U.S. patent application number 12/894525 was filed with the patent office on 2011-02-03 for pyrido[4,3-d]pyrimidin-4(3h)-one derivatives as calcium receptor antagonists.
This patent application is currently assigned to Pfizer Inc. Invention is credited to Feng C. Bi, Mary T. Didiuk, David A. Griffith, Angel Guzman-Perez, Kevin K. Liu, Daniel P. Walker.
Application Number | 20110028452 12/894525 |
Document ID | / |
Family ID | 39183043 |
Filed Date | 2011-02-03 |
United States Patent
Application |
20110028452 |
Kind Code |
A1 |
Didiuk; Mary T. ; et
al. |
February 3, 2011 |
PYRIDO[4,3-d]PYRIMIDIN-4(3H)-ONE DERIVATIVES AS CALCIUM RECEPTOR
ANTAGONISTS
Abstract
The present invention is directed to novel
pyrido[4,3-d]pyrimidin-4(3H)-one derivatives and pharmaceutically
acceptable salts thereof of structural formula I ##STR00001##
wherein the variables R.sup.1, R.sup.2, R.sup.3, R.sup.4 and
R.sup.5 are as described herein. Also provided are pharmaceutical
compositions comprising the compounds of formula I as well as
methods of treatment employing compounds of formula I to treat a
disease or disorder characterized by abnormal bone or mineral
homeostasis such as hypoparathyroidism, osteoporosis, osteopenia,
periodontal disease, Paget's disease, bone fracture,
osteoarthritis, rheumatoid arthritis, and humoral hypercalcemia of
malignancy.
Inventors: |
Didiuk; Mary T.; (Madison,
CT) ; Liu; Kevin K.; (East Lyme, CT) ;
Griffith; David A.; (Old Saybrook, CT) ;
Guzman-Perez; Angel; (Mystic, CT) ; Bi; Feng C.;
(Groton, CT) ; Walker; Daniel P.; (Chesterfield,
MO) |
Correspondence
Address: |
PFIZER INC.;PATENT DEPARTMENT
Bld 114 M/S 9114, EASTERN POINT ROAD
GROTON
CT
06340
US
|
Assignee: |
Pfizer Inc
|
Family ID: |
39183043 |
Appl. No.: |
12/894525 |
Filed: |
September 30, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11867255 |
Oct 4, 2007 |
7829572 |
|
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12894525 |
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60828162 |
Oct 4, 2006 |
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60969083 |
Aug 30, 2007 |
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Current U.S.
Class: |
514/210.21 ;
514/234.2; 514/252.16; 514/264.1; 544/117; 544/279 |
Current CPC
Class: |
C07D 471/04 20130101;
A61P 29/00 20180101; A61P 19/10 20180101; A61P 19/00 20180101; A61P
43/00 20180101; A61P 3/14 20180101; A61P 3/00 20180101; A61P 19/08
20180101; A61P 1/02 20180101; A61P 5/18 20180101; A61P 19/02
20180101; A61P 35/00 20180101 |
Class at
Publication: |
514/210.21 ;
544/279; 544/117; 514/264.1; 514/234.2; 514/252.16 |
International
Class: |
A61K 31/5377 20060101
A61K031/5377; C07D 471/04 20060101 C07D471/04; A61K 31/519 20060101
A61K031/519; A61P 5/18 20060101 A61P005/18; A61P 19/10 20060101
A61P019/10; A61P 35/00 20060101 A61P035/00; A61P 19/02 20060101
A61P019/02 |
Claims
1. A compound of formula I ##STR00144## wherein R.sup.1 is -Q or
(C.sub.1-C.sub.6)alkyl-Q; R.sup.2 is (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, aryl or heteroaryl; wherein said aryl
or heteroaryl is substituted with hydroxy and additionally said
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl, aryl or
heteroaryl within the definition of R.sup.2 is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy or hydroxy; R.sup.3
is hydrogen, (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.1-C.sub.6)alkyl-Q, aryl, heteroaryl, OR.sup.6, or
NR.sup.7R.sup.8; wherein said (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, aryl or heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl; (C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or
hydroxy; R.sup.4 and R.sup.5 are each independently hydrogen, halo,
cyano, (C.sub.1-C.sub.6)alkyl optionally substituted with one to
three fluoro, aryl, heteroaryl, or OR.sup.6; R.sup.6 at each
occurrence is independently (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl-(C.sub.1-C.sub.6)alkyl, aryl,
heteroaryl, (C.sub.1-C.sub.6)alkylaryl, or
(C.sub.1-C.sub.6)alkylheteroaryl; each of said
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl-(C.sub.1-C.sub.6)alkyl, aryl,
heteroaryl, (C.sub.1-C.sub.6)alkylaryl, or
(C.sub.1-C.sub.6)alkylheteroaryl optionally substituted with one to
three substituents independently selected from halo, hydroxy or
(C.sub.1-C.sub.3)alkyl; R.sup.7 and R.sup.8, at each occurrence,
are independently hydrogen, (C.sub.1-C.sub.6)alkyl, or
(C.sub.3-C.sub.7)cycloalkyl; or R.sup.7 and R.sup.8 taken together
with the nitrogen to which they are attached form a 3 to 7 membered
fully saturated, partially saturated or fully unsaturated ring
optionally containing one to two additional heteroatoms
independently selected from N(R.sup.9).sub.n; O or S(O).sub.p; n is
0 or 1; p is 0, 1 or 2; R.sup.9 is hydrogen or
(C.sub.1-C.sub.6)alkyl; Q, at each occurrence, is independently
aryl or heteroaryl; wherein said aryl or heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy or hydroxy; or a
pharmaceutically acceptable salt thereof.
2. A compound according to claim 1, wherein R.sup.2 is aryl or
heteroaryl, wherein said aryl or heteroaryl is substituted with
hydroxy and optionally substituted with one to three substituents
independently selected from (C.sub.1-C.sub.6)alkoxy or halo; or a
pharmaceutically acceptable salt thereof.
3. A compound according to claim 2, wherein R.sup.2 is aryl,
wherein said aryl is substituted with hydroxy and optionally
substituted with one to three substituents independently selected
from (C.sub.1-C.sub.6)alkoxy or halo; or a pharmaceutically
acceptable salt thereof.
4. A compound according to claim 1, wherein R.sup.2 is heteroaryl,
wherein said heteroaryl is substituted with hydroxy and optionally
substituted with one to three substituents independently selected
from (C.sub.1-C.sub.6)alkoxy or halo; or a pharmaceutically
acceptable salt thereof.
5. A compound according to claim 2, wherein R.sup.2 is
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl; wherein said
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl is optionally
substituted with one to two substituents independently selected
from (C.sub.1-C.sub.6)alkoxy or fluoro; or a pharmaceutically
acceptable salt thereof.
6. A compound according to claim 5, wherein R.sup.1 is
(C.sub.1-C.sub.6)alkyl-Q; and Q is phenyl optionally substituted
with one or two fluoro; or a pharmaceutically acceptable salt
thereof.
7. A compound according to claim 1, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; or a pharmaceutically acceptable salt thereof.
8. A compound according to claim 7 wherein R.sup.3 is
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl-Q, aryl, heteroaryl,
OR.sup.6, or NR.sup.7R.sup.8, wherein said (C.sub.1-C.sub.6)alkyl,
aryl or heteroaryl is optionally substituted with one to three
substituents independently selected from halo, cyano,
trifluoromethyl, trifluoromethoxy, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or hydroxy; and R.sup.4
and R.sup.5 are each hydrogen; or a pharmaceutically acceptable
salt thereof.
9. A compound according to claim 8, wherein R.sup.3 is
trifluoromethyl; or a pharmaceutically acceptable salt thereof.
10. A compound according to claim 1, wherein R.sup.3 is
trifluoromethyl; or a pharmaceutically acceptable salt thereof.
11. A compound according to claim 10, wherein R.sup.4 and R.sup.5
are each independently hydrogen or methyl; or a pharmaceutically
acceptable salt thereof.
12. A compound according to claim 11, wherein R.sup.2 is aryl or
heteroaryl, wherein said aryl or heteroaryl is substituted with
hydroxy and optionally substituted with one to three substituents
independently selected from (C.sub.1-C.sub.6)alkoxy or halo; or a
pharmaceutically acceptable salt thereof.
13. A compound according to claim 12, wherein R.sup.2 is
hydroxy-phenyl or hydroxy-pyridyl; wherein said hydroxy-phenyl or
hydroxy-pyridyl is optionally substituted with one to two
substituents independently selected from (C.sub.1-C.sub.6)alkoxy or
fluoro; or a pharmaceutically acceptable salt thereof.
14. A compound according to claim 13, wherein R.sup.1 is
(C.sub.1-C.sub.6)alkyl-Q; and Q is phenyl optionally substituted
with one or two substituents independently selected from fluoro,
methyl or methoxy; or a pharmaceutically acceptable salt
thereof.
15. A compound according to claim 11, wherein R.sup.1 is phenethyl
or 1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two fluoro; or a pharmaceutically acceptable salt
thereof.
16. A compound according to claim 11, wherein R.sup.2 is
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl, wherein said
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl is optionally
substituted with one or two substituents independently selected
from (C.sub.1-C.sub.3)alkyl or halo; or a pharmaceutically
acceptable salt thereof.
17. A compound according to claim 1 wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two substituents independently selected from halo or
(C.sub.1-C.sub.3)alkyl or (C.sub.1-C.sub.3)alkoxy; R.sup.2 is
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl, wherein said
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl is optionally
substituted with one or two substituents independently selected
from (C.sub.1-C.sub.3)alkyl or halo; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.1-C.sub.6)alkyl-Q, aryl, heteroaryl, or OR.sup.6; wherein
said (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl, aryl or
heteroaryl is optionally substituted with one to three substituents
independently selected from fluoro, cyano, trifluoromethyl,
trifluoromethoxy, (C.sub.1-C.sub.3)alkyl; (C.sub.1-C.sub.3)alkoxy,
NR.sup.7R.sup.8 or hydroxy; and R.sup.4 and R.sup.5 are each
independently hydrogen or methyl; or a pharmaceutically acceptable
salt thereof.
18. A compound selected from the group consisting of:
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyri-m-
idin-4-one;
2-(3-Fluoro-2-hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-
-d]pyrimidin-4-one;
2-(3-Hydroxy-pyridin-2-yl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]-
pyrimidin-4-one;
(R)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one;
(S)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one;
(R)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromet-
hyl-3H-pyrido[4,3-d]pyrimidin-4-one;
(R,S)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-t-
rifluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
(R)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-tri-
fluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
(S)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-tri-
fluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
2-(2-Hydroxyphenyl)-3-(2-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[-
4,3-d]pyrimidin-4-one;
2-(2-Hydroxyphenyl)-3-(3-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[-
4,3-d]pyrimidin-4-one;
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(2-fluoro-phenyl)-ethyl]-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-7-methyl-3-(phenyl-ethyl)-5-trifluoromethyl-3H-pyrid-
o[4,3-d]pyrimidin-4-one;
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(3-fluoro-phenyl)-ethyl]-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one;
3-[2-(2-Fluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluoromethy-
l-3H-pyrido[4,3-d]pyrimidin-4-one;
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
3-[2-(2,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-5-methylamino-3-phenethyl-3H-pyrido[4,3-d]pyrimidin--
4-one;
2-(2-Hydroxy-phenyl)-5-isopropylamino-3-phenethyl-3H-pyrido[4,3-d]p-
yrimidin-4-one;
2-(2-Hydroxy-phenyl)-3-phenethyl-5-pyrrolidin-1-yl-3H-pyrido[4,3-d]pyrimi-
din-4-one;
2-(2-Hydroxy-phenyl)-5-(4-methyl-piperazin-1-yl)-3-phenethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-3-phenethyl-5-piperazin-1-yl-3H-pyrido[4,3-d]pyrimid-
in-4-one;
5-Dimethylamino-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d-
]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-5-morpholin-4-yl-3-phenethyl-3H-pyrido[4,3-d]pyrimid-
in-4-one;
5-Azetidin-1-yl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d-
-pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-3-phenethyl-5-phenyl-3H-pyrido[4,3-d]pyrimidin-4-one-
;
5-Benzyl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-on-
e;
2-(2-Hydroxy-phenyl)-5-methyl-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-4-on-
e;
5-(6-Dimethylamino-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-3H-py-
rido[4,3-d]pyrimidin-4-one;
5-(6-Dimethylamino-5-methyl-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethy-
l-3H-pyrido[4,3-d]pyrimidin-4-one;
5-(6-pyrrolidine-5-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyri-
do[4,3-d]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-on-
e;
2-(2-Hydroxy-phenyl)-5-(1-methyl-cyclopropylmethoxy)-3-phenethyl-3H-pyr-
ido[4,3-d]pyrimidin-4-one;
2-(2-Hydroxy-phenyl)-3-phenethyl-5-propoxy-3H-pyrido[4,3-d]pyrimidin-4-on-
e;
5-Cyclobutyloxy-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one;
2-(2-Hydroxy-phenyl)-5-isobutoxy-3-phenethyl-3H-pyrido[4,3-d]py-
rimidin-4-one;
2-(2-Hydroxy-phenyl)-5-isopropoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-
-one;
2-(2-Hydroxy-phenyl)-3-phenethyl-5-(2,2,2-trifluoro-ethoxy)-3H-pyrid-
o[4,3-d]pyrimidin-4-one;
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(R)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(S)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one;
3-(2,3-difluorophenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)py-
rido[4,3-d]pyrimidin-4(3H)-one;
3-(5-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(2-fluoro-6-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(1-(2-fluorophenyl)butan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorome-
thyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(3-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(2-cyclopentylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one;
3-(2-cyclohexylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one;
(R)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(S)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(R,S)-2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-2-yl)ethyl)-5-(-
trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(R)-2-(3-fluoro-2-hydroxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one;
(R)-2-(1H-imidazol-2-yl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one;
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4-
,3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide;
(R)-3-(1-phenylpropan-2-yl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,-
3-d]pyrimidin-4(3H)-one;
3-phenethyl-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(-
3H)-one;
3-(2-methoxyphenethyl)-2-(2-hydroxyphenyl)-5-(trifluoromethyl)pyr-
ido[4,3-d]pyrimidin-4(3H)-one;
3-(2-fluorophenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one;
(R)-2-cyclopentyl-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido[4,3-d-
]pyrimidin-4(3H)-one;
2-isopropyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-o-
ne
2-cyclopentyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3-
H)-one;
3-(2-methoxyphenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one;
3-(2-cyclohexylethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one;
2-(3-(2-cyclohexylethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4,3--
d]pyrimidin-2-yl)benzonitrile;
2-(3-aminopyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrim-
idin-4(3H)-one;
N-(2-(4-oxo-3-phenethyl-5-(trifluoromethyl)-3,4-dihydropyrido[4,3-d]pyrim-
idin-2-yl)pyridin-3-yl)methanesulfonamide;
3-(1-(2-fluorophenyl)propan-2-yl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyr-
ido[4,3-d]pyrimidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-isopentyl-5-(trifluoromethyl)pyrido[4,3-d]pyr-
imidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluo-
romethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-4-yl)ethyl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(2-fluoro-5-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
3-(2-fluoro-3-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one;
2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydroquinazolin--
2-yl)benzonitrile;
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4-
,3-d]pyrimidin-2-yl)pyridin-3-yl)-2,2,2-trifluoroacetamide;
(R)-2,2,2-trifluoro-N-(2-(4-oxo-3-(1-phenylpropan-2-yl)-5-(trifluoromethy-
l)-3,4-dihydropyrido[4,3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide;
3-cyclohexyl-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one;
3-(cyclohexylmethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one;
3-Phenethyl-5-trifluoromethyl-2-(3-trifluoromethyl-pyridin-2-yl)-3H-pyrid-
o[4,3-d]pyrimidin-4-one;
2-(5-aminothiazol-4-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrim-
idin-4(3H)-one;
2-(2-(difluoromethyl)phenyl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]-
pyrimidin-4(3H)-one;
2-(3-(difluoromethyl)pyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one;
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(2-(trifluoromethyl)phenyl)py-
rido[4,3-d]pyrimidin-4(3H)-one; and
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(3-(trifluoromethyl)pyridin-2-
-yl)pyrido[4,3-d]pyrimidin-4(3H)-one; or a pharmaceutically
acceptable salt thereof.
19. A compound according to claim 1 which is
(R)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one or a pharmaceutically acceptable salt
thereof.
20. A compound according to claim 1 which is
(R)-3-(1-(3,4-difluorophenyl)
propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one or a
pharmaceutically acceptable salt thereof.
21. A pharmaceutical composition comprising a compound according to
claim 1 and a pharmaceutically acceptable carrier, adjuvant or
diluent.
22. A method of treating a disease or disorder selected from
hypoparathyroidism, osteoporosis, osteopenia, periodontal disease,
Paget's disease, bone fracture, osteoarthritis, rheumatoid
arthritis and humoral hypercalcemia of malignancy, the method
comprising administration to a patient in need of treatment thereof
a therapeutically effective amount of a compound according to claim
1.
23. The method according to claim 22 wherein the disease or
disorder is osteoporosis.
Description
[0001] This application is a divisional of U.S. patent application
Ser. No. 11/867,255 filed Oct. 4, 2007, which claims the benefit of
Provisional Patent Application Nos. 60/828,162 filed Oct. 4, 2006
and 60/969,083 filed Aug. 30, 2007.
FIELD OF INVENTION
[0002] The present invention is directed toward novel
pyrido[4,3-d]pyrimidin-4(3H)-one derivatives, pharmaceutical
compositions containing these compounds, methods for their use and
processes for their production. These novel
pyrido[4,3-d]pyrimidin-4(3H)-one derivatives are able to inhibit
calcium receptor activity and thus act as calcium receptor
antagonists.
BACKGROUND OF THE INVENTION
[0003] In mammals, extracellular Ca.sup.2+ is under rigid
homeostatic control with the serum calcium concentration strictly
maintained at a concentration of approximately 1.1 to 1.3 mM in a
healthy mammal. The extracellular Ca.sup.2+ homeostasis depends on
integrated regulation of Ca.sup.2+ fluxes with respect to the
intestine, kidneys and bone. The extracellular Ca.sup.2+ regulates
various processes such as blood coagulation, nerve and muscle
excitability, and normal bone homeostasis. When the Ca.sup.2+ serum
concentration decreases by 50% tetania occurs, and when the
Ca.sup.2+ serum concentration increases by 50% consciousness is
clouded, in both instances a potentially life threatening
circumstance. Extracellular Ca.sup.2+ also inhibits the secretion
of parathyroid hormone (PTH) from parathyroid cells, inhibits bone
resorption by osteoclasts, stimulates secretion of calcitonin from
C-cells and is involved in re-absorption and excretion in the
kidney.
[0004] The extracellular calcium-sensing receptor (CaSR) is a
hormone-like receptor, more particularly a plasma membrane-bound G
protein-coupled receptor (GPCR) that belongs to family 3 of the
GPCR superfamily. Family 3 of the GPCR superfamily includes
metabotropic glutamate receptors (mGluRs), .gamma.-aminobutyric
acid B-type receptors (GABA.sub.BRs) as well as putative pheromone
and taste receptors. The CaSR has a large extracellular domain
exhibiting "Venus flytrap" topology, a seven-transmembrane domain
and a relatively large cytoplasmic domain. Human CaSR consists of
1078 amino acids and shares 93% amino acid homology with bovine
CaSR. The CaSR senses and is activated by changes in extracellular
Ca.sup.2+ levels. The presence of CaSR on certain specialized cells
enables those Ca.sup.2+-sensing cells to respond to changes in
extracellular Ca.sup.2+ concentration. Examples of
Ca.sup.2+-sensing cells include the parathyroid-secreting cells of
the parathyroid gland, the calcitonin-secreting C cells of the
thyroid gland and certain cells in the kidney. In addition, the
CaSR has been found in a wide variety of other tissues including
intestine, bone, bone marrow, brain, skin, pancreas, lung and
heart.
[0005] The CaSR on the surface of parathyroid chief cells is the
primary entity that regulates secretion of PTH from parathyroid
cells. Activation of the CaSR on parathyroid chief cells by
extracellular Ca.sup.2+ suppresses PTH production and secretion,
inhibits parathyroid cellular proliferation and likely inhibits PTH
gene expression. The CaSR on the surface of the
calcitonin-secreting C cells of the thyroid gland mediate the
stimulatory action of high extracellular Ca.sup.2+ concentration on
calcitonin secretion, thereby increasing the circulating level of
the Ca.sup.2+-lowering hormone calcitonin. The CaSR is also present
in the kidney, along much of the nephrons and at the basolateral
surface in the cortical thick ascending limb. In the basolateral
surface in the cortical thick ascending limb the CaSR is thought to
mediate high Ca.sup.2+-induced inhibition of the tubular
re-absorption of Ca.sup.2+ and magnesium. A reduction of renal
cortical synthesis of 1,25(OH).sub.2 vitamin D and polyuria with
dilute urine are partially the result of hypercalcaemic activation
of the CaSR in the nephron.
[0006] PTH is the primary endocrine hormone regulating Ca.sup.2+
homeostasis in the blood and extracellular fluids. PTH, by acting
on bone and kidney cells, increases the level of Ca.sup.2+ in the
plasma. This increase in plasma Ca.sup.2+ concentration then acts
as a negative feedback signal, thereby depressing PTH secretion.
The reciprocal relationship between extracellular Ca.sup.2+ and PTH
secretion forms an important mechanism for maintaining bodily
Ca.sup.2+ homeostasis. PTH has been found to increase bone
turnover, but the overall effect on bone is dependent on temporal
changes in circulating levels of PTH. Sustained elevations in
circulating plasma PTH levels, as occurs in hyperparathyroidism,
have been found to result in a net catabolic effect on bone. By
contrast, transient increases in plasma PTH levels, achieved by
daily or near daily injection of exogenous hormone, have been found
to exhibit a net anabolic effect on bone. The effect of PTH on bone
is likely due to PTH being able to induce a rapid release of
calcium from bone and mediate other changes by acting directly on
osteoblasts and indirectly on osteoclasts. PTH affects cellular
metabolic activity, ion transport, cell shape, gene transcriptional
activity and secretion of proteases in osteoblasts. Also, PTH
stimulates the production of RANKL, a protein that plays a crucial
role in osteoclast differentiation and activity.
[0007] Various compounds are known to modulate the effects of
extracellular Ca.sup.2+ on the CaSR. Calcimimetics are agents that
act as allosteric modulators of the CaSR that increase the
sensitivity of the CaSR to activation by extracellular Ca.sup.2+.
Calcilytics, or calcium receptor antagonists, are agents that act
as modulators of the CaSR that inhibit CaSR activity. This
inhibition of the CaSR activity results in a decrease of one or
more CaSR activities that are evoked by extracellular
Ca.sup.2+.
[0008] Certain urea derivatives, such as those disclosed in PCT
International Publication WO 02/059102, are described as having
calcimimetic activity. In addition, certain phenylalkylamine
derivatives have been identified as calcimimetics. Phenylalkylamine
calcimimetic compounds include
(R)-N-(1-(3-methoxyphenyl)ethyl)-3-phenylpropan-1-amine
hydrochloride (NPS-467);
(R)-3-(2-chlorophenyl)-N-(1-(3-methoxyphenyl)ethyl)propan-1-amine
hydrochloride (NPS R-568, tecalcet hydrochloride) and
(R)-(-)-N-(1-(naphthalen-1-yl)ethyl)-3-(3-(trifluoromethyl)phenyl)propan--
1-amine hydrochloride (NPS-1493, cinacalcet hydrochloride).
Cinacalcet hydrochloride and uses thereof are disclosed in U.S.
Pat. Nos. 6,011,068; 6,031,003; 6,211,244 and 6,313,146. Cinacalcet
hydrochloride is marketed as Sensipar.RTM. and Minpara.RTM. in the
U.S. and Europe, respectively, and is indicated for the treatment
of secondary hyperparathyroidism in patients with chronic kidney
disease on dialysis and for hypercalcemia in patients with
parathyroid carcinoma.
[0009] Calcilytics, or calcium receptor antagonists, have been
described in various publications such as PCT International
Publication Nos. WO 93/04373; WO 94/18959; WO 95/11211; WO
97/37967; WO 98/44925; WO 98/45255; WO 99/51241; WO 99/51569; WO
00/45816; WO 02/14259; WO 02/38106; WO 2004/041755; and WO
2005/030746; Nemeth, E. F.; Journal of Molecular Endocrinology
(2002) 29, 15-21; Kessler, A. et al.; ChemBioChem (2004) 5, 1131;
Steddon, S. J. et al.; Lancet (2005) 365, 2237-2239; and
Shcherbakova, I.; et al.; Bioorganic & Medicinal Chemistry
Letters (2005) 15, 1557-1560.
[0010] Calcium receptor antagonists are useful in the treatment of
various disease states characterized by abnormal levels of one or
more components, e.g., polypeptides such as hormones, enzymes or
growth factors, the expression and/or secretion of which is
regulated or affected by activity at one or more CaSR. Target
diseases or disorders for calcium receptor antagonists include
diseases involving abnormal bone and mineral homeostasis. Abnormal
calcium homeostasis is characterized by one or more of the
following activities: an abnormal increase or decrease in serum
calcium; an abnormal increase or decrease in urinary excretion of
calcium; an abnormal increase or decrease in bone calcium levels
(for example, as assessed by bone mineral density measurements); an
abnormal absorption of dietary calcium; an abnormal increase or
decrease in the production and/or release of messengers which
affect serum calcium levels such as PTH and calcitonin; and an
abnormal change in the response elicited by messengers which affect
serum calcium levels.
[0011] The novel calcium receptor antagonists of this invention are
useful in the treatment of diseases associated with abnormal bone
or mineral homeostasis. Thus, these calcium receptor antagonists
are useful in the treatment of hypoparathyroidism, osteoporosis,
osteopenia, periodontal disease, bone fracture, osteoarthritis,
rheumatoid arthritis, Paget's disease, humoral hypercalcemia
associated with malignancy.
SUMMARY OF THE INVENTION
[0012] The present invention is directed towards calcium receptor
antagonist compounds, pharmaceutical compositions containing the
calcium receptor antagonist compounds and methods of treatment
employing the calcium receptor antagonist compounds.
[0013] More specifically, the present invention is directed to
calcium receptor antagonists that are
pyrido[4,3-d]pyrimidin-4(3H)-one derivatives of structural formula
I
##STR00002##
wherein R.sup.1 is -Q or (C.sub.1-C.sub.6)alkyl-Q; R.sup.2 is
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl, aryl or
heteroaryl; wherein said aryl or heteroaryl is substituted with
hydroxy and additionally said (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, aryl or heteroaryl within the
definition of R.sup.2 is optionally substituted with one to three
substituents independently selected from halo, cyano,
trifluoromethyl, trifluoromethoxy, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy or hydroxy; R.sup.3 is hydrogen,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.1-C.sub.6)alkyl-Q, aryl, heteroaryl, OR.sup.6, or
NR.sup.7R.sup.8; wherein said (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, aryl or heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl; (C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or
hydroxy; R.sup.4 and R.sup.5 are each independently hydrogen, halo,
cyano, (C.sub.1-C.sub.6)alkyl optionally substituted with one to
three fluoro, aryl, heteroaryl, or OR.sup.6; R.sup.6 at each
occurrence is independently (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl-(C.sub.1-C.sub.6)alkyl, aryl,
heteroaryl, (C.sub.1-C.sub.6)alkylaryl, or
(C.sub.1-C.sub.6)alkylheteroaryl; each of said
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl-(C.sub.1-C.sub.6)alkyl, aryl,
heteroaryl, (C.sub.1-C.sub.6)alkylaryl, or
(C.sub.1-C.sub.6)alkylheteroaryl optionally substituted with one to
three substituents independently selected from halo, hydroxy or
(C.sub.1-C.sub.3)alkyl; R.sup.7 and R.sup.8, at each occurrence,
are independently hydrogen, (C.sub.1-C.sub.6)alkyl, or
(C.sub.3-C.sub.7)cycloalkyl; or R.sup.7 and R.sup.8 taken together
with the nitrogen to which they are attached form a 3 to 7 membered
fully saturated, partially saturated or fully unsaturated ring
optionally containing one to two additional heteroatoms
independently selected from N(R.sup.9).sub.n; O or S(O).sub.p; n is
0 or 1; p is 0, 1 or 2; R.sup.9 is hydrogen or
(C.sub.1-C.sub.6)alkyl; Q, at each occurrence, is independently
aryl or heteroaryl; wherein said aryl or heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy or hydroxy; or a
pharmaceutically acceptable salt thereof.
[0014] "Halo" refers to fluoro, chloro, bromo or iodo.
[0015] "(C.sub.1-C.sub.6)alkyl" refers to a hydrocarbon group
having one to six carbon atoms joined together by single
carbon-carbon bonds. The (C.sub.1-C.sub.6)alkyl group may be
straight-chain or contain one or more branches and may be
unsubstituted or substituted as specified. Examples of
(C.sub.1-C.sub.6)alkyl groups include methyl, ethyl, n-propyl,
isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl,
(1-methyl)butyl, (2-methyl)butyl, (3-methyl)butyl,
(1,2-dimethyl)propyl, n-hexyl, (1-methyl)pentyl, (2-methyl)pentyl,
(3-methyl)pentyl, (4-methyl)pentyl, (1-ethyl)butyl, (2-ethyl)butyl,
(1,2-dimethyl)butyl, (1,3-dimethyl)butyl, (2,3-dimethyl)butyl and
the like.
[0016] "(C.sub.1-C.sub.6)alkoxy" refers to an oxygen joined to a
(C.sub.1-C.sub.6)alkyl group. The (C.sub.1-C.sub.6)alkyl group in
the (C.sub.1-C.sub.6)alkoxy moiety may be straight-chain or contain
one or more branches and may be unsubstituted or substituted as
specified. Examples of (C.sub.1-C.sub.6)alkoxy groups include
methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy,
tert-butoxy, n-pentoxy and the like. Likewise, alkyl or alkoxy
groups of differing length such as "(C.sub.1-C.sub.3)alkyl" or
"(C.sub.1-C.sub.3)alkoxy" are defined in the same manner but
limited to the number of carbons present.
[0017] "(C.sub.3-C.sub.7)cycloalkyl" refers to a saturated
carbocyclic group having three to seven carbons and encompasses
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
The (C.sub.3-C.sub.7)cycloalkyl group can be unsubstituted or
substituted as specified.
[0018] "Aryl" refers to a six to sixteen membered carbocyclic
aromatic group having at least one ring with a conjugated
pi-electron system. The aryl group can have conjugated or fused
rings and can be unsubstituted or substituted as specified.
Examples of aryl groups include phenyl, naphthalenyl, anthracenyl,
phenanthrenyl, azulenyl and biphenyl.
[0019] "Heteroaryl" refers to a five to sixteen membered aromatic
group with at least one ring with a conjugated pi-electron system
and containing one to four heteroatoms such as N, O or S. The
hetroaryl group can have conjugated or fused rings and can be
unsubstituted or substituted as specified. Examples of heteroaryl
groups include thienyl, furanyl, pyrrolyl, pyrazolyl, imidazoyl,
oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl,
oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyridizinyl,
pyrimidinyl, pyrazinyl, triazinyl, indolyl, isoindolyl,
indolizinyl, benzofuranyl, benzothienyl, indazolyl, benzimidazoyl,
benzthiazolyl, purinyl, quinolizinyl, quinolinyl, isoquinolinyl,
cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl,
naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl,
phenothiazinyl or phenoxazinyl.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1 is a chart depicting plasma PTH levels from the time
of intravenous injection to 480 minutes following intravenous
injection of 1 mg/kg of the compound of Example 1,
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one), in normal rats.
DETAILED DESCRIPTION OF THE INVENTION
[0021] The present invention provides novel
pyrido[4,3-d]pyrimidin-4(3H)-one derivatives and pharmaceutically
acceptable salts thereof of structural formula I
##STR00003##
wherein the variables R.sup.1, R.sup.2, R.sup.3, R.sup.4 and
R.sup.5 are described hereinabove. The pharmaceutically acceptable
salts of the compounds of formula I include the acid addition and
base salts (including disalts) thereof.
[0022] Suitable acid addition salts are formed from acids which
form non-toxic salts. Examples include the acetate, aspartate,
benzoate, besylate, bicarbonate/carbonate, bisulphate/sulphate,
borate, camsylate, citrate, edisylate, esylate, formate, fumarate,
gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate,
hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide,
isethionate, lactate, malate, maleate, malonate, mesylate,
methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate,
orotate, oxalate, palmitate, pamoate, phosphate/hydrogen
phosphate/dihydrogen phosphate, saccharate, stearate, succinate,
tartrate, tosylate and trifluoroacetate salts.
[0023] Suitable base salts are formed from bases which form
non-toxic salts. Examples include the aluminium, arginine,
benzathine, calcium, choline, diethylamine, diolamine, glycine,
lysine, magnesium, meglumine, olamine, potassium, sodium,
tromethamine and zinc salts. For a review on suitable salts, see
"Handbook of Pharmaceutical Salts: Properties, Selection, and Use"
by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).
[0024] A pharmaceutically acceptable salt of a compound of formula
I may be readily prepared by mixing together solutions of the
compound of formula I and the desired acid or base, as appropriate.
The salt may precipitate from solution and be collected by
filtration or may be recovered by evaporation of the solvent. The
degree of ionization in the salt may vary from completely ionized
to almost non-ionized.
[0025] The compounds of the invention include compounds of formula
I as hereinbefore defined, polymorphs, and isomers thereof
(including optical, geometric and tautomeric isomers) as
hereinafter defined and isotopically-labeled compounds of formula
I.
[0026] The compounds of the present invention may be administered
as prodrugs. Thus certain derivatives of compounds of formula I
which may have little or no pharmacological activity themselves
can, when administered into or onto the body, be converted into
compounds of formula I having the desired activity, for example, by
hydrolytic cleavage. Such derivatives are referred to as
`prodrugs`. Further information on the use of prodrugs may be found
in `Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium
Series (T Higuchi and W Stella) and `Bioreversible Carriers in Drug
Design`, Pergamon Press, 1987 (ed. E B Roche, American
Pharmaceutical Association).
[0027] Prodrugs can, for example, be produced by replacing
appropriate functionalities present in the compounds of formula I
with certain moieties known to those skilled in the art as
`pro-moieties` as described, for example, in "Design of Prodrugs"
by H Bundgaard (Elsevier, 1985).
[0028] Some examples of such prodrugs include:
(i) where the compound of formula I contains a carboxylic acid
functionality (--COOH), an ester thereof, for example, replacement
of the hydrogen with (C.sub.1-C.sub.8)alkyl; (ii) where the
compound of formula I contains an alcohol functionality (--OH), an
ether thereof, for example, replacement of the hydrogen with
(C.sub.1-C.sub.6)alkanoyloxymethyl; and (iii) where the compound of
formula I contains a primary or secondary amino functionality
(--NH.sub.2 or --NHR where R is not H), an amide thereof, for
example, replacement of one or both hydrogens with
(C.sub.1-C.sub.10)alkanoyl. Further examples of replacement groups
in accordance with the foregoing examples and examples of other
prodrug types may be found in the aforementioned references.
Finally, certain compounds of formula I may themselves act as
prodrugs of other compounds of formula I.
[0029] Compounds of formula I containing one or more asymmetric
carbon atoms can exist as two or more stereoisomers. Where a
compound of formula I contains an alkenyl or alkenylene group,
geometric cis/trans (or Z/E) isomers are possible. Where the
compound contains, for example, a keto or oxime group or an
aromatic moiety, tautomeric isomerism (`tautomerism`) can occur. It
follows that a single compound may exhibit more than one type of
isomerism.
[0030] Included within the scope of the claimed compounds of
formula I are all stereoisomers, geometric isomers and tautomeric
forms of the compounds of formula I, including compounds exhibiting
more than one type of isomerism, and mixtures of one or more
thereof. Also included are acid addition or base salts wherein the
counterion is optically active, for example, D-lactate or L-lysine,
or racemic, for example, DL-tartrate or DL-arginine. Cis/trans
isomers may be separated by conventional techniques well known to
those skilled in the art, for example, chromatography and
fractional crystallisation. Conventional techniques for the
preparation/isolation of individual enantiomers include chiral
synthesis from a suitable optically pure precursor or resolution of
the racemate (or the racemate of a salt or derivative) using, for
example, chiral high pressure liquid chromatography (HPLC).
[0031] Alternatively, the racemate (or a racemic precursor) may be
reacted with a suitable optically active compound, for example, an
alcohol, or, in the case where the compound of formula (I) contains
an acidic or basic moiety, an acid or base such as tartaric acid or
1-phenylethylamine. The resulting diastereomeric mixture may be
separated by chromatography and/or fractional crystallization and
one or both of the diastereoisomers converted to the corresponding
pure enantiomer(s) by means well known to a skilled person. Chiral
compounds of the invention (and chiral precursors thereof) may be
obtained in enantiomerically-enriched form using chromatography,
typically HPLC, on an asymmetric resin with a mobile phase
consisting of a hydrocarbon, typically heptane or hexane,
containing from 0 to 50% isopropanol, typically from 2 to 20%, and
from 0 to 5% of an alkylamine, typically 0.1% diethylamine.
Concentration of the eluate affords the enriched mixture. Mixtures
of stereoisomers may be separated by conventional techniques known
to those skilled in the art. [see, for example, "Stereochemistry of
Organic Compounds" by E L Eliel (Wiley, New York, 1994).]
[0032] The present invention includes all pharmaceutically
acceptable isotopically-labelled compounds of formula I wherein one
or more atoms are replaced by atoms having the same atomic number,
but an atomic mass or mass number different from the atomic mass or
mass number usually found in nature.
[0033] Examples of isotopes suitable for inclusion in the compounds
of the invention include isotopes of hydrogen, such as .sup.2H and
.sup.3H, carbon, such as .sup.11C, .sup.13C and .sup.14C, chlorine,
such as .sup.38Cl, fluorine, such as .sup.18F, iodine, such as
.sup.123I and .sup.125I, nitrogen, such as .sup.13N and .sup.15N,
oxygen, such as .sup.15O, .sup.17O and .sup.18O, phosphorus, such
as .sup.32P, and sulfur, such as .sup.35S. Certain
isotopically-labelled compounds of formula I, for example, those
incorporating a radioactive isotope, are useful in drug and/or
substrate tissue distribution studies. The radioactive isotopes
tritium, i.e. .sup.3H, and carbon-14, i.e. .sup.14C, are
particularly useful for this purpose in view of their ease of
incorporation and ready means of detection. Substitution with
heavier isotopes such as deuterium, i.e. .sup.2H, may afford
certain therapeutic advantages resulting from greater metabolic
stability, for example, increased in vivo half-life or reduced
dosage requirements, and hence may be preferred in some
circumstances. Substitution with positron emitting isotopes, such
as .sup.11C, .sup.18F, .sup.15O and .sup.13N, can be useful in
Positron Emission Topography (PET) studies for examining substrate
receptor occupancy. Isotopically-labeled compounds of formula (I)
can generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described in
the accompanying Examples and Preparations using an appropriate
isotopically-labeled reagents in place of the non-labeled reagent
previously employed.
[0034] A preferred embodiment of the present invention are
compounds of Formula I as described above wherein R.sup.2 is aryl
or heteroaryl, wherein said aryl or heteroaryl is substituted with
hydroxy and optionally substituted with one to three substituents
independently selected from (C.sub.1-C.sub.6)alkoxy or halo; or a
pharmaceutically acceptable salt thereof. Another preferred
embodiment of this invention is a compound of Formula I, wherein
R.sup.2 is 2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl; wherein said
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl is optionally
substituted with one to two substituents independently selected
from (C.sub.1-C.sub.6)alkoxy or fluoro; or a pharmaceutically
acceptable salt thereof. Yet another preferred embodiment of the
present invention is a compound of Formula I as described above
wherein R.sup.1 is (C.sub.1-C.sub.6)alkyl-Q; and Q is phenyl
optionally substituted with one or two fluoro; or a
pharmaceutically acceptable salt thereof.
[0035] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; or a pharmaceutically acceptable salt thereof.
[0036] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkyl-Q,
aryl, heteroaryl, OR.sup.6, or NR.sup.7R.sup.8, wherein said
(C.sub.1-C.sub.6)alkyl, aryl or heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or
hydroxy; and R.sup.4 and R.sup.5 are each hydrogen; or a
pharmaceutically acceptable salt thereof.
[0037] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is trifluoromethyl; and R.sup.4 and R.sup.5 are each
hydrogen; or a pharmaceutically acceptable salt thereof.
[0038] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is (C.sub.1-C.sub.6)alkyl; and R.sup.4 and R.sup.5
are each hydrogen; or a pharmaceutically acceptable salt
thereof.
[0039] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is (C.sub.1-C.sub.6)alkyl-Q; and R.sup.4 and R.sup.5
are each hydrogen; or a pharmaceutically acceptable salt
thereof.
[0040] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is aryl, wherein said aryl is optionally substituted
with one to three substituents independently selected from halo,
cyano, trifluoromethyl, trifluoromethoxy, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or hydroxy; and R.sup.4
and R.sup.5 are each hydrogen; or a pharmaceutically acceptable
salt thereof.
[0041] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is heteroaryl; wherein said heteroaryl is optionally
substituted with one to three substituents independently selected
from halo, cyano, trifluoromethyl, trifluoromethoxy,
(C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy, NR.sup.7R.sup.8 or
hydroxy; and R.sup.4 and R.sup.5 are each hydrogen; or a
pharmaceutically acceptable salt thereof.
[0042] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is OR.sup.6; and R.sup.4 and R.sup.5 are each
hydrogen; or a pharmaceutically acceptable salt thereof.
[0043] Another preferred embodiment of the present invention is a
compound of Formula I, wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; and R.sup.2 is 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl, wherein said 2-hydroxy-phenyl or
3-hydroxy-pyridin-2-yl is optionally substituted with one or two
substituents independently selected from (C.sub.1-C.sub.3)alkyl or
halo; R.sup.3 is NR.sup.7R.sup.8; and R.sup.4 and R.sup.5 are each
hydrogen; or a pharmaceutically acceptable salt thereof.
[0044] A preferred embodiment of the present invention is a
compound of Formula I wherein R.sup.3 is trifluoromethyl; or a
pharmaceutically acceptable salt thereof. Another preferred
embodiment of the present invention is a compound of Formula I
wherein R.sup.3 is trifluoromethyl; and R.sup.4 and R.sup.5 are
each independently hydrogen or methyl; or a pharmaceutically
acceptable salt thereof. Yet another preferred embodiment of the
present invention is a compound of Formula I wherein R.sup.2 is
aryl or heteroaryl, wherein said aryl or heteroaryl is substituted
with hydroxy and optionally substituted with one to three
substituents independently selected from (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkoxy or fluoro; R.sup.3 is trifluoromethyl; and
R.sup.4 and R.sup.5 are each independently hydrogen or methyl; or a
pharmaceutically acceptable salt thereof. Yet another preferred
embodiment of the present invention is a compound of Formula I
wherein R.sup.2 is hydroxy-phenyl or hydroxy-pyridyl, wherein said
hydroxy-phenyl or hydroxy-pyridyl is optionally substituted with
one to three substituents independently selected from methyl,
methoxy or fluoro; R.sup.3 is trifluoromethyl; and R.sup.4 and
R.sup.5 are each independently hydrogen or methyl; or a
pharmaceutically acceptable salt thereof.
[0045] Another preferred embodiment of the present invention is a
compound of Formula I wherein R.sup.1 is (C.sub.1-C.sub.6)alkyl-Q;
Q is phenyl optionally substituted with one or two substituents
independently selected from fluoro, methyl or methoxy; R.sup.2 is
hydroxy-phenyl or hydroxy-pyridyl, wherein said hydroxy-phenyl or
hydroxy-pyridyl is optionally substituted with one to three
substituents independently selected from methyl, methoxy or fluoro;
R.sup.3 is trifluoromethyl; and R.sup.4 and R.sup.5 are each
independently hydrogen or methyl; or a pharmaceutically acceptable
salt thereof. Still another preferred embodiment of the present
invention is a compound of Formula I wherein R.sup.1 is phenethyl
or 1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two fluoro; R.sup.3 is trifluoromethyl; and R.sup.4 and
R.sup.5 are each independently hydrogen or methyl; or a
pharmaceutically acceptable salt thereof.
[0046] Another preferred embodiment of the present invention is a
compound of Formula I wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two halo; R.sup.2 is hydroxy-phenyl or hydroxy-pyridyl,
wherein said hydroxy-phenyl or hydroxy-pyridyl is optionally
substituted with one or two substituents independently selected
from (C.sub.1-C.sub.3)alkyl or halo; R.sup.3 is trifluoromethyl;
and R.sup.4 and R.sup.5 are each independently hydrogen or methyl;
or a pharmaceutically acceptable salt thereof.
[0047] Another preferred embodiment of the present invention is a
compound of Formula I wherein R.sup.1 is phenethyl or
1-methyl-2-(phenyl)ethyl, wherein said phenethyl or
1-methyl-2-(phenyl)ethyl is optionally substituted on phenyl with
one or two substituents independently selected from halo,
(C.sub.1-C.sub.3)alkyl or (C.sub.1-C.sub.3)alkoxy; R.sup.2 is
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl, wherein said
2-hydroxy-phenyl or 3-hydroxy-pyridin-2-yl is optionally
substituted with one or two substituents independently selected
from (C.sub.1-C.sub.3)alkyl or halo; R.sup.3 is
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl,
(C.sub.1-C.sub.6)alkyl-Q, aryl, heteroaryl, or OR.sup.6; wherein
said (C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.7)cycloalkyl, aryl or
heteroaryl is optionally substituted with one to three substituents
independently selected from fluoro, cyano, trifluoromethyl,
trifluoromethoxy, (C.sub.1-C.sub.3)alkyl; (C.sub.1-C.sub.3)alkoxy,
NR.sup.7R.sup.8 or hydroxy; and R.sup.4 and R.sup.5 are each
independently hydrogen or methyl; or a pharmaceutically acceptable
salt thereof.
[0048] Another preferred embodiment of the present invention is a
compound selected from the group consisting of: [0049]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyri-m-
id in-4-one; [0050]
2-(3-Fluoro-2-hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-
-d]pyrimidin-4-one; [0051]
2-(3-Hydroxy-pyridin-2-yl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]-
pyrimidin-4-one; [0052]
(R)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one; [0053]
(S)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H-
-pyrido[4,3-d]pyrimidin-4-one; [0054]
(R)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromet-
hyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0055]
(R,S)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-t-
rifluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0056]
(R)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-tri-
fluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0057]
(S)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-tri-
fluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0058]
2-(2-Hydroxyphenyl)-3-(2-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[-
4,3-d]pyrimidin-4-one; [0059]
2-(2-Hydroxyphenyl)-3-(3-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[-
4,3-d]pyrimidin-4-one; [0060]
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(2-fluoro-phenyl)-ethyl]-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0061]
2-(2-Hydroxy-phenyl)-7-methyl-3-(phenyl-ethyl)-5-trifluoromethyl-3H-pyrid-
o[4,3-d]pyrimidin-4-one; [0062]
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(3-fluoro-phenyl)-ethyl]-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0063]
3-[2-(2-Fluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluoromethy-
l-3H-pyrido[4,3-d]pyrimidin-4-one; [0064]
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0065]
3-[2-(2,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0066]
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorom-
ethyl-3H-pyrido[4,3-d]pyrimidin-4-one; [0067]
2-(2-Hydroxy-phenyl)-5-methylamino-3-phenethyl-3H-pyrido[4,3-d]pyrimidin--
4-one; [0068]
2-(2-Hydroxy-phenyl)-5-isopropylamino-3-phenethyl-3H-pyrido[4,3-d]pyrimid-
in-4-one; [0069]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-pyrrolidin-1-yl-3H-pyrido[4,3-d]pyrimi-
din-4-one; [0070]
2-(2-Hydroxy-phenyl)-5-(4-methyl-piperazin-1-yl)-3-phenethyl-3H-pyrido[4,-
3-d]pyrimidin-4-one; [0071]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-piperazin-1-yl-3H-pyrido[4,3-d]pyrimid-
in-4-one; [0072]
5-Dimethylamino-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-
n-4-one; [0073]
2-(2-Hydroxy-phenyl)-5-morpholin-4-yl-3-phenethyl-3H-pyrido[4,3-d]pyrimid-
in-4-one; [0074]
5-Azetidin-1-yl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d-pyrimidi-
n-4-one; [0075]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-phenyl-3H-pyrido[4,3-d]pyrimidin-4-one-
; [0076]
5-Benzyl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimid-
in-4-one; [0077]
2-(2-Hydroxy-phenyl)-5-methyl-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-4-one;
[0078]
5-(6-Dimethylamino-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl--
3H-pyrido[4,3-d]pyrimidin-4-one; [0079]
5-(6-Dimethylamino-5-methyl-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethy-
l-3H-pyrido[4,3-d]pyrimidin-4-one; [0080]
5-(6-pyrrolidine-5-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyri-
do[4,3-d]pyrimidin-4-one; [0081]
2-(2-Hydroxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-on-
e; [0082]
2-(2-Hydroxy-phenyl)-5-(1-methyl-cyclopropylmethoxy)-3-phenethyl-
-3H-pyrido[4,3-d]pyrimidin-4-one; [0083]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-propoxy-3H-pyrido[4,3-d]pyrimidin-4-on-
e; [0084]
5-Cyclobutyloxy-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d-
]pyrimidin-4-one; [0085]
2-(2-Hydroxy-phenyl)-5-isobutoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4--
one; [0086]
2-(2-Hydroxy-phenyl)-5-isopropoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-
-one; [0087]
2-(2-Hydroxy-phenyl)-3-phenethyl-5-(2,2,2-trifluoro-ethoxy)-3H-pyrido[4,3-
-d]pyrimidin-4-one; [0088]
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0089]
(R)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0090]
(S)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0091]
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0092]
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0093]
3-(2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one; [0094]
3-(2,3-difluorophenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)py-
rido[4,3-d]pyrimidin-4(3H)-one; [0095]
3-(5-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0096]
3-(2-fluoro-6-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0097]
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0098]
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0099]
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluoro-
methyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0100]
3-(1-(2-fluorophenyl)butan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorome-
thyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0101]
3-(3-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0102]
3-(2-cyclopentylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one; [0103]
3-(2-cyclohexylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one; [0104]
(R)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0105]
(S)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0106]
(R,S)-2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-2-yl)ethyl)-5-(-
trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0107]
(R)-2-(3-fluoro-2-hydroxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0108]
(R)-2-(1H-imidazol-2-yl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrid-
o[4,3-d]pyrimidin-4(3H)-one; [0109]
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4-
,3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide; [0110]
(R)-3-(1-phenylpropan-2-yl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,-
3-d]pyrimidin-4(3H)-one; [0111]
3-phenethyl-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(-
3H)-one; [0112]
3-(2-methoxyphenethyl)-2-(2-hydroxyphenyl)-5-(trifluoromethyl)pyrido[4,3--
d]pyrimidin-4(3H)-one; [0113]
3-(2-fluorophenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one; [0114]
(R)-2-cyclopentyl-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido[4,3-d-
]pyrimidin-4(3H)-one; [0115]
2-isopropyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-o-
ne [0116]
2-cyclopentyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimi-
din-4(3H)-one; [0117]
3-(2-methoxyphenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]p-
yrimidin-4(3H)-one; [0118]
3-(2-cyclohexylethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one; [0119]
2-(3-(2-cyclohexylethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4,3--
d]pyrimidin-2-yl)benzonitrile; [0120]
2-(3-aminopyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrim-
idin-4(3H)-one; [0121]
N-(2-(4-oxo-3-phenethyl-5-(trifluoromethyl)-3,4-dihydropyrido[4,3-d]pyrim-
idin-2-yl)pyridin-3-yl)methanesulfonamide; [0122]
3-(1-(2-fluorophenyl)propan-2-yl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyr-
ido[4,3-d]pyrimidin-4(3H)-one; [0123]
2-(3-hydroxypyridin-2-yl)-3-isopentyl-5-(trifluoromethyl)pyrido[4,3-d]pyr-
imidin-4(3H)-one; [0124]
2-(3-hydroxypyridin-2-yl)-3-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluo-
romethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0125]
2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-4-yl)ethyl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0126]
3-(2-fluoro-5-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0127]
3-(2-fluoro-3-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one; [0128]
2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydroquinazolin--
2-yl)benzonitrile [0129]
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4-
,3-d]pyrimidin-2-yl)pyridin-3-yl)-2,2,2-trifluoroacetamide; [0130]
(R)-2,2,2-trifluoro-N-(2-(4-oxo-3-(1-phenylpropan-2-yl)-5-(trifluoromethy-
l)-3,4-dihydropyrido[4,3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide;
[0131]
3-cyclohexyl-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one; [0132]
3-(cyclohexylmethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one; [0133]
3-Phenethyl-5-trifluoromethyl-2-(3-trifluoromethyl-pyridin-2-yl)-3H-pyrid-
o[4,3-d]pyrimidin-4-one; [0134]
2-(5-aminothiazol-4-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrim-
idin-4(3H)-one; [0135]
2-(2-(difluoromethyl)phenyl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]-
pyrimidin-4(3H)-one; [0136]
2-(3-(difluoromethyl)pyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one; [0137]
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(2-(trifluoromethyl)phenyl)py-
rido[4,3-d]pyrimidin-4(3H)-one; and [0138]
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(3-(trifluoromethyl)pyridin-2-
-yl)pyrido[4,3-d]pyrimidin-4(3H)-one; or a pharmaceutically
acceptable salt thereof.
[0139] Another embodiment of the present invention is a
pharmaceutical composition comprising a compound according to
Formula I as described in any of the preceding embodiments
hereinabove and a pharmaceutically acceptable carrier, adjuvant or
diluent.
[0140] Another embodiment of the present invention are novel
intermediate compounds 4-Amino-2-(trifluoromethyl)nicotinic acid or
4-Amino-2-chloronicotinic acid; or a pharmaceutically acceptable
salt thereof.
[0141] Another embodiment of the present invention is a method of
treating a disease or disorder characterized by abnormal bone or
mineral homeostasis which comprises the administration to a patient
in need of treatment thereof a therapeutically effective amount of
a compound according to Formula I as described in any of the
preceding embodiments hereinabove. A preferred embodiment of the
present invention is the method according to the preceding
embodiment wherein the disease or disorder characterized by
abnormal bone or mineral homeostasis is selected from the group
consisting of osteoporosis, osteopenia, periodontal disease,
Paget's disease, bone fracture, osteoarthritis, rheumatoid
arthritis, and humoral hypercalcemia of malignancy. Yet another
preferred embodiment is the method according to the preceding
embodiment wherein the disease or disorder characterized by
abnormal bone or mineral homeostasis is osteoporosis.
[0142] The following reaction schemes, Reaction Schemes I-IV,
depict methods of synthesis for compounds of formula I. In the
following general methods for preparation of the compounds of
formula I the variables R.sup.1, R.sup.2, R.sup.3, R.sup.4, and
R.sup.5, are as previously defined for a compound of the formula I
unless otherwise stated. The Reaction Schemes herein described are
intended to provide a general description of the methodology
employed in the preparation of many of the Examples given. However,
it will be evident from the detailed descriptions given in the
Experimental section that the modes of preparation employed extend
further than the general procedures described herein. In
particular, it is noted that the compounds prepared according to
these Schemes may be modified further to provide new Examples
within the scope of this invention. The reagents and intermediates
used in the following examples are either commercially available or
can be prepared according to standard literature procedures by
those skilled in the art of organic synthesis.
[0143] Reaction Scheme I, below, depicts the synthesis of
intermediates of formula V which are useful for preparing compounds
of formula I. Treatment of an appropriately substituted pyridine of
formula IX with an appropriate base, such as lithium
diisopropylethylamide, in an appropriate solvent, such as THF, at
approximately -65.degree. C. to -78.degree. C. followed by addition
of CO.sub.2 and acidification upon workup provides the
corresponding nicotinic acid of formula VIII. Treatment of the
appropriately substituted nicotinic acid of formula VIII with an
appropriate base, such as n-butyl lithium and lithium
2,2,6,6-tetramethylpiperidide (LiTMP), followed by trapping with an
appropriate halogen source, such as hexachloroethane, and followed
by esterification, with diazomethane for example, provides the
corresponding chlorinated pyridine ester of formula VII. The
chlorinated pyridine ester VII is then treated with an appropriate
nitrogen source, such as azide anion (i.e. sodium azide) in an
appropriate solvent, such as DMF, typically at 40.degree. C. to
50.degree. C. for 1 to 24 hours, to provide the azido pyridine
ester of formula VI [Cf: Kiyama, Chem. Pharm. Bull. 1995, 43,
450.]. Reduction of the azide group in the compound of formula VI
to the corresponding amine can be carried out using several
procedures known in the art, such as catalytic hydrogenation,
treatment with aqueous hydriodic acid at ambient temperature or
treatment with triphenylphosphine. Saponification of the ester can
then be carried out in a variety of ways such as aqueous lithium
hydroxide in an appropriate solvent such as dioxane:MeOH:water
(3:2:1) at a temperature of 40.degree. C. to 100.degree. C.,
preferably at approximately 85.degree. C. for 1 to 24 hours to
provide the corresponding nicotinic acid of formula V. It is to be
appreciated that the group R.sup.3 in the compounds of formulae IX,
VIII, VII, VI and V can have the same meaning as defined for group
R.sup.3 in compounds of formula I or alternatively can be a
functionality that can be converted to a group R.sup.3 as defined
for compounds of formula I. For example, R.sup.3 in formulae IX,
VIII, VII, VI and V can be a halo group, such as chloro, which can
then be further converted, for example, as shown in Reaction Scheme
VI, below to a group R.sup.3 as defined for a compound of formula
I.
##STR00004##
[0144] Reaction Scheme II, below, depicts the preparation of
compounds of Formula I. Formula IV compounds are prepared by
reacting the corresponding acid of Formula V with a variety of
primary amines of general formula R.sup.1NH.sub.2 under standard
amide coupling conditions for a period between 4 to 24 hours.
Formula III compounds are prepared by reaction of Formula IV
compounds with an appropriate aldehyde, R.sup.2'CHO, in the
presence of a catalytic amount of an acid, such as
p-toluenesulfonic acid, in a suitable reaction inert solvent such
as acetonitrile or toluene at a temperature between 55.degree. C.
to 100.degree. C., typically 80.degree. C., for a period between 1
to 24 hours. The R.sup.2' moiety in the aldehyde of formula
R.sup.2'CHO is one in which a protected hydroxy group typically
exists, such as a methoxy or benzyloxy, which can subsequently be
deprotected to the free hydroxy present in group R.sup.2 in the
compound of Formula I. Compounds of Formula III are then treated
with an oxidizing agent such as potassium permanganate in a polar
solvent such as acetone at a temperature between 20.degree. C. to
50.degree. C., typically ambient temperature, to provide compounds
of Formula II after a period of 1 to 24 hours. Compounds of Formula
II which contain a protected hydroxy group such as a suitable
methyl ether moiety or benzyloxy moiety, may be converted to the
free hydroxy (such as a phenolic hydroxy group) by standard
deprotection conditions.
##STR00005##
[0145] In the reaction schemes described herein it is to be
understood that hydroxy groups in intermediates useful for
preparing compounds of Formula I may be protected by other
conventional protecting groups known to those skilled in the art.
For example, intermediates containing a hydroxy group may be
protected as the corresponding benzyloxy ether and subsequently
deprotected by hydrogenation to provide the free hydroxy
derivative. Suitable protecting groups and methods for their
removal are illustrated in Protective Groups in Organic Synthesis,
3.sup.rd Ed., Theodora W. Greene, and Peter G. M. Wuts (John Wiley
& Sons, 1999). For example, in cases where the compound of
formula II contains a suitable methyl ether moiety, the compound of
Formula II is dissolved in a suitable solvent such as methylene
chloride at a temperature between -78.degree. C. to 10.degree. C.
followed by the addition of boron trichloride or boron tribromide.
The reaction mixture is stirred at 10.degree. C. to 50.degree. C.
for 1 to 24 hours to provide the desired deprotected compound of
Formula I. Similarly, compounds of formula II containing a suitable
benzyloxy moiety can be deprotected to provide compounds of formula
I by hydrogenation, such as H-cube hydrogenation (Pd/C cartridge)
carried out at ambient temperature using methanol as solvent.
##STR00006##
[0146] Reaction Scheme III depicts Formula IVa compounds prepared
by reacting the corresponding acid of Formula Va with a variety of
primary amines of general formula R.sup.1NH.sub.2 under standard
amide coupling conditions for a period between 4 to 24 hours.
Formula IIIa compounds may be prepared by reaction of Formula IVa
compounds with an appropriate aldehyde, R.sup.2'CHO, in the
presence of a catalytic amount of an acid, such as
p-toluenesulfonic acid, in a suitable reaction inert solvent such
as acetonitrile or toluene at a temperature between 55.degree. C.
to 100.degree. C., typically 80.degree. C., for a period between 1
to 24 hours. The R.sup.2' moiety in the aldehyde of formula
R.sup.2'CHO is one in which a protected hydroxy group exists, such
as a methoxy or benzyloxy, which can subsequently be deprotected to
the free hydroxy present in group R.sup.2 in the compound of
formula I. Compounds of Formula IIIa are then treated with an
oxidizing agent such as potassium permanganate in a polar solvent
such as acetone at a temperature between 20.degree. C. to
50.degree. C., typically ambient to provide compounds of Formula
IIa after a period of 1 to 24 hours.
##STR00007##
[0147] Reaction Scheme IV depicts the conversion of functional
groups at the R.sup.3 position of various intermediates useful in
preparing compounds of Formula I. Formula IIa compounds, wherein
R.sup.3 is chloro, are converted to Formula IIb compounds, wherein
R.sup.3 is iodo, by reaction with a reagent such as sodium iodide
and trimethylsilyl chloride in the dark in an inert solvent such as
acetonitrile or propionitrile at a temperature between 55.degree.
C. to 120.degree. C., typically 100.degree. C., for a period
between 1 to 24 hours. The compounds of Formula IIc are prepared by
reacting the compounds of Formula IIb with
methyl-2,2-difluoro-2-(fluoro-sulfonyl)acetate and copper iodide in
a mixture of solvent such as HMPA and DMF at a temperature between
55.degree. C. to 120.degree. C., typically 80.degree. C., for a
period between 1 to 24 hours. The R.sup.2' group in the compounds
of formulae IIa, IIb and IIc is one in which a protected hydroxy
group exists, such as a methoxy or benzyloxy, which can
subsequently be deprotected to the free hydroxy present in group
R.sup.2 of the corresponding compound within formula I.
##STR00008##
[0148] Reaction Scheme V depicts the conversion of a methyl ether
moiety in certain intermediates of formulae VI or VI' wherein
R.sup.2 is 2-methoxyphenyl, to a free phenolic hydroxy moiety in
compounds of Formula Ia or Ia'. Compounds of Formula VI (or VI' in
which R.sup.3 is CF.sub.3) which contain a methyl ether moiety may
be converted to the free phenol by dissolving the compounds of
Formula VI or VI' in a suitable solvent such as methylene chloride
at a temperature between -78.degree. C. to 10.degree. C. followed
by the addition of boron trichloride or boron tribromide. The
reaction mixture is stirred at 10.degree. C. to 50.degree. C. for 1
to 24 hours to provide the desired compounds, such as those of
Formula Ia or Ia'.
[0149] Reaction Scheme VI, below, depicts three reactions showing
the conversion of chloro derivative IIa or IIa' to alkoxy (Ib,
wherein R.sup.3 is OR.sup.6), amino (Ic, wherein R.sup.3 is
NR.sup.7R.sup.8) and carbon-linked derivatives (Id, wherein R.sup.3
results from an appropriate boronic acid derivative such as an
aryl, heteroaryl or (C.sub.1-C.sub.6)alkyl boronic acid),
respectively. Compounds of Formula IIa may be converted to
compounds of Formula Ib by treatment of the corresponding alcohol,
R.sup.6OH, in the presence of a suitable base such as sodium
hydride in an inert solvent such as DMF or THF at 20.degree. C. to
80.degree. C. for 6 to 24 hours, followed by deprotection as
necessary to provide compounds of Formula Ib.
[0150] Compounds of Formula IIa', in which the hydroxy moiety in
R.sup.2 is in deprotected form, may be reacted with an appropriate
amine of formula R.sup.7R.sup.8NH in a solvent such as methanol or
ethanol at 20.degree. C. to 80.degree. C. for 1 to 24 hours to
provide compounds of Formula Ic. Compounds of Formula IIa may also
be treated with an appropriate corresponding boronic acid
derivative of formula R.sup.3B(OH).sub.2 in the presence of a
catalyst such as palladium tetrakis(triphenylphosphine) in a
mixture of aqueous sodium carbonate and a polar solvent such as
dioxane in a microwave for 1 to 25 minutes at 50.degree. C. to
150.degree. C., followed by deprotection as necessary to provide
compounds of Formula Id.
##STR00009##
[0151] The term "patient in need of treatment thereof" means humans
and other animals who have or are at risk of having a disease or
disorder characterized by abnormal bone or mineral homeostasis. The
"patient in need of treatment thereof" may have or be at risk of
having a disease or disorder characterized by abnormal bone or
mineral homeostasis selected from the group consisting of
osteoporosis, osteopenia, periodontal disease, Paget's disease,
bone fracture, osteoarthritis, rheumatoid arthritis, and humoral
hypercalcemia of malignancy. As certain of the conditions being
treated have a higher incidence in females a preferred patient is a
female, and particularly a postmenopausal female human.
[0152] The term "treating", "treat" or "treatment" as used herein
includes preventative (e.g., prophylactic), palliative, adjuvant
and curative treatment. For example, the treatment of osteoporosis,
as used herein means that a patient having osteoporosis or at risk
of having osteoporosis can be treated according to the methods
described herein. For patients undergoing preventative treatment, a
resulting reduction in the incidence of the disease state being
preventively treated is the measurable outcome of the preventative
treatment.
[0153] The present invention provides methods of treating
osteopenia and osteoporosis by administering to a patient in need
thereof a therapeutically effective amount of a compound of formula
I. Osteopenia is a thinning of the bones, but less than is seen
with osteoporosis and is the stage before true osteoporosis. The
World Health Organization has developed diagnostic categories based
on bone mass density (BMD) to indicate if a person has normal
bones, has osteopenia or has osteoporosis. Normal bone density is
within one standard deviation (+1 or -1) of the young adult mean
bone density. Osteopenia (low bone mass) is defined as bone density
of 1 to 2.5 standard deviations below the young adult mean (-1 to
-2.5), and osteoporosis is defined as a bone density that is 2.5
standard deviations or more below the young adult mean
(>-2.5).
[0154] The present invention provides methods of treating bone
fractures by administering to a patient in need thereof a
therapeutically effective amount of a compound of formula I. Bone
fractures can be a fracture to any bone in the body, and hip
fracture being of particular concern. Hip fracture has a
significant impact on medical resources and patient morbidity and
mortality. Few patients admitted with a hip fracture are considered
for prophylactic measures aimed at the reduction of further
fracture risk. Currently, 10-13% of patients will later sustain a
second hip fracture. Of patients who suffered a second hip
fracture, fewer patients maintained their ability to walk
independently after the second fracture than did so after the first
(53 and 91% respectively, P<0.0005). Pearse E. O. et al.,
Injury, 2003, 34(7), 518-521. Following second hip fracture,
patients' level of mobility determined their future social
independence. Older patients and those with a history of multiple
falls had a shorter time interval between fractures. Second hip
fracture has a significant further impact on patients' mobility and
social independence. It is therefore desirable to have new methods
for the treatment of bone fractures including hip fracture.
[0155] The compounds of Formula I can be administered together with
additional agents which are useful for treating a disease or
disorder characterized by abnormal bone or mineral homeostasis.
Particularly contemplated additional agents include calcium
receptor antagonists other than those of Formula I, selective
estrogen receptor modulators (SERMs), bisphosphonates, parathyroid
hormone (PTH) and fragments and analogues thereof, estrogens,
calcitonins, synthetic steroids, synthetic isoflavones, vitamin D
analogues, vitamin K analogues, strontium salts, cathepsin K
inhibitors, .alpha..sub.v.beta..sub.3 integrin (vitronectin)
antagonists, prostaglandin (PGE2) receptor agonists and receptor
activator of nuclear factor .kappa.B ligand (RANKL) inhibitors.
[0156] Additional calcium receptor antagonists that can be used
together with compounds of Formula I in the methods and
compositions of this invention include those described in PCT
International Publication Nos. WO 93/04373; WO 94/18959; WO
95/11211; WO 97/37967; WO 98/44925; WO 98/45255; WO 99/51241; WO
99/51569; WO 00/45816; WO 02/14259; WO 02/38106; WO 2004/041755;
and WO 2005/030746; Nemeth, E. F.; Journal of Molecular
Endocrinology (2002) 29, 15-21; Kessler, A. et al.; ChemBioChem
(2004) 5, 1131; Steddon, S. J. et al.; Lancet (2005) 365,
2237-2239; and Shcherbakova, I.; et al.; Bioorganic & Medicinal
Chemistry Letters (2005) 15, 1557-1560. Specific calcilytic
compounds that can be used together with compounds of Formula I in
the methods and compositions of this invention include NPS-2143 and
423562.
[0157] SERMs that can be used together with compounds of Formula I
in the methods and compositions of this invention include, but are
not limited to, lasofoxifene (Oporia.RTM.), raloxifene
(Evista.RTM.), arzoxifene, bazedoxifene, ospemifene, Chiesi's
CHF-4227 and Prostrakan's PSK-3471. Bisphosphonates that can be
used together with compounds of Formula I in the methods and
compositions of this invention include, but are not limited to,
tiludronate (Skelid.RTM.), clondronate (Bonefos.RTM.), etidronate
(Didronel.RTM.), alendronate (Fosamax.RTM.), risedronate
(Actonel.RTM.), ibandronate (Boniva.RTM.), zoledronate
(Zometa.RTM.), minodronate (Onobis.RTM.), neridronate and
pamidronate.
[0158] In humans, PTH is an 84 amino acid polypeptide produced by
the parathyroid gland that controls serum calcium levels through
its action on various cells. Several N-terminal amino acids
fragments of PTH, including the 1-31, 1-34 and 1-38 fragments
(PTH-related proteins; "PTHrP") are considered biologically
equivalent to the full length hormone. Parathyroid hormone (PTH)
and fragments and analogues thereof that can be used together with
compounds of Formula I in the methods and compositions of this
invention include, but are not limited to, the full length PTH
(such as PTH 1-84, Preos.RTM./Preotact.RTM., Unigene's 768974, Bone
Medical's BN-003), the 1-31 (such as Zelos Therapeutics'
Ostabolin-C), 1-34 (such as teriparatide, Forteo.RTM., or Ipsen's
BIM-44058) or 1-38 fragments.
[0159] Estrogens that can be used together with compounds of
Formula I in the methods and compositions of this invention
include, but are not limited to, estradiol, conjugated equine
estrogens (Wyeth's Premarin.RTM.) or other estrogens.
[0160] Calcitonin is a 32 amino-acid peptide hormone produced by
the thyroid gland which inhibits osteoclast activity by binding to
calcitonin receptors on the surface of those cells. Calcitonins
that can be used together with compounds of Formula I in the
methods and compositions of this invention include, but are not
limited to, human calcitonin or salmon or eel calcitonins. The
calcitonins may be used as injectable or intranasal formulations
such as Miacalcin.RTM., Miacalcic.RTM., Calcitonia.RTM.,
Fortical.RTM. or Elcitonin.RTM. or as oral formulations such as
Novartis' SMC-021, Bone Medical's BN-002 (Capsitonin.RTM.) or
Nobex's NCT-025 (Oratonin.RTM.).
[0161] Synthetic steroids that can be used together with compounds
of Formula I in the methods and compositions of this invention
include, but are not limited to, mixed estrogen and progesterone
agonists such as tibolone which is marketed as Livial.RTM..
Synthetic isoflavones are chemically synthesized derivatives of
plant isoflavones, such as phytoestrogens extracted from soy
products. A synthetic isoflavone that can be used together with
compounds of Formula I in the methods and compositions of this
invention includes, but is not limited to, ipraflavone which is
marketed by Takeda as Iprosten.RTM. and Osten.RTM..
[0162] Vitamin D analogues are compounds that act by binding to the
nuclear vitamin D receptor in osteoblasts. Vitamin D analogues that
can be used together with compounds of Formula I in the methods and
compositions of this invention include, but are not limited to,
Chugai's ED-71 and Deltanoid's 2MD.
[0163] A strontium salt that can be used together with compounds of
Formula I in the methods and compositions of this invention
includes, but is not limited to, strontium ranelate (Servier's
Protelos.RTM.). Cathepsin K inhibitors that can be used together
with compounds of Formula I in the methods and compositions of this
invention include, but are not limited to, Novartis's AAE-581,
balicatib, GlaxoSmithKline's SB-462795 and Merck's c-3578. An
.alpha..sub.v.beta..sub.3 integrin (vitronectin) antagonist that
can be used together with compounds of Formula I in the methods and
compositions of this invention includes, but is not limited to,
Merck's MRL-123.
[0164] Prostaglandin E2 (PGE2) receptor agonists that can be used
together with compounds of Formula I in the methods and
compositions of this invention include, but are not limited to,
PGE2 subtype 2 (EP2) receptor agonists, such as
(3-{[4-Tert-butyl-benzyl)-(pyridine-3-sulfonyl)-amino]-methyl}-phenoxy)-a-
cetic acid, or a pharmaceutically acceptable salt thereof or PGE2
subtype 4 (EP4) receptor agonists, such as ONO-4819. A receptor
activator of nuclear factor .kappa.B ligand (RANKL) inhibitor that
can be used together with compounds of Formula I in the methods and
compositions of this invention includes, but is not limited to,
Amgen's RANKL antibody AMG-162.
[0165] Specific combinations of particular interest include
compounds of Formula I and lasofoxifene or compounds of Formula I
and
(3-{[4-Tert-butyl-benzyl)-(pyridine-3-sulfonyl)-amino]-methyl}-phenoxy)-a-
cetic acid, or a pharmaceutically acceptable salt thereof.
[0166] Compounds of the invention intended for pharmaceutical use
may be administered as crystalline or amorphous products. The
compounds may be obtained, for example, as solid plugs, powders, or
films by methods such as precipitation, crystallization, freeze
drying, spray drying, or evaporative drying. Microwave or radio
frequency drying may be used for this purpose.
[0167] For the above-mentioned therapeutic uses, the dosage
administered will, of course, vary with the compound employed, the
mode of administration, the treatment desired and the disorder
indicated. The total daily dosage of the compound of formula
I/salt/solvate (active ingredient) will, generally, be in the range
from 1 mg to 1 gram, preferably 1 mg to 250 mg, more preferably 10
mg to 100 mg. The total daily dose may be administered in single or
divided doses. The present invention also encompasses sustained
release compositions.
[0168] The pharmaceutical composition may, for example, be in a
form suitable for oral administration as a tablet, capsule, pill,
powder, sustained release formulations, solution, suspension, for
parenteral injection as a sterile solution, suspension or emulsion,
for topical administration as an ointment or cream or for rectal
administration as a suppository. The pharmaceutical composition may
be in unit dosage forms suitable for single administration of
precise dosages. The pharmaceutical composition will include a
conventional pharmaceutical carrier or excipient and a compound
according to the invention as an active ingredient. In addition, it
may include other medicinal or pharmaceutical agents, carriers,
adjuvants, etc. The dissolution rate of poorly water-soluble
compounds may be enhanced by the use of a spray-dried dispersion,
such as those described by Takeuchi, H., et al. in "Enhancement of
the dissolution rate of a poorly water-soluble drug (tolbutamide)
by a spray-drying solvent depostion method and disintegrants" J.
Pharm. Pharmacol., 39, 769-773 (1987).
[0169] Exemplary parenteral administration forms include solutions
or suspensions of active compounds in sterile aqueous solutions,
for example, aqueous propylene glycol or dextrose solutions. Such
dosage forms can be suitably buffered, if desired.
[0170] Suitable pharmaceutical carriers include inert diluents or
fillers, water and various organic solvents. The pharmaceutical
compositions may, if desired, contain additional ingredients such
as flavorings, binders, excipients and the like. Thus for oral
administration, tablets containing various excipients, such as
citric acid may be employed together with various disintegrants
such as starch, alginic acid and certain complex silicates and with
binding agents such as sucrose, gelatin and acacia. Additionally,
lubricating agents such as magnesium stearate, sodium lauryl
sulfate and talc are often useful for tableting purposes. Solid
compositions of a similar type may also be employed in soft and
hard filled gelatin capsules. Preferred materials, therefor,
include lactose or milk sugar and high molecular weight
polyethylene glycols. When aqueous suspensions or elixirs are
desired for oral administration the active compound therein may be
combined with various sweetening or flavoring agents, coloring
matters or dyes and, if desired, emulsifying agents or suspending
agents, together with diluents such as water, ethanol, propylene
glycol, glycerin, or combinations thereof. Methods of preparing
various pharmaceutical compositions with a specific amount of
active compound are known, or will be apparent, to those skilled in
this art. For examples, see Remington's Pharmaceutical Sciences,
Mack Publishing Company, Easter, Pa., 15th Edition (1975).
[0171] Dosage regimens may be adjusted to provide the optimum
desired response. For example, a single bolus may be administered,
several divided doses may be administered over time or the dose may
be proportionally reduced or increased as indicated by the
exigencies of the therapeutic situation. It is especially
advantageous to formulate 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 mammalian subjects 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 (a) the unique characteristics of the chemotherapeutic
agent and the particular therapeutic or prophylactic effect to be
achieved, and (b) the limitations inherent in the art of
compounding such an active compound for the treatment of
sensitivity in individuals. Thus, the skilled artisan would
appreciate, based upon the disclosure provided herein, that the
dose and dosing regimen is adjusted in accordance with methods
well-known in the therapeutic arts. That is, the maximum tolerable
dose can be readily established, and the effective amount providing
a detectable therapeutic benefit to a patient may also be
determined, as can the temporal requirements for administering each
agent to provide a detectable therapeutic benefit to the patient.
Accordingly, while certain dose and administration regimens are
exemplified herein, these examples in no way limit the dose and
administration regimen that may be provided to a patient in
practicing the present invention.
[0172] It is to be noted that dosage values may vary with the type
and severity of the condition to be alleviated, and may include
single or multiple doses. It is to be further understood that for
any particular subject, specific dosage regimens should be adjusted
over time according to the individual need and the professional
judgment of the person administering or supervising the
administration of the compositions, and that dosage ranges set
forth herein are exemplary only and are not intended to limit the
scope or practice of the claimed composition. For example, doses
may be adjusted based on pharmacokinetic or pharmacodynamic
parameters, which may include clinical effects such as toxic
effects and/or laboratory values. Thus, the present invention
encompasses intra-patient dose-escalation as determined by the
skilled artisan. Determining appropriate dosages and regiments for
administration of the chemotherapeutic agent are well-known in the
relevant art and would be understood to be encompassed by the
skilled artisan once provided the teachings disclosed herein.
[0173] A pharmaceutical composition of the invention may be
prepared, packaged, or sold in bulk, as a single unit dose, or as a
plurality of single unit doses. As used herein, a "unit dose" is
discrete amount of the pharmaceutical composition comprising a
predetermined amount of the active ingredient. The amount of the
active ingredient is generally equal to the dosage of the active
ingredient which would be administered to a subject or a convenient
fraction of such a dosage such as, for example, one-half or
one-third of such a dosage.
[0174] The relative amounts of the active ingredient, the
pharmaceutically acceptable carrier, and any additional ingredients
in a pharmaceutical composition of the invention will vary,
depending upon the identity, size, and condition of the subject
treated and further depending upon the route by which the
composition is to be administered. By way of example, the
composition may comprise between 0.1% and 100% (w/w) active
ingredient.
[0175] In addition to the active ingredient, a pharmaceutical
composition of the invention may further comprise one or more
additional pharmaceutically active agents. Particularly
contemplated additional agents include selective estrogen receptor
modulators (SERMs), bisphosphonates, parathyroid hormone (PTH) and
fragments and analogues thereof, estrogens, calcitonins, synthetic
steroids, synthetic isoflavones, vitamin D analogues, vitamin K
analogues, strontium salts, cathepsin K inhibitors,
.alpha..sub.v.beta..sub.3 integrin (vitronectin) antagonists,
prostaglandin (PGE2) receptor agonists and receptor activator of
nuclear factor .kappa.B ligand (RANKL) inhibitors, such as those
described hereinabove.
[0176] Controlled- or sustained-release formulations of a
pharmaceutical composition of the invention may be made using
conventional technology.
[0177] As used herein, "parenteral administration" of a
pharmaceutical composition includes any route of administration
characterized by physical breaching of a tissue of a subject and
administration of the pharmaceutical composition through the breach
in the tissue. Parenteral administration thus includes, but is not
limited to, administration of a pharmaceutical composition by
injection of the composition, by application of the composition
through a surgical incision, by application of the composition
through a tissue-penetrating non-surgical wound, and the like. In
particular, parenteral administration is contemplated to include,
but is not limited to, subcutaneous, intraperitoneal,
intramuscular, intrasternal injection, and kidney dialytic infusion
techniques.
[0178] Formulations of a pharmaceutical composition suitable for
parenteral administration comprise the active ingredient combined
with a pharmaceutically acceptable carrier, such as sterile water
or sterile isotonic saline. Such formulations may be prepared,
packaged, or sold in a form suitable for bolus administration or
for continuous administration. Injectable formulations may be
prepared, packaged, or sold in unit dosage form, such as in ampules
or in multi-dose containers containing a preservative. Formulations
for parenteral administration include, but are not limited to,
suspensions, solutions, emulsions in oily or aqueous vehicles,
pastes, and implantable sustained-release or biodegradable
formulations as discussed below. Such formulations may further
comprise one or more additional ingredients including, but not
limited to, suspending, stabilizing, or dispersing agents. In one
embodiment of a formulation for parenteral administration, the
active ingredient is provided in dry (i.e. powder or granular) form
for reconstitution with a suitable vehicle (e.g. sterile
pyrogen-free water) prior to parenteral administration of the
reconstituted composition.
[0179] A composition of the present invention can be administered
by a variety of methods known in the art. The route and/or mode of
administration vary depending upon the desired results. The active
compounds can be prepared with carriers that protect the compound
against rapid release, such as a controlled release formulation,
including implants, transdermal patches, and microencapsulated
delivery systems. Biodegradable, biocompatible polymers can be
used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic
acid, collagen, polyorthoesters, and polylactic acid. Many methods
for the preparation of such formulations are described by e.g.,
Sustained and Controlled Release Drug Delivery Systems, J. R.
Robinson, ed., Marcel Dekker, Inc., New York, (1978).
Pharmaceutical compositions are preferably manufactured under GMP
conditions.
[0180] The pharmaceutical compositions may be prepared, packaged,
or sold in the form of a sterile injectable aqueous or oily
suspension or solution. This suspension or solution may be
formulated according to the known art, and may comprise, in
addition to the active ingredient, additional ingredients such as
the dispersing agents, wetting agents, or suspending agents
described herein. Such sterile injectable formulations may be
prepared using a non-toxic parenterally-acceptable diluent or
solvent, such as water or 1,3-butane diol, for example. Other
acceptable diluents and solvents include, but are not limited to,
Ringer's solution, isotonic sodium chloride solution, and fixed
oils such as synthetic mono- or di-glycerides. Other
parentally-administrable formulations which are useful include
those which comprise the active ingredient in microcrystalline
form, in a liposomal preparation, or as a component of a
biodegradable polymer systems. Compositions for sustained release
or implantation may comprise pharmaceutically acceptable polymeric
or hydrophobic materials such as an emulsion, an ion exchange
resin, a sparingly soluble polymer, or a sparingly soluble
salt.
[0181] While the precise dosage administered of each active
ingredient will vary depending upon any number of factors,
including but not limited to, the type of animal and type of
disease state being treated, the age of the animal and the route(s)
of administration. The amounts of various CaR antagonist compounds
of formula I to be administered can be determined by standard
procedures taking into account factors such as the compound
IC.sub.50, EC.sub.50, the biological half-life of the compound, the
age, size and weight of the patient, and the type of condition or
symptom associated with the patient. The importance of these and
other factors to be considered are known to those of ordinary skill
in the art.
[0182] PTH secretion can be measured using techniques known in the
art (see, e.g., U.S. Pat. No. 6,031,003, hereby incorporated by
reference). For example, PTH secretion can be measured by first
suspending cells in parathyroid cell buffer containing 0.5 mM
CaCl.sub.2 and 0.1% bovine serum albumin. Incubations can be
performed in plastic tubes (Falcon 2058) containing 0.3 mL of the
cell suspension with or without small volumes of CaCl.sub.2 and/or
organic polycations. After incubation at 37.degree. C., typically
30 minutes, the tubes can then be placed on ice and the cells
pelleted at 2.degree. C. Samples of the supernatant should then be
brought to pH 4.5 with acetic acid and, if needed, stored at
-70.degree. C. The amount of PTH in bovine cell supernatants can be
determined by a homologous radioimmunoassay using GW-1 antibody or
its equivalent at a final dilution of 1/45,000. 1251-PTH (65-84;
INCSTAR, Stillwater, Minn.) can be used as tracer and fractions
separated by dextran-activated charcoal. Counting of samples and
data reduction can be performed on a Packard Cobra 5005 gamma
counter. For testing PTH levels in human cell supernatants, a
commercially available radioimmunoassay kit (INS-PTH; Nichols
Institute, Los Angeles, Calif.) which recognizes intact and
N-terminal human PTH is preferable because GW-1 antibody recognizes
human PTH poorly.
[0183] In addition, specific assays useful for evaluating the
compounds of Formula I include the FLIPR Assay for Evaluating the
Potency and Selectivity of Test Compounds; Assay for Evaluating the
Effects of Test Compounds on Endogenous PTH Secretion; Evaluation
of Effects of Test Compounds on PTH Secretion In Vivo; Effect of
Calcium Receptor Antagonist Compound of Formula I on Body Weight,
Body Composition and Bone Density in the Aged Intact and
Ovariectomized Female Rat; and Fracture Healing Assays as described
below.
FLIPR Assay for Evaluating the Potency and Selectivity of Test
Compounds
[0184] Human kidney cell (HEK 293) expressing the calcium receptor
(CasR) are used to detect antagonists of the receptor using
Fluorometric imaging plate reader (FLIPR, Molecular Devices,
Sunnyvale Calif.). Receptor activation by extracellular calcium
results in the release of calcium from intracellular stores into
the cytosol. A fluorescent indicator (Fluo-4) is internalized by
the cells from growth media and interacts with calcium released
into the cytosol to provide a means of quantifying intracellular
Ca.sup.2+ levels and receptor agonism/antagonism. Fluorescence
intensity is detected by the FLIPR CCD camera and traced as a
function of time. Potential antagonists are identified by their
ability to decrease this fluorescent response.
[0185] To determine the IC.sub.50 values cells are loaded with
Fluo4 (2.05 mM Fluo-4, 0.04% pluronic acid, 2.6 mM probenecid in
90% DMEM high glucose, 10% dialyzed Fetal Bovine Serum, 1.times.
Pen Strep, 1.times. L-Glutamine, 3 ug/ml Puromycin, 27.5 nM
Methotrexate) for 1 hour at 37.degree. C. Prior to the addition of
test compound cells are washed with a 10 mM HEPES buffer solution.
The test compound, for example the compound of Example 1, is added
at various doses (from 1 .quadrature.M to 3 nM) and pre-incubated
with cells for 30 minutes followed by stimulation of the CasR by
the addition of 1.7 mM Ca.sup.2+. IC.sub.50 values are based on the
ability of the cells to inhibit the Ca.sup.2+ induced increase in
intracellular Ca.sup.2+. Fluorescence signal is read 42 seconds
after the stimulation of the CasR by the addition of 1.7 mM
Ca.sup.2+.
Assay for Evaluating the Effects of Test Compounds on Endogenous
PTH Secretion
[0186] Adult male or female Sprague-Dawley rats (Charles River
Laboratories, Wilmington, Mass.) with jugular vein catheter are
used in this assay. The test compounds at various doses are given
to the animals by various routes of administration including
subcutaneous injection, or intraveneous injection. Serum or plasma
PTH concentrations are examined before and after dosing at various
times using a commercially available rat intact PTH ELISA kit
(Immutopics, Inc. San Clemente, Calif. Cat. #60-2500).
Evaluation of Effects of Test Compounds on PTH Secretion In
Vivo
[0187] Overnight fasted male Sprague-Dawley rats (250 g) with
jugular vein catheter are used in this study. Whole blood sample is
collected from each animal prior to compound treatment for
measuring baseline PTH concentrations. The test animals are then
given a single dose of the tested compound at 1 mg/kg in glycerol
formal: 2% DMSO by intravenous administration via jugular vein.
Whole blood samples are collected at 2, 5, 15, 30 and 45 minutes,
and 1, 2, 3, 4, 6 and 8 hours after dosing. Plasma samples are
obtained by centrifugation and PTH concentrations are determined
using a commercially available rat intact PTH ELISA kit
(Immutopics, Inc. San Clemente, Calif. Cat. #60-2500). A
significant burst of PTH was seen following the treatment with the
tested compound. The elevated PTH secretion induced by the tested
compound was peak at 2 minutes and returned to baseline level at 30
minutes after dosing (FIG. 1).
Effect of Calcium Receptor Antagonist Compound of Formula I on Bad
Weight, Body Composition and Bone Density in the Aged Intact and
Ovariectomized Female Rat
[0188] The purpose of this study is to test the effects of test
compositions comprising compounds of Formula I in aged intact or
ovariectomized (OVX) female rat model. In the following protocol
the compound of Formula I can be administered as a pharmaceutically
acceptable salt or prodrug thereof.
Study Protocol
[0189] Sprague-Dawley female rats are sham-operated or OVX at 18
months of age, while a group of rats is necropsied at day 0 to
serve as baseline controls. One day post-surgery, the rats are
treated with either vehicle or test compound of Formula I, or a
combination of test compound of Formula I and other active agent
test compound for 59 days. The vehicle or test compound of Formula
I is administered either orally, by oral gavage, or by subcutaneous
injection (s.c.), with the test compound being administered at a
therapeutically effective dose.
[0190] All rats are given s.c. injection of 10 mg/kg of calcein
(Sigma, St. Louis, Mo.) for fluorescent bone label 2 and 12 days
before necropsy. On the day of necropsy, all rats under
ketamine/xylazine anesthesia are weighed and undergoe dual-energy
X-ray absorptiometry (DXA, QDR-4500/W, Hologic Inc., Waltham,
Mass.) equipped with Rat Whole Body Scan software for lean and fat
body mass determination. The rats are necropsied, then autopsied
and blood is obtained by cardiac puncture. The distal femoral
metaphysis and femoral shafts from each rat are analyzed by
peripheral quantitative computerized tomography (pQCT), and
volumetric total, trabecular and cortical bone mineral content and
density are determined.
[0191] Peripheral Quantitative Computerized Tomography (pQCT)
Analysis: Excised femurs are scanned by a pQCT X-ray machine
(Stratec XCT Research M, Norland Medical Systems, Fort Atkinson,
Wis.) with software version 5.40. A 1 millimeter (mm) thick cross
section of the femur metaphysis is taken at 5.0 mm (proximal
femoral metaphysis, a primary cancellous bone site) and 13 mm
(femoral shafts, a cortical bone site) proximal from the distal end
with a voxel size of 0.10 mm. Cortical bone is defined and analyzed
using contour mode 2 and cortical mode 4. An outer threshold
setting of 340 mg/cm.sup.3 is used to distinguish the cortical
shell from soft tissue and an inner threshold of 529 mg/cm.sup.3 to
distinguish cortical bone along the endocortical surface.
Trabecular bone is determined using peel mode 4 with a threshold of
655 mg/cm.sup.3 to distinguish (sub)cortical from cancellous bone.
An additional concentric peel of 1% of the defined cancellous bone
is used to ensure that (sub)cortical bone was eliminated from the
analysis. Volumetric content, density, and area are determined for
both trabecular and cortical bone (Jamsa T. et al., Bone
23:155-161, 1998; Ke, H. Z. et al., Journal of Bone and Mineral
Research, 16:765-773, 2001).
[0192] The experimental groups for the protocol are as follows:
Group I: Baseline controls
Group II: Sham+Vehicle
Group III: OVX+Vehicle
Group IV: OVX+Test Compound of Formula I (in Vehicle)
Group V: OVX+Test Compound of Formula I and Additional Active
Agent
[0193] Note: Group V only employed when it is desired to test a
combination of a compound of Formula I and an additional active
agent.
Fracture Healing Assays
[0194] Assay for Effects on Fracture Healing after Systemic
Administration
[0195] Fracture Technique: Sprague-Dawley rats at 3 months of age
are anesthetized with Ketamine. A 1 cm incision is made on the
anteromedial aspect of the proximal part of the right tibia or
femur. The following describes the tibial surgical technique. The
incision is carried through to the bone, and a 1 mm hole is drilled
4 mm proximal to the distal aspect of the tibial tuberosity 2 mm
medial to the anterior ridge. Intramedullary nailing is performed
with a 0.8 mm stainless steel tube (maximum load 36.3 N, maximum
stiffness 61.8 N/mm, tested under the same conditions as the
bones). No reaming of the medullary canal is performed. A
standardized closed fracture is produced 2 mm above the
tibiofibular junction by three-point bending using specially
designed adjustable forceps with blunt jaws. To minimize soft
tissue damage, care is taken not to displace the fracture. The skin
is closed with monofilament nylon sutures. The operation is
performed under sterile conditions. Radiographs of all fractures
are taken immediately after nailing, and rats with fractures
outside the specified diaphyseal area or with displaced nails are
excluded. The remaining animals are divided randomly into the
following groups with 10-12 animals per each subgroup per time
point for testing the fracture healing. The first group receives
daily gavage of vehicle (water: 100% Ethanol=95:5) at 1 ml/rat,
while the others receive daily gavage from 0.01 to 100 mg/kg/day of
the compound of Formula I to be tested (1 ml/rat) for 10, 20, 40
and 80 days.
[0196] At 10, 20, 40 and 80 days, 10-12 rats from each group are
anesthetized with Ketamine and sacrificed by exsanguination. Both
tibiofibular bones are removed by dissection and all soft tissue is
stripped. Bones from 5-6 rats for each group are stored in 70%
ethanol for histological analysis, and bones from another 5-6 rats
for each group are stored in a buffered Ringer's solution
(+4.degree. C., pH 7.4) for radiographs and biomechanical testing
which is performed.
[0197] Histological Analysis: The methods for histologic analysis
of fractured bone have been previously published by Mosekilde and
Bak (The Effects of Growth Hormone on Fracture Healing in Rats: A
Histological Description. Bone, 14:19-27, 1993). Briefly, the
fracture site is sawed 8 mm to each side of the fracture line,
embedded undecalcified in methymethacrylate, and cut frontals
sections on a Reichert-Jung Polycut microtome in 8 .mu.m thick.
Masson-Trichrome stained mid-frontal sections (including both tibia
and fibula) are used for visualization of the cellular and tissue
response to fracture healing with and without treatment. Sirius red
stained sections are used to demonstrate the characteristics of the
callus structure and to differentiate between woven bone and
lamellar bone at the fracture site. The following measurements are
performed: (1) fracture gap--measured as the shortest distance
between the cortical bone ends in the fracture, (2) callus length
and callus diameter, (3) total bone volume area of callus, (4) bony
tissue per tissue area inside the callus area, (5) fibrous tissue
in the callus, and (6) cartilage area in the callus.
[0198] Biomechanical Analysis: The methods for biomechanical
analysis have been previously published by Bak and Andreassen (The
Effects of Aging on Fracture Healing in Rats. Calcif Tissue Int
45:292-297, 1989). Briefly, radiographs of all fractures are taken
prior to the biomechanical test. The mechanical properties of the
healing fractures are analyzed by a destructive three- or
four-point bending procedure. Maximum load, stiffness, energy at
maximum load, deflection at maximum load, and maximum stress are
determined.
[0199] A calcium receptor antagonist may be usefully combined with
another pharmacologically active compound, or with two or more
other pharmacologically active compounds, particularly in the
treatment of osteoporosis. For example, a calcium receptor
antagonist, particularly a compound of the formula I, or a
pharmaceutically acceptable salt or solvate thereof, as defined
above, may be administered simultaneously, sequentially or
separately in combination with one or more agents selected from:
selective estrogen receptor modulators (SERMs), bisphosphonates,
parathyroid hormone (PTH) and fragments and analogues thereof,
estrogens, calcitonins, synthetic steroids, synthetic isoflavones,
vitamin D analogues, vitamin K analogues, strontium salts,
cathepsin K inhibitors, .alpha..sub.v.beta..sub.3 integrin
(vitronectin) antagonists, prostaglandin (PGE2) receptor agonists
and receptor activator of nuclear factor .kappa.B ligand (RANKL)
inhibitors as described hereinabove.
[0200] The following non-limiting Preparations and Examples
illustrate the preparation of compounds of the present
invention.
.sup.1H Nuclear magnetic resonance (NMR) spectra were in all cases
consistent with the proposed structures. Characteristic chemical
shifts (.delta.) are given in parts-per-million downfield from
tetramethylsilane using conventional abbreviations for designation
of major peaks: e.g. s, singlet; d, doublet; t, triplet; q,
quartet; m, multiplet; br, broad. The mass spectra (m/z) were
recorded using either electrospray ionisation (ESI) or atmospheric
pressure chemical ionisation (APCI). The following abbreviations
have been used for common solvents and various reagents:
CDCl.sub.3, deuterochloroform; DIEA, diisopropylethylamine; DMF,
dimethylformamide; CD.sub.3OD, deuteromethanol; D.sub.6-DMSO,
deuterodimethylsulfoxide; HBTU,
O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate; HCl, hydrochloric acid; HI, hydroiodic acid;
HMPA, hexamethylphosphoramide; KMnO.sub.4, potassium permanganate;
MgSO.sub.4, magnesium sulfate; NaHCO.sub.3, sodium bicarbonate;
NaHSO.sub.3, sodium bisulfite; NaOH, sodium hydroxide;
Na.sub.2SO.sub.4, sodium sulfate; NH.sub.4Cl, ammonium chloride;
TEA, triethylamine; THF, tetrahydrofuran. `Ammonia` refers to a
concentrated solution of ammonia in water possessing a specific
gravity of 0.88. Where thin layer chromatography (TLC) has been
used it refers to silica gel TLC using silica gel 60 F.sub.254
plates, R.sub.f is the distance traveled by a compound divided by
the distance traveled by the solvent front on a TLC plate. HPLC
refers to high performance liquid chromatography.
[0201] The following specific examples are included for
illustrative purposes and are not to be construed as a limitation
to this disclosure.
Preparation of Intermediates
Preparation of 2,4-Dichloronicotinic acid
##STR00010##
[0203] To a stirring solution of diisopropyl ethyl amine (11.1 ml,
81.08 mmol) in THF (50 ml) was added dropwise a solution of BuLi
(1.46 M, 43.3 ml, 73.65 mmol) in hexane below -65.degree. C. and
the mixture was stirred for 40 minutes. To this solution was added
dropwise 2,4-dichloropyridine (10 g, 67.57 mmol) in THF (15 mL) at
-78.degree. C. and stirred for 30 minutes. Carbon dioxide generated
from freshly crushed dry ice was passed through CaCl.sub.2 guard
tube and then charged into the reaction mixture for 10 minutes and
the reaction mixture was slowly allowed to come to room
temperature. The solvent was evaporated under reduced pressure and
dissolved in a minimum volume of water. The aqueous layer was
washed with water and acidified to pH 4 with conc. HCl. It was then
extracted with ethyl acetate, the organic layer was washed with
brine and dried over sodium sulfate. The organic solvent was
removed under reduced pressure to provide 2,4-dichloronicotinic
acid (10.6 g, 82%) as off white solid.
[0204] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 14.88-14.54 (br
s, 1H), 8.46 (d, J=5.3 Hz, 1H), 7.74 (d, J=5.5 Hz, 1H). FIA MS
[M+H]: 191.8
Preparation of Methyl 2,4-dichloronicotinate
##STR00011##
[0205] Nitrosomethyl urea (8 g, 78.16 mmol), taken in diethyl ether
(30 ml) was cooled to 0.degree. C. and 25% aqueous KOH solution was
added slowly under cooling. The ether layer was collected, dried
over KOH and added dropwise to a stirring solution of
2,4-Dichloronicotinic acid (3 g, 15.62 mmol) in methanol (5 mL) at
0.degree. C. The reaction mixture was allowed to come to room
temperature within 1 hour. The organic solvent was removed under
reduced pressure and the crude residue was purified by column
chromatography (5-10% EtOAc in hexane) to obtain the pure methyl
2,4-dichloronicotinate (3.00 g, 99%) as colorless oil. .sup.1H NMR
(400 MHz, CDCl.sub.3): .delta. 8.34 (d, J=5.3 Hz, 1H), 7.33 (d,
J=5.4 Hz, 1H), 3.99 (s, 3H).
Preparation of Methyl 4-azido-2-chloronicotinate
##STR00012##
[0207] Methyl 2,4-dichloronicotinate (20 g, 96.67 mmol) afforded
methyl 4-azido-2-chloronicotinate (15 g, 73%) as cream colored
solid following the literature procedure, J. Prakt. Chem, 2000;
342, 33-39 (reaction of Methyl 2,4-dichloronicotinate with
NaN.sub.3 in DMF at 50.degree. C. followed by aqueous workup).
.sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.37 (d, J=5.52 Hz,
1H), 7.06 (d, J=5.6 Hz, 1H), 3.96 (s, 3H). FIA MS [M+H]: 213.1
(small peak). IR (KBr): 2126, 1741 cm.sup.-1
Preparation of Methyl 4-amino-2-chloronicotinate
##STR00013##
[0209] Methyl 4-azido-2-chloronicotinate (15 g, 70.75 mmol)
produced methyl 4-amino-2-chloronicotinate (11.2 g, 85%) as light
yellow solid following the literature procedure, Tetrahedron
letters, 2002; 43, 6629-6631 (reduction of the azido group in
methyl 4-azido-2-chloronicotinate to the corresponding amine by
stirring methyl 4-azido-2-chloronicotinate in aqueous HI at room
temperature). .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.81 (d,
J=5.79 Hz, 1H), 6.76 (br s, 2H), 6.62 (d, J=5.81 Hz, 1H), 3.82 (s,
3H). FIA MS [M+H]: 186.9
Preparation of 4-amino-2-chloronicotinic acid
##STR00014##
[0211] Methyl 4-amino-2-chloronicotinate (21.0 g, 112.9 mmol) and
LiOH (10.3 g, 247.5 mmol) was taken in a mixture of dioxane:
MeOH:water (3:2:1, ml) and the reaction mixture was heated to
85.degree. C. for 2 hours. The solvent was evaporated under reduced
pressure, the residue was dissolved in minimum volume of water and
acidified (up to pH 4) with saturated citric acid. The aqueous
solution was concentrated until precipitation just started. The
mixture was allowed to stand overnight at 10.degree. C. for
complete precipitation. The resultant white solid was collected by
filtration and recrystallized from isopropyl alcohol-hexane to
afford 4-amino-2-chloronicotinic acid (13 g, 66% yield). .sup.1H
NMR (400 MHz, DMSO-d.sub.6): .delta. 7.59 (d, 1H), 6.47 (d, 1H),
6.37 (brs, 2H).
Preparation of 4-Chloro-2-(trifluoromethyl)nicotinic acid
##STR00015##
[0213] To a stirred solution of 2,2,6,6-tetramethylpiperidine (21.0
mL, 124 mmol) in dry THF (200 mL) in a -78.degree. C. bath is added
n-Butyl lithium (66.2 mL of a 2.5M solution in hexanes, 166 mmol).
The solution is stirred at -78.degree. C. for 30 minutes, followed
by the addition of a solution of 2-(trifluoromethyl)nicotinic acid
(7.90 g, 41.4 mmol) in THF (35 mL) via canula. The solution is
stirred at -78.degree. C. for 20 minutes, followed by warming to
-50.degree. C. for 1 hour (to provide the corresponding lithiated
pyridine). In a separate flask, hexachloroethane (29.4 g, 124 mmol)
is dissolved in THF (200 mL) and cooled to -15.degree. C. and
stirred rapidly. The solution containing the lithiated pyridine is
added to the hexachloroethane solution via canula. After complete
addition, the mixture is allowed to ward to room temperature. Water
(ca 200 mL) is added, followed by removal of the tetrathydrofuran
in vacuo. The remaining aqueous residue is extracted with
ether-pentane (1:1, 100 mL). The organic layer was discarded. The
aqueous layer is acidified with hydrochloric acid (ca. 50 mL of a
1.0 M solution). The aqueous layer is extracted with ethyl acetate.
The organic layer is dried over anhydrous magnesium sulfate,
filtered and concentrated in vacuo to afford 8.75 g (94%) of the
title compound as a tan solid: .sup.1H NMR (400 MHz, CD.sub.3OD),
8.63 (d, 1H), 7.79 (d, 1H). LC/MS (M+1)=226.1; [see: Schlosser et
al., Eur. J. Org. Chem. 2003, 1559].
Preparation of Methyl 4-chloro-2-(trifluoromethyl)nicotinate
##STR00016##
[0215] To a stirred solution of
4-chloro-2-(trifluoromethyl)nicotinic acid (8.75 g, 38.8 mmol) in
ethyl acetate (100 mL) in an ice bath is added a solution of
diazomethane [generated from 12 g (116 mmol) of
N-nitroso-N-methylurea] in diethyl ether (200 mL). The solution is
stirred for 15 minutes, followed by the addition of acetic acid
(until effervescence stops). The solution is concentrated in vacuo
to afford 9.75 g (99%) of the title compound as a brown oil:
.sup.1H NMR (400 MHz, CD.sub.3OD), 8.686 (d, 1H), 7.828 (d, 1H),
3.937 (s, 3H). LC/MS (M+1)=240.1
Preparation of Methyl 4-azido-2-(trifluoromethyl)nicotinate
##STR00017##
[0216] To a stirred solution of methyl
4-chloro-2-(trifluoromethyl)nicotinate (9.75 g, 40.7 mmol) in DMF
(300 mL) is added sodium azide (21.2 g, 326 mmol). The solution is
warmed to 45.degree. C. for 3 hours and then to 50.degree. C. for 1
hour. The reaction is diluted with water (300 mL) and extracted
with ether-pentane (1:1, ca. 1 L). The organic layer is back
extracted with water (3.times.). The organic layer is dried over
anhydrous magnesium sulfate, filtered and concentrated in vacuo to
afford 9.50 g (95%) of the title compound as a brown oil: .sup.1H
NMR (400 MHz, CD.sub.3OD), 8.651 (d, 1H), 7.603 (d, 1H), 3.878 (s,
3H).
Preparation of Methyl 4-amino-2-(trifluoromethyl)nicotinate and
4-amino-2-(trifluoromethyl)nicotinic acid
##STR00018##
[0218] Cold (0.degree. C. ice bath) concentrated hydriodic acid
(182 mL of a 57% aqueous solution) is added to methyl
4-azido-2-(trifluoromethyl)nicotinate (7.50 g, 30.5 mmol) in a
flask. The mixture is stirred at 0.degree. C. until TLC (ethyl
acetate-hexanes, 75:25) shows complete consumption of starting
material. The mixture is poured over an ice cold solution of
potassium hydroxide (95 g of solid KOH dissolved in 300 mL of
water). The mixture is extracted with diethyl ether. The ethereal
layer is washed with water, dried over anhydrous magnesium sulfate,
filtered and concentrated in vacuo to afford a yellow solid. The
solid is dissolved in a minimal amount of dichloromethane and
passed through a silica gel plug, eluting with ethyl
acetate-hexanes (10:90). The eluant is concentrated to afford 3.42
g (51%) of methyl 4-amino-2-(trifluoromethyl)nicotinate as an
off-white solid. .sup.1HNMR (400 MHz, CD.sub.3OD), 8.11 (d, 1H),
6.93 (d, 1H), 3.878 (s, 3H). LC/MS (M+1)=221.2
[0219] The remaining aqueous layer from above is acidified with
concentrated hydrochloric acid to pH 2. The aqueous layer is
extracted with ethyl acetate (16.times.). The organic layer is
washed with water, dried over anhydrous magnesium sulfate, filtered
and concentrated in vacuo to provide an orange solid. The orange
solid is triturated in diethyl ether to afford 2.5 g (40%) of
4-amino-2-(trifluoromethyl)nicotinic acid as a yellow solid.
.sup.1H NMR (400 MHz, CD.sub.3OD), 8.09 (d, 1H), 6.93 (d, 1H),
LC/MS (M+1)=207.1.
Preparation of 4-Amino-2-(trifluoromethyl)nicotinic acid
##STR00019##
[0221] To a stirred solution of methyl
4-amino-2-(trifluoromethyl)nicotinate (1.97 g, 8.97 mmol) in
dioxane/methanol/water (3:2:1 ratio by volume) is added lithium
hydroxide (0.365 g, 15.2 mmol). The mixture was heated at
85.degree. C. for 8 hours. The reaction mixture was cooled to room
temperature and conc. HCl was added dropwise to adjust the pH to
2-3. Saturated NaCl solution was added. The aqueous layer is
extracted with ethyl acetate (12.times.). The organic layer is
washed with water, dried over anhydrous magnesium sulfate, filtered
and concentrated in vacuo to afford 1.75 g (95%) of
4-amino-2-(trifluoromethyl)nicotinic acid as a yellow solid.
.sup.1H NMR (400 MHz, CD.sub.3OD), 8.09 (d, 1H), 6.93 (d, 1H),
LC/MS (M+1)=207.1.
Preparation of ethyl 2-amino-2-cyanoacetate
##STR00020##
[0223] To a stirred solution of the ethyl
(hydroxyimino)cyanoacetate (10 g, 70.4 mmol) in water and sodium
bicarbonate saturated solution (80 mL) was added sodium dithionite
(34.3 g, 197 mmol). The reaction was heated up to 35.degree. C. for
3 h. The solution was then saturated with sodium chloride and
extracted with ethyl acetate (3.times.200 mL). The combined organic
phases were dried over sodium sulfate and concentrated, giving an
oil of ethyl 2-amino-2-cyanoacetate (1.6 g). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.42 (s, 1H), 4.32 (q, 2H), 1.33 (t, 3H).
Preparation of ethyl 2-cyano-2-formamidoacetate
##STR00021##
[0225] To ethyl 2-amino-2-cyanoacetate (800 mg, 6.24 mmol) in
dichloromethane (5 mL) was added cyanomethyl formate (531 mg, 6.24
mmol) in dichloromethane (5 mL) dropwise at 0.degree. C. The
reaction was warmed up to room temperature for twelve hours. The
reaction was concentrated and purified by silica gel chromatography
to give an oil of ethyl 2-cyano-2-formamidoacetate (500 mg). NMR
spectrum shows a mixture of starting material and desired product
in a 1:1 ratio. The product was dissolved in methylene chloride (10
mL). To this solution was added cyanomethyl formate (797 mg, 9.37
mmol) and the reaction was stirred for twelve more hours. The
reaction was concentrated and purified with silica gel
chromatography to give an oil of ethyl 2-cyano-2-formamidoacetate
(460 mg). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.30 (s, 1H),
6.61 (s, 1H), 5.55 (d, 2H), 4.36 (q, 2H), 1.35 (t, 3H).
Preparation of ethyl 5-aminothiazole-4-carboxylate
##STR00022##
[0227] A solution of ethyl 2-cyano-2-formamidoacetate (480 mg, 3.07
mmol) and Lawesson's reagent (746 mg, 1.84 mmol) in toluene (5 mL)
was heated up to 80.degree. C. for overnight. The reaction was then
diluted with sodium carbonate aqueous solution (20 mL) and
methylene chloride (50 mL). Organic phase was washed with brine (20
mL), dried over sodium sulfate and concentrated. Purified with
silica gel chromatography to give a solid material of ethyl
5-aminothiazole-4-carboxylate (120 mg). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.86 (s, 1H), 5.98 (s, br, 2H), 4.38 (q, 2H),
1.41 (t, 3H). MS m/z 173.2 (M+H).sup.+.
Preparation of ethyl
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carboxylate
##STR00023##
[0228] A mixture of ethyl 5-aminothiazole-4-carboxylate (91.3 mg,
0.53 mmol), scandium (III) triflate (2.61 mg, 0.005 mmol) and
2,5-hexanedione (1.5 mL) was heated up to 180.degree. C. for 1 h by
microwave. The reaction was then concentrated and purified with
silica gel chromatography to give a tan solid of ethyl
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carboxylate (104 mg).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.82 (s, 1H), 5.92 (s,
2H), 4.25 (q, 2H), 2.01 (s, 6H), 1.20 (t, 3H). MS m/z 251.3
(M+H).sup.+.
Preparation of ethyl
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carboxylate
##STR00024##
[0230] To ethyl
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carboxylate (104 mg,
0.415 mmol) in THF was added 1M DIBAL/THF (1.24 mL, 1.24 mmol)
dropwise slowly at -78.degree. C. and stirred for 1 h. The reaction
mixture was warmed up to room temperature slowly and stirred for
twelve hours. The reaction was then quenched with ice water (5 mL)
and diluted with ethyl acetate (100 mL) and 2N NaOH (20 mL). The
organic phase was then washed with brine (10 mL), dried over sodium
sulfate and concentrated, giving a semi-solid of ethyl
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carboxylate (86 mg).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.81 (s, 1H), 5.90 (s,
2H), 4.43 (s, 2H), 2.00 (s, 6H). MS m/z 209.3 (M+H).sup.+.
Preparation of
5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carbaldehyde
##STR00025##
[0232] To a solution of
(5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazol-4-yl)methanol (86 mg, 0.41
mmol) in methylene chloride was added manganese dioxide (359 mg,
4.13 mmol). The reaction was warmed up to reflux for 3 h. The crude
mixture was purified with silica gel chromatography to give an oil
of 5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carbaldehyde (17 mg).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.72 (s, 1H), 8.87 (s,
1H), 5.98 (s, 2H), 2.05 (s, 6H). MS m/z 207.2 (M+H).sup.+.
2-(5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazol-4-yl)-3-phenethyl-5-(trifluorome-
thyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00026##
[0234] A solution of
4-amino-N-phenethyl-2-(trifluoromethyl)nicotinamide (25.5 mg, 0.082
mmol), 5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazole-4-carbaldehyde (17
mg, 0.082 mmol) and TsOH (1.4 mg, 0.008 mmol) in toluene was heated
at reflux overnight. The reaction was then filtered and to the
crude mixture was added manganese oxide. The reaction was heated at
110.degree. C. for 4 h. The reaction mixture was filtered,
concentrated and purified with silica gel chromatography providing
a solid,
2-(5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazol-4-yl)-3-phenethyl-5-(trifluorom-
ethyl)pyrido[4,3-d]pyrimidin-4(3H)-one (7 mg). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 9.02 (s, 1H), 8.76 (d, 1H), 7.69 (m, 1H),
7.51 (m, 1H), 7.48 (d, 1H), 7.32 (m, 5H), 4.20 (m, 2H), 3.32 (m,
2H), 2.07 (s, 6H). MS m/z 496.5 (M+H).sup.+.
Preparation of methyl 2-(difluoromethyl)benzoate
##STR00027##
[0236] A solution of methyl 2-formylbenzoate (1.0 g, 6.09 mmol) and
bis(2-methoxyethyl)amino-sulfur trifluoride (4.04 g, 18.3 mmol) in
methylene chloride (10 mL) was heated at reflux for twelve hours.
The reaction was concentrated, diluted with ethyl acetate (200 mL)
and water (100 mL). Sodium bicarbonate solid was used to neutralize
the mixture slowly to pH of 8. The organic phase was then separated
and washed with brine (50 mL), dried over sodium sulfate and
concentrated. The residue was purified with silica gel
chromatography to give methyl 2-(difluoromethyl)benzoate as an oil
(700 mg). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.02 (d, 1H),
7.82 (d, 1H), 7.64 (t, 1H), 7.52 (m, 1H). 7.39 (t, 1H), 3.93 (s,
3H).
Preparation of (2-(difluoromethyl)phenyl)methanol
##STR00028##
[0238] To a solution of methyl 2-(difluoromethyl)benzoate (700 mg,
3.76 mmol) in THF was added 1M lithium aluminum hydride in THF
(5.64 mL, 11.3 mmol) dropwise at room temperature for 1 h. The
reaction was then poured into ice water (100 mL) and diluted with
ethyl acetate (200 mL) and 2N sodium hydroxide aqueous solution
(100 mL). The organic phase was then separated and washed with
brine (50 mL), dried over sodium sulfate and concentrated, to give
(2-(difluoromethyl)phenyl)methanol as an oil (480 mg).
[0239] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.56 (d, 1H), 7.41
(m, 3H), 6.93 (t, 1H), 4.82 (s, 2H).
Preparation of 2-(difluoromethyl)benzaldehyde
##STR00029##
[0241] A mixture of (2-(difluoromethyl)phenyl)methanol (100 mg,
0.632 mmol) and manganese oxide (275 mg, 3.16 mmol) in
dichloromethane was stirred at room temperature for twelve hours
and then at 45.degree. C. for 1 h. The reaction was filtered
through celite and concentrated, to give
2-(difluoromethyl)benzaldehyde as an oil (99 mg). .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 10.2 (s, 1H), 7.94 (d, 1H), 7.81 (d, 1H),
7.70 (m, 2H), 6.93 (t, 1H).
Preparation of 2-methylnicotinaldehyde
##STR00030##
[0243] A mixture of (2-methylpyridin-3-yl)methanol (1.0 g, 8.12
mmol) in dichloromethane (10 mL) and manganese oxide (7.06 g, 81.2
mmol) was stirred at room temperature for sixty hours and then
heated at 45.degree. C. for 4 h. The reaction was filtered through
celite and a plug of silica gel (5.times.7 cm) using ethyl acetate
as eluent. The filtrate was then concentrated to give 2-methyl
nicotinaldehyde as an oil (680 mg).
[0244] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.32 (s, 1H),
8.67 (dd, 1H), 8.09 (dd, 1H), 7.31 (dd, 1H), 2.88 (s, 3H).
Preparation of 3-(difluoromethyl)-2-methylpyridine
##STR00031##
[0246] A solution of 2-methylnicotinaldehyde (680 mg, 5.61 mmol)
and bis(2-methoxyethyl)amino-sulfur trifluoride (3.72 g, 16.8 mmol)
in dichloroethane (10 mL) was warmed at reflux overnight. The
reaction was concentrated, diluted with ethyl acetate (200 mL) and
water (100 mL). Sodium bicarbonate solid was used to neutralize the
mixture slowly to pH of 8. The organic phase was then separated and
washed with brine (50 mL), dried over sodium sulfate and
concentrated. The residue was purified with flash chromatography
(25+S Biotage, heptane/ethyl acetate=100:0 to 80:20), to give
3-(difluoromethyl)-2-methylpyridine as an oil (110 mg). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.58 (d, 1H), 7.81 (d, 1H), 7.22 (m,
1H), 6.77 (t, 1H), 2.64 (s, 3H).
Preparation of 3-(difluoromethyl)picolinaldehyde
##STR00032##
[0248] A mixture of 3-(difluoromethyl)-2-methylpyridine (110 mg,
0.768 mmol) and selenium oxide (102 mg, 0.922 mmol) in 1,4-dioxane
(3 mL) was warmed at reflux for twelve hours. The reaction mixture
was then concentrated and purified with flash chromatography (12+S
Biotage, heptane/ethyl acetate=1:0 to 1:1), to give
3-(difluoromethyl)picolinaldehyde as an oil (11 mg). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 10.1 (s, 1H), 8.90 (d, 1H), 8.22 (d,
1H), 7.64 (m, 1H), 7.59 (t, 1H).
Preparation of 1,2-difluoro-4-(2-nitroprop-1-enyl)benzene
##STR00033##
[0250] 3,4-difluorobenzaldehyde (54 g, 380 mmol), ammonium acetate
(19 g, 247 mmol), and 4A molecular sieves (54 g) were combined in
nitroethane (810 mL) and heated at reflux for 24 hours. The
reaction was filtered, and concentrated in vacuo. The resultant
mixture was partitioned between ethyl acetate (500 mL) and water
(300 mL). The organic phase was separated, washed with brine, dried
over sodium sulfate, filtered and concentrated in vacuo. The
residue was cooled to 4.degree. C. for 2 hours then ethanol (100
mL) was added to give yellow crystals which were collected by
filtration. (57.2 g 75.6% yield). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.0 (s, 1H), 7.15-7.25 (m, 2H), 3.7 (t, 1H), 2.4 (s,
3H).
Preparation of 1-(3,4-difluorophenyl)propan-2-amine
hydrochloride
##STR00034##
[0251] (E,Z)-1,2-difluoro-4-(2-nitroprop-1-enyl)benzene (2.52 g,
12.65 mmol) was dissolved in anhydrous THF (30 mL) and cooled in an
ice bath. To this was added a 1.0M THF solution of lithium aluminum
hydride (38 mL, 38 mmol). The reaction was stirred at room
temperature for 15 minutes then heated at reflux for 2 hours. The
reaction mixture was cooled to room temperature, and worked up by
the successive addition of water (1.5 mL), 15% NaOH (1.5 mL), and
water (4.5 mL). The reaction mixture was stirred to granulate a
white precipitate which was filtered. The filtrate was diluted with
water (50 mL), extracted with ethyl acetate (2.times.100 mL). The
organic phase was dried over sodium sulfate, filtered, and
concentrated in vacuo. The residue was redissolved in ethyl acetate
(25 mL), to which was added 4N HCl/dioxane solution (7 mL), and the
resulting mixture was stirred at room temperature for 10 minutes.
This mixture was concentrated in vacuo and triturated with hexanes
to afford the product as a colorless solid (2.32 g, 88% yield).
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.2 (m, 2H), 7.05 (m,
1H), 3.5 (m, 1H), 2.8-3.0 (dd, 2H), 1.25 (d, 3H). MS m/z 172.2
(M+H).sup.+.
Preparation of
4-amino-N-(1-(3,4-difluorophenyl)propan-2-yl)-2-(trifluoromethyl)nicotina-
mide
##STR00035##
[0253] 4-amino-2-(trifluoromethyl)nicotinic acid (4.4 g, 18.14
mmol) was suspended in a mixture of anhydrous DMF (100 mL) and
anhydrous dichloromethane (200 mL). To this was added triethylamine
(6 g, 60 mmol), followed by 1-(3,4-difluorophenyl)propan-2-amine
(3.75 g, 21.90 mmol). HBTU (9.63 g, 25.4 mmol) was added and the
mixture was allowed to stir at room temperature overnight. The
reaction mixture was concentrated in vacuo to remove
dichloromethane. The resultant DMF solution was poured into water
and extracted twice with ethyl acetate. The organic layer was
washed successively with 1N NaOH and water, then dried over
magnesium sulfate, filtered and concentrated in vacuo to give an
oil. This oil was purified by silica gel column chromatography
using 20-40% acetone/hexanes as eluant to provide the product as a
colorless solid (6.57 g, 55% yield). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.2 (d, 1H), 7.1 (m, 2H), 6.95 (m, 1H), 6.7 (d,
1H), 4.07 (m, 1H), 4.9 (bd, 2H), 4.4 (m, 1H), 2.7-2.9 (dd, 2H), 1.2
(d, 2H). MS m/z 360.2 (M+H).sup.+.
Preparation of (3-(benzyloxy)pyridin-2-yl)methanol
##STR00036##
[0255] 2-hydroxymethyl-3-hydroxypyridine (100.3 g, 620.7 mmol) was
dissolved in acetone (1 L). Potassium carbonate (180 g, 1.3 mol)
was added followed by benzyl bromide (127 g, 745 mmol). The mixture
was heated at reflux for 48 hours. The resultant mixture was cooled
to room temperature and filtered through celite. The filter cake
was washed with acetone (1 L). The combined filtrate was
concentrated in vacuo to provide an orange oil. This oil was
purified by silica gel column chromatography using 10-40% ethyl
acetate/hexanes as eluant. The resultant tan solid was
recrystallized from hexanes to provide the product (88.2 g, 66%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.2 (t, 1H), 7.4
(m, 5H), 7.15 (d, 2H), 5.1 (s, 2H), 4.8 (d, 2H), 4.3 (t, 1H).
Preparation of 3-(benzyloxy)picolinaldehyde
##STR00037##
[0257] (3-(benzyloxy)pyridin-2-yl)methanol (87 g, 400 mmol) was
dissolved in anhydrous dioxane (80 mL). Manganese dioxide (351 g)
was added and the mixture was heated for 2 hours at 80.degree.
C.
The reaction mixture was filtered through celite and the filter
cake was washed with ethyl acetate (300 mL). The filtrate was
concentrated in vacuo to yield a brown oil which solidified upon
standing. (84.12 g, 98% yield). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.4 (d, 1H), 7.25-7.45 (m, 8H), 5.25 (s, 2H).
Preparation of
2-(3-(benzyloxy)pyridin-2-yl)-3-(1-(3,4-difluorophenyl)propan-2-yl)-5-(tr-
ifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one
##STR00038##
[0259]
4-amino-N-(1-(3,4-difluorophenyl)propan-2-yl)-2-(trifluoromethyl)ni-
cotinamide (3.61 g, 10.05 mmol) and 3-(benzyloxy)picolinaldehyde
(2.57 g, 12.1 mmol) were combined along with a catalytic amount of
4-toluenesulfonic acid (0.03 g, 0.20 mmol) in anhydrous toluene
(150 mL). This was heated at reflux overnight in a round bottom
flask fitted with a Dean-Stark trap. The resultant dark solution
was cooled to room temperature, concentrated in vacuo to a dark
brown oil. This oil was purified by silica gel column
chromatography using 30% acetone/hexanes as eluant to provide the
product diastereomers as an oil (4.61 g, 82% yield).
[0260] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.2 (m, 1H), 8.0
(m, 1H), 7.3-7.6 (m, 7H), 5.15 (d, 2H), 3.0 (dd, 1H), 2.8 (m, 1H),
1.25 (d, 3H). MS m/z 555.2 (M+H).sup.+.
Preparation of
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one
##STR00039##
[0262]
2-(3-(benzyloxy)pyridin-2-yl)-3-(1-(3,4-difluorophenyl)propan-2-yl)-
-5-(trifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one
(4.61 g, 8.31 mmol) was dissolved in absolute ethanol (150 mL). To
this was added 10% palladium on carbon (0.50 g) and the mixture was
hydrogenated on a Parr shaker at 45 PSI for 2.5 hours. The reaction
was filtered through celite, and the filter cake washed with
ethanol. The combined filtrates were concentrated in vacuo to an
oil. This oil was purified by silica gel column chromatography
using 20%-40% acetone/hexanes as eluant to yield the product
diastereomers as a pale yellow foam (3.86 g, 77% yield).
[0263] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.0 (t, 1H), 7.8
(t, 1H), 7-7.2 (m, 5H), 4.8 (m, 1H), 4.5 (m, 1H), 2.8-3.1 (dd, 2H),
1.3 (d, 3H), 1.0 (d, 3H). MS m/z 465.3 (M+H).sup.+.
Preparation of
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00040##
[0265]
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(-
trifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one (27.3
g, 58.8 mmol) was dissolved in 4-methyl-2-pentanone (500 mL). To
this was added manganese dioxide (76.7 g, 882 mmol) and the mixture
was heated to 90.degree. C. for 1 hour. The reaction mixture was
cooled and filtered through celite. The filter cake was washed with
ethyl acetate. Combined filtrates were concentrated in vacuo to a
pale yellow solid (8.35 g). The solid was slurried in hexanes and
the resultant colorless solid was collected. The filtrate was
concentrated and purified by silica gel column chromatography using
20-30% acetone/hexanes as eluant to yield additional product (3.47
g, 43.5% total yield).
Preparation of
(R)-4-amino-N-(1-phenylpropan-2-yl)-2-(trifluoromethyl)nicotinamide
##STR00041##
[0267] 4-amino-2-(trifluoromethyl)nicotinic acid (15 g, 61.83 mmol)
was suspended in a mixture of anhydrous DMF (300 mL) and anhydrous
dichloromethane (500 mL). To this was added triethylamine (30.4 mL,
216 mmol), followed by L-amphetamine hydrochloride (12.7 g, 74.2
mmol). HBTU (30.5 g, 80.4 mmol) was added and the mixture was
allowed to stir at room temperature overnight. The reaction mixture
was concentrated in vacuo to remove dichloromethane. The resultant
DMF solution was poured into water, extracted with ethyl acetate (1
L). The organic layer was washed successively with 1N NaOH,
saturated sodium bicarbonate, water, dried over sodium sulfate,
filtered and concentrated in vacuo to give an oil. This oil was
purified by silica gel column chromatography using 40-70% ethyl
acetate/heptanes as eluant to provide the product as a colorless
foam (18.3 g, 92% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.2 (d, 1H), 7.2-7.4 (m, 5H), 6.6 (d, 1H), 5.8 (d, 1H), 4.7 (bs,
2H), 4.5 (m, 1H), 2.9 (m, 2H), 1.25 (d, 3H). MS m/z 324.3
(M+H).sup.+.
Preparation of
2-(2-methoxyphenyl)-3-((R)-1-phenylpropan-2-yl)-5-(trifluoromethyl)-2,3-d-
ihydropyrido[4,3-d]pyrimidin-4(1H)-one
##STR00042##
[0269]
(R)-4-amino-N-(1-phenylpropan-2-yl)-2-(trifluoromethyl)nicotinamide
(16.23 g, 50.2 mmol) and ortho-anisaldehyde (8.2 g, 60.2 mmol) were
combined along with a catalytic amount of 4-toluenesulfonic acid
(0.174 g, 1.0 mmol) in anhydrous toluene (550 mL). This mixture was
heated at reflux overnight in a round bottom flask fitted with a
Dean-Stark trap. The resultant dark solution was cooled to room
temperature, then concentrated in vacuo to a dark brown paste. This
paste was suspended in ethyl acetate, and the resultant solids were
collected by filtration to provide the product (6.94 g). The
filtrate was concentrated in vacuo then purified by silica gel
column chromatography using 40-60% ethyl acetate/hexanes as eluant
to give 12.1 g of additional product diastereomers (84% total
yield). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.19 (d, 1H),
7.1-7.3 (m, 7H), 6.9 (d, 1H), 6.8 (t, 1H), 6.5 (d, 1H), 5.95 (d,
1H), 5.7 (d, 1H), 4.8 (m, 1H), 3.95 (s, 3H), 3.0 (dd, 1H), 2.4 (dd,
1H), 1.3 (d, 3H). MS m/z 442.4 (M+H).sup.+.
Preparation of
(R)-2-(2-methoxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one
##STR00043##
[0271]
2-(2-methoxyphenyl)-3-((R)-1-phenylpropan-2-yl)-5-(trifluoromethyl)-
-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one (19.04 g, 43.13 mmol)
was dissolved in acetone (800 mL). To this was added a 5% aqueous
solution of potassium permanganate (34.1 g KMnO.sub.4 in 700 mL
water). The resultant mixture was stirred at room temperature
overnight. The reaction was quenched by pouring it into a 10%
aqueous solution of sodium bisulfite (3 L). The resulting aqueous
solution was extracted with ethyl acetate (3.times.700 mL). The
organic phase was washed with brine, dried over magnesium sulfate,
filtered and concentrated in vacuo to provide an oil. This oil was
purified by flash chromatography using 25-50% ethyl
acetate/heptanes as eluant to provide the product as a solid. This
solid was recrystallized from ethyl acetate/hexanes to yield a
colorless solid (12.5 g). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.8 (d, 1H), 7.65 (d, 1H), 7.4 (t, 1H), 7.15 (m, 7H), 6.95 (t, 1H),
6.8 (d, 1H), 6.35 (d, 1H), 4.2 (m, 1H), 3.8 (s, 3H), 3.6 (dd, 1H),
2.9 (dd, 1H), 1.75 (d, 3H). MS m/z 440.3 (M+H).sup.+.
Example 1
Preparation of
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one)
Preparation 1a: 4-Amino-2-chloro-N-phenethyl-nicotinamide
##STR00044##
[0273] 4-Amino-2-chloro-nicotinic acid (1 g, 5.795 mmol) was
dissolved in DMF (30 mL) and dichloromethane (50 mL).
Phenethylamine (0.87 mL, 6.954 mmol, 1.2 equiv.) was added followed
by TEA (1.21 mL, 8.693 mmol, 1.5 equiv.) and HBTU (2.64 g, 6.954
mmol, 1.2 equiv.) and the reaction stirred at 20.degree. C. for 12
hours. The reaction mixture was then diluted with dichloromethane
and the aqueous layer was extracted with 20% isopropyl
alcohol/dichloromethane. The combined organic portions were washed
with 0.5N NaOH solution, NH.sub.4Cl solution, water, brine, dried
over anhydrous MgSO.sub.4, filtered and concentrated. The crude
material was purified by column chromatography using 2:1 ethyl
acetate:hexane to provide 1.23 g (77%) of product.
Preparation 1b:
5-Chloro-2-(2-methoxy-phenyl)-3-phenethyl-2,3-dihydro-1H-pyrido[4,3-d]pyr-
imidin-4-one
##STR00045##
[0275] To a solution of 4-amino-2-chloro-N-phenethyl-nicotinamide
(808 mg, 2.93 mmol) in 8 mL of acetonitrile at 20.degree. C. was
added o-anisaldehyde (399 mg, 2.93 mmol) and a catalytic amount of
p-toluenesulfonic acid (10 mg, 0.059 mmol). The resulting solution
was then heated to reflux under Dean-Stark trap for 12 hours. The
reaction mixture was cooled to 20.degree. C. and diluted with ethyl
acetate. The mixture was then partitioned between ethyl acetate and
aqueous sodium bicarbonate. The organic phase was washed 3.times.
with NaHCO.sub.3, 1.times. with water, 1.times. with brine and then
dried over anhydrous Na.sub.2SO.sub.4, filtered and evaporated to
dryness. 1.08 g of pale yellow sticky solid was obtained. The
resulting crude material was used without further purification.
Preparation 1c:
5-Chloro-2-(2-methoxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00046##
[0277]
5-Chloro-2-(2-methoxy-phenyl)-3-phenethyl-2,3-dihydro-1H-pyrido[4,3-
-d]pyrimidin-4-one (1.0 g, 2.54 mmol) was placed in a flask,
acetone (10 mL) was added and the mixture stirred vigorously at
20.degree. C. A 5% aqueous solution of potassium permanganate
(KMnO.sub.4) was added (50.8 mmol, 2 equiv.). The reaction progress
was monitored by HPLC. The reaction was stirred for 12 hours.
Another 2 equiv. of KMnO.sub.4 was added and reaction stirred at
ambient temperature for 4 hours until reaction was complete
according to HPLC. The reaction was then quenched with 10% aqueous
NaHSO.sub.3, filtered by gravity filtration and partitioned between
ethyl acetate and water. The aqueous layer was extracted 3.times.
with ethyl acetate, the combined organic layers washed 1.times.
with water, 1.times. brine and then dried over anhydrous
Na.sub.2SO.sub.4. Filtration and evaporation of the solvent gave
398 mg of tan solid. The original filter cake was suspended in
dichloromethane and sonicated for 5 minutes, then repeated two more
times. The suspensions were filtered through a nylon filter,
combined and evaporated to dryness to give additional 282 mg of
product for combined total crude yield of 680 mg (68%) of a tan
solid. The product was used further without purification.
Preparation 1d:
5-Iodo-2-(2-methoxy-phenyl-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00047##
[0279] A mixture of
5-chloro-2-(2-methoxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
(300 mg, 0.766 mmol), trimethylsilyl chloride (0.097 mL, 0.766
mmol) and sodium iodide (344 mg, 2.297 mmol, 3 equiv.) in
propionitrile (7.66 mL) was heated to reflux under nitrogen in the
dark for 4 hours. TLC showed no starting material remaining. The
reaction mixture was cooled to ambient temperature and poured into
1N NaOH (30 mL). The resulting suspension was extracted with ethyl
acetate (3.times.20 mL), the combined extracts were washed with
water, brine and dried, filtered and concentrated in vacuo to give
a brown oil. The crude product was purified by column
chromatography using 10% ethyl acetate in dichloromethane to
provide 235 mg (64%) of product.
Preparation 1e:
2-(2-Methoxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one
##STR00048##
[0281] In a flame dried 2-dram vial, a mixture of
5-iodo-2-(2-methoxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
(67 mg, 0.138 mmol), methyl 2,2-difluoro-2-(fluoro-sulfonyl)acetate
(0.035 mL, 0.277 mmol, 2 equiv.), copper iodide (32 mg, 0.167 mmol,
1.2 equiv), HMPA (0.048 mL, 0.277 mmol, 2 equiv) in DMF (2 mL) was
stirred under nitrogen at 80.degree. C. for 12 hrs. The reaction
mixture was then cooled to ambient temperature, diluted with
dichloromethane and washed 3.times. with water. The organic phase
was then dried over anhydrous MgSO.sub.4, filtered and
concentrated. The crude material was purified on preparative TLC in
50:50 ethyl acetate:hexane to provide 16.8 mg of product (29%).
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyri-mi-
din-4-one)
##STR00049##
[0282]
2-(2-Methoxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]-
pyrimidin-4-one (8.4 mg, 0.0197 mmol) was dissolved in
dichloromethane (0.5 mL), the mixture cooled to 0.degree. C. and a
solution of boron trichloride was added (1M in hexane, 0.16 mL, 8
equiv.). The reaction mixture was then heated at 40.degree. C.
overnight. The reaction mixture was then cooled to ambient
temperature, diluted with methanol (2 mL) and then heated to
70.degree. C. for 1 hour. The solvent was evaporated and the
residue partitioned between methylene chloride and saturated
NaHCO.sub.3 solution. The aqueous layer was extracted 3.times. with
dichloromethane. The pooled organic extracts were washed with
water, brine, dried, filtered and concentrated. The crude product
was purified using preparative TLC in 50:50 ethyl acetate:hexane to
provide 5 mg (62%) of final product.
[0283] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.82 (d, 1H), 7.67
(d, 1H), 7.41 (t, 1H), 7.27 (m, 1H), 7.15 (m, 3H), 7.05 (m, 2H),
6.86 (m, 2H), 4.38 (t, 2H), 2.94 (t, 2H). MS m/z 412.3
(M+H).sup.+.
[0284] Alternatively,
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one) can be prepared using
4-amino-2-(trifluoromethyl)nicotinic acid as the starting
material.
Preparation 1f:
4-Amino-N-phenethyl-2-(trifluoromethyl)nicotinamide
##STR00050##
[0285] 4-Amino-2-(trifluoromethyl)nicotinic acid (150 mg, 0.78
mmol) was dissolved in DMF (4 mL) and dichloromethane (6 mL).
Phenethylamine (106 mg, 0.87 mmol, 1.2 equiv.) was added followed
by TEA (0.15 mL, 1.1 mmol, 1.5 equiv.) and HBTU (331 mg, 0.87 mmol,
1.2 equiv.) and the reaction stirred at 20.degree. C. for 12 hours.
The reaction mixture was then diluted with dichloromethane and the
aqueous layer was extracted with dichloromethane. The combined
organic portions were washed with 0.5N NaOH solution, NH.sub.4Cl
solution, water, brine, dried over anhydrous MgSO.sub.4, filtered
and concentrated. The crude material was purified by column
chromatography using 2:1 ethyl acetate:hexane to provide 79 mg
(36%) of product. The product can be converted to
2-(2-Hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]pyri-m-
id in-4-one) by using the title compound above, Preparation 1f, and
following the procedures described in Preparation 1b (condensation
with an appropriate aldehyde R.sup.2'CHO), Preparation 1c
(oxidation with KMnO.sub.4) and the deprotection step using
BCl.sub.3 (when converting a methoxy moiety in R.sup.2' to the
hydroxy in R.sup.2) as described above.
[0286] The following title compounds may be prepared in a manner
analogous to Example 1 using Preparation 1a,
4-Amino-2-chloro-N-phenethyl-nicotinamide or
4-amino-2-chloro-6-methylnicotinic acid or Preparation 1f,
4-Amino-N-phenethyl-2-(trifluoromethyl)nicotinamide, or other
appropriate nicotinamide derivative with the corresponding aldehyde
of formula R.sup.2'CHO (it is to be understood that a methoxy group
or benzyloxy group in the R.sup.2' moiety can be deprotected to the
corresponding hydroxy group to provide the R.sup.2 moiety in the
compound of formula I).
Example 2
2-(3-Fluoro-2-hydroxy-phenyl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3--
d]pyrimidin-4-one
##STR00051##
[0288] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.90 (d, 1H), 7.75
(d, 1H), 7.25 (t, 3H), 7.05 (m, 3H), 6.90 (m, 2H), 4.39 (m, 2H),
2.94 (m, 2H). MS m/z 430.3 (M+H).sup.+.
Example 3
2-(3-Hydroxy-pyridin-2-yl)-3-phenethyl-5-trifluoromethyl-3H-pyrido[4,3-d]p-
yrimidin-4-one
##STR00052##
[0290] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.95 (d, 1H), 8.30
(d, 1H), 7.75 (d, 1H), 7.45 (m, 3H), 7.25 (m, 5H), 4.99 (m, 2H),
3.33 (m, 2H). MS m/z 413.2 (M+H).sup.+.
Example 4a
(R)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H--
pyrido[4,3-d]pyrimidin-4-one
##STR00053##
[0292]
(R)-2-(2-methoxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)-
pyrido[4,3-d]pyrimidin-4(3H)-one (6.6 g, 15.02 mmol) was dissolved
in anhydrous dichloromethane (75 mL) and cooled to 0.degree. C. in
an ice bath. To this was added a 1M dichloromethane solution of
boron trichloride (31.5 mL, 31.5 mmol) slowly to maintain
temperature. After addition, the reaction mixture was stirred for
2-5 minutes. The reaction mixture was transferred via canulla to a
0.degree. C. aqueous solution of diethanolamine (10.3 g, 97.6 mmol
in 150 mL water) with stirring. The reaction mixture was allowed to
come to room temperature and stir for 1 hour. The reaction mixture
was diluted with dichloromethane, and the layers separated. The
organic layer was washed with brine, dried over magnesium sulfate,
filtered, and concentrated in vacuo to provide a foam. This foam
was purified by silica gel column chromatography using 20-40% ethyl
acetate/hexanes as eluant to give a colorless foam which was then
crystallized from ethyl acetate/hexanes to provide the product (4.2
g). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.79 (d, 1H), 7.6 (d,
1H), 7.45 (m, 1H), 7.00 (m, 6H), 6.90 (m, 2H), 3.60 (m, 2H), 3.10
(m, 2H), 1.99 (t, 3H). MS m/z 426.3 (M+H).sup.+.
Example 4b
(S)-2-(2-Hydroxy-phenyl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluoromethyl-3H--
pyrido[4,3-d]pyrimidin-4-one
##STR00054##
[0293] Prepared analogous to Example 4a starting from
(S)-2-(2-methoxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.79 (d, 1H), 7.6 (d, 1H), 7.45 (m, 1H), 7.00 (m, 6H), 6.90
(m, 2H), 3.60 (m, 2H), 3.10 (m, 2H), 1.99 (t, 3H). MS m/z 426.3
(M+H).sup.+.
Example 5
(R)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-phenyl-ethyl)-5-trifluorometh-
yl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00055##
[0295] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.78 (d, 1H), 8.31
(dd, 1H), 7.49 (d, 1H), 7.41 (m, 2H), 7.02 (m, 2H), 6.88 (m, 3H),
5.31 (m, 1H), 3.62 (dd, 1H), 3.18 (dd, 1H), 1.89 (d, 3H). MS m/z
427.4 (M+1).
Example 6
(R,S)-2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-tr-
ifluoromethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00056##
[0297] .sup.1H NMR (400 MHz, CD.sub.3OD), 8.784 (d, 1H), 8.10 (dd,
1H), 7.701 (d, 1H), 7.40 (m, 2H), 7.14 (m, 1H), 6.97 (t, 1H), 6.890
(m 2H), 4.23 (m, 1H), 3.42 (dd, 2H), 3.29 (dd, 2H), 1.59 (d,
3H)
[0298] MS m/z (M+1) 445.4
[0299] The racemic compound of Example 6 can be separated into its
single enantiomer components (Examples 6a and 6b) by preparative
chromatography on a Chiralpak.TM. AS (10 cm.times.50 cm) column
(Daicel (U.S.A.) Chemical Industries, Ltd, Fort Lee, N.J. 07024,
U.S.A.) using Heptane/EtOH (90/10) as mobile phase at a flow rate
of 475 mL/min.
Example 6a
Enantiomer 1:
2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-trifluo-
romethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00057##
[0301] .sup.1H NMR (400 MHz, CD.sub.3OD), 8.78 (d, 1H), 8.10 (dd,
1H), 7.70 (d, 1H), 7.40 (m, 2H), 7.12 (m, 1H), 6.96 (dt, 1H), 6.87
(m, 2H), 4.28 (m, 1H), 3.34 (m, 1H), 3.30 (m, 1H), 1.59 (d, 2H)
[0302] LC/MS (M+1)=445.3
Example 6b
Enantiomer 2:
2-(3-Hydroxy-pyridin-2-yl)-3-(1-methyl-2-(2-fluorophenyl)ethyl)-5-trifluo-
romethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00058##
[0304] .sup.1H NMR (400 MHz, CD.sub.3OD), 8.78 (d, 1H), 8.10 (dd,
1H), 7.70 (d, 1H), 7.40 (m, 2H), 7.12 (m, 1H), 6.96 (dt, 1H), 6.87
(m, 2H), 4.28 (m, 1H), 3.34 (m, 1H), 3.30 (m, 1H), 1.59 (d, 2H)
[0305] LC/MS (M+1)=445.3.
Example 7
2-(2-Hydroxyphenyl)-3-(2-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[4-
,3-d]pyrimidin-4-one
##STR00059##
[0307] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.8 (d, 1H), 7.75
(d, 1H), 7.4 (m, 1H), 7.2 (m, 1H), 6.8-7 (m, 6H), 4.2 (t, 2H), 2.95
(t, 2H). MS m/z 430.1 (M+H).sup.+.
Example 8
2-(2-Hydroxyphenyl)-3-(3-fluorophenyl)ethyl)-5-trifluoromethyl-3H-pyrido[4-
,3-d]pyrimidin-4-one
##STR00060##
[0309] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.8 (d, 1H), 7.75
(d, 1H), 7.45 (t, 1H), 7.15 (m, 2H), 7 (m, 2H), 6.9 (m, 1H), 6.65
(d, 1H), 6.55 (d, 1H), 4.15 (t, 2H), 2.9 (t, 2H). MS m/z 430.1
(M+H).sup.+.
Example 9
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(2-fluoro-phenyl)-ethyl]-5-trifluoromet-
hyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00061##
[0311] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.85 (d, 1H), 7.8
(d, 1H), 7.35 (m, 1H), 7.2 (m, 1H), 7 (m, 2H), 6.9 (m, 1H), 6.65
(d, 1H), 6.6 (d, 1H), 4.15 (t, 2H), 2.9 (t, 2H). MS m/z 448.2
(M+H).sup.+.
Example 10
2-(2-Hydroxy-phenyl)-7-methyl-3-(phenyl-ethyl)-5-trifluoromethyl-3H-pyrido-
[4,3-d]pyrimidin-4-one
##STR00062##
[0313] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.6 (s, 1H), 7.45
(m, 1H), 7.30 (m, 1H), 7.12 (m, 5H), 6.90 (m, 2H), 4.44 (m, 2H),
2.99 (m, 2H), 2.80 (s, 3H). MS m/z 426.3 (M+H).sup.+.
Example 11
2-(3-Fluoro-2-hydroxy-phenyl)-3-[2-(3-fluoro-phenyl)-ethyl]-5-trifluoromet-
hyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00063##
[0315] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.8 (d, 1H), 7.75
(d, 1H), 7.25 (m, 1H), 7.2 (m, 1H), 6.8-7 (m, 4H), 6.75 (d, 1H),
4.2 (t, 2H), 2.95 (t, 2H). MS m/z 448.2 (M+H).sup.+.
Example 12
3-[2-(2-Fluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluoromethyl-
-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00064##
[0317] .sup.1H NMR (400 MHz, (CD.sub.3).sub.2SO) .delta. 9.89 (s,
1H), 8.95 (d, 1H), 8.25 (d, 1H), 7.95 (d, 1H), 7.50 (m, 2H), 7.25
(m, 1H), 7.00 (m, 3H), 4.11 (m, 2H), 2.99 (m, 2H). MS m/z 431.1
(M+H).sup.+.
Example 13
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00065##
[0319] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 12.66 (brs, 1H),
8.86 (d, 1H), 8.30 (d, 1H), 7.66 (d, 1H), 7.44 (m, 2H), 7.10-6.95
(m, 3H), 4.84 (t, 2H), 3.24 (t, 2H). MS (LC-MS) 449.2
(M+H).sup.+.
Example 14
3-[2-(2,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00066##
[0321] .sup.1H NMR (400 MHz, CDCl.sub.3) rotomeric mixture .delta.
12.40 (brs), 8.85 (d), 8.82 (d), 8.25 (m), 7.68 (d), 7.65 (d), 7.37
(m), 7.15 (q), 7.06 (m), 6.71 (m), 4.98 (t), 4.20 (t), 3.23 (t),
3.13 (t).
[0322] MS (LC-MS) 449.2 (M+H).sup.+.
Example 15
3-[2-(3,4-Difluoro-phenyl)-ethyl]-2-(3-hydroxy-pyridin-2-yl)-5-trifluorome-
thyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00067##
[0324] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.83 (d, 1H), 7.84
(brs, 1H), 7.67 (d, 1H), 7.44 (dt, 1H), 7.28 (dd, 1H), 7.04 (t,
1H), 7.02 (d, 1H), 6.91 (q, 1H), 6.59 (m, 2H), 4.34 (t, 2H), 2.88
(t, 2H).
[0325] MS (LC-MS) 448.2 (M+H).sup.+.
Example 16
2-(2-Hydroxy-phenyl)-5-methylamino-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-
-one
##STR00068##
[0327]
5-Chloro-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-
-4-one, (obtained from Example 1, Preparation 1c and deprotection
1f), (0.0048 mmol, 1 equiv.) was dissolved in methanol (0.5 mL)
then 500 uL of methylamine (2M solution in methanol) was added. The
reaction mixture was then heated to 50.degree. C. for 48 hour. The
solvent was removed and crude products purified by preparative HPLC
or preparative TLC (80% ethyl acetate in hexane).
[0328] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.17 (d, 1H), 7.40
(t, 1H), 7.15 (m, 3H), 7.05 (d, 1H), 6.94 (m, 2H), 6.80 (m, 2H),
6.59 (d, 1H), 4.10 (t, 2H), 3.08 (s, 3H), 2.85 (t, 2H). MS m/z
373.2 (M+H).sup.+.
[0329] The following title compounds (Examples 17-23) may be
prepared in a manner analogous to Example 16 using procedures from
Example 16 with the corresponding amine followed by deprotection
conditions employing BCl.sub.3 found in the final step of Example
1.
Example 17
2-(2-Hydroxy-phenyl)-5-isopropylamino-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-
n-4-one
##STR00069##
[0331] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.00 (m, 1H), 8.17
(d, 1H), 7.40 (t, 1H), 7.15 (m, 3H), 7.05 (d, 1H), 6.94 (m, 2H),
6.80 (m, 2H), 6.59 (d, 1H), 4.30 (m, 1H), 4.09 (t, 2H), 2.85 (t,
2H), 1.32 (d, 6H). MS m/z 401.3 (M+H).sup.+.
Example 18
2-(2-Hydroxy-phenyl)-3-phenethyl-5-pyrrolidin-1-yl-3H-pyrido[4,3-d]pyrimid-
in-4-one
##STR00070##
[0333] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.12 (d, 1H), 7.41
(t, 1H), 7.14 (m, 4H), 6.95 (m, 2H), 6.85 (m, 2H), 6.70 (d, 1H),
4.12 (t, 2H), 3.51 (t, 4H), 2.81 (t, 2H), 1.97 (m, 4H). MS m/z
413.2 (M+H).sup.+.
Example 19
2-(2-Hydroxy-phenyl)-5-(4-methyl-piperazin-1-yl)-3-phenethyl-3H-pyrido[4,3-
-d]pyrimidin-4-one
##STR00071##
[0335] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.27 (d, 1H), 7.4
(m, 1H), 7.11-7.15 (m, 4H), 6.94 (m, 2H), 6.88 (d, 1H), 6.82 (m,
2H), 4.15 (t, 2H), 3.46 (bs, 4H), 2.80 (t, 2H), 2.65 (t, 4H), 2.36
(S, 3H). MS m/z 440.1 (M+H).sup.+.
Example 20
2-(2-Hydroxy-phenyl)-3-phenethyl-5-piperazin-1-yl-3H-pyrido[4,3-d]pyrimidi-
n-4-one
##STR00072##
[0337] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.8.26 (d, 1H), 7.41
(t, 1H), 7.14-7.10 (m, 4H), 6.92-6.96 (m, 2H), 6.89 (d, 1H),
6.83-6.81 (m, 2H), 4.16 (t, 2H), 3.42 (t, 4H), 3.04 (t, 4H),
2.79-2.83 (m, 2H). MS m/z 428.2 (M+H).sup.+.
Example 21
5-Dimethylamino-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-
-4-one
##STR00073##
[0339] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.19 (d, 1H),
7.39-7.43 (m, 1H), 7.15-7.09 (m, 4H), 6.98-6.94 (m, 2H), 6.83-6.80
(m, 2H), 6.75 (d, 1H), 4.11 (t, 2H), 3.07 (s, 6H), 2.82 (t, 2H). MS
m/z 387.1 (M+H).sup.+.
Example 22
2-(2-Hydroxy-phenyl)-5-morpholin-4-yl-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-
n-4-one
##STR00074##
[0341] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.28 (d, 1H),
7.44-7.39 (m, 1H), 7.15-7.11 (m, 4H), 6.99-6.95 (m, 2H), 6.89 (d,
1H), 6.83-6.81 (m, 2H), 4.14 (t, 2H), 3.86 (t, 4H), 3.41 (t, 4H),
2.81 (t, 2H). MS m/z 429.2 (M+H).sup.+.
Example 23
5-Azetidin-1-yl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d-pyrimidin-
-4-one
##STR00075##
[0343] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.19 (d, 1H), 7.4
(t, 1H), 7.15-7.13 (m, 3H), 7.07-7.04 (m, 1H), 6.95-6.93 (m, 2H),
6.82-6.8 (m, 2H), 6.71 (d, 1H), 4.26 (t, 4H), 4.09 (t, 2H), 2.82
(t, 2H), 2.37 (q, 2H). MS m/z 399.1 (M+H).sup.+.
Example 24
Preparation of
2-(2-Hydroxy-phenyl)-3-phenethyl-5-phenyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00076##
[0345]
5-Chloro-2-(2-methoxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-
-4-one, phenylboronic acid (1.1 equiv.), palladium
tetrakis(triphenylphosphine) (7 mol %) and 2N aqueous
Na.sub.2CO.sub.3 (4 equiv.) were combined in dioxane (0.5 mL) and
heated in a microwave for 15 minutes to 140.degree. C. The reaction
mixture was cooled to room temperature, diluted with water and
extracted (3.times.) with ethyl acetate. The combined organic
extracts were washed with brine, dried over anhydrous MgSO.sub.4
and filtered. The crude products were purified on preparative TLC
using 1:1 ethyl acetate:hexane. The desired product can be obtained
by using the methyl ether deprotection conditions employing
BCl.sub.3 and isolation conditions found in the final step of
Example 1. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.90 (s, 1H),
8.78 (d, 1H), 7.53 (m, 2H), 7.46 (m, 3H), 7.37 (t, 1H), 7.29 (d,
1H), 7.13 (m, 3H), 6.97 (m, 2H), 6.77 (m, 2H), 4.30 (t, 2H), 2.79
(t, 2H). MS m/z 420.4 (M+H).sup.+.
[0346] The following title compounds (Examples 25-29) may be
prepared in a manner analogous to Example 24 using procedures from
Example 24 with the corresponding boronic acid followed by
deprotection conditions employing BCl.sub.3 and isolation
conditions found in the final step of Example 1.
Example 25
5-Benzyl-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00077##
[0348] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.40 (s, 1H), 7.58
(d, 1H), 7.40-7.26 (m, 4H), 7.24 (m, 2H), 7.12 (m, 5H), 6.96 (m,
2H), 6.74 (m, 2H), 4.87 (s, 2H), 4.29 (t, 2H), 2.81 (t, 2H).
[0349] MS m/z 434.3 (M+H).sup.+.
Example 26
2-(2-Hydroxy-phenyl)-5-methyl-3-phenethyl-3H-pyrido[4,3-d]pyrimidi-4-one
##STR00078##
[0351] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.44 (d, 1H), 7.42
(m, 2H), 7.30-7.10 (m, 5H), 7.00 (m, 1H); 6.82 (m, 2H), 4.30 (t,
2H), 3.16 (s, 3H), 2.89 (t, 2H). MS m/z 358.3 (M+H).sup.+.
Example 27
5-(6-Dimethylamino-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrid-
o[4,3-d]pyrimidin-4-one
##STR00079##
[0353] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72 (d, 1H), 8.30
(s, 1H), 7.72 (d, 1H), 7.46 (m, 2H), 7.15 (m, 4H), 7.00 (m, 2H),
6.80 (m, 2H), 6.73 (d, 1H), 4.08 (t, 2H), 3.19 (s, 6H), 2.80 (t,
2H). MS m/z 464.4 (M+H).sup.+.
Example 28
5-(6-Dimethylamino-5-methyl-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-
-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00080##
[0355] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.77 (d, 1H), 8.20
(s, 1H), 7.66 (s, 1H), 7.54 (d, 1H), 7.45 (m, 1H), 7.19 (d, 1H),
7.12 (m, 3H), 6.99 (m, 2H), 6.78 (m, 2H), 4.09 (t, 2H), 2.87 (s,
6H), 2.80 (t, 2H), 2.40 (s, 3H). MS m/z 478.4 (M+H).sup.+.
Example 29
5-(6-pyrrolidine-5-pyridin-3-yl)-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrid-
o[4,3-d]pyrimidin-4-one
##STR00081##
[0357] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72 (d, 1H), 8.28
(s, 1H), 7.72 (d, 1H), 7.45 (m, 2H), 7.19 (d, 1H), 7.15 (m, 3H),
6.99 (m, 2H), 6.79 (m, 2H), 6.58 (d, 1H), 4.09 (t, 2H), 3.55 (t,
4H), 2.80 (t, 2H), 2.07 (m, 4H). MS m/z 490.4 (M+H).sup.+.
Example 30
Preparation of
2-(2-Hydroxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-on-
e
Preparation of
2-(2-Benzyloxy-phenyl)-5-chloro-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-o-
ne
##STR00082##
[0359]
5-Chloro-2-(2-methoxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-
-4-one was subjected to the same deprotection conditions found in
the final step of Example 1 to provide the desired
5-chloro-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one-
. A mixture of
5-chloro-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
(88 mg, 0.233 mmol), potassium carbonate (64 mg, 0.466 mmol, 2
equiv.) and benzyl bromide (0.029 mL, 0.245 mmol, 1.05 equiv.) in 2
mL of acetone was heated to reflux overnight. The reaction mixture
was cooled to RT, filtered through celite, concentrated and the
residue purified on preparative TLC plate using 1:1 ethyl
acetate:heptane to give 95 mg (87%) of the desired product.
Preparation of
2-(2-Benzyloxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4--
one
##STR00083##
[0361] A mixture of
2-(2-benzyloxy-phenyl)-5-chloro-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-o-
ne, sodium hydride (60% suspension in mineral oil) (1.5 equiv.),
and corresponding alcohol (1.5 equiv.) in THF was heated at
70.degree. C. overnight. The reaction was cooled to RT, solvent
removed, water and ethyl acetate were added and phases separated.
The aqueous phase was washed with ethyl acetate (2.times.), organic
portions combined, washed with water, brine, dried over anhydrous
MgSO.sub.4, filtered and concentrated. The residue was purified on
preparative TLC plate using 1:1 ethyl acetate:heptane.
2-(2-Hydroxy-phenyl-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00084##
[0363]
2-(2-Benzyloxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one dissolved in methanol (0.05M) was subjected to H-cube
hydrogenation (room temperature, Pd/C cartridge). Solvent was then
removed and product purified on preparative TLC plate to afford the
desired
2-(2-Hydroxy-phenyl)-5-methoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimi-
din-4-one.
[0364] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (d, 1H), 7.42
(t, 1H), 7.16 (m, 3H), 7.05 (m, 2H), 6.95 (m, 2H), 6.80 (m, 2H),
4.10 (m, 5H), 2.86 (t, 2H). MS m/z 374.2 (M+H).sup.+.
[0365] The following title compounds (Examples 31-36) may be
prepared in a manner analogous to Example 30 using procedures from
Example 30 with the corresponding alcohol followed by H-cube
hydrogenation.
Example 31
2-(2-Hydroxy-phenyl)-5-(1-methyl-cyclopropylmethoxy)-3-phenethyl-3H-pyrido-
[4,3-d]pyrimidin-4-one
##STR00085##
[0367] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (d, 1H), 7.45
(t, 1H), 7.19 (m, 3H), 7.12 (d, 1H), 7.05 (d, 1H), 7.00 (m, 2H),
6.83 (m, 2H), 4.39 (s, 2H), 4.12 (t, 2H), 2.91 (t, 2H), 1.32 (s,
3H), 0.70 (t, 2H), 0.47 (t, 2H). MS m/z 428.2 (M+H).sup.+.
Example 32
2-(2-Hydroxy-phenyl)-3-phenethyl-5-propoxy-3H-pyrido[4,3-d]pyrimidin-4-one
##STR00086##
[0369] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (d, 1H), 7.45
(t, 1H), 7.19 (m, 3H), 7.12 (d, 1H), 7.05 (d, 1H), 7.00 (m, 2H),
6.83 (m, 2H), 4.52 (t, 2H), 4.12 (t, 2H), 2.91 (t, 2H), 1.93 (m,
2H), 1.12 (t, 3H). MS m/z 402.2 (M+H).sup.+.
Example 33
5-Cyclobutyloxy-2-(2-hydroxy-phenyl)-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-
-4-one
##STR00087##
[0371] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (d, 1H), 7.45
(t, 1H), 7.19 (m, 3H), 7.12 (d, 1H), 7.05 (d, 1H), 7.00 (m, 2H),
6.83 (m, 2H), 5.41 (m, 1H), 4.12 (t, 2H), 2.91 (t, 2H), 2.55 (m,
2H), 2.36 (m, 2H), 1.93 (m, 1H), 1.76 (m, 1H). MS m/z 414.2
(M+H).sup.+.
Example 34
2-(2-Hydroxy-phenyl)-5-isobutoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4-o-
ne
##STR00088##
[0373] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (d, 1H), 7.45
(t, 1H), 7.19 (m, 3H), 7.12 (d, 1H), 7.05 (d, 1H), 7.00 (m, 2H),
6.83 (m, 2H), 4.33 (d, 2H), 4.12 (t, 2H), 2.91 (t, 2H), 2.28 (m,
1H), 1.12 (d, 6H). MS m/z 416.3 (M+H).sup.+.
Example 35
2-(2-Hydroxy-phenyl)-5-isopropoxy-3-phenethyl-3H-pyrido[4,3-d]pyrimidin-4--
one
##STR00089##
[0375] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34 (d, 1H), 7.46
(t, 1H), 7.19 (m, 3H), 7.10 (d, 1H), 7.04 (d, 1H), 7.00 (m, 2H),
6.82 (m, 2H), 5.61 (m, 1H), 4.12 (t, 2H), 2.90 (t, 2H), 1.50 (d,
6H). MS m/z 402.3 (M+H).sup.+.
Example 36
2-(2-Hydroxy-phenyl)-3-phenethyl-5-(2,2,2-trifluoro-ethoxy)-3H-pyrido[4,3--
d]pyrimidin-4-one
##STR00090##
[0377] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36 (d, 1H), 7.45
(t, 1H), 7.15 (m, 5H), 7.00 (m, 2H), 6.83 (m, 2H), 5.10 (q, 2H),
4.15 (m, 2H), 2.90 (t, 2H). MS m/z 442.2 (M+H).sup.+.
Example 37
Preparation of
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00091##
[0379]
2-(3-(benzyloxy)pyridin-2-yl)-3-(1-(3,4-difluorophenyl)propan-2-yl)-
-5-(trifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one
(12.1 grams, 21.8 mmol) prepared by the same reactions conditions
described above was dissolved in 300 mL of ethanol and added to a
parr bottle that contained 10% Pd(C). The mixture was subjected to
hydrogenation condition (45 psi) for two hours. At this time, the
reaction mixture was filtered through Celite and washed with
ethanol. The fitrate was concentrated in vacuo and purified by
silica gel chromatography to afford a pale yellow foam of
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-
-(trifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one.
##STR00092##
[0380]
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(-
trifluoromethyl)-2,3-dihydropyrido[4,3-d]pyrimidin-4(1H)-one (5.90
g, 12.7 mmol) was dissolved in 500 mL of 4-methyl-2-pentanone. To
this solution, manganese dioxide was added (16.6 g, 191 mmol) and
the mixture was heated to 90.degree. C. for 1 hour. The reaction
was cooled, filtered through Celite and washed with
CH.sub.2CL.sub.2. The filtrate was collected and concentrated in
vacuo as a yellow solid which was purified by silica gel
chromatography to afford the racemic mixture
3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one.
[0381] The racemic compound of Example 37 can be separated into its
single enantiomer components (Examples 37a and 37b) by preparative
chromatography on a Chiralcel OD-H (2.1 cm.times.25 cm) column
(Daicel (U.S.A.) Chemical Industries, Ltd, Fort Lee, N.J. 07024,
U.S.A.) using MeOH/CHCL.sub.3 (3/1) as mobile phase at a flow rate
of 65 g/min.
Example 37a
(R)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00093##
[0383] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.4 (s, 1H), 8.85
(d, 1H), 8.39 (d, 1H), 7.60 (d, 1H), 7.42 (m, 2H), 6.99 (m, 2H),
6.67 (brs, 1H), 5.24 (m, 1H), 3.63 (m, 1H), 3.24 (m, 1H), 1.90 (d,
3H).
[0384] MS m/z 463.3 (M+H).sup.+.
Example 37b
(S)-3-(1-(3,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)
5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00094##
[0386] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.4 (s, 1H), 8.85
(d, 1H), 8.39 (d, 1H), 7.60 (d, 1H), 7.42 (m, 2H), 6.99 (m, 2H),
6.67 (brs, 1H), 5.24 (m, 1H), 3.63 (m, 1H), 3.24 (m, 1H), 1.90 (d,
3H).
[0387] MS m/z 463.3 (M+H).sup.+.
Example 38a
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(triflu-
oromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00095##
[0389] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.90 (d, 1H), 8.30
(d, 1H), 7.60 (d, 1H), 7.42 (m, 2H), 6.99 (m, 1H), 6.66 (m, 2H),
5.65 (m, 1H), 3.63 (m, 1H), 3.20 (m, 1H), 1.90 (d, 3H).
[0390] MS m/z 463.2 (M+H).sup.+.
Example 38b
3-(1-(2,4-difluorophenyl)propan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluo-
romethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00096##
[0392] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.90 (d, 1H), 8.30
(d, 1H), 7.60 (d, 1H), 7.42 (m, 2H), 6.99 (m, 1H), 6.66 (m, 2H),
5.65 (m, 1H), 3.63 (m, 1H), 3.20 (m, 1H), 1.90 (d, 3H).
[0393] MS m/z 463.2 (M+H).sup.+.
Example 39
3-(2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one
##STR00097##
[0395] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.64 (s, 1H),
8.87 (d, 1H), 8.17 (dd, 1H), 7.87 (d, 1H), 7.46 (dq, 2H), 7.10 (dt,
1H), 6.81 (t, 1H), 6.79 (t, 1H), 6.721 (t, 1H), 3.97 (t, 2H), 3.45
(s, 3H), 2.75 (t, 2H). MS m/z 443.1 (M+H).sup.+.
Example 40
3-(2,3-difluorophenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyr-
ido[4,3-d]pyrimidin-4(3H)-one
##STR00098##
[0397] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.85 (d, 1H), 8.28
(m, 1H), 7.65 (d, 1H), 7.37 (m, 2H), 6.95 (m, 4H), 5.01 (t, 2H),
3.31 (t, 2H). MS m/z 449.1 (M+H).sup.+.
Example 41
3-(5-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00099##
[0399] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.90 (d, 1H), 8.20
(d, 1H), 7.70 (d, 1H), 7.38 (m, 2H), 6.80 (m, 1H), 6.56 (m, 2H),
3.40 (s, 3H), 3.01 (m, 2H). MS m/z 461.2 (M+H).sup.+.
Example 42
3-(2-fluoro-6-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00100##
[0401] MS m/z 461 (M+H).sup.+.
Example 43
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluorom-
ethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00101##
[0403] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (d, 1H), 8.10
(m, 1H), 7.70 (d, 1H), 7.40 (m, 2H), 7.10 (t, 1H), 6.90 (d, 1H),
6.70 (d, 1H), 6.61 (t, 1H), 4.45 (m, 1H), 3.40 (s, 3H), 3.39 (m,
1H) 3.01 (m, 2H), 1.75 (d, 3H). MS m/z 457.3 (M+H).sup.+.
Example 43a
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluorom-
ethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00102##
[0405] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (d, 1H), 8.10
(m, 1H), 7.70 (d, 1H), 7.40 (m, 2H), 7.10 (t, 1H), 6.90 (d, 1H),
6.70 (d, 1H), 6.61 (t, 1H), 4.45 (m, 1H), 3.40 (s, 3H), 3.39 (m,
1H) 3.01 (m, 2H), 1.75 (d, 3H). MS m/z 457.3 (M+H).sup.+.
Example 43b
2-(3-hydroxypyridin-2-yl)-3-(1-(2-methoxyphenyl)propan-2-yl)-5-(trifluorom-
ethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00103##
[0407] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (d, 1H), 8.10
(m, 1H), 7.70 (d, 1H), 7.40 (m, 2H), 7.10 (t, 1H), 6.90 (d, 1H),
6.70 (d, 1H), 6.61 (t, 1H), 4.45 (m, 1H), 3.40 (s, 3H), 3.39 (m,
1H) 3.01 (m, 2H), 1.75 (d, 3H). MS m/z 457.3 (M+H).sup.+.
Example 44
3-(1-(2-fluorophenyl)butan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromet-
hyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00104##
[0409] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.75 (d, 1H), 8.07
(brs, 1H), 7.51 (d, 1H), 7.36 (t, 2H), 7.07 (m, 2H), 6.95 (m, 2H),
6.84 (m, 2H), 6.76 (t, 1H), 4.48 (m, 1H), 3.48 (dd, 1H), 3.24 (dd,
1H), 2.41 (m, 1H), 2.29 (m, 1H), 0.91 (t, 3H). MS m/z 458.3
(M+H).sup.+.
Example 45
3-(3-fluoro-2-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00105##
[0411] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (d, 1H), 8.18
(d, 1H), 7.80 (d, 1H), 7.45 (m, 2H), 6.99 (m, 1H), 6.85 (m, 1H),
6.79 (d, 1H), 4.25 (m, 2H), 3.60 (s, 3H), 2.99 (m, 2H). MS m/z
461.2 (M+H).sup.+.
Example 46
3-(2-cyclopentylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one
##STR00106##
[0413] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.64 (s, 1H),
8.87 (d, 1H), 8.15 (dd, 1H), 7.86 (d, 1H), 7.43 (m, 2H), 3.77 (brm,
2H), 1.48 (brm, 5H), 1.34 (brm, 4H), 0.69 (brm, 2H). MS m/z 405.2
(M+H).sup.+.
Example 47
3-(2-cyclohexylethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[-
4,3-d]pyrimidin-4(3H)-one
##STR00107##
[0415] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.62 (s, 1H),
8.87 (d, 1H), 8.15 (dd, 1H), 7.86 (d, 1H), 7.43 (m, 2H), 3.81 (m,
2H), 1.48 (brm, 5H), 1.50-1.25 (m, 7H), 1.00 (m, 4H), 0.58 (m, 2H).
MS m/z 419.2 (M+H).sup.+.
Example 48a
(R)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00108##
[0417] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.61 (s, 1H),
8.85 (d, 1H), 8.12 (dd, 1H), 7.83 (d, 1H), 7.39 (m, 2H), 3.97 (brs,
1H), 1.82 (m, 1H), 1.72 (m, 1H), 1.46 (m, 2H), 1.42 (d, 3H), 1.35
(m, 2H), 1.17-0.93 (m, 5H), 0.66 (m, 1H), 0.56 (m, 1H). MS m/z
433.3 (M+H).sup.+.
Example 48b
(S)-3-(1-cyclohexylpropan-2-yl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00109##
[0419] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.61 (s, 1H),
8.85 (d, 1H), 8.12 (dd, 1H), 7.83 (d, 1H), 7.39 (m, 2H), 3.97 (brs,
1H), 1.82 (m, 1H), 1.72 (m, 1H), 1.46 (m, 2H), 1.42 (d, 3H), 1.35
(m, 2H), 1.17-0.93 (m, 5H), 0.66 (m, 1H), 0.56 (m, 1H). MS m/z
433.3 (M+H).sup.+.
Example 49
(R,S)-2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-2-yl)ethyl)-5-(t-
rifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00110##
[0421] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.61 (s, 1H),
8.86 (d, 1H), 8.15 (dd, 1H), 7.85 (d, 1H), 7.41 (m, 2H), 3.94 (m,
1H), 3.79 (m, 1H), 3.56 (d, 1H), 3.10 (t, 1H), 3.02 (m, 1H), 1.57
(m, 3H), 1.27 (m, 3H), 1.07 (m, 4H), 0.93 (m, 1H). MS m/z 421.3
(M+H).sup.+.
Example 50
(R)-2-(3-fluoro-2-hydroxyphenyl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethy-
l)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00111##
[0423] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.80 (d, 1H), 7.60
(d, 1H), 7.2 (m, 5H), 6.90 (m, 1H), 6.80 (m, 1H), 6.67 (brs, 1H),
4.44 (m, 1H), 3.63 (m, 1H), 3.10 (m, 1H), 1.90 (d, 3H). MS m/z
444.3 (M+H).sup.+.
Example 51
(R)-2-(1H-imidazol-2-yl)-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido-
[4,3-d]pyrimidin-4(3H)-one
##STR00112##
[0425] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.79 (d, 1H), 7.58
(d, 1H), 7.4 (s, 1H), 7.30 (d, 1H), 7.10 (m, 4H), 6.98 (m, 1H),
3.70 (m, 1H), 3.30 (m, 1H), 1.90 (d, 3H). MS m/z 400.2
(M+H).sup.+.
Example 52
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4,-
3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide
##STR00113##
[0427] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.40 (brs, 1H),
8.98 (d, 1H), 8.80 (d, 1H), 8.45 (m, 1H), 7.67 (d, 1H), 7.44 (m,
1H), 7.10 (m, 1H), 7.00 (m, 1H), 6.99 (m, 1H), 6.85 (m, 1H), 4.60
(m, 2H), 3.05 (m, 2H), 2.10 (s, 3H). MS m/z 472.1 (M+H).sup.+.
Example 53
(R)-3-(1-phenylpropan-2-yl)-2-(thiazol-4-yl)-5(trifluoromethyl)pyrido[4,3--
d]pyrimidin-4(3H)-one
##STR00114##
[0429] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.99 (s, 1H), 8.89
(d, 1H), 7.60 (d, 1H), 7.59 (s, 1H), 7.10 (m, 3H), 6.98 (m, 2H),
4.61 (m, 1H), 3.61 (m, 2H), 3.10 (m, 2H), 1.90 (d, 3H). MS m/z
417.2 (M+H).sup.+.
Example 54
3-phenethyl-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3-
H)-one
##STR00115##
[0431] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.99 (s, 1H), 8.89
(d, 1H), 8.01 (s, 1H), 7.70 (d, 1H), 7.22 (m, 5H), 4.60 (m, 2H),
3.15 (m, 2H), 3.24 (m, 1H).
Example 55
3-(2-methoxyphenethyl)-2-(2-hydroxyphenyl)-5-(trifluoromethyl)pyrido[4,3-d-
]pyrimidin-4(3H)-one
##STR00116##
[0433] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.82 (d, 1H), 7.65
(d, 1H), 7.40 (m, 1H), 7.20 (m, 2H), 6.90 (m, 2H), 6.79 (d, 1H),
6.65 (m, 2H), 4.60 (t, 2H), 3.40 (s, 3H), 3.01 (t, 2H). MS m/z
442.3 (M+H).sup.+.
Example 56
3-(2-fluorophenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyr-
imidin-4(3H)-one
##STR00117##
[0435] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.99 (s, 1H), 8.89
(d, 1H), 7.90 (s, 1H), 7.79 (d, 1H), 7.22 (m, 5H), 4.68 (m, 2H),
3.15 (m, 2H). MS m/z 421.3 (M+H).sup.+.
Example 57
(R)-2-cyclopentyl-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl)pyrido[4,3-d]-
pyrimidin-4(3H)-one
##STR00118##
[0437] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.79 (d, 1H), 7.59
(d, 1H), 7.2 (m, 5H), 4.67 (brd, 1H), 3.69 (m, 1H), 3.30 (m, 1H),
3.0 (brm, 1H), 1.90 (m, 8H), 1.89 (d, 3H). MS m/z 402.3
(M+H).sup.+.
Example 58
2-isopropyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)-on-
e
##STR00119##
[0439] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.80 (d, 1H), 8.65
(d, 1H), 7.35 (m, 5H), 4.42 (m, 2H), 3.13 (m, 2H), 2.99 (m, 1H),
1.90 (d, 6H). MS m/z 362.3 (M+H).sup.+.
Example 59
2-cyclopentyl-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimidin-4(3H)--
one
##STR00120##
[0441] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.79 (d, 1H), 8.65
(d, 1H), 7.35 (m, 5H), 4.40 (m, 2H), 3.10 (m, 2H), 2.00-1.6 (m,
8H). MS m/z 388.3 (M+H).sup.+.
Example 60
3-(2-methoxyphenethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]py-
rimidin-4(3H)-one
##STR00121##
[0443] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.89 (t, 2H), 8.70
(d, 1H), 7.45 (s, 1H), 7.10 (m, 1H), 6.79 (m, 1H), 6.70 (m, 1H),
6.60 (d, 1H), 4.80 (m, 2H), 3.40 (s, 3H), 3.0 (t, 2H), MS m/z 433.4
(M+H).sup.+.
Example 61
3-(2-cyclohexylethyl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyr-
imidin-4(3H)-one
##STR00122##
[0445] .sup.1H NMR (500 MHz, CDCl.sub.3) .quadrature. ppm 0.78-0.95
(m, 2H), 1.07-1.25 (m, 3H), 1.24-1.34 (m, 1H), 1.56-1.74 (m, 7H),
4.35-4.52 (m, 2H), 7.75 (d, 1H), 8.22 (d, 1H), 8.86 (d, 1H), 8.98
(d, 1H). MS m/z 409.5 (M+H).sup.+.
Example 62
2-(3-(2-cyclohexylethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4,3-d-
]pyrimidin-2-yl)benzonitrile
##STR00123##
[0447] .sup.1H NMR (500 MHz, CDCl.sub.3) .quadrature. (ppm)
0.67-0.80 (m, 2H), 1.03-1.20 (m, 4H), 1.44 (d, 2H), 1.50-1.57 (m,
2H), 1.57-1.65 (m, 3H), 3.87-4.02 (m, 2H), 7.64 (d, 1H), 7.70-7.81
(m, 2H), 7.85 (t, 1H), 7.93 (d, 1H), 8.91 (d, 1H). MS m/z 427.5
(M+H).sup.+.
Example 63
2-(3-aminopyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyrimi-
din-4(3H)-one
##STR00124##
[0449] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.84 (d, 1H), 8.17
(q, 1H), 7.69 (d, 1H), 7.27 (m, 1H), 7.18 (m, 4H), 7.06 (m, 2H),
4.68 (s, br, 2H), 4.38 (m, 2H), 3.11 (m, 2H). MS m/z 412.4
(M+H).sup.+.
Example 64
N-(2-(4-oxo-3-phenethyl-5-(trifluoromethyl)-3,4-dihydropyrido[4,3-d]pyrimi-
din-2-yl)pyridin-3-yl)methanesulfonamide
##STR00125##
[0451] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.94 (s, 1H), 8.90
(d, 1H), 8.55 (dd, 1H), 8.12 (dd, 1H), 7.70 (d, 1H), 7.52 (dd, 1H),
7.22 (m, 3H), 7.07 (m, 2H), 4.44 (m, 2H), 3.19 (m, 2H), 3.11 (s,
3H). MS m/z 490.5 (M+H).sup.+.
Example 65
3-(1-(2-fluorophenyl)propan-2-yl)-2-(thiazol-4-yl)-5-(trifluoromethyl)pyri-
do[4,3-d]pyrimidin-4(3H)-one
##STR00126##
[0453] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.99 (s, 1H), 8.89
(d, 1H), 7.65 (d, 1H), 7.59 (s, 1H), 7.18 (m, 1H), 6.99 (m, 1H),
6.86 (m, 2H), 4.88 (m, 1H), 3.60 (m, 1H), 3.05 (m, 1H), 1.90 (d,
3H).
[0454] MS m/z 435.2 (M+H).sup.+.
Example 66
2-(3-hydroxypyridin-2-yl)-3-isopentyl-5-(trifluoromethyl)pyrido[4,3-d]pyri-
midin-4(3H)-one
##STR00127##
[0456] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.63 (s, 1H),
8.87 (d, 1H), 8.16 (m, 1H), 7.86 (d, 1H), 7.42 (m, 2H), 3.79 (t,
2H), 1.40-1.28 (m, 3H), 0.58 (d, 6H). MS m/z 379.2 (M+H).sup.+.
Example 67
2-(3-hydroxypyridin-2-yl)-3-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluor-
omethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00128##
[0458] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.66 (s, 1H),
8.87 (d, 1H), 8.16 (dd, 1H), 7.86 (d, 1H), 7.42 (m, 2H), 3.79 (d,
2H), 3.64 (dd, 2H), 3.03 (dt, 2H), 1.76 (m, 1H), 1.24 (d, 2H), 0.94
(dq, 2H). MS m/z 407.3 (M+H).sup.+.
Example 68
2-(3-hydroxypyridin-2-yl)-3-(2-(tetrahydro-2H-pyran-4-yl)ethyl)-5-(trifluo-
romethyl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00129##
[0460] .sup.1H NMR (400 MHz, dmso-d.sub.6) .delta. 10.68 (s, 1H),
8.91 (d, 1H), 8.19 (dd, 1H), 7.80 (d, 1H), 7.47 (m, 2H), 3.86 (t,
2H), 3.66 (dd, 2H), 3.12 (t, 2H), 1.46 (q, 2H), 1.26 (m, 3H), 0.86
(dq, 2H). MS m/z 421.2 (M+H).sup.+.
Example 69
3-(2-fluoro-5-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00130##
[0462] MS m/z 461 (M+H).sup.+.
Example 70
3-(2-fluoro-3-methoxyphenethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluorometh-
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00131##
[0464] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.90 (d, 1H), 8.20
(d, 1H), 7.89 (d, 1H), 7.56 (m, 2H), 6.90 (t, 1H), 6.76 (m, 1H),
6.56 (d, 1H), 4.30 (m, 2H), 3.64 (s, 3H), 2.99 (m, 2H). MS m/z
461.2 (M+H).sup.+.
Example 71
2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydroquinazolin-2-
-yl)benzonitrile
##STR00132##
[0466] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.68 (t, 1H), 7.64
(t, 1H), 7.25 (m, 6H), 7.14 (m, 1H), 7.01 (t, 1H), 6.96 (t, 1H),
4.32 (t, 2H), 3.06 (t, 2H). MS m/z 438.0 (M+H).sup.+.
Example 72
N-(2-(3-(2-fluorophenethyl)-4-oxo-5-(trifluoromethyl)-3,4-dihydropyrido[4,-
3-d]pyrimidin-2-yl)pyridin-3-yl)-2,2,2-trifluoroacetamide
##STR00133##
[0468] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.45 (s, 1H),
9.99 (d, 1H), 8.79 (d, 1H), 8.55 (d, 1H), 7.76 (d, 1H), 7.50 (m,
1H), 7.15 (m, 1H), 6.90 (m, 1H), 6.70 (m, 1H), 4.99 (m, 2H), 3.05
(m, 2H). MS m/z 526.3 (M+H).sup.+.
Example 73
(R)-2,2,2-trifluoro-N-(2-(4-oxo-3-(1-phenylpropan-2-yl)-5-(trifluoromethyl-
)-3,4-dihydropyrido[4,3-d]pyrimidin-2-yl)pyridin-3-yl)acetamide
##STR00134##
[0470] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.65 (s, 1H),
9.99 (d, 1H), 8.79 (d, 1H), 8.60 (d, 1H), 7.66 (m, 1H), 7.44 (d,
1H), 7.00 (m, 2H), 6.79 (m, 2H), 4.69 (m, 1H), 3.70 (m, 1H), 2.95
(m, 1H), 2.01 (d, 3H). MS m/z 520.3 (M+H).sup.+.
Example 74
3-cyclohexyl-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[4,3-d]pyr-
imidin-4(3H)-one
##STR00135##
[0472] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.99 (d, 1H), 8.25
(d, 1H), 7.77 (d, 1H), 7.43 (m, 2H), 4.80 (m, 1H), 2.65 (m, 2H),)
1.50-1.25 (m, 8H). MS m/z 391.3 (M+H).sup.+.
Example 75
3-(cyclohexylmethyl)-2-(3-hydroxypyridin-2-yl)-5-(trifluoromethyl)pyrido[4-
,3-d]pyrimidin-4(3H)-one
##STR00136##
[0474] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 12.39 (brs, 1H),
8.99 (d, 1H), 8.30 (d, 1H), 7.70 (d, 1H), 7.43 (m, 2H), 5.0 (brs,
2H), 1.50-1.25 (m, 10H). MS m/z 405.4 (M+H).sup.+.
Example 76
3-Phenethyl-5-trifluoromethyl-2-(3-trifluoromethyl-pyridin-2-yl)-3H-pyrido-
[4,3-d]pyrimidin-4-one
##STR00137##
[0476] .sup.1H NMR (400 MHz, CD.sub.3OD).quadrature. 9.02 (d, 1H);
8.91-8.92 (d, 1H); 8.25-8.27 (d, 1H); 7.78-7.79, (m, 2H); 7.22-7.23
(m, 3H); 7.0-7.02 (d, 2H); 3.95 (br, 2H), 3.10-3.14 (t, 2H). MS m/z
465.5 (M+H).sup.+.
Example 77
##STR00138##
[0478] .sup.1H NMR (400 MHz, CD.sub.3OD) .quadrature. 8.88 (d, 1H);
8.87 (s, 1H); 8.22-8.24 (d, 1H); 7.73, 7.72, 7.69 (t, 2H); 7.20,
7.19 (d, 3H); 6.98, 6.96 (d, 2H); 3.88 (m, 1H); 3.64 (br, 1H), 3.36
(br, 1H); 1.59 (s, 3H); 1.55 (br, 1H). LC/MS 479.5 (M+H).sup.+.
Example 78
2-(5-aminothiazol-4-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]pyridi-
min-4(3H)-one
##STR00139##
[0480] To a solution of
2-(5-(2,5-dimethyl-1H-pyrrol-1-yl)thiazol-4-yl)-3-phenethyl-5-(trifluorom-
ethyl)pyrido[4,3-d]pyrimidin-4(3H)-one (7 mg, 0.014 mmol) in
ethanol was added hydroxylamine hydrochloride (9.82 mg, 0.14 mmol)
and the mixture was heated up to 80.degree. C. for 60 hours. The
reaction was then concentrated and purified with flash
chromatography providing an off-white solid (1.5 mg). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.72 (d, 1H), 8.02 (s, 1H), 7.52 (d,
1H), 7.30 (m, 5H), 6.93 (s, br, 2H), 4.89 (t, 2H), 3.17 (t, 2H). MS
m/z 418.3 (M+H).sup.+.
Example 79
2-(2-(difluoromethyl)phenyl)-3-phenethyl-5-(trifluoromethyl)pyrido[4,3-d]p-
yrimidin-4(3H)-one
##STR00140##
[0482] Prepared analogous to Example 1 starting from
2-(difluoromethyl)benzaldehyde and
4-Amino-N-phenethyl-2-(trifluoromethyl)nicotinamide. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.88 (d, 1H), 7.77 (d, 1H), 7.72 (d,
1H), 7.69 (t, 1H), 7.61 (t, 1H), 7.19 (m, 4H), 6.89 (m, 2H), 6.77
(t, 1H), 4.28 (m, 1H), 3.78 (m, 1H), 2.95 (m, 2H). MS m/z 446.4
(M+H).sup.+.
Example 80
2-(3-(difluoromethyl)pyridin-2-yl)-3-phenethyl-5-(trifluoromethyl)pyrido[4-
,3-d]pyrimidin-4(3H)-one
##STR00141##
[0484] Prepared analogous to Example 1 starting from
2-(difluoromethyl)picolinaldehyde and
4-Amino-N-phenethyl-2-(trifluoromethyl)nicotinamide. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.88 (d, 1H), 8.19 (d, 1H), 7.72 (d,
1H), 7.67 (m, 1H), 7.19 (m, 4H), 7.02 (m, 2H), 4.13 (m, 2H), 3.12
(m, 2H). MS m/z 447.5 (M+H).sup.+.
Example 81
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(2-(trifluoromethyl)phenyl)pyr-
ido[4,3-d]pyrimidin-4(3H)-one
##STR00142##
[0486] .sup.1H NMR (500 MHz, CDCl.sub.3) .quadrature. ppm 0.55-0.71
(m, 1H) 0.73-0.86 (m, 1H) 1.03-1.19 (m, 3H) 1.24-1.41 (m, 2H)
1.46-1.67 (m, 5H) 1.68-1.83 (m, 1H) 3.38-3.52 (m, 1H) 4.07-4.25 (m,
1H) 7.54 (d, 1H) 7.69-7.84 (m, 3H) 7.90 (d, 1H) 8.89 (d, 1H). MS
m/z 470.5 (M+H).sup.+.
Example 82
3-(2-cyclohexylethyl)-5-(trifluoromethyl)-2-(3-(trifluoromethyl)pyridin-2--
yl)pyrido[4,3-d]pyrimidin-4(3H)-one
##STR00143##
[0488] .sup.1H NMR (500 MHz, CDCl.sub.3) .quadrature. ppm 0.66-0.82
(m, 2H), 1.02-1.20 (m, 4H), 1.46 (d, 2H), 1.54-1.71 (m, 5H), 3.86
(br. s., 2H), 7.72 (dd, 1H), 7.76 (d, 1H), 8.25 (d, 1H), 8.89 (d,
1H), 8.96 (d, 1H). MS m/z 471.5 (M+H).sup.+.
[0489] The following table provides FLIPR IC.sub.50 data for the
specified Examples. The IC.sub.50s are reported as micromolar
concentration with n being the number of times the particular
compound was assayed.
TABLE-US-00001 TABLE of FLIPR assay data FLIPR IC.sub.50 Example n=
(uM) 1 6 0.05 2 6 0.62 3 4 0.39 4a 11 0.06 4b 3 0.16 5 4 0.02 6 4
0.05 6a 5 0.05 6b 3 1.40 7 6 0.13 8 5 0.09 9 6 0.29 10 6 0.12 11 5
0.46 12 12 0.25 13 8 0.07 14 8 0.17 15 4 0.16 16 3 10.8 17 2 11.0
18 5 12.3 19 1 >100 20 1 >100 21 2 13.8 22 1 >100 23 3
13.7 24 1 2.60 25 1 3.01 26 1 2.99 27 1 2.26 28 1 4.83 29 4 0.49 30
6 1.17 31 3 1.93 32 5 0.57 33 5 0.52 34 3 1.47 35 6 0.38 36 2 0.60
37a 11 .008 37b 3 2.12 38a 3 0.02 38b 3 4.69 39 8 0.11 40 3 0.46 41
7 0.25 42 3 1.12 43 2 1.12 43a 2 0.53 43b 4 61.3 44 4 2.44 45 2
1.87 46 3 0.29 47 6 0.06 48a 2 0.16 48b 2 1.69 49 2 0.48 50 8 0.03
51 4 0.11 52 2 0.23 53 6 0.26 54 4 0.71 55 5 0.72 56 6 1.54 57 2
1.67 58 4 4.68 59 4 7.79 60 4 23.2 61 2 0.50 62 2 4.63 63 2 0.77 64
2 61.8 65 4 10.5 65a 4 0.32 66 2 36.2 67 1 >1.00 68 1 >1.00
69 2 66.9 70 2 61.9 71 4 0.42 72 4 0.94 73 4 0.20 74 2 47.8 75 2
10000 76 3 6.30 77 7 0.70 78 2 9.83 79 3 0.39 80 2 0.30 81 2 7.89
82 2 2.53
[0490] All documents cited in this application, including
scientific publications, patents and patent applications, are
hereby incorporated by reference in their entirety.
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