopped by the addition of
non-radioactive SAM (10 .mu.L) to a final concentration of 600
.mu.M, which dilutes the .sup.3H-SAM to a level where its
incorporation into the chicken erythrocyte oligonucleosome
substrate is no longer detectable. 50 .mu.L of the reaction in the
384-well polypropylene plate was then transferred to a 384-well
Flashplate and the chicken erythrocyte nucleosomes were immobilized
to the surface of the plate, which was then washed three times with
0.1% Tween20 in a Biotek ELx405 plate washer. The plates were then
read in a PerkinElmer TopCount platereader to measure the quantity
of .sup.3H-labeled chicken erythrocyte oligonucleosome bound to the
Flashplate surface, measured as disintegrations per minute (dpm) or
alternatively, referred to as counts per minute (cpm).
% Inhibition Calculation ##EQU00001## % inh = 100 - ( dpm cmpd -
dpm min dpm max - dpm min ) .times. 100 ##EQU00001.2##
[2309] Where dpm=disintegrations per minute, cmpd=signal in assay
well, and min and max are the respective minimum and maximum signal
controls.
Four - parameter IC 50 fit ##EQU00002## Y = Bottom + ( Top - Bottom
) 1 + ( X IC 50 ) Hill Coefficient ##EQU00002.2##
[2310] Where top and bottom are the normally allowed to float, but
may be fixed at 100 or 0 respectively in a 3-parameter fit. The
Hill Coefficient normally allowed to float but may also be fixed at
1 in a 3-parameter fit. Y is the % inhibition and X is the compound
concentration.
[2311] IC.sub.50 values for the PRC2 enzyme assays on peptide
substrates (e.g., EZH2 wild type and Y641F) are presented in Table
3 below.
[2312] WSU-DLCL2 Methylation Assay
[2313] WSU-DLCL2 suspension cells were purchased from DSMZ (German
Collection of Microorganisms and Cell Cultures, Braunschweig,
Germany). RPMI/Glutamax Medium, Penicillin-Streptomycin, Heat
Inactivated Fetal Bovine Serum, and D-PBS were purchased from Life
Technologies, Grand Island, N.Y., USA. Extraction Buffer and
Neutralization Buffer (5.times.) were purchased from Active Motif,
Carlsbad, Calif., USA. Rabbit anti-Histone H3 antibody was
purchased from Abcam, Cambridge, Mass., USA. Rabbit anti-H3K27me3
and HRP-conjugated anti-rabbit-IgG were purchased from Cell
Signaling Technology, Danvers, Mass., USA. TMB "Super Sensitive"
substrate was sourced from BioFX Laboratories, Owings Mills, Md.,
USA. IgG-free Bovine Serum Albumin was purchased from Jackson
ImmunoResearch, West Grove, Pa., USA. PBS with Tween
(10.times.PBST) was purchased from KPL, Gaithersburg, Md., USA.
Sulfuric Acid was purchased from Ricca Chemical, Arlington, Tex.,
USA. Immulon ELISA plates were purchased from Thermo, Rochester,
N.Y., USA. V-bottom cell culture plates were purchased from Corning
Inc., Corning, N.Y., USA. V-bottom polypropylene plates were
purchased from Greiner Bio-One, Monroe, N.C., USA.
[2314] WSU-DLCL2 suspension cells were maintained in growth medium
(RPMI 1640 supplemented with 10% v/v heat inactivated fetal bovine
serum and 100 units/mL penicillin-streptomycin) and cultured at
37.degree. C. under 5% CO.sub.2. Under assay conditions, cells were
incubated in Assay Medium (RPMI 1640 supplemented with 20% v/v heat
inactivated fetal bovine serum and 100 units/mL
penicillin-streptomycin) at 37.degree. C. under 5% CO.sub.2 on a
plate shaker.
[2315] WSU-DLCL2 cells were seeded in assay medium at a
concentration of 50,000 cells per mL to a 96-well V-bottom cell
culture plate with 200 .mu.L per well. Compound (1 .mu.L) from 96
well source plates was added directly to V-bottom cell plate.
Plates were incubated on a titer-plate shaker at 37.degree. C., 5%
CO2 for 96 hours. After four days of incubation, plates were spun
at 241.times.g for five minutes and medium was aspirated gently
from each well of cell plate without disturbing cell pellet. Pellet
was resuspended in 200 .mu.L DPBS and plates were spun again at
241.times.g for five minutes. The supernatant was aspirated and
cold (4.degree. C.) Extraction buffer (100 .mu.L) was added per
well. Plates were incubated at 4.degree. C. on orbital shaker for
two hours. Plates were spun at 3427.times.g.times.10 minutes.
Supernatant (80 .mu.L per well) was transferred to its respective
well in 96 well V-bottom polypropylene plate. Neutralization Buffer
5.times. (20 per well) was added to V-bottom polypropylene plate
containing supernatant. V-bottom polypropylene plates containing
crude histone preparation (CHP) were incubated on orbital
shaker.times. five minutes. Crude Histone Preparations were added
(2 .mu.L per well) to each respective well into duplicate 96 well
ELISA plates containing 100 .mu.L Coating Buffer (1.times.PBS+BSA
0.05% w/v). Plates were sealed and incubated overnight at 4.degree.
C. The following day, plates were washed three times with 300 .mu.L
per well 1.times.PBST. Wells were blocked for two hours with 300
.mu.L per well ELISA Diluent ((PBS (1.times.) BSA (2% w/v) and
Tween20 (0.05% v/v)). Plates were washed three times with
1.times.PBST. For the Histone H3 detection plate, 100 per well were
added of anti-Histone-H3 antibody (Abcam, ab1791) diluted 1:10,000
in ELISA Diluent. For H3K27 trimethylation detection plate, 100
.mu.L per well were added of anti-H3K27me3 diluted 1:2000 in ELISA
diluent. Plates were incubated for 90 minutes at room temperature.
Plates were washed three times with 300 .mu.L 1.times.PBST per
well. For Histone H3 detection, 100 .mu.L of HRP-conjugated
anti-rabbit IgG antibody diluted to 1:6000 in ELISA diluent was
added per well. For H3K27me3 detection, 100 .mu.L of HRP conjugated
anti-rabbit IgG antibody diluted to 1:4000 in ELISA diluent was
added per well. Plates were incubated at room temperature for 90
minutes. Plates were washed four times with 1.times.PBST 300 .mu.L
per well. TMB substrate 100 .mu.L was added per well. Histone H3
plates were incubated for five minutes at room temperature.
H3K27me3 plates were incubated for 10 minutes at room temperature.
The reaction was stopped with sulfuric acid 1N (100 .mu.L per
well). Absorbance for each plate was read at 450 nm.
[2316] First, the ratio for each well is determined by:
( H 3 K 27 me 3 OD 450 Histone H 3 OD 450 ) ##EQU00003##
[2317] Each plate included eight control wells of DMSO only
treatment (Minimum Inhibition) as well as eight control wells for
maximum inhibition (Background wells).
[2318] The average of the ratio values for each control type was
calculated and used to determine the percent inhibition for each
test well in the plate. Test compound was serially diluted
three-fold in DMSO for a total of ten test concentrations,
beginning at 25 .mu.M. Percent inhibition was determined and
IC.sub.50 curves were generated using duplicate wells per
concentration of compound. IC.sub.50 values for this assay are
presented in Table 3 below.
Percent Inhibition = 100 - ( ( ( Individual Test Sample Ratio ) - (
Background Avg Ratio ) ( Minimum Inhibition Ratio ) - ( Background
Average Ratio ) ) * 100 ) ##EQU00004##
Cell Proliferation Analysis
[2319] WSU-DLCL2 suspension cells were purchased from DSMZ (German
Collection of Microorganisms and Cell Cultures, Braunschweig,
Germany). RPMI/Glutamax Medium, Penicillin-Streptomycin, Heat
Inactivated Fetal Bovine Serum were purchased from Life
Technologies, Grand Island, N.Y., USA. V-bottom polypropylene
384-well plates were purchased from Greiner Bio-One, Monroe, N.C.,
USA. Cell culture 384-well white opaque plates were purchased from
Perkin Elmer, Waltham, Mass., USA. Cell-Titer Glo.RTM. was
purchased from Promega Corporation, Madison, Wis., USA. SpectraMax
M5 plate reader was purchased from Molecular Devices LLC,
Sunnyvale, Calif., USA.
[2320] WSU-DLCL2 suspension cells were maintained in growth medium
(RPMI 1640 supplemented with 10% v/v heat inactivated fetal bovine
serum and cultured at 37.degree. C. under 5% CO.sub.2. Under assay
conditions, cells were incubated in Assay Medium (RPMI 1640
supplemented with 20% v/v heat inactivated fetal bovine serum and
100 units/mL penicillin-streptomycin) at 37.degree. C. under 5%
CO.sub.2.
[2321] For the assessment of the effect of compounds on the
proliferation of the WSU-DLCL2 cell line, exponentially growing
cells were plated in 384-well white opaque plates at a density of
1250 cell/ml in a final volume of 50 .mu.l of assay medium. A
compound source plate was prepared by performing triplicate
nine-point 3-fold serial dilutions in DMSO, beginning at 10 mM
(final top concentration of compound in the assay was 20 .mu.M and
the DMSO was 0.2%). A 100 nL aliquot from the compound stock plate
was added to its respective well in the cell plate. The 100%
inhibition control consisted of cells treated with 200 nM final
concentration of staurosporine and the 0% inhibition control
consisted of DMSO treated cells. After addition of compounds, assay
plates were incubated for 6 days at 37.degree. C., 5% CO.sub.2,
relative humidity>90% for 6 days. Cell viability was measured by
quantization of ATP present in the cell cultures, adding 35 .mu.l
of Cell Titer Glo.RTM. reagent to the cell plates. Luminescence was
read in the SpectraMax M5. The concentration inhibiting cell
viability by 50% was determined using a 4-parametric fit of the
normalized dose response curves. IC.sub.50 values for this assay
are also presented in Table 3 below.
TABLE-US-00004 TABLE 3 Mutant H3K27Me3 WSU Y641F ELISA
proliferation WT EZH2 IC.sub.50 IC.sub.50 IC.sub.50 Cpd # IC.sub.50
(.mu.M) (.mu.M) (.mu.M) (.mu.M) 1 0.0199 3.09728 2 0.02348 0.62109
3 0.04011 4 0.02231 0.54412 5 0.02389 2.19558 6 0.02703 0.50577 7
0.03317 8 0.03622 0.02506 9 0.03501 10 0.03978 11 0.04682 12
0.05026 13 0.06396 14 0.04353 15 0.07173 16 0.10471 17 0.08438 18
0.1603 19 0.10845 20 0.16251 21 0.17104 22 0.20549 23 0.17755 24
0.22919 25 0.23 26 0.29374 27 0.48381 28 0.54889 29 0.62173 30
0.62615 31 0.6324 32 0.63441 33 0.65963 35 0.82442 36 0.91712 37
1.00379 38 1.0545 39 1.07137 40 1.09168 41 1.13408 42 2.59043 43
2.67651 44 4.32369 45 25.22746 46 26.53597 47 39.133 48 0.24998 49
3.59887 50 0.30813 51 0.37599 52 0.04814 53 0.41267 54 0.50073 55
0.01626 3.91279 56 0.01519 0.21216 57 0.02987 7.72974 58 0.05639 59
0.03149 60 0.05852 61 0.01585 0.33187 62 0.09429 63 0.05709 64
0.02472 2.06774 65 1.07044 66 1.71172 67 3.61905 68 1.3267 69
4.71695 70 0.90608 71 1.21512 72 1.01436 73 4.14125 74 0.07113 75
0.3843 76 1.24461 77 0.14901 78 0.14362 79 0.02465 0.02116 80
1.64878 82 4.20192 83 0.18011 0.09576 84 0.11391 0.10243 85 0.10754
0.07626 86 2.57252 87 0.09019 0.10057 88 0.03758 0.02817 89 0.07342
0.08298 90 0.0998 0.1002 91 0.02271 0.0176 2.5696 92 0.01655
0.02341 1.23721 93 1.18036 1.08352 94 3.80992 5.5015 95 0.35125
0.33255 96 0.38143 0.40305 97 0.02024 0.0292 2.4644 98 0.0174
0.03732 1.75896 99 0.01481 0.01028 3.86397 100 0.03203 0.01966 101
0.00711 0.00717 1.07296 102 0.01545 0.00727 0.61272 103 0.072
0.06724 104 0.03077 0.0318 9.05638 105 0.01728 0.01229 6.47052 106
0.2446 0.19119 107 0.05396 0.10057 108 0.02003 0.0175 4.42418 109
0.01112 0.01094 0.44691 110 0.04792 0.06226 111 0.05328 0.02875 112
0.0587 0.02666 113 0.18895 0.17503 114 0.1051 0.11091 115 0.06117
0.05267 116 0.06276 0.0452 117 0.02807 0.02213 118 0.01759 0.01615
8.60536 119 0.00951 0.00748 1.40617 120 0.00911 0.01085 0.36889 121
0.01385 0.0133 0.5035 122 0.01511 0.01932 0.18096 123 0.09153
0.06041 124 0.16779 0.11113 125 0.04721 0.0315 126 0.04784 0.03472
127 0.01691 0.01923 1.50101 128 0.04911 0.0438 129 0.11098 0.08998
130 0.01722 0.01508 2.3697 131 0.02287 0.00778 1.65122 132 0.06337
0.0717 133 0.04829 0.03059 134 0.02562 0.02189 1.08958 135 0.04667
0.02555 4.14394 136 0.11517 0.0957 137 0.01602 0.00604 3.29434 138
0.03495 0.04493 >20.0 uM 139 0.0204 0.01392 1.79898 140 1.66626
0.91482 141 0.06632 0.18147 142 0.02672 0.02005 3.72097 143 0.13711
0.05828 144 2.294 1.3667 145 0.10444 0.07112 146 0.32313 0.28854
147 0.14779 0.25593 148 0.10411 0.0887 149 0.04469 0.03168 150
0.02894 0.02965 0.57037 151 0.01594 0.0157 0.52874 152 0.02585
0.02197 1.40549 153 0.08502 0.09128 154 0.013 0.01062 1.16887 155
0.02133 0.02385 0.46034 156 0.0588 0.01936 157 0.00315 0.02001
0.71676 158 0.00901 0.00955 0.65779 159 0.02579 0.02495 0.58237 160
0.01766 0.02255 6.6023 161 0.01969 0.01299 1.59243 162 0.01532
0.01819 1.18391 163 0.46447 0.26959 164 0.0546 0.06284 165 0.02452
0.01895 0.84174 166 0.01495 0.01835 0.69533 167 0.0131 0.00987
0.10896 168 0.00808 0.00611 0.82333 169 0.09611 0.1054 170 0.01635
0.01145 4.16823 171 0.07652 0.0652 172 0.65963 173 1.13408 174
0.0199 3.09728 175 15.49647 176 35.36685 177 4.13196 178 4.13196
179 2.875 180 2.31858 181 1.18357 182 0.13256 183 7.67584 184
1.07044 186 1.64878 187 2.57252 188 0.15624 0.16162 189 3.80992
5.5015 190 0.2839 0.21448 191 0.0109 0.00548 0.43262 192 0.13004
0.09414 193 0.0822 0.05193 194 0.01771 0.00919 0.41697 195 0.01654
0.01113 1.45306 196 0.01817 0.01479 0.35933 197 0.01822 0.00537
1.05176 198 0.20387 0.13976 199 1.38563 0.99179 200 0.00624 0.00596
0.2826 201 1.33026 1.11031 202 0.04811 0.03758 3.12395 203 0.02138
0.0142 0.47667 204 0.22175 0.21976 205 0.0077 0.00479 0.82074 206
0.01284 0.01106 0.63322 207 0.10198 0.11905 >20.0 uM 208 1.31405
1.19521 209 1.487 1.18544 210 1.26697 211 0.00679 0.0147 0.54832
212 1.05201 0.98451 213 0.05325 0.0438 14.71083 214 0.07475 0.06722
2.45231 215 0.01551 0.02167 0.80033 216 0.02489 0.04884 1.54705 217
0.18259 0.34753 218 0.02083 0.02903 1.27027 219 0.1748 0.16449 220
0.0205 0.04357 1.04201 221 0.0191 0.01616 1.11978 222 0.00078
0.00225 0.18533 0.13712 223 0.0136 0.02689 1.54605 224 0.03257
0.03334 1.76879 225 0.00695 0.01342 0.20778 226 0.03426 0.06624
0.23132 227 0.00365 0.00622 0.21515 228 0.0144 0.02015 1.27869 229
0.02235 0.02847 1.54848 230 0.34423 0.25475 231 0.29712 0.15945
>20.0 uM 232 0.15485 0.0881 8.04526 233 1.42344 0.5799 234
1.97267 1.62693 235 0.0632 0.05138 3.79895 236 0.61213 0.83099
14.62129 237 2.18156 1.47293 238 1.40428 0.80506 239 4.95504
4.77826 240 6.94031 6.8594 241 6.57787 242 0.1998 0.12679 8.4057
243 0.06318 6.47336 244 0.03749 3.06427 245 0.33295
246 247 0.01787 0.02136 3.19941 248 0.03458 0.0601 4.57081 249
11.45613 250 0.15959 0.20744 4.58295 251 13.11386 252 >20.0 uM
253 0.45669 0.28936 14.42495 254 255 0.01095 0.009 1.63338 256
0.00648 0.00738 1.69149 257 0.02224 0.07406 4.49758 258 1.90754 259
0.58066 0.60207 >20.0 uM 260 0.01282 0.01164 1.08525 261 0.01426
0.02069 0.9055 262 0.00961 0.00523 1.89119 263 0.03631 0.02181
4.89099 264 0.04094 0.04072 8.72536 265 0.02507 0.01843 5.09483 266
0.19847 0.08014 8.72547 267 0.16128 >20.0 uM 268 0.00131 0.0024
0.38531 269 0.09984 0.11108 14.2318 270 9.47417 >10.0 uM 271
0.05109 0.02565 5.32683 272 0.11472 0.06925 273 0.00328 0.00537
0.28489 274 7.76353 >10.0 uM >20.0 uM 275 0.90643 3.32337
>20.0 uM 276 0.00152 0.00407 0.18228 277 0.0023 0.00264 2.47797
278 0.00493 0.01117 0.61255 279 0.01143 0.01999 0.78715 280 0.00139
0.00556 0.43228 281 >10.0 uM >10.0 uM 282 0.00492 0.00985
0.21356 283 0.00316 0.0177 0.27638 284 0.008 0.0119 0.26282 285
0.00211 0.00506 0.21307 286 0.00122 0.00272 0.46721 287 0.00882
0.01141 1.35674 288 0.00694 0.00732 0.53484 289 0.14621 0.09339 290
0.00743 0.53732 291 0.00951 0.71974 292 0.01046 1.21021 293 0.01737
0.01158 1.29406 294 0.02412 2.16178 295 0.01161 0.01056 2.51445 296
0.00655 0.01825 4.33357 297 0.03337 0.02251 11.2951 298 0.0371
0.04303 6.54579 299 0.00324 0.00307 0.25937 300 0.02277 0.01709
4.94656 301 0.00497 0.00507 302 0.01217 0.01607 1.97477 303 0.03293
0.04119 3.89541 304 0.01722 0.0126 2.14418 305 0.00254 0.00785
0.28787 306 0.00942 0.00922 0.17284 307 0.0402 0.10188 3.85437 308
0.00997 0.01169 0.16744 309 0.00817 310 0.00596 0.60994 311 0.00323
0.16549 312 0.01507 313 0.00818 314 2.90826 4.7092 315 0.00973
0.00922 316 0.01202 1.40268 317 1.37527 1.96382 318 1.06978 0.91775
9.7596 319 0.0108 0.01137 2.95728 320 0.01694 0.01579 0.17566 321
0.00783 0.01144 322 0.03101 0.02239 3.59961 323 0.05148 0.03445
4.15894 324 0.40349 0.15525 16.80211 325 0.13585 0.21534 13.3829
326 0.0206 0.0095 1.43256 327 0.23134 0.09407 3.11887 328 0.84894
>20.0 uM 329 0.00755 0.00371 0.88119 330 0.03576 0.0368 1.20313
331 0.03891 0.01805 0.19105 332 0.27993 0.45426 >20.0 uM 333
0.30257 0.54098 >20.0 uM 334 0.05384 0.11977 335 0.01004 0.0027
0.97124 336 0.03144 0.02075 337 4.32339 3.56167 >20.0 uM 338
0.49234 0.19347 >20.0 uM 339 0.00858 0.00734 0.97312 340 0.01056
0.00412 1.59354 341 0.00592 0.00552 2.28944 342 0.03972 0.01592
1.9859 343 0.01081 0.00912 0.63289 344 0.00623 0.00396 1.30672 345
0.02228 0.01819 1.2346 346 0.00428 0.00245 0.12639 347 0.00654
0.00328 0.13526 348 0.00566 0.00363 0.32865 349 0.00872 0.00153 350
0.01244 0.00357 0.38084 351 0.00911 0.00138 4.85151 352 0.13195
5.54765 353 0.00792 0.00463 0.14599 354 0.01312 0.01039 0.14232
0.15832 355 0.08966 0.06081 356 0.03629 0.00721 357 0.00437 0.00456
0.14928 358 0.17315 0.06636 359 0.00729 0.00554 360 0.00883 0.00841
361 0.00661 0.00423 362 0.01388 0.00491 363 0.00394 0.00206 364
0.00943 0.00687 365 0.00609 0.00665 0.23299 366 0.03483 0.02934
>25.0 uM 367 0.33336 0.16784 >25.0 uM 368 0.02799 0.03855
>25.0 uM 369 0.07146 0.02755 3.7881 370 0.02998 0.01113 371
0.01181 0.00537 372 0.00801 0.0081 0.3659 373 0.00694 0.00369 374
0.00319 0.00831 375 0.01661 0.04182 376 0.01641 0.01977 377 0.07647
0.02552 378 0.00296 0.0031 >25.0 uM 379 0.00516 0.00322 >25.0
uM 380 0.02778 0.04196 >25.0 uM 381 0.05627 0.05243 >25.0 uM
382 0.00341 0.02167 4.16185 383 0.03974 0.04428 >25.0 uM 384
0.00206 0.00156 1.53716 385 0.06573 0.03511 >25.0 uM 386 2.42146
1.15144 387 0.74351 0.39771 388 0.00728 0.00396 2.29835 389 0.01564
0.0177 390 0.01251 0.01037 1.32773 391 0.04809 0.06404 >25.0 uM
392 0.048 0.039 393 0.12932 0.10995 394 0.29421 0.23058 395 0.30434
0.17359 396 4.42445 6.6822 397 3.37221 2.39173 398 2.61756 2.00394
399 >10.0 uM 8.31018 400 0.025 0.018 401 0.01466 0.01426 402
>10.0 uM 8.0921 403 0.53602 0.58786 404 2.27552 1.40203 405
0.172 0.107 406 0.017 0.053 407 0.0861 0.04069 408 0.1656 0.0936
409 0.45616 0.3404 410 2.28273 2.85881 411 1.19988 1.40119 412
0.22183 0.21475 413 0.76484 0.64167 414 0.00692 0.0084 415 1.5555
0.89979 416 0.016 0.015
Example 47
Derivation of the Lowest Cytotoxic Concentration (LCC)
[2322] It is well established that cellular proliferation proceeds
through cell division that results in a doubling of the number of
cells after division, relative to the number of cells prior to
division. Under a fixed set of environmental conditions (e.g., pH,
ionic strength, temperature, cell density, medium content of
proteins and growth factors, and the like) cells will proliferate
by consecutive doubling (i.e., division) according to the following
equation, provided that sufficient nutrients and other required
factors are available.
N t = N 0 .times. 2 t t D ( A .1 ) ##EQU00005##
where N.sub.t is the cell number at a time point (t) after
initiation of the observation period, N.sub.0 is the cell number at
the initiation of the observation period, t is the time after
initiation of the observation period and t.sub.D is the time
interval required for cell doubling, also referred to as the
doubling time. Equation A.1 can be converted into the more
convenient form of an exponential equation in base e, taking
advantage of the equality, 0.693=ln(2).
N t = N 0 0.693 t t D ( A .2 ) ##EQU00006##
[2323] The rate constant for cell proliferation (k.sub.p) is
inversely related to the doubling time as follows.
k p = 0.693 t D ( A .3 ) ##EQU00007##
[2324] Combining equation A.2 and A.3 yields,
N.sub.t=N.sub.0e.sup.k.sup.p.sup.t (A.4)
[2325] Thus, according to equation A.4 cell number is expected to
increase exponentially with time (FIG. 1 A) during the early period
of cell growth referred to as log-phase growth. Exponential
equations like equation A.4 can be linearized by taking the natural
logarithm of each side.
ln(N.sub.t)=ln(N.sub.0)+k.sub.pt (A.5)
[2326] Thus a plot of ln(N.sub.t) as a function of time is expected
to yield an ascending straight line with slope equal to k.sub.p and
y-intercept equal to ln(N.sub.0), as illustrated in FIG. 1 B.
[2327] Changes in environmental conditions can result in a change
in the rate of cellular proliferation that is quantifiable as
changes in the proliferation rate constant k.sub.p. Among
conditions that may result in a change in proliferation rate is the
introduction to the system of an antiproliferative compound at the
initiation of the observation period (i.e., at t=0). When an
antiproliferative compound has an immediate impact on cell
proliferation, one expects that plots of ln(N.sub.t) as a function
of time will continue to be linear at all compound concentrations,
with diminishing values of k.sub.p at increasing concentrations of
compound.
[2328] Depending on the mechanistic basis of antiproliferative
action, some compounds may not immediately effect a change in
proliferation rate. Instead, there may be a period of latency
before the impact of the compound is realized. In such cases a plot
of ln(N.sub.t) as a function of time will appear biphasic, and a
time point at which the impact of the compound begins can be
identified as the breakpoint between phases (FIG. 2). Regardless of
whether a compound's impact on proliferation is immediate or begins
after a latency period, the rate constant for proliferation at each
compound concentration is best defined by the slope of the
ln(N.sub.t) vs. time curve from the time point at which compound
impact begins to the end of the observation period of the
experiment.
[2329] A compound applied to growing cells may affect the observed
proliferation in one of two general ways: by inhibiting further
cell division (cytostasis) or by cell killing (cytotoxicity). If a
compound is cytostatic, increasing concentration of compound will
reduce the value of k.sub.p until there is no further cell
division. At this point, the rate of cell growth, and therefore the
value of k.sub.p, will be zero. If, on the other hand, the compound
is cytotoxic, then the value of k.sub.p will be composed of two
rate constants: a rate constant for continued cell growth in the
presence of the compound (k.sub.g) and a rate constant for cell
killing by the compound (k.sub.d). The overall rate constant for
proliferation at a fixed concentration of compound will thus be the
difference between the absolute values of these opposing rate
constants.
k.sub.p=|k.sub.g|-|k.sub.d| (A.6)
[2330] At compound concentrations for which the rate of cell growth
exceeds that of cell killing, the value of k.sub.p will have a
positive value (i.e., k.sub.p>0). At compound concentrations for
which the rate of cell growth is less than that for cell killing,
the value of k.sub.p will have a negative value (i.e.,
k.sub.p<0) and the cell number will decrease with time,
indicative of robust cytotoxicity. When k.sub.g exactly matches
k.sub.d then the overall proliferation rate constant, k.sub.p, will
have a value of zero. We can thus define the lowest cytotoxic
concentration (LCC) as that concentration of compound that results
in a value of k.sub.p equal to zero, because any concentration
greater than this will result in clearly observable cytotoxicity.
Nota bene: at concentrations below the LCC there is likely to be
cell killing occurring, but at a rate that is less than that of
residual cell proliferation. The treatment here is not intended to
define the biological details of compound action. Rather, the goal
here is to merely define a practical parameter with which to
objectively quantify the concentration of compound at which the
rate of cell killing exceeds new cell growth. Indeed, the LCC
represents a breakpoint or critical concentration above which frank
cytotoxicity is observed, rather than a cytotoxic concentration per
se. In this regard, the LCC can be viewed similar to other physical
breakpoint metrics, such as the critical micelle concentration
(CMC) used to define the concentration of lipid, detergent or other
surfactant species above which all molecules incorporate into
micellar structures.
[2331] Traditionally, the impact of antiproliferative compounds on
cell growth has been most commonly quantified by the IC.sub.50
value, which is defined as that concentration of compound that
reduces the rate of cell proliferation to one half that observed in
the absence of compound (i.e., for the vehicle or solvent control
sample; FIG. 2). The IC.sub.50, however, does not allow the
investigator to differentiate between cytostatic and cytotoxic
compounds. The LCC, in contrast, readily allows one to make such a
differentiation and to further quantify the concentration at which
the transition to robust cytotoxic behavior occurs.
[2332] If one limits the observation time window to between the
start of impact (as defined above and in FIG. 2) and the end of the
experiment, then the data will generally fit well to a linear
equation when plotted as ln(N.sub.t) as a function of time (vide
supra). From fits of this type, the value of k.sub.p can be
determined at each concentration of compound tested. A replot of
the value of k.sub.p as a function of compound concentration ([I])
will have the form of a descending isotherm, with a maximum value
at [I]=0 of k.sub.max (defined by the vehicle or solvent control
sample) and a minimum value at infinite compound concentration of
k.sub.m (FIG. 3).
k p = ( k max - k min ) 1 + [ I ] I mid + k min ( A .7 )
##EQU00008##
where I.sub.mid is the concentration of compound yielding a value
of k.sub.p that is midway between the values of k.sub.max and
k.sub.min (note that the value of I.sub.mid is not the same as the
IC.sub.50, except in the case of a complete and purely cytostatic
compound). Thus, fitting the replot data to equation A.7 provides
estimates of k.sub.max, k.sub.min and I.sub.mid. If a compound is
cytostatic (as defined here), the value of k.sub.min cannot be less
than zero. For cytotoxic compounds, k.sub.min will be less than
zero and the absolute value of k.sub.min will relate directly to
the effectiveness of the compound in killing cells.
[2333] The fitted values derived from equation A.7 can also be used
to determine the value of the LCC. By definition, when [I]=LCC,
k.sub.p=0. Thus, under these conditions equation A.7 becomes.
0 = ( k max - k min ) 1 + LCC I mid + k min ( A .8 )
##EQU00009##
[2334] Algebraic rearrangement of equation A.8 yields an equation
for the LCC.
LCC = I mid [ ( k max - k min - k min ) - 1 ] ( A .9 )
##EQU00010##
[2335] This analysis is simple to implement with nonlinear curve
fitting software and may be applied during cellular assays of
compound activity throughout the drug discovery and development
process. In this manner, the LCC may provide a valuable metric for
the assessment of compound SAR (structure-activity
relationship).
[2336] Table 4 below provides LCC and IC.sub.50 data for certain
compounds of the invention on WSU-DLCL2 cells.
TABLE-US-00005 TABLE 4 WSU-DLCL2 WSU-DLCL2 Cpd# 11-day LCC (.mu.M)
11-day IC50 (.mu.M) 8 0.207 0.076 56 0.442 0.0097 88 1.27 0.035 101
2.59 0.25 120 0.343 0.006 121 0.0597 0.0094 222 0.0268 0.0043 261
0.556 0.051 277 0.037 0.0024 354 0.043 0.013 357 0.026 0.0095
Example 48
In Vivo Assay
Mice
[2337] Female Fox Chase SCID.RTM. Mice
(CB17/Icr-Prkdcscid/IcrIcoCrl, Charles River Laboratories) or
athymic nude mice (Crl:NU(Ncr)-Foxnlnu, Charles River Laboratories)
were 8 weeks old and had a body-weight (BW) range of 16.0-21.1 g on
D1 of the study. The animals were fed ad libitum water (reverse
osmosis 1 ppm Cl) and NIH 31 Modified and Irradiated Lab Diet.RTM.
consisting of 18.0% crude protein, 5.0% crude fat, and 5.0% crude
fiber. The mice were housed on irradiated Enrich-o'cobs.TM. bedding
in static microisolators on a 12-hour light cycle at 20-22.degree.
C. (68-72.degree. F.) and 40-60% humidity. All procedures comply
with the recommendations of the Guide for Care and Use of
Laboratory Animals with respect to restraint, husbandry, surgical
procedures, feed and fluid regulation, and veterinary care.
Tumor Cell Culture
[2338] Human lymphoma cell lines line were obtained from different
sources (ATCC, DSMZ) and maintained at Piedmont as suspension
cultures in RPMI-1640 medium containing 100 units/mL penicillin G
sodium salt, 100 g/mL streptomycin, and 25 g/mL gentamicin. The
medium was supplemented with 10% fetal bovine serum and 2 mM
glutamine. The cells were cultured in tissue culture flasks in a
humidified incubator at 37.degree. C., in an atmosphere of 5% CO2
and 95% air.
In Vivo Tumor Implantation
[2339] Human lymphoma cell lines were harvested during mid-log
phase growth, and resuspended in PBS with 50% Matrigel.TM. (BD
Biosciences). Each mouse received 1.times.10.sup.7 cells (0.2 mL
cell suspension) subcutaneously in the right flank. Tumors were
calipered in two dimensions to monitor growth as the mean volume
approached the desired 80-120 mm.sup.3 range. Tumor size, in
mm.sup.3, was calculated from:
Tumor Volume = w 2 .times. l 2 ##EQU00011##
where w=width and l=length, in mm, of the tumor. Tumor weight can
be estimated with the assumption that 1 mg is equivalent to 1 mm3
of tumor volume. After 10-30 days (depending on the cell line used)
mice with 108-126 mm.sup.3 tumors were sorted into eight groups
with mean tumor volumes of 117-119 mm3.
Test Articles
[2340] Test compounds were stored at room temperature and protected
from light. On each treatment day, a fresh compound formulations
were prepared by suspending the powders in 0.5% sodium
carboxymethylcellulose (NaCMC) and 0.1% Tween.RTM. 80 in deionized
water. The EM10 vehicle, 0.5% NaCMC and 0.1% Tween.RTM. 80 in
deionized water, Was used to treat the control groups at the same
schedules. Formulations were stored away from light at 4.degree. C.
prior to administration.
Treatment Plan
[2341] Mice were treated at compound doses ranging from 1-1000
mg/kg and at TID (3 times a day every 8 h), BID (twice a day every
12 h) or QD (once a day) schedules for various amount of days by
oral gavage or injections via the intravenous, intraperitoneal or
subcutaneous routes. Each dose was delivered in a volume of 0.2
mL/20 g mouse (10 mL/kg), and adjusted for the last recorded weight
of individual animals. The maximal treatment length was 28
days.
Median Tumor Volume (MTV) and Tumor Growth Inhibition (TGI)
Analysis
[2342] Treatment efficacy was determined on the last treatment day.
MTV(n), the median tumor volume for the number of animals, n,
evaluable on the last day, was determined for each group. Percent
tumor growth inhibition (% TGI) can be defined several ways. First,
the difference between the MTV(n) of the designated control group
and the MTV(n) of the drug-treated group is expressed as a
percentage of the MTV(n) of the control group:
% TGI = ( MTV ( n ) control - MTV ( n ) treated MTV ( n ) control )
.times. 100 ##EQU00012##
[2343] Another way of calculating % TGI is taking the change of the
tumor size from day 1 to day n into account with n being the last
treatment day.
% TGI = ( .DELTA. MTV ( n ) control - .DELTA. MTV ( n ) treated
.DELTA. MTV ( n ) control ) .times. 100 ##EQU00013## .DELTA. MTV
control = MTV ( n ) control - MTV ( 1 ) control ##EQU00013.2##
.DELTA. MTV treated = MTV ( n ) treated - MTV ( 1 ) treated
##EQU00013.3##
Tumor Growth Delay Analysis
[2344] Alternatively, mice were kept alive after the last treatment
day for tumor growth delay analysis. Tumors were callipered
twice-weekly and each test animal was euthanized when its neoplasm
reached the endpoint volume of 2000 mm.sup.3 or on the
pre-specified last day of the study, whichever came first. The
time-to-endpoint (TTE) for each mouse was calculated from the
following equation:
TTE ( days ) = log 10 ( endpoint volume , mm 3 ) - b m
##EQU00014##
where b is the intercept and m is the slope of the line obtained by
linear regression of a log-transformed tumor growth data set. The
data sets were composed of the first observation that exceeded the
study endpoint volume and the three consecutive observations that
immediately preceded the attainment of the endpoint volume. Animals
that did not reach the volume endpoint were assigned a TTE value
equal to the last day of the study (prespecified). Any animal
classified as a treatment-related (TR) death was to be assigned a
TTE value equal to the day of death. Any animal classified as a
nontreatment-related (NTR) death was excluded from TTE calculations
and all further analyses.
[2345] Treatment outcome was determined from tumor growth delay
(TGD), defined as the increase in the median TTE in a treatment
group compared to the control group:
TGD=T-C
expressed in days, or as a percentage of the median TTE of the
control group:
% TGD = T - C C .times. 100 ##EQU00015##
where: T=median TTE for a treatment group C=median TTE for the
control group
Toxicity
[2346] Animals were weighed daily on Days 1-5, and then twice
weekly until the completion of the study. The mice were examined
frequently for overt signs of any adverse, treatment related side
effects, which were documented. Acceptable toxicity for the maximum
tolerated dose (MTD) was defined as a group mean BW loss of less
than 20% during the test, and not more than 10% mortality due to TR
deaths. A death was to be classified as TR if it was attributable
to treatment side effects as evidenced by clinical signs and/or
necropsy, or due to unknown causes during the dosing period. A
death was to be classified as NTR if there was evidence that the
death was unrelated to treatment side effects. NTR deaths during
the dosing interval would typically be categorized as NTRa (due to
an accident or human error) or NTRm (due to necropsy-confirmed
tumor dissemination by invasion and/or metastasis). Orally treated
animals that die from unknown causes during the dosing period may
be classified as NTRu when group performance does not support a TR
classification and necropsy, to rule out a dosing error, is not
feasible.
Sampling
[2347] On several days during the study mice were sampled in a
pre-specified fashion. Sampling included non-terminal bleeds (0.25
mL) from the mandibular vein without anesthesia and full volume
blood collection via terminal cardiac puncture under CO.sub.2
anesthesia. Blood samples were processed for plasma, with K2-EDTA
as anti-coagulant. The plasma samples were frozen at -80.degree. C.
and stored prior to bioanalysis of compound levels.
[2348] Tumors were harvested from specified mice under RNAse free
conditions and bisected. A 2 mm thick slice from one half of each
tumor was formalin-fixed for 24 h and transferred to 70% ethanol.
The fixed tumor tissues were paraffin embedded. The remaining tumor
tissue from each animal was snap frozen in liquid N.sub.2 and
pulverized with a mortar and pestle.
[2349] Specified mice were sampled for the surrogate tissues
including spleen, skin, bone marrow, and whiskers. Each tissue was
isolated and fixed and/or snap frozen.
Statistical and Graphical Analyses
[2350] All statistical and graphical analyses were performed with
Prism 3.03 (GraphPad) for
[2351] Windows. Several analyses methods were applied. Median D29
tumor volumes were compared with the Kruskal-Wallis test, and a
post hoc Dunn's multiple comparison test. These tests were
performed three times.
[2352] The two-tailed statistical analyses were conducted at
P=0.05. Prism reports results as non-significant (ns) at P>0.05,
significant (symbolized by "*") at 0.01<P<0.05, very
significant ("**") at 0.001<P<0.01 and extremely significant
("***") at P<0.001.
[2353] To test statistical significance between the control and
treated groups over the whole treatment time course, either a
repeated measures ANOVA test followed by Dunnets multiple
comparison post test or a 2 way ANOVA test was employed.
[2354] For graphical representations s "box and whiskers" diagram
was constructed to show the distribution of individual tumor
volumes for each group. The box represents the 25th to 75th
percentile of observations, the horizontal line corresponds to the
median value, and the "whiskers" indicate the maximum and minimum
values. Median or mean (.+-.SEM) tumor volumes were graphed on a
semilog or linear plot as functions of time. Group mean BW changes
during the study were plotted as percent change, .+-.SEM, from
D1.
[2355] A scatter plot was constructed to show TTE values, by group.
The TTE plot includes NTR deaths, which are excluded from all other
graphical analyses. When an animal exited the study because of
tumor size, the final tumor volume recorded for the animal was
included with the data used to calculate the median volume at
subsequent time points. The percentage of animals in each group
remaining in the study versus time was presented in a Kaplan-Meier
survival plot.
Histone Extraction
[2356] For isolation of histones, 60-90 mg tumor tissue was
homogenized in 1.5 ml nuclear extraction buffer (10 mM Tris-HCl, 10
mM MgCl2, 25 mM KCl, 1% Triton X-100, 8.6% Sucrose, plus a Roche
protease inhibitor tablet 1836145) and incubated on ice for 5
minutes. Nuclei were collected by centrifugation at 600 g for 5
minutes at 4.degree. C. and washed once in PBS. Supernatant was
removed and histones extracted for one hour, with vortexing every
15 minutes, with 0.4 N cold sulfuric acid. Extracts were clarified
by centrifugation at 10000 g for 10 minutes at 4.degree. C. and
transferred to a fresh microcentrifuge tube containing 10.times.
volume of ice cold acetone. Histones were precipitated at
-20.degree. C. for 2 hours-overnight, pelleted by centrifugation at
10000 g for 10 minutes and resuspended in water.
Western Blot Analysis
[2357] Protein concentrations for acid extracted histones were
determined by BCA assay (Pierce). 400-800 ng of each lysate was
fractionated on 10-20% Tris-Glycine gel (Biorad), transferred using
iBlot (7 minutes on program 3, using Nitrocellulose transfer
stacks), and probed with the following antibodies in Odyssey
blocking buffer: rabbit anti-H3K27me3 (CST 9733; 1:20000 dilution)
and mouse anti-Total H3 (CST 3638; 1:20000 dilution). Following
primary Ab incubation, membranes were probed with IRDye 800CW
Donkey-anti-mouse IgG (LiCOR #926-32212) and Alexa Fluor 680
goat-anti-rabbit IgG (Invitrogen #A-21076) secondary Ab and imaged
using the LiCOR Odyssey system.
ELISA
[2358] Histones were prepared in equivalent concentrations in
coating buffer (PBS+0.05% BSA) yielding 0.5 ng/ul of sample, and
100 ul of sample or standard was added in duplicate to 2 96-well
ELISA plates (Thermo Labsystems, Immulon 4HBX #3885). The plates
were sealed and incubated overnight at 4.degree. C. The following
day, plates were washed 3.times. with 300 ul/well PBST (PBS+0.05%
Tween 20; 10.times.PBST, KPL #51-14-02) on a Bio Tek plate washer.
Plates were blocked with 300 ul/well of diluent (PBS+2% BSA+0.05%
Tween 20), incubated at RT for 2 hours, and washed 3.times. with
PBST. All antibodies were diluted in diluent. 100 ul/well of
anti-H3K27me3 (CST #9733, 50% glycerol stock 1:1,000) or anti-total
H3 (Abcam ab1791, 50% glycerol 1:10,000) was added to each plate.
Plates were incubated for 90 min at RT and washed 3.times. with
PBST. 100 ul/well of anti-Rb-IgG-HRP (Cell Signaling Technology,
7074) was added 1:2,000 to the H3K27Me3 plate and 1:6,000 to the H3
plate and incubated for 90 min at RT. Plates were washed 4.times.
with PBST. For detection, 100 ul/well of TMB substrate (BioFx
Laboratories, #TMBS) was added and plates incubated in the dark at
RT for 5 min. Reaction was stopped with 100 ul/well 1N
H.sub.2SO.sub.4. Absorbance at 450 nm was read on SpectaMax M5
Microplate reader.
[2359] The in vivo results for Compound 222 were shown in FIGS. 4
and 5. FIG. 4 shows tumor growth of WSU-DLCL2 xenograft bearing
mice treated with Compound 222 over 27 days. Tumor growth
inhibition was observed at all 3 doses 100 mg/kg (b.i.d., 27 days),
200 mg/kg (b.i.d., 27 days) and 400 mg/kg (400 mg/kg b.i.d. 7 days,
0 mg/kg for 7 days and 300 mg/kg b.i.d., 13 days). FIG. 5 shows
global H3K27me3 methylation in WSU-DLCL2 tumors from mice treated
with compound 222 or vehicle for 27 days. This figure shows a
reduction in the H3K27Me3 mark for each of the dose groups.
INCORPORATION BY REFERENCE
[2360] The entire disclosure of each of the patent documents and
scientific articles referred to herein is incorporated by reference
for all purposes.
EQUIVALENTS
[2361] The invention can be embodied in other specific forms
without departing from the spirit or essential characteristics
thereof. The foregoing embodiments are therefore to be considered
in all respects illustrative rather than limiting on the invention
described herein. Scope of the invention is thus indicated by the
appended claims rather than by the foregoing description, and all
changes that come within the meaning and range of equivalency of
the claims are intended to be embraced therein.
Sequence CWU 1
1
2126PRTArtificial SequenceChemically synthesized polypeptide 1Ala
Thr Lys Ala Ala Arg Lys Ser Ala Pro Ala Thr Gly Gly Val Lys 1 5 10
15 Lys Pro His Arg Tyr Arg Pro Gly Gly Lys 20 25 226PRTArtificial
SequenceChemically synthesized polypeptide 2Ala Thr Lys Ala Ala Arg
Lys Ser Ala Pro Ala Thr Gly Gly Val Lys 1 5 10 15 Lys Pro His Arg
Tyr Arg Pro Gly Gly Lys 20 25
* * * * *
References