U.S. patent application number 15/541040 was filed with the patent office on 2018-05-03 for p2x7 modulators.
This patent application is currently assigned to Janssen Pharmaceutica NV. The applicant listed for this patent is Jansssen Pharmaceutica NV. Invention is credited to Jose Ignacio ANDRES GIL, Christa C. Chrovian, Akinola Soyode Johnson, Michael A. Letavic, Jason C. Rech, Dale A. Rudolph.
Application Number | 20180118749 15/541040 |
Document ID | / |
Family ID | 54062820 |
Filed Date | 2018-05-03 |
United States Patent
Application |
20180118749 |
Kind Code |
A1 |
ANDRES GIL; Jose Ignacio ;
et al. |
May 3, 2018 |
P2X7 MODULATORS
Abstract
The present invention is directed to compounds of Formula (I),
which includes enantiomer and diasteromers thereof: These compounds
are suitable for use in the treatment of diseases associated with
P2X7 receptor activity such as diseases of the autoimmune and
inflammatory system, diseases of the nervous and neuro-immune
system, diseases involved with neuroinflammation of the Central
Nervous System (CNS) or diseases of the cardiovascular, metabolic,
gastrointestinal and urogenital systems. ##STR00001##
Inventors: |
ANDRES GIL; Jose Ignacio;
(Madrid, ES) ; Chrovian; Christa C.; (La Jolla,
CA) ; Letavic; Michael A.; (San Diego, CA) ;
Rech; Jason C.; (San Diego, CA) ; Rudolph; Dale
A.; (San Diego, CA) ; Johnson; Akinola Soyode;
(San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Jansssen Pharmaceutica NV |
Beerse |
|
BE |
|
|
Assignee: |
Janssen Pharmaceutica NV
Beerse
BE
|
Family ID: |
54062820 |
Appl. No.: |
15/541040 |
Filed: |
August 26, 2015 |
PCT Filed: |
August 26, 2015 |
PCT NO: |
PCT/US2015/046852 |
371 Date: |
June 30, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62049727 |
Sep 12, 2014 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 25/08 20180101;
A61P 25/16 20180101; A61P 33/00 20180101; A61P 25/04 20180101; A61P
11/00 20180101; A61P 11/06 20180101; A61P 25/18 20180101; A61P
13/10 20180101; A61P 31/04 20180101; A61P 7/02 20180101; A61P 25/24
20180101; A61P 35/00 20180101; A61P 13/12 20180101; A61P 3/10
20180101; C07D 487/04 20130101; A61P 9/12 20180101; A61P 25/22
20180101; A61P 17/06 20180101; A61P 27/06 20180101; A61P 31/00
20180101; A61P 17/00 20180101; A61P 17/10 20180101; A61P 19/10
20180101; A61P 25/28 20180101; A61P 1/00 20180101; A61P 37/08
20180101; A61P 19/02 20180101; A61P 25/00 20180101 |
International
Class: |
C07D 487/04 20060101
C07D487/04; A61P 27/06 20060101 A61P027/06; A61P 13/12 20060101
A61P013/12; A61P 33/00 20060101 A61P033/00; A61P 31/04 20060101
A61P031/04; A61P 35/00 20060101 A61P035/00; A61P 17/10 20060101
A61P017/10; A61P 19/02 20060101 A61P019/02; A61P 13/10 20060101
A61P013/10; A61P 17/06 20060101 A61P017/06; A61P 31/00 20060101
A61P031/00; A61P 17/00 20060101 A61P017/00; A61P 11/06 20060101
A61P011/06; A61P 11/00 20060101 A61P011/00; A61P 37/08 20060101
A61P037/08; A61P 25/04 20060101 A61P025/04; A61P 25/00 20060101
A61P025/00; A61P 25/24 20060101 A61P025/24; A61P 25/18 20060101
A61P025/18; A61P 25/22 20060101 A61P025/22; A61P 25/28 20060101
A61P025/28; A61P 25/08 20060101 A61P025/08; A61P 25/16 20060101
A61P025/16; A61P 3/10 20060101 A61P003/10; A61P 7/02 20060101
A61P007/02; A61P 1/00 20060101 A61P001/00; A61P 9/12 20060101
A61P009/12; A61P 19/10 20060101 A61P019/10 |
Claims
1. A compound of Formula (I): ##STR00225## and enantiomers or
diastereomers thereof; and pharmaceutically acceptable salts
thereof; wherein: R.sup.a is ##STR00226## R.sup.1 is halo or
C.sub.1-C.sub.3alkyl; R.sup.2 is independently selected from the
group consisting of: H, halo, and C.sub.1-C.sub.3perhaloalkyl;
R.sup.3 is H or halo; R.sup.4 is halo, R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; R.sup.b is independently selected from
the group consisting of: ##STR00227## Wherein: R.sup.6, R.sup.9,
R.sup.10, and R.sup.12are independently H or halo; R.sup.7 and
R.sup.13 are independently selected from the group consisting of:
H, halo and OC.sub.1-C.sub.3alkyl; R.sup.8 is independently
selected from the group consisting of: H, halo, OH and
OC.sub.1-C.sub.3alkyl; R.sup.11 is independently selected from the
group consisting of: H, halo and C.sub.1-C.sub.3perhaloalkyl;
R.sup.c is selected from the group consisting of: ##STR00228##
R.sup.d and R.sup.e are independently H or C.sub.1-C.sub.3alkyl;
and provided that at least one of R.sup.c, R.sup.d and R.sup.e are
not H.
2. A compound as claimed in claim 1 wherein, R.sup.a is
##STR00229##
3. A compound as claimed in claim 1 wherein, R.sup.a is
##STR00230##
4. A compound as claimed in claim 1 wherein, R.sup.a is
##STR00231## and R.sup.1 is halo.
5. A compound as claimed in claim 1 wherein R.sup.a is ##STR00232##
and R.sup.1 is C.sub.1-C.sub.3alkyl.
6. A compound as claimed in claim 1 wherein R.sup.a is ##STR00233##
and R.sup.2 is C.sub.1-C.sub.3perhaloalkyl.
7. A compound as claimed in claim 1 wherein R.sup.a is ##STR00234##
and R.sup.2 is halo.
8. A compound as claimed in claim 1 wherein R.sup.a is ##STR00235##
and R.sup.3 is H.
9. A compound as claimed in claim 1 wherein R.sup.a is ##STR00236##
R.sup.1 is halo, R.sup.2 is C.sub.1-C.sub.3perhaloalkyl, and
R.sup.3 is H.
10. A compound as claimed in claim 1 wherein R.sup.a is
##STR00237## and R.sup.1, R.sup.2, and R.sup.3 are halo.
11. A compound as claimed in claim 1 wherein R.sup.a is
##STR00238## R.sup.1 and R.sup.3 are halo and R.sup.2 is H.
12. A compound as claimed in claim 1 wherein R.sup.a is
##STR00239## R.sup.1 and R.sup.2 are halo and R.sup.3 is H.
13. A compound as claimed in claim 1 wherein R.sup.a is
##STR00240## R.sup.4 is halo and R.sup.5 is
C.sub.1-C.sub.3perhaloalkyl.
14. A compound as claimed in claim 1 wherein R.sup.b is
independently selected from the group consisting of:
##STR00241##
15. A compound as claimed in claim 1 wherein R.sup.b is
independently selected from the group consisting of:
##STR00242##
16. A compound as claimed in claim 1 wherein R.sup.b is
independently selected from the group consisting of:
C.sub.3-C.sub.6 cycloalkyl and C.sub.1-C.sub.4 alkyl.
17. A compound as claimed in claim 1 wherein R.sup.b is
##STR00243##
18. A compound as claimed in claim 1 wherein R.sup.b is
##STR00244##
19. A compound as claimed in claim 1 wherein R.sup.b is
##STR00245##
20. A compound as claimed in claim 1 wherein R.sup.b is
##STR00246## R.sup.6 and R.sup.7 are H and R.sup.8 is
OCH.sub.3.
21. A compound as claimed in claim 1 wherein R.sup.b is
##STR00247## and R.sup.6, R.sup.7 and R.sup.8 are H.
22. A compound as claimed in claim 1 wherein R.sup.b is
##STR00248## and R.sup.8 is OH, and R.sup.6 and R.sup.7 are H.
23. A compound as claimed in claim 1 wherein R.sup.b is
##STR00249##
24. A compound as claimed in claim 1 wherein R.sup.b is
##STR00250##
25. A compound as claimed in claim 1 wherein R.sup.b is
##STR00251## R.sup.9, R.sup.10 and R.sup.12 are H and R.sup.11 is
F.
26. A compound as claimed in claim 1 wherein R.sup.b is
##STR00252##
27. A compound as claimed in claim 1 wherein R.sup.b is
##STR00253##
28. A compound as claimed in claim 1 wherein R.sup.c is H or
CH.sub.3.
29. A compound as claimed in claim 1 wherein R.sup.c is selected
from the group consisting of: ##STR00254##
30. A compound as claimed in claim 1 wherein R.sup.c is
##STR00255##
31. A compound as claimed in claim 1 wherein R.sup.c is:
##STR00256##
32. A compound as claimed in claim 1 wherein R.sup.d is
CH.sub.3.
33. A compound as claimed in claim 1 wherein R.sup.e is
CH.sub.3.
34. A compound as claimed in claim 1 wherein R.sup.c is CH.sub.3
and R.sup.d and R.sup.e are H.
35. A compound as claimed in claim 1 wherein R.sup.d is CH.sub.3
and R.sup.c and R.sup.e are H.
36. A compound as claimed in claim 1 wherein R.sup.e is CH.sub.3
and R.sup.c and R.sup.d are H.
37. A compound as claimed in claim 1 wherein R.sup.a is
##STR00257## R.sup.1 and R.sup.2 are Cl, R.sup.c is CH.sub.3,
R.sup.b is ##STR00258## and R.sup.d, R.sup.e, R.sup.3, R.sup.6,
R.sup.7 and R.sup.8 are H.
38. A compound as claimed in claim 1 wherein R.sup.a is
##STR00259## R.sup.1 and R.sup.2 are Cl, R.sup.d is CH.sub.3,
R.sup.b ##STR00260## is R.sup.c, R.sup.e, R.sup.3, R.sup.9,
R.sup.19 and R.sup.12 are H and R.sup.11 is F.
39. A compound as claimed in claim 1 wherein R.sup.a is
##STR00261## R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is
CH.sub.3, R.sup.b is ##STR00262## R.sup.c, R.sup.e, R.sup.3,
R.sup.9, R.sup.19 and R.sup.12 are H and R.sup.11 is F.
40. A compound as claimed in claim 1 wherein R.sup.a is
##STR00263## R.sup.1 and R.sup.2 are Cl, R.sup.d is CH.sub.3,
R.sup.b is ##STR00264## R.sup.8 is OCH.sub.3, and R.sup.c, R.sup.e,
R.sup.3, R.sup.6, and R.sup.7 are H.
41. A compound as claimed in claim 1 wherein R.sup.a is
##STR00265## R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is
CH.sub.3, R.sup.c is ##STR00266## R.sup.b is ##STR00267## and
R.sup.d, R.sup.e, and R.sup.3 are H.
42. A compound as claimed in claim 1 wherein R.sup.a is
##STR00268## R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is
CH.sub.3, R.sup.c is ##STR00269## R.sup.b is ##STR00270## and
R.sup.d, R.sup.e, and R.sup.3 are H.
43. A compound as claimed in claim 1 wherein R.sup.a is
##STR00271## R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is
CH.sub.3, R.sup.c is ##STR00272## R.sup.b is ##STR00273## and
R.sup.d, R.sup.e, and R.sup.3 are H.
44. A compound selected from the group consisting of:
(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]triazol-
o[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
2-chloro-3-(trifluoromethyl)phenyl)(5-methyl-3-phenyl-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2,3-dichlorophenyl)(5-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]-
pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-5-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2,3-dichlorophenyl)(3-(4-fluorophenyl)-5-methyl-5,6-dihydro-[1,2,4]triaz-
olo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]triaz-
olo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4-
]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]-
pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,-
3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,-
3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-methyl-3-(pyridin-2-yl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-fluoro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-di-
hydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-8-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(4-(trifluoromethyl)p-
henyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(3-fluoro-4-(trifluoromethyl)phenyl)-6-methyl--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluoro-4-(trifluoromethyl)p-
henyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methano-
ne;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(3,4,5-trifluoroph-
enyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichlorophenyl)(5-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]triazol-
o[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(4-(trifluoromethyl)pheny-
l)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyrazin-2-yl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-chlorophenyl)-8-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2-fluorophenyl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluorophenyl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,3-difluorophenyl)-6-methyl--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3,4-difluorophenyl)-6-methyl--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,4-difluorophenyl)-6-methyl--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,2,4-
]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(6-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyra-
zin-7(8H)-yl)methanone;
(3,4-difluoro-2-methylphenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazo-
lo[4,3-a]pyrazin-7(8H)-yl)methanone;
(2-chloro-4-fluorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4-
,3-a]pyrazin-7(8H)-yl)methanone;
(2,3-dichloropyridin-4-yl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[-
4,3-a]pyrazin-7(8H)-yl)methanone;
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl-
)(2,3-dichlorophenyl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclohexyl-8-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl-
)(2,3-dichloro-4-fluorophenyl)methanone;
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-y-
l)(2,3-dichlorophenyl)methanone;
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-y-
l)(2,3-dichloro-4-fluorophenyl)methanone;
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-fluoropyridin-2-yl)-6-methy-
l-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
((S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyridin-2-yl)-6-met-
hyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(5-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyrimidin-2-yl)-6-me-
thyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(1-hydroxyethyl)-8-phenyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(-
8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone;
(S)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(-
8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-di-
hydro-[[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-di-
hydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluorom-
ethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluorom-
ethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-(trifluorometh-
yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-(trifluorometh-
yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-fluorophenyl)-
-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-fluorophenyl)-
-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazi-
n-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone;
(S)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazi-
n-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone;
(R)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone; (.+-.)-(8-(1
H-pyrazol-5-yl)-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyr-
azin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(pyridin-3-yl)-3-(trifluoro-
methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(R)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-8-phenyl-3-(trifluor-
omethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(.+-.)-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyraz-
in-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-hydroxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone;
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.11C]methoxypyridin-2-yl)-6-methyl-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone and
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.18F]fluoropyridin-2-yl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone.
45. A pharmaceutical composition, comprising: (a) a therapeutically
effective amount of at least one compound independently selected
from compounds of Formula (I): ##STR00274## and enantiomers or
diastereomers thereof; and pharmaceutically acceptable salts
thereof; wherein: R.sup.a is ##STR00275## R.sup.1 is halo or
C.sub.1-C.sub.3alkyl; R.sup.2 is independently selected from the
group consisting of: H, halo, and C.sub.1-C.sub.3perhaloalkyl;
R.sup.3 is H or halo; R.sup.4 is halo, R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; R.sup.b is independently selected from
the group consisting of: ##STR00276## Wherein: R.sup.6, R.sup.9,
R.sup.10, R.sup.12, R.sup.14 are independently H or halo; R.sup.7
and R.sup.13 are independently selected from the group consisting
of: H, halo and OC.sub.1-C.sub.3alkyl; R.sup.8 is independently
selected from the group consisting of: H, halo, OH and
OC.sub.1-C.sub.3alkyl; R.sup.11 is independently selected from the
group consisting of: H, halo and C.sub.1-C.sub.3perhaloalkyl;
R.sup.c is selected from the group consisting of: ##STR00277##
R.sup.d and R.sup.e are independently H or C.sub.1-C.sub.3alkyl;
and provided that at least one of R.sup.c, R.sup.d and R.sup.e are
not H; and (b) at least one pharmaceutically acceptable
excipient.
46. A pharmaceutical composition comprising a therapeutically
effective amount of at least one compound of claim 44 and at least
one pharmaceutically acceptable excipient.
47. A method of treating a subject suffering from or diagnosed with
a disease, disorder, or medical condition mediated by P2X7 receptor
activity, comprising administering to a subject in need of such
treatment an effective amount of at least one compound selected
from compounds of Formula (I): ##STR00278## and enantiomers or
diastereomers thereof; and pharmaceutically acceptable salts
thereof; wherein: R.sup.a is ##STR00279## R.sup.1 is halo or
C.sub.1-C.sub.3alkyl; R.sup.2 is independently selected from the
group consisting of: H, halo, and C.sub.1-C.sub.3perhaloalkyl;
R.sup.3 is H or halo; R.sup.4 is halo, R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; R.sup.b is independently selected from
the group consisting of: ##STR00280## Wherein: R.sup.6, R.sup.9,
R.sup.10, R.sup.12, R.sup.14 are independently H or halo; R.sup.7
and R.sup.13 are independently selected from the group consisting
of: H, halo and OC.sub.1-C.sub.3alkyl; R.sup.8 is independently
selected from the group consisting of: H, halo, OH and
OC.sub.1-C.sub.3alkyl; R.sup.11 is independently selected from the
group consisting of: H, halo and C.sub.1-C.sub.3perhaloalkyl;
R.sup.c is selected from the group consisting of: ##STR00281##
R.sup.d and R.sup.e are independently H or C.sub.1-C.sub.3alkyl;
and provided that at least one of R.sup.c, R.sup.d and R.sup.e are
not H.
48. A method according to claim 47, wherein the disease, disorder,
or medical condition is selected from the group consisting of:
diseases of the autoimmune and inflammatory system; diseases of the
nervous and neuro-immune system; diseases involved with, and
without, neuroinflammation of the Central Nervous System (CNS);
diseases of the cardiovascular, metabolic, gastrointestinal and
urogenital systems; skeletal disorders, diseases involving the
secretory function of exocrine glands and glaucoma,
Glomerulonephritis, Chaga's Disease, chlamydia, neuroblastoma,
Tuberculosis, Polycystic Kidney Disease, cancer, and acne.
49. A method according to claim 47 wherein the disease, disorder,
or medical condition is selected from the group consisting of:
rheumatoid arthritis, osteoarthritis, interstitial cystitis,
psoriasis, septic shock, sepsis, allergic dermatitis, asthma,
allergic asthma, mild to severe asthma, and steroid resistant
asthma, idiopathic pulmonary fibrosis, allergic rhinitis, chronic
obstructive pulmonary disease and airway hyper-responsivenes; acute
and chronic pain,neuropathic pain, inflammatory pain, migraine,
spontaneous pain, opioid induced pain, diabetic neuropathy,
postherpetic neuralgia, low back pain, chemotherapy-induced
neuropathic pain, fibromyalgia; mood disorders, major depression,
major depressive disorder, treatment resistant depression, bipolar
disorder, anxious depression, anxiety, cognition, sleep disorders,
multiple sclerosis, epileptic seizures, Parkinson's disease,
schizophrenia, Alzheimer's disease, Huntington's disease,
Amyotrophic Lateral Sclerosis, autism, spinal cord injury and
cerebral ischemia/traumatic brain injury, and stress-related
disorders; diabetes, diabetes mellitus, thrombosis, irritable bowel
disease, irritable bowel syndrome, Crohn's disease, cardiovascular
diseases (examples of cardiovascular disease include hypertension,
myocardial infarction, ischemic heart disease, ischemia, ureteric
obstruction, lower urinary tract syndrome, lower urinary tract
dysfunction such as incontinence, and disease after cardiac
transplantation, osteoporosis/osteopetrosis, diseases involving the
secretory function of exocrine glands, glaucoma,
Glomerulonephritis, Chaga's Disease, chlamydia, neuroblastoma,
Tuberculosis, Polycystic Kidney Disease, cancer, and acne.
50. A method according to claim 48, wherein the disease, disorder,
or medical condition is diseases of the autoimmune and inflammatory
system.
51. A method according to claim 50, wherein the diseases of the
autoimmune and inflammatory system selected from the group
consisting of rheumatoid arthritis, osteoarthritis, interstitial
cystitis, psoriasis, septic shock, sepsis, allergic dermatitis,
asthma, idiopathic pulmonary fibrosis, allergic rhinitis, chronic
obstructive pulmonary disease and airway hyper-responsivenes
52. The method of claim 48, wherein the disease, disorder or
medical condition is a disease involved with, and without,
neuroinflammation of the Central Nervous System (CNS).
53. A method according to claim 52, wherein the diseases involved
with, and without, neuroinflammation of the Central Nervous System
(CNS) is selected from the group consisting of: mood disorders,
cognition, sleep disorders, multiple sclerosis, epileptic seizures,
Parkinson's disease, schizophrenia, Alzheimer's disease,
Huntington's disease, Amyotrophic Lateral Sclerosis, autism, spinal
cord injury and cerebral ischemia/traumatic brain injury, and
stress-related disorders
54. The method of claim 53, wherein wherein the mood disorder
selected from the group consisting of: major depression, major
depressive disorder, treatment resistant depression, bipolar
disorder, anxious depression, and anxiety.
55. The method of claim 54, wherein wherein the mood disorder is
treatment resistant depression.
Description
FIELD OF THE INVENTION
[0001] The present invention is related to compounds having P2X7
modulating properties, pharmaceutical compositions comprising these
compounds, chemical processes for preparing these compounds and
their use in the treatment of diseases associated with P2X7
receptor activity in animals, in particular humans.
BACKGROUND OF THE INVENTION
[0002] The P2X7 receptor is a ligand-gated ion channel and is
present on a variety of cell types, largely those known to be
involved in the inflammatory and/or immune process, specifically,
macrophages and monocytes in the periphery and predominantly in
glial cells (microglia and astrocytes) of the CNS. (Duan and Neary,
Glia 2006, 54, 738-746; Skaper et al., FASEB J 2009, 24, 337-345;
Surprenant and North, Annu. Rev. Physiol. 2009, 71, 333-359).
Activation of the P2X7 receptor by extracellular nucleotides, in
particular adenosine triphosphate, leads to the release of
proinflammatory cytokines IL-1.beta. and IL-18 (Muller, et. Al. Am.
J. Respir. Cell Mol. Biol. 2011, 44, 456-464), giant cell formation
(macrophages/microglial cells), degranulation (mast cells) and
L-selectin shedding (lymphocytes) (Ferrari et al., J. Immunol.
2006, 176, 3877-3883; Surprenant and North, Annu. Rev. Physiol.
2009, 71, 333-359). P2X7 receptors are also located on
antigen-presenting cells (keratinocytes, salivary acinar cells
(parotid cells)), hepatocytes, erythrocytes, erythroleukaemic
cells, monocytes, fibroblasts, bone marrow cells, neurones, and
renal mesangial cells.
[0003] The importance of P2X7 in the nervous system arises
primarily from experiments using P2X7 knockout mice. These mice
demonstrate the role of P2X7 in the development and maintenance of
pain, as these mice are protected from the development of both
adjuvant-induced inflammatory pain and partial nerve
ligation-induced neuropathic pain (Chessell et al., Pain 2005, 114,
386-396). In addition, P2X7 knockout mice also exhibit an
anti-depressant phenotype based on reduced immobility in forced
swim and tail suspension tests (Basso et al., Behav. Brain Res.
2009, 198, 83-90.). Moreover, the P2X7 pathway is linked to the
release of the pro-inflammatory cytokine, IL-1.beta., which has
been linked to precipitation of mood disorders in humans (Dantzer,
Immunol. Allergy Clin. North Am. 2009, 29, 247-264; Capuron and
Miller, Pharmacol. Ther. 2011, 130, 226-238). In addition, in
murine models of Alzheimer's disease, P2X7 was upregulated around
amyloid plaques indicating a role of this target in such pathology
as well (Parvathenani et al., J. Biol. Chem. 2003, 278,
13309-13317).
[0004] Several reviews on small molecule inhibitors of P2X7 which
have been published are: Guile, S. D., et al., J. Med. Chem, 2009,
52, 3123-3141; Gunosewoyo, H. and Kassiou, M., Exp Opin, 2010, 20,
625-646.
[0005] In view of the clinical importance of P2X7, the
identification of compounds that modulate P2X7 receptor function
represents an attractive avenue into the development of new
therapeutic agents. Such compounds are provided herein.
SUMMARY OF THE INVENTION
[0006] The invention is directed to the general and preferred
embodiments defined, respectively, by the independent and dependent
claims appended hereto, which are incorporated by reference herein.
One aspect of this invention concerns compounds of Formula (I):
##STR00002## [0007] and enantiomers or diastereomers thereof;
[0008] and pharmaceutically acceptable salts thereof; [0009]
wherein: [0010] R.sup.a is
[0010] ##STR00003## [0011] R.sup.1 is halo or C.sub.1-C.sub.3alkyl;
[0012] R.sup.2 is independently selected from the group consisting
of: H, halo, and C.sub.1-C.sub.3perhaloalkyl; [0013] R.sup.3 is H
or halo; [0014] R.sup.4 is halo, [0015] R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; [0016] R.sup.b is independently
selected from the group consisting of:
[0016] ##STR00004## [0017] Wherein: [0018] R.sup.6, R.sup.9,
R.sup.10, R.sup.12, R.sup.14 are independently H or halo; [0019]
R.sup.7, R.sup.8, R.sup.13 is independently selected from the group
consisting of: H, halo and OC.sub.1-C.sub.3alkyl; [0020] R.sup.11
is independently selected from the group consisting of: H, halo and
C.sub.1-C.sub.3perhaloalkyl; [0021] R.sup.c is selected from the
group consisting of:
[0021] ##STR00005## [0022] R.sup.d and R.sup.e are independently H
or C.sub.1-C.sub.3alkyl; and [0023] provided that at least one of
R.sup.c, R.sup.d and R.sup.e are not H.
[0024] Further embodiments are provided by pharmaceutically
acceptable salts of compounds of Formulas (I), pharmaceutically
acceptable prodrugs of compounds of Formula (I), and
pharmaceutically active metabolites of compounds of Formula
(I).
[0025] In certain embodiments, the compounds of Formula (I) are
compounds selected from those species described or exemplified in
the detailed description below.
[0026] In a further aspect, the invention relates to enantiomers
and diastereomers of the compounds of Formula I, as well as the
pharmaceutically acceptable salts.
[0027] In a further aspect, the invention relates to pharmaceutical
compositions for treating a disease, disorder, or medical condition
mediated by P2X7 receptor activity, comprising an effective amount
of at least one compound selected from compounds of Formula (I),
pharmaceutically acceptable salts of compounds of Formula (I),
pharmaceutically acceptable prodrugs of compounds of Formula (I),
and pharmaceutically active metabolites of Formula (I).
[0028] Pharmaceutical compositions according to the invention may
further comprise one or more pharmaceutically acceptable
excipients.
[0029] In another aspect, the chemical embodiments of the present
invention are useful as P2X7 receptor modulators. Thus, the
invention is directed to a method for modulating P2X7 receptor
activity, including when such receptor is in a subject, comprising
exposing P2X7 receptor to an effective amount of at least one
compound selected from compounds of Formula (I), pharmaceutically
acceptable salts of compounds of Formula (I), pharmaceutically
acceptable prodrugs of compounds of Formula (I), and
pharmaceutically active metabolites of compounds of Formula
(I).
[0030] In another aspect, the invention is directed to a method of
treating a subject suffering from, or diagnosed with a disease,
disorder, or medical condition mediated by P2X7 receptor activity,
comprising administering to the subject in need of such treatment
an effective amount of at least one compound selected from
compounds of Formula (I), pharmaceutically acceptable salts of
compounds of Formula (I), pharmaceutically acceptable prodrugs of
compounds of Formula (I), and pharmaceutically active metabolites
of compounds of Formula (I). Additional embodiments of methods of
treatment are set forth in the detailed description.
[0031] In another aspect, the method of studying isotopically
labeled compounds in metabolic studies (preferably with .sup.14C),
reaction kinetic studies (with, for example .sup.2H or .sup.3H),
detection or imaging techniques [such as positron emission
tomography (PET) or single-photon emission computed tomography
(SPECT)] including drug or substrate tissue distribution assays, or
in radioactive treatment of patients. For example, an .sup.18F or
.sup.11C labeled compound may be particularly preferred for PET or
SPECT studies.
[0032] An object of the present invention is to overcome or
ameliorate at least one of the disadvantages of the conventional
methodologies and/or prior art, or to provide a useful alternative
thereto.
[0033] Additional embodiments, features, and advantages of the
invention will be apparent from the following detailed description
and through practice of the invention.
[0034] Additional embodiments of this invention include methods of
making compounds of Formula (I), pharmaceutically acceptable salts
of compounds of Formula (I), pharmaceutically acceptable prodrugs
of compounds of Formula (I), and pharmaceutically active
metabolites of Formula (I).
DETAILED DESCRIPTION OF THE INVENTION
[0035] A compound of Formula (I):
##STR00006## [0036] and enantiomers or diastereomers thereof;
[0037] and pharmaceutically acceptable salts thereof; [0038]
wherein: [0039] R.sup.a is
[0039] ##STR00007## [0040] R.sup.1 is halo or C.sub.1-C.sub.3alkyl;
[0041] R.sup.2 is independently selected from the group consisting
of: H, halo, and C.sub.1-C.sub.3perhaloalkyl; [0042] R.sup.3 is H
or halo; [0043] R.sup.4 is halo, [0044] R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; [0045] R.sup.b is independently
selected from the group consisting of:
[0045] ##STR00008## [0046] Wherein: [0047] R.sup.6, R.sup.9,
R.sup.10, R.sup.12, R.sup.14 are independently H or halo; [0048]
R.sup.7, R.sup.8, R.sup.13 is independently selected from the group
consisting of: H, halo and OC.sub.1-C.sub.3alkyl; [0049] R.sup.11
is independently selected from the group consisting of: H, halo and
C.sub.1-C.sub.3perhaloalkyl; [0050] R.sup.c is selected from the
group consisting of:
[0050] ##STR00009## [0051] R.sup.d and R.sup.e are independently H
or C.sub.1-C.sub.3alkyl; and [0052] provided that at least one of
R.sup.c, R.sup.d and R.sup.e are not H. [0053] A further embodiment
of the current invention is a compound of Formula (I) wherein
R.sup.a is
[0053] ##STR00010## [0054] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.a is
[0054] ##STR00011## [0055] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.a is
##STR00012##
[0055] and R.sup.1 is halo. [0056] A further embodiment of the
current invention is a compound of Formula (I) wherein R.sup.a
is
##STR00013##
[0056] and R.sup.1 is C.sub.1-C.sub.3alkyl. [0057] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.a is
##STR00014##
[0057] and R.sup.2 is C.sub.1-C.sub.3perhaloalkyl. [0058] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.a is
##STR00015##
[0058] and R.sup.2 is halo. [0059] A further embodiment of the
current invention is a compound of Formula (I) wherein R.sup.a
is
##STR00016##
[0059] and R.sup.3 is H.
[0060] A further embodiment of the current invention is a compound
of Formula (I) wherein R.sup.a is
##STR00017##
[0060] R.sup.1 is halo, R.sup.2 is C.sub.1-C.sub.3perhaloalkyl, and
R.sup.3 is H. [0061] A further embodiment of the current invention
is a compound of Formula (I) wherein R.sup.a is
##STR00018##
[0061] and R.sup.1, R.sup.2, and R.sup.3 are halo. [0062] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.a is
##STR00019##
[0062] R.sup.1 and R.sup.3 are halo and R.sup.2 is H. [0063] A
further embodiment of the current invention is a compound of
Formula (I) wherein R.sup.a is
##STR00020##
[0063] R.sup.1 and R.sup.2 are halo and R.sup.3 is H. [0064] A
further embodiment of the current invention is a compound of
Formula (I) wherein R.sup.a is
##STR00021##
[0064] R.sup.4 is halo and R.sup.5 is C.sub.1-C.sub.3perhaloalkyl.
[0065] A further embodiment of the current invention is a compound
of Formula (I) wherein R.sup.b is independently selected from the
group consisting of:
[0065] ##STR00022## [0066] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
independently selected from the group consisting of:
[0066] ##STR00023## [0067] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
independently selected from the group consisting of:
[0067] ##STR00024## [0068] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
[0068] ##STR00025## [0069] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
[0069] ##STR00026## [0070] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
[0070] ##STR00027## [0071] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
##STR00028##
[0071] R.sup.6 and R.sup.7 are H and R.sup.8 is OCH.sub.3. [0072] A
further embodiment of the current invention is a compound of
Formula (I) wherein R.sup.b is
##STR00029##
[0072] and R.sup.6, R.sup.7 and R.sup.8 are H. [0073] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.b is
[0073] ##STR00030## [0074] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
[0074] ##STR00031## [0075] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
##STR00032##
[0075] R.sup.9, R.sup.10 and R.sup.12 are H and R.sup.11 is F.
[0076] A further embodiment of the current invention is a compound
of Formula (I) wherein R.sup.b is
[0076] ##STR00033## [0077] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.b is
[0077] ##STR00034## [0078] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.c is H or
CH.sub.3. [0079] A further embodiment of the current invention is a
compound of Formula (I) wherein R.sup.c is selected from the group
consisting of:
[0079] ##STR00035## [0080] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.c is:
[0080] ##STR00036## [0081] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.c is:
[0081] ##STR00037## [0082] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.d is CH.sub.3.
[0083] A further embodiment of the current invention is a compound
of Formula (I) wherein R.sup.e is CH.sub.3. [0084] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.c is CH.sub.3 and R.sup.d and R.sup.e are H. [0085] A
further embodiment of the current invention is a compound of
Formula (I) wherein R.sup.d is CH.sub.3 and R.sup.c and R.sup.e are
H. [0086] A further embodiment of the current invention is a
compound of Formula (I) wherein R.sup.e is CH.sub.3 and R.sup.c and
R.sup.d are H. [0087] A further embodiment of the current invention
is a compound of Formula (I) wherein R.sup.a is
##STR00038##
[0087] R.sup.1 and R.sup.2 are Cl, R.sup.c is CH.sub.3, R.sup.b
is
##STR00039##
and R.sup.d, R.sup.e, R.sup.3, R.sup.6, R.sup.7 and R.sup.8 are H.
[0088] A further embodiment of the current invention is a compound
of Formula (I) wherein R.sup.a is
##STR00040##
[0088] R.sup.1 and R.sup.2 are Cl, R.sup.d is CH.sub.3, R.sup.b
is
##STR00041##
and R.sup.c, R.sup.e, R.sup.3, R.sup.9, R.sup.10 and R.sup.12 are H
and R.sup.11 is F. [0089] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.a is
##STR00042##
[0089] R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is CH.sub.3,
R.sup.b is
##STR00043##
and R.sup.c, R.sup.e, R.sup.3, R.sup.9, R.sup.19 and R.sup.12 are H
and R.sup.11 is F. [0090] A further embodiment of the current
invention is a compound of Formula (I) wherein R.sup.a is
##STR00044##
[0090] R.sup.1 and R.sup.2 are Cl, R.sup.d is CH.sub.3, R.sup.b
is
##STR00045##
and R.sup.8 is OCH.sub.3, R.sup.c, R.sup.e, R.sup.3, R.sup.6, and
R.sup.7 are H. [0091] A further embodiment of the current invention
is a compound of Formula (I) wherein R.sup.a is
##STR00046##
[0091] R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is CH.sub.3,
R.sup.c is
##STR00047##
R.sup.b is
##STR00048##
[0092] and R.sup.d, R.sup.e, and R.sup.3, are H. [0093] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.a is
##STR00049##
[0093] R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is CH.sub.3,
R.sup.c is
##STR00050##
R.sup.b is
##STR00051##
[0094] and R.sup.d, R.sup.e, and R.sup.3, are H. [0095] A further
embodiment of the current invention is a compound of Formula (I)
wherein R.sup.a is
##STR00052##
[0095] R.sup.1 is Cl, and R.sup.2 is CF.sub.3, R.sup.d is CH.sub.3,
R.sup.c is
##STR00053##
R.sup.b is
##STR00054##
[0096] and R.sup.d, R.sup.e, and R.sup.3, are H. [0097] A further
embodiment of the current invention is a compound as shown below in
Table 1.
TABLE-US-00001 [0097] TABLE 1
(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
2-chloro-3-(trifluoromethyl)phenyl)(5-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichlorophenyl)(5-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-5-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
:(2,3-dichlorophenyl)(3-(4-fluorophenyl)-5-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-methyl-3-(pyridin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-fluoro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-8-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(4-
(trifluoromethyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(S)-(2,3-dichlorophenyl)(3-(3-fluoro-4-(trifluoromethyl)phenyl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluoro-4-
(trifluoromethyl)phenyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(3,4,5-
trifluorophenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichlorophenyl)(5-methyl-3-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(4-
(trifluoromethyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyrazin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-chlorophenyl)-8-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2-fluorophenyl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,3-difluorophenyl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone.
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3,4-difluorophenyl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,4-difluorophenyl)-6-methyl-
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(6-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)methanone
(3,4-difluoro-2-methylphenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2-chloro-4-fluorophenyl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(2,3-dichloropyridin-4-yl)(6-methyl-3-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)(2,3-dichlorophenyl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclohexyl-8-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)(2,3-dichloro-4-fluorophenyl)methanone
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)(2,3-dichlorophenyl)methanone
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)(2,3-dichloro-4-fluorophenyl)methanone
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-fluoropyridin-2-yl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
((S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyridin-2-yl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(S)-(2,3-dichlorophenyl)(3-(5-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5-
,6- dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyrimidin-2-yl)-6-
methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(1-hydroxyethyl)-8-phenyl-5,-
6- dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
(S)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyriclin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyriclin-2-yl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(S*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
(S*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,-
6- dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-
fluorophenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-fluorophenyl)-
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone
(S)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-
a]pyrazin-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone
(R)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(R)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(8-(1H-pyrazol-5-yl)-3-(trifluoromethyl)-5,6-dihydro-
[1,2,4]thazolo[4,3-a]pyrazin-7(8H)-yl)(2-chloro-3-
(trifluoromethyl)phenyl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(pyridin-3-yl)-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
7(8H)-yl)methanone
(R)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-8-phenyl-3-
(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-
yl)methanone
(.+-.)-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]thazolo[4,3-
a]pyrazin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone.
S)-(2,3-dichlorophenyl)(3-(4-hydroxypyridin-2-yl)-6-methyl-5,6-dihydro-
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.11C]methoxypyridin-2-yl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.18F]fluoropyridin-2-yl)-6-methyl-5,6-
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0098] An additional embodiment of the invention is a
pharmaceutical composition comprising and effective amount of at
least one compound in Table 1 and at least one pharmaceutically
acceptable excipient.
[0099] Also within the scope of the invention are enantiomers and
diastereomers of the compounds of Formula I. Also within the scope
of the invention are the pharmaceutically acceptable salts of the
compounds of Formula I, as well as the pharmaceutically acceptable
salts of the enantiomers and diastereomers of the compounds of
Formula I. Also within the scope of the invention are isotopic
variations of compounds of Formula I, such as, e.g., deuterated
compounds of Formula I.
[0100] An additional embodiment of the invention is a method of
treating a subject suffering from or diagnosed with a disease,
disorder, or medical condition mediated by P2X7 receptor activity,
comprising administering to a subject in need of such treatment an
effective amount of at least one compound selected from compounds
of Formula (I):
##STR00055##
and enantiomers or diastereomers thereof;
[0101] and pharmaceutically acceptable salts thereof;
wherein:
[0102] R.sup.a is
##STR00056## [0103] R.sup.1 is halo or C.sub.1-C.sub.3alkyl; [0104]
R.sup.2 is independently selected from the group consisting of: H,
halo, and C.sub.1-C.sub.3perhaloalkyl; [0105] R.sup.3 is H or halo;
[0106] R.sup.4 is halo, [0107] R.sup.5 is halo or
C.sub.1-C.sub.3perhaloalkyl; [0108] R.sup.b is independently
selected from the group consisting of:
##STR00057##
[0109] Wherein: [0110] R.sup.6, R.sup.9, R.sup.10, R.sup.12,
R.sup.14 are independently H or halo; [0111] R.sup.7, R.sup.8,
R.sup.13 is independently selected from the group consisting of: H,
halo and OC.sub.1-C.sub.3alkyl; [0112] R.sup.11 is independently
selected from the group consisting of: H, halo and
C.sub.1-C.sub.3perhaloalkyl; [0113] R.sup.c is selected from the
group consisting of:
[0113] ##STR00058## [0114] R.sup.d and R.sup.e are independently H
or C.sub.1-C.sub.3alkyl; and [0115] provided that at least one of
R.sup.c, R.sup.d and R.sup.e are not H.
[0116] In preferred embodiments of the inventive method, the
disease, disorder, or medical condition is selected from: diseases
of the autoimmune and inflammatory system (Arulkumaran, N. et al.
Expert Opin. Invetig Drugs, 2011, July; 20(7):897-915) [examples of
diseases of the autoimmune and inflammatory system include
rheumatoid arthritis, osteoarthritis, interstitial cystitis
(Martins J P, et. al., Br J Pharmacol. 2012 January;
165(1):183-96), psoriasis (Killeen, M. E., et al., J Immunol. 2013
Apr. 15; 190(8):4324-36), septic shock, sepsis, allergic
dermatitis, asthma (examples of asthma include allergic asthma,
mild to severe asthma, and steroid resistant asthma), idiopathic
pulmonary fibrosis, allergic rhinitis, chronic obstructive
pulmonary disease and airway hyper-responsiveness]; diseases of the
nervous and neuro-immune system [examples of diseases of the
nervous and neuro-immune system include acute and chronic pain
(examples of acute and chronic pain include neuropathic pain,
inflammatory pain, migraine, spontaneous pain (examples of
spontaneous pain include opioid induced pain, diabetic neuropathy,
postherpetic neuralgia, low back pain, chemotherapy-induced
neuropathic pain, fibromyalgia) (Romagnoli, R, et. al., Expert
Opin. Ther. Targets, 2008, 12(5), 647-661)], and diseases involved
with, and without, neuroinflammation of the Central Nervous System
(CNS) [examples of diseases involved with, and without,
neuroinflammation of the Central Nervous System (CNS) include mood
disorders (examples of mood disorders include major depression,
major depressive disorder, treatment resistant depression, bipolar
disorder, anxious depression, anxiety) (Friedle, S A, et. al.,
Recent Patents on CNS Drug Discovery, 2010, 5, 35-45, Romagnoli, R,
et. al., 2008), cognition, sleep disorders, multiple sclerosis
(Sharp A J, et. al., J Neuroinflammation. 2008 Aug. 8; 5:33,
Oyanguren-Desez O, et. al., Cell Calcium. 2011 November;
50(5):468-72, Grygorowicz T, et. al., Neurochem Int. 2010 December;
57(7):823-9), epileptic seizures (Engel T, et. al., FASEB J. 2012
April; 26(4):1616-28, Kim J E, et. al. Neurol Res. 2009 November;
31(9):982-8, Avignone E, et. al., J Neurosci. 2008 Sep. 10;
28(37):9133-44), Parkinson's disease (Marcellino D, et. al., J
Neural Transm. 2010 June; 117(6):681-7), schizophrenia, Alzheimer's
disease (Diaz-Hernandez J I, et. al., Neurobiol Aging. 2012 August;
33(8):1816-28, Delarasse C, J Biol Chem. 2011 Jan. 28;
286(4):2596-606, Sanz J M, et. al., J Immunol. 2009 Apr. 1;
182(7):4378-85), Huntington's disease (Diaz-Hernandez M, et. Al.,
FASEB J. 2009 June; 23(6):1893-906), Amyotrophic Lateral Sclerosis,
autism, spinal cord injury,cerebral ischemia/traumatic brain injury
(Chu K, et. al., J Neuroinflammation. 2012 Apr. 18; 9:69, Arbeloa
J, et. al, Neurobiol Dis. 2012 March; 45(3):954-61) and
stress-related disorders].
[0117] In addition, P2X7 intervention may be beneficial in diseases
of the cardiovascular, metabolic, gastrointestinal and urogenital
systems [examples of diseases of the cardiovascular, metabolic,
gastrointestinal and urogenital systems include diabetes
(Arterioscler Thromb Vasc Biol. 2004 July; 24(7):1240-5, J Cell
Physiol. 2013 January; 228(1):120-9), diabetes mellitus, thrombosis
(Furlan-Freguia C, et. al., J Clin Invest. 2011 July;
121(7):2932-44, Vergani, A. et al., Diabetes, 2013, 62, 1665-1675),
irritable bowel disease, irritable bowel syndrome, (J Immunol. 2011
Aug. 1; 187(3):1467-74. Epub 2011 Jun. 22), Crohn's disease,
cardiovascular diseases (examples of cardiovascular disease include
hypertension (Ji X, et. al., Am J Physiol Renal Physiol. 2012
October; 303(8):F1207-15), myocardial infarction, ischemic heart
disease, ischemia) ureteric obstruction, lower urinary tract
syndrome (Br J Pharmacol. 2012 January; 165(1):183-96), lower
urinary tract dysfunction such as incontinence, and disease after
cardiac transplant (Vergani, A. et al., Circulation. 2013;
127:463-475)].
[0118] P2X7 antagonism may also present a novel therapeutic
strategy for skeletal disorders, (examples of skeletal disorders
include osteoporosis/osteopetrosis) and may also modulate secretory
function of exocrine glands.
[0119] It is also hypothesized that modulation of the P2X7 receptor
may also be beneficial in conditions such as: glaucoma,
Glomerulonephritis, Chaga's Disease, chlamydia, neuroblastoma,
Tuberculosis, Polycystic Kidney Disease, cancer, and acne
(Thiboutot, D. M. J Investigative Dermatology, 2014, 134,
595-597).
[0120] An additional embodiment of the invention is a method of
treating a subject suffering from or diagnosed with a disease,
disorder, or medical condition mediated by P2X7 receptor activity,
wherein the disease, disorder, or medical condition is selected
from the group consisting of: diseases of the autoimmune and
inflammatory system [examples of diseases of the autoimmune and
inflammatory system include rheumatoid arthritis, osteoarthritis,
interstitial cystitis, psoriasis, septic shock, sepsis, allergic
dermatitis, asthma (examples of asthma include allergic asthma,
mild to severe asthma, and steroid resistant asthma), idiopathic
pulmonary fibrosis, allergic rhinitis, chronic obstructive
pulmonary disease and airway hyper-responsivenes]; diseases of the
nervous and neuro-immune system [examples of diseases of the
nervous and neuro-immune system include acute and chronic pain
(examples of acute and chronic pain include neuropathic pain,
inflammatory pain, migraine, spontaneous pain (examples of
spontaneous pain include opioid induced pain, diabetic neuropathy,
postherpetic neuralgia, low back pain, chemotherapy-induced
neuropathic pain, fibromyalgia)]; diseases involved with, and
without, neuroinflammation of the Central Nervous System (CNS)
[examples of diseases involved with, and without, neuroinflammation
of the Central Nervous System (CNS) include mood disorders
(examples of mood disorders include major depression, major
depressive disorder, treatment resistant depression, bipolar
disorder, anxious depression, anxiety), cognition, sleep disorders,
multiple sclerosis, epileptic seizures, Parkinson's disease,
schizophrenia, Alzheimer's disease, Huntington's disease,
Amyotrophic Lateral Sclerosis, autism, spinal cord injury and
cerebral ischemia/traumatic brain injury, and stress-related
disorders]; diseases of the cardiovascular, metabolic,
gastrointestinal and urogenital systems [examples of diseases of
the cardiovascular, metabolic, gastrointestinal and urogenital
systems include diabetes, diabetes mellitus, thrombosis, irritable
bowel disease, irritable bowel syndrome, Crohn's disease,
cardiovascular diseases (examples of cardiovascular disease include
hypertension, myocardial infarction, ischemic heart disease,
ischemia) ureteric obstruction, lower urinary tract syndrome, lower
urinary tract dysfunction such as incontinence, and disease after
cardiac transplantation]; skeletal disorders, (examples of skeletal
disorders include osteoporosis/osteopetrosis) and diseases
involving the secretory function of exocrine glands and diseases
such as glaucoma, Glomerulonephritis, Chaga's Disease, chlamydia,
neuroblastoma, Tuberculosis, Polycystic Kidney Disease, cancer, and
acne.
[0121] An additional embodiment of the invention is a method of
treating a subject suffering from or diagnosed with a disease,
disorder, or medical condition mediated by P2X7 receptor activity
wherein the disease, disorder or medical condition is a disease
involved with, and without, neuroinflammation of the Central
Nervous System (CNS).
[0122] An additional embodiment of the invention is a method of
treating a subject suffering from or diagnosed with a disease
involved with, and without, neuroinflammation of the Central
Nervous System (CNS) wherein the disease, disorder or medical
condition is a mood disorder.
[0123] An additional embodiment of the invention is a method of
treating a subject suffering from a mood disorder wherein the mood
disorder is treatment resistant depression.
[0124] Additional embodiments, features, and advantages of the
invention will be apparent from the following detailed description
and through practice of the invention.
[0125] The invention may be more fully appreciated by reference to
the following description, including the following glossary of
terms and the concluding examples. For the sake of brevity, the
disclosures of the publications, including patents, cited in this
specification are herein incorporated by reference.
[0126] As used herein, the terms "including", "containing" and
"comprising" are used herein in their open, non-limiting sense.
[0127] The term "alkyl" refers to a straight- or branched-chain
alkyl group having from 1 to 12 carbon atoms in the chain. Examples
of alkyl groups include methyl (Me, which also may be structurally
depicted by the symbol, "/"), ethyl (Et), n-propyl, isopropyl,
butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl, isopentyl,
tert-pentyl, hexyl, isohexyl, and groups that in light of the
ordinary skill in the art and the teachings provided herein would
be considered equivalent to any one of the foregoing examples. The
term C.sub.1-C.sub.3 alkyl as used here refers to a straight- or
branched-chain alkyl group having from 1 to 3 carbon atoms in the
chain. The term C.sub.1-C.sub.4 alkyl as used here refers to a
straight- or branched-chain alkyl group having from 1 to 4 carbon
atoms in the chain.
[0128] The term "alkoxy" includes a straight chain or branched
alkyl group with a terminal oxygen linking the alkyl group to the
rest of the molecule. Alkoxy includes methoxy, ethoxy, propoxy,
isopropoxy, butoxy, t-butoxy, pentoxy and so on.
[0129] The term "alkalkoxy" refers to the group alkyl-O-alkyl,
where alkyl is defined above. Such groups include methylenemethoxy
(--CH.sub.2OCH.sub.3) and ethylenemethoxy
(--CH.sub.2CH.sub.2OCH.sub.3).
[0130] The terms "hydroxyl" and "hydroxy" refer to an --OH
group.
[0131] The term "cycloalkyl" refers to a saturated carbocycle
having from 3 to 6 ring atoms per carbocycle. Illustrative examples
of cycloalkyl groups include the following entities, in the form of
properly bonded moieties:
##STR00059##
The term "C.sub.3-C.sub.4 cycloalkyl" as used here refers to a
saturated carbocycle having from 3 to 4 ring atoms.
[0132] A "heterocycloalkyl" refers to a monocyclic ring structure
that is saturated and has from 4 to 6 ring atoms per ring structure
selected from carbon atoms and one nitrogen atom. Illustrative
entities, in the form of properly bonded moieties, include:
##STR00060##
[0133] The term "aryl" refers to a monocyclic, aromatic carbocycle
(ring structure having ring atoms that are all carbon) having 6
atoms per ring. (Carbon atoms in the aryl groups are sp.sup.2
hybridized.)
[0134] The term "phenyl" represents the following moiety:
##STR00061##
[0135] The term "heteroaryl" refers to a monocyclic or fused
bicyclic heterocycle (ring structure having ring atoms selected
from carbon atoms and up to four heteroatoms selected from
nitrogen, oxygen, and sulfur) having from 3 to 9 ring atoms per
heterocycle. Illustrative examples of heteroaryl groups include the
following entities, in the form of properly bonded moieties:
##STR00062##
[0136] Those skilled in the art will recognize that the species of
heteroaryl, cycloalkyl, aryl and heterocycloalkyl groups listed or
illustrated above are not exhaustive, and that additional species
within the scope of these defined terms may also be selected.
[0137] The term "cyano" refers to the group --CN.
[0138] The term "halo" represents chloro, fluoro, bromo or
iodo.
[0139] The term "perhaloalkyl" refers to a straight- or
branched-chain alkyl group having from 1 to 4 carbon atoms in the
chain optionally substituting hydrogens with halogens. Examples of
perhaloalkyl groups include trifluoromethyl (CF.sub.3),
difluoromethyl (CF.sub.2H), monofluoromethyl (CH.sub.2F),
pentafluoroethyl (CF.sub.2CF.sub.3), tetrafluoroethyl
(CHFCF.sub.3),monofluoroethyl (CH.sub.2CH.sub.2F), trifluoroethyl
(CH.sub.2CF.sub.3), tetrafluorotrifluoromethylethyl
(--CF(CF.sub.3).sub.2), and groups that in light of the ordinary
skill in the art and the teachings provided herein would be
considered equivalent to any one of the foregoing examples.
[0140] The term "perhaloalkoxy" refers to a straight- or
branched-chain alkoxy group having from 1 to 4 carbon atoms in the
chain optionally substituting hydrogens with halogens. Examples of
perhaloalkoxy groups include trifluoromethoxy (OCF.sub.3),
difluoromethoxy (OCF.sub.2H), monofluoromethoxy (OCH.sub.2F),
momofluoroethoxy (OCH.sub.2CH.sub.2F), pentafluoroethoxy
(OCF.sub.2CF.sub.3), tetrafluoroethoxy (OCHFCF.sub.3),
trifluoroethoxy (OCH.sub.2CF.sub.3),
tetrafluorotrifluoromethylethoxy (--OCF(CF.sub.3).sub.2), and
groups that in light of the ordinary skill in the art and the
teachings provided herein would be considered equivalent to any one
of the foregoing examples.
[0141] The term "substituted" means that the specified group or
moiety bears one or more substituents. The term "unsubstituted"
means that the specified group bears no substituents. The term
"optionally substituted" means that the specified group is
unsubstituted or substituted by one or more substituents. Where the
term "substituted" is used to describe a structural system, the
substitution is meant to occur at any valency-allowed position on
the system. In cases where a specified moiety or group is not
expressly noted as being optionally substituted or substituted with
any specified substituent, it is understood that such a moiety or
group is intended to be unsubstituted.
[0142] The terms "para", "meta", and "ortho" have the meanings as
understood in the art. Thus, for example, a fully substituted
phenyl group has substituents at both "ortho"(o) positions adjacent
to the point of attachment of the phenyl ring, both "meta" (m)
positions, and the one "para" (p) position across from the point of
attachment. To further clarify the position of substituents on the
phenyl ring, the 2 different ortho positions will be designated as
ortho and ortho' and the 2 different meta positions as meta and
meta' as illustrated below.
##STR00063##
[0143] When referring to substituents on a pyridyl group, the terms
"para", "meta", and "ortho" refer to the placement of a substituent
relative to the point of attachment of the pyridyl ring. For
example the structure below is described as 4-pyridyl with the X
substituent in the ortho position and the Y substituent in the meta
position:
##STR00064##
[0144] To provide a more concise description, some of the
quantitative expressions given herein are not qualified with the
term "about". It is understood that, whether the term "about" is
used explicitly or not, every quantity given herein is meant to
refer to the actual given value, and it is also meant to refer to
the approximation to such given value that would reasonably be
inferred based on the ordinary skill in the art, including
equivalents and approximations due to the experimental and/or
measurement conditions for such given value. Whenever a yield is
given as a percentage, such yield refers to a mass of the entity
for which the yield is given with respect to the maximum amount of
the same entity that could be obtained under the particular
stoichiometric conditions. Concentrations that are given as
percentages refer to mass ratios, unless indicated differently.
[0145] The terms "buffered" solution or "buffer" solution are used
herein interchangeably according to their standard meaning.
Buffered solutions are used to control the pH of a medium, and
their choice, use, and function is known to those of ordinary skill
in the art. See, for example, G. D. Considine, ed., Van Nostrand's
Encyclopedia of Chemistry, p. 261, 5.sup.th ed. (2005), describing,
inter alia, buffer solutions and how the concentrations of the
buffer constituents relate to the pH of the buffer. For example, a
buffered solution is obtained by adding MgSO.sub.4 and NaHCO.sub.3
to a solution in a 10:1 w/w ratio to maintain the pH of the
solution at about 7.5.
[0146] Any formula given herein is intended to represent compounds
having structures depicted by the structural formula as well as
certain variations or forms. In particular, compounds of any
formula given herein may have asymmetric centers and therefore
exist in different enantiomeric forms. All optical isomers of the
compounds of the general formula, and mixtures thereof, are
considered within the scope of the formula. Thus, any formula given
herein is intended to represent a racemate, one or more
enantiomeric forms, one or more diastereomeric forms, one or more
atropisomeric forms, and mixtures thereof. Furthermore, certain
structures may exist as geometric isomers (i.e., cis and trans
isomers), as tautomers, or as atropisomers.
[0147] It is also to be understood that compounds that have the
same molecular formula but differ in the nature or sequence of
bonding of their atoms or the arrangement of their atoms in space
are termed "isomers." Isomers that differ in the arrangement of
their atoms in space are termed "."
[0148] Stereoisomers that are not mirror images of one another are
termed "diastereomers" and those that are non-superimposable mirror
images of each other are termed "enantiomers." When a compound has
an asymmetric center, for example, it is bonded to four different
groups, and a pair of enantiomers is possible. An enantiomer can be
characterized by the absolute configuration of its asymmetric
center and is described by the R-and S-sequencing rules of Cahn and
Prelog, or by the manner in which the molecule rotates the plane of
polarized light and designated as dextrorotatory or levorotatory
(i.e., as (+)- or (-)-isomers respectively). A chiral compound can
exist as either an individual enantiomer or as a mixture thereof. A
mixture containing equal proportions of the enantiomers is called a
"racemic mixture."
[0149] "Tautomers" refer to compounds that are interchangeable
forms of a particular compound structure, and that vary in the
displacement of hydrogen atoms and electrons. Thus, two structures
may be in equilibrium through the movement of .pi. electrons and an
atom (usually H). For example, enols and ketones are tautomers
because they are rapidly interconverted by treatment with either
acid or base. Another example of tautomerism is the aci-and
nitro-forms of phenyl nitromethane, that are likewise formed by
treatment with acid or base.
[0150] Tautomeric forms may be relevant to the attainment of the
optimal chemical reactivity and biological activity of a compound
of interest.
[0151] Compounds of the invention may also exist as "rotamers,"
that is, conformational isomers that occur when the rotation
leading to different conformations is hindered, resulting in a
rotational energy barrier to be overcome to convert from one
conformational isomer to another.
[0152] The compounds of this invention may possess one or more
asymmetric centers; such compounds can therefore be produced as
individual (R)- or (S)-stereoisomers or as mixtures thereof.
[0153] Unless indicated otherwise, the description or naming of a
particular compound in the specification and claims is intended to
include both individual enantiomers and mixtures, racemic or
otherwise, thereof. The methods for the determination of
stereochemistry and the separation of stereoisomers are well-known
in the art.
[0154] Certain examples contain chemical structures that are
depicted as an absolute enantiomer but are intended to indicate
enatiopure material that is of unknown configuration. In these
cases (R*) or (S*) is used in the name to indicate that the
absolute stereochemistry of the corresponding stereocenter is
unknown. Thus, a compound designated as (R*) refers to an
enantiopure compound with an absolute configuration of either (R)
or (S). In cases where the absolute stereochemistry has been
confirmed, the structures are named using (R) and (S).
[0155] The symbols and are used as meaning the same spatial
arrangement in chemical structures shown herein. Analogously, the
symbols and are used as meaning the same spatial arrangement in
chemical structures shown herein.
[0156] Additionally, any formula given herein is intended to refer
also to hydrates, solvates, and polymorphs of such compounds, and
mixtures thereof, even if such forms are not listed explicitly.
Certain compounds of Formula (I) or pharmaceutically acceptable
salts of compounds of Formula (I) may be obtained as solvates.
Solvates include those formed from the interaction or complexation
of compounds of the invention with one or more solvents, either in
solution or as a solid or crystalline form. In some embodiments,
the solvent is water and the solvates are hydrates. In addition,
certain crystalline forms of compounds of Formula (I) or
pharmaceutically acceptable salts of compounds of Formula (I) may
be obtained as co-crystals. In certain embodiments of the
invention, compounds of Formula (I) were obtained in a crystalline
form. In other embodiments, crystalline forms of compounds of
Formula (I) were cubic in nature. In other embodiments,
pharmaceutically acceptable salts of compounds of Formula (I) were
obtained in a crystalline form. In still other embodiments,
compounds of Formula (I) were obtained in one of several
polymorphic forms, as a mixture of crystalline forms, as a
polymorphic form, or as an amorphous form. In other embodiments,
compounds of Formula (I) convert in solution between one or more
crystalline forms and/or polymorphic forms.
[0157] Reference to a compound herein stands for a reference to any
one of: (a) the actually recited form of such compound, and (b) any
of the forms of such compound in the medium in which the compound
is being considered when named. For example, reference herein to a
compound such as R--COOH, encompasses reference to any one of, for
example, R--COOH.sub.(s), R--COOH.sub.(sol), and
R--COO.sup.-.sub.(sol). In this example, R--COOH.sub.(s) refers to
the solid compound, as it could be for example in a tablet or some
other solid pharmaceutical composition or preparation;
R--COOH.sub.(sol) refers to the undissociated form of the compound
in a solvent; and R--COO.sup.-.sub.(sol) refers to the dissociated
form of the compound in a solvent, such as the dissociated form of
the compound in an aqueous environment, whether such dissociated
form derives from R--COOH, from a salt thereof, or from any other
entity that yields R--COO.sup.- upon dissociation in the medium
being considered. In another example, an expression such as
"exposing an entity to compound of formula R--COOH" refers to the
exposure of such entity to the form, or forms, of the compound
R--COOH that exists, or exist, in the medium in which such exposure
takes place. In still another example, an expression such as
"reacting an entity with a compound of formula R--COOH" refers to
the reacting of (a) such entity in the chemically relevant form, or
forms, of such entity that exists, or exist, in the medium in which
such reacting takes place, with (b) the chemically relevant form,
or forms, of the compound R--COOH that exists, or exist, in the
medium in which such reacting takes place. In this regard, if such
entity is for example in an aqueous environment, it is understood
that the compound R--COOH is in such same medium, and therefore the
entity is being exposed to species such as R--COOH.sub.(aq) and/or
R--COO.sup.-.sub.(aq), where the subscript "(aq)" stands for
"aqueous" according to its conventional meaning in chemistry and
biochemistry. A carboxylic acid functional group has been chosen in
these nomenclature examples; this choice is not intended, however,
as a limitation but it is merely an illustration. It is understood
that analogous examples can be provided in terms of other
functional groups, including but not limited to hydroxyl, basic
nitrogen members, such as those in amines, and any other group that
interacts or transforms according to known manners in the medium
that contains the compound. Such interactions and transformations
include, but are not limited to, dissociation, association,
tautomerism, solvolysis, including hydrolysis, solvation, including
hydration, protonation, and deprotonation. No further examples in
this regard are provided herein because these interactions and
transformations in a given medium are known by any one of ordinary
skill in the art.
[0158] In another example, a zwitterionic compound is encompassed
herein by referring to a compound that is known to form a
zwitterion, even if it is not explicitly named in its zwitterionic
form. Terms such as zwitterion, zwitterions, and their synonyms
zwitterionic compound(s) are standard IUPAC-endorsed names that are
well known and part of standard sets of defined scientific names.
In this regard, the name zwitterion is assigned the name
identification CHEBI:27369 by the Chemical Entities of Biological
Interest (ChEBI) dictionary of molecular entities. As generally
well known, a zwitterion or zwitterionic compound is a neutral
compound that has formal unit charges of opposite sign. Sometimes
these compounds are referred to by the term "inner salts". Other
sources refer to these compounds as "dipolar ions", although the
latter term is regarded by still other sources as a misnomer. As a
specific example, aminoethanoic acid (the amino acid glycine) has
the formula H.sub.2NCH.sub.2COOH, and it exists in some media (in
this case in neutral media) in the form of the zwitterion
.sup.+H.sub.3NCH.sub.2COO.sup.-. Zwitterions, zwitterionic
compounds, inner salts and dipolar ions in the known and well
established meanings of these terms are within the scope of this
invention, as would in any case be so appreciated by those of
ordinary skill in the art. Because there is no need to name each
and every embodiment that would be recognized by those of ordinary
skill in the art, no structures of the zwitterionic compounds that
are associated with the compounds of this invention are given
explicitly herein. They are, however, part of the embodiments of
this invention. No further examples in this regard are provided
herein because the interactions and transformations in a given
medium that lead to the various forms of a given compound are known
by any one of ordinary skill in the art.
[0159] Any formula given herein is also intended to represent
unlabeled forms as well as isotopically labeled forms of the
compounds. Isotopically labeled compounds have structures depicted
by the formulas given herein except that one or more atoms are
replaced by an atom having a selected atomic mass or mass number.
Examples of isotopes that can be incorporated into compounds of the
invention include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorus, sulfur, fluorine, chlorine, and iodine such as .sup.2H,
.sup.3H, .sup.11C, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
.sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F, .sup.36Cl,
.sup.125I, respectively. Such isotopically labeled compounds are
useful in metabolic studies (preferably with .sup.14C), reaction
kinetic studies (with, for example .sup.2H or .sup.3H), detection
or imaging techniques [such as positron emission tomography (PET)
or single-photon emission computed tomography (SPECT)] including
drug or substrate tissue distribution assays, or in radioactive
treatment of patients. In particular, an .sup.18F or .sup.11C
labeled compound may be particularly preferred for PET or SPECT
studies. Further, 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. Isotopically
labeled compounds of this invention and prodrugs thereof can
generally be prepared by carrying out the procedures disclosed in
the schemes or in the examples and preparations described below by
substituting a readily available isotopically labeled reagent for a
non-isotopically labeled reagent.
[0160] When referring to any formula given herein, the selection of
a particular moiety from a list of possible species for a specified
variable is not intended to define the same choice of the species
for the variable appearing elsewhere. In other words, where a
variable appears more than once, the choice of the species from a
specified list is independent of the choice of the species for the
same variable elsewhere in the formula, unless stated
otherwise.
[0161] According to the foregoing interpretive considerations on
assignments and nomenclature, it is understood that explicit
reference herein to a set implies, where chemically meaningful and
unless indicated otherwise, independent reference to embodiments of
such set, and reference to each and every one of the possible
embodiments of subsets of the set referred to explicitly.
[0162] The invention includes also pharmaceutically acceptable
salts of the compounds of Formula (I), preferably of those
described above and of the specific compounds exemplified herein,
and methods of treatment using such salts.
[0163] The term "pharmaceutically acceptable" means approved or
approvable by a regulatory agency of Federal or a state government
or the corresponding agency in countries other than the United
States, or that is listed in the U.S. Pharmcopoeia or other
generally recognized pharmacopoeia for use in animals, and more
particularly, in humans.
[0164] A "pharmaceutically acceptable salt" is intended to mean a
salt of a free acid or base of compounds represented by Formula (I)
that are non-toxic, biologically tolerable, or otherwise
biologically suitable for administration to the subject. It should
possess the desired pharmacological activity of the parent
compound. See, generally, G. S. Paulekuhn, et al., "Trends in
Active Pharmaceutical Ingredient Salt Selection based on Analysis
of the Orange Book Database", J. Med. Chem., 2007, 50:6665-72, S.
M. Berge, et al., "Pharmaceutical Salts", J Pharm Sci., 1977,
66:1-19, and Handbook of Pharmaceutical Salts, Properties,
Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA,
Zurich, 2002. Examples of pharmaceutically acceptable salts are
those that are pharmacologically effective and suitable for contact
with the tissues of patients without undue toxicity, irritation, or
allergic response. A compound of Formula (I) may possess a
sufficiently acidic group, a sufficiently basic group, or both
types of functional groups, and accordingly react with a number of
inorganic or organic bases, and inorganic and organic acids, to
form a pharmaceutically acceptable salt.
[0165] Examples of pharmaceutically acceptable salts include
sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,
phosphates, monohydrogen-phosphates, dihydrogenphosphates,
metaphosphates, pyrophosphates, chlorides, bromides, iodides,
acetates, propionates, decanoates, caprylates, acrylates, formates,
isobutyrates, caproates, heptanoates, propiolates, oxalates,
malonates, succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methyl benzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, sulfonates, xylenesulfonates,
phenylacetates, phenylpropionates, phenylbutyrates, citrates,
lactates, .gamma.-hydroxybutyrates, glycolates, tartrates,
methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,
naphthalene-2-sulfonates, and mandelates.
[0166] When the compounds of Formula (I) contain a basic nitrogen,
the desired pharmaceutically acceptable salt may be prepared by any
suitable method available in the art. For example, treatment of the
free base with an inorganic acid, such as hydrochloric acid,
hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric
acid, phosphoric acid, and the like, or with an organic acid, such
as acetic acid, phenylacetic acid, propionic acid, stearic acid,
lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid,
isethionic acid, succinic acid, valeric acid, fumaric acid, malonic
acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid,
oleic acid, palmitic acid, lauric acid, a pyranosidyl acid, such as
glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such
as mandelic acid, citric acid, or tartaric acid, an amino acid,
such as aspartic acid, glutaric acid or glutamic acid, an aromatic
acid, such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid,
or cinnamic acid, a sulfonic acid, such as laurylsulfonic acid,
p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid,
any compatible mixture of acids such as those given as examples
herein, and any other acid and mixture thereof that are regarded as
equivalents or acceptable substitutes in light of the ordinary
level of skill in this technology.
[0167] The invention may be more fully appreciated by reference to
the following description, including the following glossary of
terms and the concluding examples. For the sake of brevity, the
disclosures of the publications, including patents, cited in this
specification are herein incorporated by reference.
[0168] As used herein, the terms "including", "containing" and
"comprising" are used herein in their open, non-limiting sense.
[0169] When the compound of Formula (I) is an acid, such as a
carboxylic acid or sulfonic acid, the desired pharmaceutically
acceptable salt may be prepared by any suitable method, for
example, treatment of the free acid with an inorganic or organic
base, such as an amine (primary, secondary or tertiary), an alkali
metal hydroxide, alkaline earth metal hydroxide, any compatible
mixture of bases such as those given as examples herein, and any
other base and mixture thereof that are regarded as equivalents or
acceptable substitutes in light of the ordinary level of skill in
this technology. Illustrative examples of suitable salts include
organic salts derived from amino acids, such as
N-methyl-D-glucamine, lysine, choline, glycine and arginine,
ammonia, carbonates, bicarbonates, primary, secondary, and tertiary
amines, and cyclic amines, such as tromethamine, benzylamines,
pyrrolidines, piperidine, morpholine, and piperazine, and inorganic
salts derived from sodium, calcium, potassium, magnesium,
manganese, iron, copper, zinc, aluminum, and lithium.
[0170] The invention also relates to pharmaceutically acceptable
prodrugs of the compounds of Formula (I), and treatment methods
employing such pharmaceutically acceptable prodrugs. The term
"prodrug" means a precursor of a designated compound that,
following administration to a subject, yields the compound in vivo
via a chemical or physiological process such as solvolysis or
enzymatic cleavage, or under physiological conditions (e.g., a
prodrug on being brought to physiological pH is converted to the
compound of Formula (I). A "pharmaceutically acceptable prodrug" is
a prodrug that is non-toxic, biologically tolerable, and otherwise
biologically suitable for administration to the subject.
Illustrative procedures for the selection and preparation of
suitable prodrug derivatives are described, for example, in "Design
of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.
[0171] Exemplary prodrugs include compounds having an amino acid
residue, or a polypeptide chain of two or more (e.g., two, three or
four) amino acid residues, covalently joined through an amide or
ester bond to a free amino, hydroxyl, or carboxylic acid group of a
compound of Formula (I, IIa or IIb). Examples of amino acid
residues include the twenty naturally occurring amino acids,
commonly designated by three letter symbols, as well as
4-hydroxyproline, hydroxylysine, demosine, isodemosine,
3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
citrulline homocysteine, homoserine, ornithine and methionine
sulfone.
[0172] Additional types of prodrugs may be produced, for instance,
by derivatizing free carboxyl groups of structures of Formula (I)
as amides or alkyl esters. Examples of amides include those derived
from ammonia, primary C.sub.1-6alkyl amines and secondary
di(C.sub.1-6alkyl) amines. Secondary amines include 5- or
6-membered heterocycloalkyl or heteroaryl ring moieties. Examples
of amides include those that are derived from ammonia,
C.sub.1-3alkyl primary amines, and di(C.sub.1-2alkyl)amines.
Examples of esters of the invention include C.sub.1-7alkyl,
C.sub.5-7cycloalkyl, phenyl, and phenyl(C.sub.1-6alkyl) esters.
Preferred esters include methyl esters. Prodrugs may also be
prepared by derivatizing free hydroxy groups using groups including
hemisuccinates, phosphate esters, dimethylaminoacetates, and
phosphoryloxymethyloxycarbonyls, following procedures such as those
outlined in Fleisher et al., Adv. Drug Delivery Rev. 1996, 19,
115-130. Carbamate derivatives of hydroxy and amino groups may also
yield prodrugs. Carbonate derivatives, sulfonate esters, and
sulfate esters of hydroxy groups may also provide prodrugs.
Derivatization of hydroxy groups as (acyloxy)methyl and
(acyloxy)ethyl ethers, wherein the acyl group may be an alkyl
ester, optionally substituted with one or more ether, amine, or
carboxylic acid functionalities, or where the acyl group is an
amino acid ester as described above, is also useful to yield
prodrugs. Prodrugs of this type may be prepared as described in
Robinson et al., J Med Chem. 1996, 39 (1), 10-18. Free amines can
also be derivatized as amides, sulfonamides or phosphonamides. All
of these prodrug moieties may incorporate groups including ether,
amine, and carboxylic acid functionalities.
[0173] The present invention also relates to pharmaceutically
active metabolites of the compounds of Formula (I), which may also
be used in the methods of the invention. A "pharmaceutically active
metabolite" means a pharmacologically active product of metabolism
in the body of a compound of Formula (I, IIa or IIb) or salt
thereof. Prodrugs and active metabolites of a compound may be
determined using routine techniques known or available in the art.
See, e.g., Bertolini, et al., J Med Chem. 1997, 40, 2011-2016;
Shan, et al., J Pharm Sci. 1997, 86 (7), 765-767; Bagshawe, Drug
Dev Res. 1995, 34, 220-230; Bodor, Adv Drug Res. 1984, 13, 224-331;
Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen,
Design and Application of Prodrugs, Drug Design and Development
(Krogsgaard-Larsen, et al., eds., Harwood Academic Publishers,
1991).
[0174] The compounds of Formula (I) and their pharmaceutically
acceptable salts, pharmaceutically acceptable prodrugs, and
pharmaceutically active metabolites of the present invention are
useful as modulators of the P2X7 receptor in the methods of the
invention. As such modulators, the compounds may act as
antagonists, agonists, or inverse agonists. The term "modulators"
include both inhibitors and activators, where "inhibitors" refer to
compounds that decrease, prevent, inactivate, desensitize, or
down-regulate the P2X7 receptor expression or activity, and
"activators" are compounds that increase, activate, facilitate,
sensitize, or up-regulate P2X7 receptor expression or activity.
[0175] The term "treat", "treatment" or "treating", as used herein,
is intended to refer to administration of an active agent or
composition of the invention to a subject for the purpose of
affecting a therapeutic or prophylactic benefit through modulation
of P2X7 receptor activity. Treating includes reversing,
ameliorating, alleviating, inhibiting the progress of, lessening
the severity of, or preventing a disease, disorder, or condition,
or one or more symptoms of such disease, disorder or condition
mediated through modulation of P2X7 receptor activity. The term
"subject" refers to a mammalian patient in need of such treatment,
such as a human.
[0176] Accordingly, the invention relates to methods of using the
compounds described herein to treat subjects diagnosed with or
suffering from a disease, disorder, or condition mediated by P2X7
receptor activity, such as: diseases of the autoimmune and
inflammatory system [examples of diseases of the autoimmune and
inflammatory system include rheumatoid arthritis, osteoarthritis,
interstitial cystitis, psoriasis, septic shock, sepsis, allergic
dermatitis, asthma (examples of asthma include allergic asthma,
mild to severe asthma, and steroid resistant asthma), idiopathic
pulmonary fibrosis, allergic rhinitis, chronic obstructive
pulmonary disease and airway hyper-responsivenes]; diseases of the
nervous and neuro-immune system [examples of diseases of the
nervous and neuro-immune system include acute and chronic pain
(examples of acute and chronic pain include neuropathic pain,
inflammatory pain, migraine, spontaneous pain (examples of
spontaneous pain include opioid induced pain, diabetic neuropathy,
postherpetic neuralgia, low back pain, chemotherapy-induced
neuropathic pain, fibromyalgia)]; diseases involved with, and
without, neuroinflammation of the Central Nervous System (CNS)
[examples of diseases involved with, and without, neuroinflammation
of the Central Nervous System (CNS) include mood disorders
(examples of mood disorders include major depression, major
depressive disorder, treatment resistant depression, bipolar
disorder, anxious depression, anxiety), cognition, sleep disorders,
multiple sclerosis, epileptic seizures, Parkinson's disease,
schizophrenia, Alzheimer's disease, Huntington's disease,
Amyotrophic Lateral Sclerosis, autism, spinal cord injury and
cerebral ischemia/traumatic brain injury, and stress-related
disorders]; diseases of the cardiovascular, metabolic,
gastrointestinal and urogenital systems [examples of diseases of
the cardiovascular, metabolic, gastrointestinal and urogenital
systems include diabetes, diabetes mellitus, thrombosis, irritable
bowel disease, irritable bowel syndrome, Crohn's disease,
cardiovascular diseases (examples of cardiovascular disease include
hypertension, myocardial infarction, ischemic heart disease,
ischemia) ureteric obstruction, lower urinary tract syndrome, lower
urinary tract dysfunction such as incontinence, and disease after
cardiac transplantation]; skeletal disorders, (examples of skeletal
disorders include osteoporosis/osteopetrosis) and diseases
involving the secretory function of exocrine glands and diseases
such as glaucoma, Glomerulonephritis, Chaga's Disease, chlamydia,
neuroblastoma, Tuberculosis, Polycystic Kidney Disease, cancer, and
acne.
[0177] In treatment methods according to the invention, an
effective amount of a pharmaceutical agent according to the
invention is administered to a subject suffering from or diagnosed
as having such a disease, disorder, or condition. An "effective
amount" means an amount or dose sufficient to generally bring about
the desired therapeutic or prophylactic benefit in patients in need
of such treatment for the designated disease, disorder, or
condition. Effective amounts or doses of the compounds of the
present invention may be ascertained by routine methods such as
modeling, dose escalation studies or clinical trials, and by taking
into consideration routine factors, e.g., the mode or route of
administration or drug delivery, the pharmacokinetics of the
compound, the severity and course of the disease, disorder, or
condition, the subject's previous or ongoing therapy, the subject's
health status and response to drugs, and the judgment of the
treating physician. An example of a dose is in the range of from
about 0.001 to about 200 mg of compound per kg of subject's body
weight per day, preferably about 0.05 to 100 mg/kg/day, or about 1
to 35 mg/kg/day, in single or divided dosage units (e.g., BID, TID,
QID). For a 70-kg human, an illustrative range for a suitable
dosage amount is from about 0.05 to about 7 g/day, or about 0.2 to
about 2.5 g/day.
[0178] Once improvement of the patient's disease, disorder, or
condition has occurred, the dose may be adjusted for preventative
or maintenance treatment. For example, the dosage or the frequency
of administration, or both, may be reduced as a function of the
symptoms, to a level at which the desired therapeutic or
prophylactic effect is maintained. Of course, if symptoms have been
alleviated to an appropriate level, treatment may cease. Patients
may, however, require intermittent treatment on a long-term basis
upon any recurrence of symptoms.
[0179] In addition, the active agents of the invention may be used
in combination with additional active ingredients in the treatment
of the above conditions. The additional active ingredients may be
coadministered separately with an active agent of compounds of
Tables 1 or included with such an agent in a pharmaceutical
composition according to the invention. In an exemplary embodiment,
additional active ingredients are those that are known or
discovered to be effective in the treatment of conditions,
disorders, or diseases mediated by P2X7 activity, such as another
P2X7 modulator or a compound active against another target
associated with the particular condition, disorder, or disease. The
combination may serve to increase efficacy (e.g., by including in
the combination a compound potentiating the potency or
effectiveness of an active agent according to the invention),
decrease one or more side effects, or decrease the required dose of
the active agent according to the invention.
[0180] The active agents of the invention are used, alone or in
combination with one or more additional active ingredients, to
formulate pharmaceutical compositions of the invention. A
pharmaceutical composition of the invention comprises: (a) an
effective amount of at least one active agent in accordance with
the invention; and (b) a pharmaceutically acceptable excipient.
[0181] A "pharmaceutically acceptable excipient" refers to a
substance that is non-toxic, biologically tolerable, and otherwise
biologically suitable for administration to a subject, such as an
inert substance, added to a pharmacological composition or
otherwise used as a vehicle, carrier, or diluent to facilitate
administration of an agent and that is compatible therewith.
Examples of excipients include calcium carbonate, calcium
phosphate, various sugars and types of starch, cellulose
derivatives, gelatin, vegetable oils, and polyethylene glycols.
[0182] Delivery forms of the pharmaceutical compositions containing
one or more dosage units of the active agents may be prepared using
suitable pharmaceutical excipients and compounding techniques known
or that become available to those skilled in the art. The
compositions may be administered in the inventive methods by a
suitable route of delivery, e.g., oral, parenteral, rectal,
topical, or ocular routes, or by inhalation.
[0183] The preparation may be in the form of tablets, capsules,
sachets, dragees, powders, granules, lozenges, powders for
reconstitution, liquid preparations, or suppositories. Preferably,
the compositions are formulated for intravenous infusion, topical
administration, or oral administration.
[0184] For oral administration, the compounds of the invention can
be provided in the form of tablets or capsules, or as a solution,
emulsion, or suspension. To prepare the oral compositions, the
compounds may be formulated to yield a dosage of, e.g., from about
0.05 to about 100 mg/kg daily, or from about 0.05 to about 35 mg/kg
daily, or from about 0.1 to about 10 mg/kg daily. For example, a
total daily dosage of about 5 mg to 5 g daily may be accomplished
by dosing once, twice, three, or four times per day.
[0185] Oral tablets may include a compound according to the
invention mixed with pharmaceutically acceptable excipients such as
inert diluents, disintegrating agents, binding agents, lubricating
agents, sweetening agents, flavoring agents, coloring agents and
preservative agents. Suitable inert fillers include sodium and
calcium carbonate, sodium and calcium phosphate, lactose, starch,
sugar, glucose, methyl cellulose, magnesium stearate, mannitol,
sorbitol, and the like. Exemplary liquid oral excipients include
ethanol, glycerol, water, and the like. Starch,
polyvinyl-pyrrolidone (PVP), sodium starch glycolate,
microcrystalline cellulose, and alginic acid are suitable
disintegrating agents. Binding agents may include starch and
gelatin. The lubricating agent, if present, may be magnesium
stearate, stearic acid or talc. If desired, the tablets may be
coated with a material such as glyceryl monostearate or glyceryl
distearate to delay absorption in the gastrointestinal tract, or
may be coated with an enteric coating.
[0186] Capsules for oral administration include hard and soft
gelatin capsules. To prepare hard gelatin capsules, compounds of
the invention may be mixed with a solid, semi-solid, or liquid
diluent. Soft gelatin capsules may be prepared by mixing the
compound of the invention with water, an oil such as peanut oil or
olive oil, liquid paraffin, a mixture of mono and di-glycerides of
short chain fatty acids, polyethylene glycol 400, or propylene
glycol.
[0187] Liquids for oral administration may be in the form of
suspensions, solutions, emulsions or syrups or may be lyophilized
or presented as a dry product for reconstitution with water or
other suitable vehicle before use. Such liquid compositions may
optionally contain: pharmaceutically-acceptable excipients such as
suspending agents (for example, sorbitol, methyl cellulose, sodium
alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose,
aluminum stearate gel and the like); non-aqueous vehicles, e.g.,
oil (for example, almond oil or fractionated coconut oil),
propylene glycol, ethyl alcohol, or water; preservatives (for
example, methyl or propyl p-hydroxybenzoate or sorbic acid);
wetting agents such as lecithin; and, if desired, flavoring or
coloring agents.
[0188] The active agents of this invention may also be administered
by non-oral routes. For example, the compositions may be formulated
for rectal administration as a suppository. For parenteral use,
including intravenous, intramuscular, intraperitoneal, or
subcutaneous routes, the compounds of the invention may be provided
in sterile aqueous solutions or suspensions, buffered to an
appropriate pH and isotonicity or in parenterally acceptable oil.
Suitable aqueous vehicles include Ringer's solution and isotonic
sodium chloride. Such forms will be presented in unit-dose form
such as ampules or disposable injection devices, in multi-dose
forms such as vials from which the appropriate dose may be
withdrawn, or in a solid form or pre-concentrate that can be used
to prepare an injectable formulation. Illustrative infusion doses
may range from about 1 to 1000 .mu.g/kg/minute of compound, admixed
with a pharmaceutical carrier over a period ranging from several
minutes to several days.
[0189] For topical administration, the compounds may be mixed with
a pharmaceutical carrier at a concentration of about 0.1% to about
10% of drug to vehicle. Another mode of administering the compounds
of the invention may utilize a patch formulation to affect
transdermal delivery.
[0190] Compounds of the invention may alternatively be administered
in methods of this invention by inhalation, via the nasal or oral
routes, e.g., in a spray formulation also containing a suitable
carrier.
Schemes
[0191] The group PG represents a protecting group. One skilled in
the art will select the appropriate protecting group compatible
with the desired reactions. The protecting groups may be removed at
a convenient subsequent stage using methods known from the art.
Alternatively, it may be necessary to employ, in the place of the
ultimately desired substituent, a suitable group that may be
carried through the reaction scheme and replaced as appropriate
with the desired substituent. Such compounds, precursors, or
prodrugs are also within the scope of the invention. Examples of
preferred protecting groups include; carbamates, benzyl and
substituted benzyl groups. Especially preferred protecting groups
are tert-butyloxycarbonyl and benzyl.
##STR00065##
[0192] Compound IA can be converted to compound IIA by reaction
with Lawesson's reagent, in a solvent such as THF, diethyl ether or
DCM. This reaction may be performed at room temperature or heated
overnight at or near the boiling point of the solvent.
[0193] Compound IIA may be converted to amine IIIA by treatment
with an alkylating agent such as trimethyloxonium tetrafluoroborate
or methyl iodide in a solvent such as DCM or DMF, at a temperature
of between room temperature and 40.degree. C. for between 1 and 48
hours.
##STR00066##
##STR00067##
[0194] Compound IVA may be converted to compound VA by treatment
with hydrazine monohydrate in a solvent such as an alcohol, DCM or
DMF at a temperature near room temperature for from 1 to 25 hours.
Compound VIA may be converted to compound VIIA by treatment with an
appropriate acylating agent such as oxalyl chloride in the presence
of a catalyst such as DMF in a solvent such as DCM or DMF for from
1 to 8 hours. Compound VIIA may then also be converted to compound
VA by treatment with hydrazine monohydrate in a solvent such as an
alcohol, DCM or DMF at a temperature near room temperature for from
1 to 12 hours. Additionally compound VIAB may be converted to
compound VIIAB by treatment with an appropriate acylating agent
such as oxalyl chloride in the presence of a catalyst such as DMF
in a solvent such as DCM or DMF for from 1 to 8 hours. If compounds
of type IVA, VIA or VIAB are not commercially available, one
skilled in the art will realize there are numerous methods for
synthesizing these compounds. These may include hydrolysis of the
corresponding nitrile to afford VIA followed by esterification to
give IVA. The nitrile in turn can be obtained from a cross-coupling
reaction with a suitable halogen containing compound. Hydrolysis of
the corresponding nitrile to could also afford VIAB. Or VIA or VIAB
can be directly formed from the halogen compound via metal halogen
exchange followed by quenching with CO.sub.2. VIA or VIAB can also
be formed by oxidation of a suitable methyl substituted compound
with a reagent, such as, KMnO.sub.4 and then IVA may be formed by
subsequent esterification of VIA. These compounds can also be
formed by oxidation of an appropriately substituted hydroxymethyl
compound in either one or two steps to afford VIA or VIAB.
##STR00068##
[0195] Compound IIIA may be converted to compound VIIIA by the
addition of compound VA and a suitable base such as potassium
t-butoxide in an alcohol solvent such as methanol. This reaction
can be performed at a temperature from room temperature to
120.degree. C. for from 30 minutes to 48 hours. Compound VIIIA can
then be converted to compound IXA by addition of a suitable acid
such as HCl or TFA, preferably TFA in a solvent such as DCM, DCE or
dioxane. This reaction can be performed at a temperature from room
temperature to 50.degree. C. for from 30 minutes to 24 hours.
EXAMPLES
[0196] Exemplary compounds useful in methods of the invention will
now be described by reference to the illustrative synthetic schemes
for their general preparation below and the specific examples that
follow. Artisans will recognize that, to obtain the various
compounds herein, starting materials may be suitably selected so
that the ultimately desired substituents will be carried through
the reaction scheme with or without protection as appropriate to
yield the desired product. Alternatively, it may be necessary or
desirable to employ, in the place of the ultimately desired
substituent, a suitable group that may be carried through the
reaction scheme and replaced as appropriate with the desired
substituent. Unless otherwise specified, the variables are as
defined above in reference to Formula (I, IIa and IIb). Reactions
may be performed between the melting point and the reflux
temperature of the solvent, and preferably between 0.degree. C. and
the reflux temperature of the solvent. Reactions may be heated
employing conventional heating or microwave heating. Reactions may
also be conducted in sealed pressure vessels above the normal
reflux temperature of the solvent.
[0197] In obtaining the compounds described in the examples below
and the corresponding analytical data, the following experimental
and analytical protocols were followed unless otherwise
indicated.
[0198] Unless otherwise stated, reaction mixtures were magnetically
stirred at room temperature (rt) under a nitrogen atmosphere. Where
solutions were "dried," they were generally dried over a drying
agent such as Na.sub.2SO.sub.4 or MgSO.sub.4. Where mixtures,
solutions, and extracts were "concentrated", they were typically
concentrated on a rotary evaporator under reduced pressure.
Reactions under microwave irradiation conditions were carried out
in a Biotage Initiator or CEM Corporation Discover instrument.
Hydrogenations on the H-cube were run by passing solvent containing
reactant through a catalyst cartridge on an H-Cube hydrogenation
apparatus at a pressure of 15 to 100 bar and a flow rate of 1 to 30
ml/min.
[0199] Normal-phase silica gel column chromatography (sgc) was
performed on silica gel (SiO.sub.2) using prepackaged cartridges,
eluting with 2 M NH.sub.3/MeOH in CH.sub.2Cl.sub.2 unless otherwise
indicated.
[0200] Preparative reverse-phase high performance liquid
chromatography (HPLC) was performed on a Agilent HPLC with an
Xterra Prep RID.sub.18 (5 .mu.m, 30.times.100 mm, or 50.times.150
mm) column, and a gradient of 10 to 99% acetonitrile/water (20 mM
NH.sub.4OH) over 12 to 18 min, and a flow rate of 30 or 80 mL/min,
unless otherwise indicated.
[0201] Mass spectra (MS) were obtained on an Agilent series 1100
MSD using electrospray ionization (ESI) in positive mode unless
otherwise indicated. Calculated (calcd.) mass corresponds to the
exact mass.
[0202] Nuclear magnetic resonance (NMR) spectra were obtained on
Bruker model DRX spectrometers. The format of the .sup.1H NMR data
below is: chemical shift in ppm downfield of the tetramethylsilane
reference (multiplicity, coupling constant J in Hz,
integration).
[0203] A notation of (.+-.) or R/S indicates that the product is a
racemic mixture of enantiomers and/or diastereomers. A notation of,
for example, (2S, 3R) indicates that product stereochemistry
depicted is based on the known stereochemistry of similar compounds
and/or reactions. A notation of, for example, (2S*, 3R*) indicates
that the product is a pure and single diastereomer but the absolute
stereochemistry is not established and relative stereochemistry is
shown.
[0204] Chemical names were generated using Chem Draw Ultra 6.0.2
(CambridgeSoft Corp., Cambridge, Mass.).
[0205] Abbreviations and acronyms used herein include the
following:
TABLE-US-00002 Term Acronym/Abbreviation Reverse Phase
High-pressure liquid HPLC or RP HPLC chromatography Tetrahydrofuran
THF tert-Butylcarbamoyl Boc, BOC Dichloromethane DCM
Trifluoroacetic acid TFA N,N-Dimethylformamide DMF Methanol MeOH
Ethanol EtOH Isopropanol IPA, iPrOH n-butanol n-BuOH Acetonitrile
ACN, MeCN Ethyl Acetate EtOAc, or EA Triethylamine TEA
1-Ethyl-3-(3- EDCI dimethylaminopropyl)carbodiimide Dimethyl
sulfoxide DMSO Hexane HEX Supercritical fluid chromatography SFC
Sodium Acetate NaOAc Room Temperature RT, rt
Example 1
(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]triazolo-
[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00069##
[0206] Example 1
Step a: tert-Butyl 2-methyl-3-oxopiperazine-1-carboxylate
[0207] To 3-methylpiperazin-2-one (1.08 g, 9.33 mmol) in 1:1
THF/H.sub.2O (45 mL) was added Na.sub.2CO.sub.3 (2.08 g, 19.60
mmol) and BOC-anhydride (2.24 g, 10.27 mmol). The reaction was
allowed to stir for 4 h, then extracted with DCM. The combined
organics were washed 1.times. with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo (2.00 g, 99%).
MS (ESI) mass calcd. C.sub.10H.sub.18N.sub.2O.sub.3, 214.13; m/z
found 429.0 [2M+H].sup.+, 159.0 [M+H-tBu].sup.+. 1H NMR (500 MHz,
CDCl.sub.3): 8.05 (s, 1H), 4.42 (s, 2H), 3.69-3.61 (m, 2H), 2.63
(s, 2H), 1.49 (s, 9H).
Example 1
Step b: tert-Butyl
8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H-
)-carboxylate
[0208] To a solution of the product of Example 1, step a (230 mg,
1.08 mmol) in DCM (5 mL) was added trimethyloxonium
tetrafluoroborate (194 mg, 1.25 mmol). The reagent slowly dissolved
and after stirring overnight all of the starting material was
consumed. To this solution was added 2-picolinyl hydrazide (181 mg,
1.29 mmol). After 24 h the reaction was concentrated in vacuo and
dissolved in dioxane (2 mL) and saturated aqueous NaHCO.sub.3
solution (2 mL). The mixture was heated for 3 h at 90.degree. C.
and the dioxane was removed in vacuo and the aqueous layer
extracted with DCM and EtOAc. The combined organic extracts were
dried over Na.sub.2SO.sub.4 filtered and concentrated in vacuo.
Chromatography on SiO.sub.2 eluting with IPA/EtOAc afforded the
title compound (150 mg, 44%). MS (ESI) mass calcd.
C.sub.16H.sub.21N.sub.5O.sub.2, 315.17; m/z found 316.0
[M+H].sup.-.
Example 1
Step c:
8-methyl-3-(pyridin-2-yl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a-
]pyrazine
[0209] To the product of Example 1, step b (150 mg, 0.48 mmol) in
DCM (2 mL) was added TFA (0.48 mL). After stirring 3 h, the
reaction was concentrated in vacuo. The residue was redissolved in
DCM and treated with Dowex 550A resin. The resin was removed by
filtration and concentration afforded a white solid. Chromatography
on SiO.sub.2 eluting with 2M NH.sub.3 in MeOH/DCM afforded the
desired compound (100 mg, 98%). MS (ESI) mass calcd.
C.sub.11H.sub.13N.sub.5, 215.12; m/z found 216.0 [M+H].sup.+.
Example 1
Step d:
7-[(2,3-Dichlorophenyl)carbonyl]-8-methyl-3-pyridin-2-yl-5,6,7,8-t-
etrahydro[1,2,4]triazolo[4,3-a]pyrazine
[0210] To a solution of the product of Example 1, step c (83 mg,
0.39 mmol) in DCM (4 mL) was added 2,4-dichlorobenzoic acid (74 mg,
0.39 mmol) followed by EDCI (111 mg, 0.58 mmol), HOBt (36 mg, 0.27
mmol) and TEA (0.11 mL, 0.77 mmol). The mixture was stirred
overnight and then loaded directly on a column. Chromatography on
SiO.sub.2 eluting with EtOAc/Hex afforded impure material.
Purification of this material on a Prep Agilent system with a
XBridge C18 OBD 50.times.100 mm column eluting with 5 to 99% 0.05%
NH.sub.4OH in H.sub.2O/ACN over 17 min afforded the desired product
(51 mg, 34%). MS (ESI) mass calcd.
C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.07; m/z found 387.9
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.68-8.52 (m, 1H), 8.38-8.28
(m, 1H), 7.88-7.80 (m, 1H), 7.60-7.53 (m, 1H), 7.39-7.27 (m, 3H),
6.24-6.17 (m, 0.5H), 5.22-4.91 (m, 2H), 4.44-4.10 (m, 1H),
3.73-3.32 (m, 1.5H), 1.84-1.57 (m, 3H).
Example 2
(R)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00070##
[0212] Example 2, absolute configuration unknown, was obtained by
chiral separation of Example 1 utilizing SFC.
[0213] Stationary Phase: Amycoat 5 .mu.m 250.times.30 mm
(L.times.I.D.) at 40.degree. C.
[0214] Mobile Phase: 25.5 mL/min EtOH with 0.2% isopropylamine,
59.5 mL/min CO.sub.2
[0215] Detection: UV 254 nm.
[0216] Example 2 was the second compound off the column (16 mg). MS
(ESI) mass calcd. C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.07; m/z
found 388.1 [M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.67-8.53 (m,
1H), 8.37-8.29 (m, 1H), 7.87-7.80 (m, 1H), 7.59-7.54 (m, 1H),
7.38-7.23 (m, 3H), 6.24-6.17 (m, 0.5H), 5.23-4.89 (m, 2H),
4.43-4.10 (m, 1H), 3.74-3.61 (m, 1H), 3.59-3.32 (m, 0.5H),
1.85-1.55 (m, 3H).
Example 3
(S)-(2,3-dichlorophenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00071##
[0218] Example 3, absolute configuration unknown, was obtained by
chiral separation of Example 1 utilizing SFC.
[0219] Stationary Phase: Amycoat 5 .mu.m 250.times.30 mm
(L.times.I.D.) at 40.degree. C.
[0220] Mobile Phase: 25.5 mL/min EtOH with 0.2% isopropylamine,
59.5 mL/min CO.sub.2
[0221] Detection: UV 254 nm.
[0222] Example 3 was the first compound off the column (16 mg). MS
(ESI) mass calcd. C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.07; m/z
found 388.1 [M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.68-8.53 (m,
1H), 8.37-8.29 (m, 1H), 7.88-7.81 (m, 1H), 7.60-7.54 (m, 1H),
7.38-7.23 (m, 3H), 6.23-6.17 (m, 0.5H), 5.22-4.91 (m, 2H),
4.43-4.10 (m, 1H), 3.73-3.63 (m, 1H), 3.59-3.32 (m, 0.5H),
1.84-1.52 (m, 3H).
[0223] Examples 4-11 can be made in a manner analogous to Example
1, substituting the appropriate starting materials for each
step.
Example 4
(2-chloro-3-(trifluoromethyl)phenyl)(5-methyl-3-phenyl-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00072##
[0224] Example 5
(2,3-dichlorophenyl)(5-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]p-
yrazin-7(8H)-yl)methanone
##STR00073##
[0225] Example 6
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-5-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00074##
[0226] Example 7
(2,3-dichlorophenyl)(3-(4-fluorophenyl)-5-methyl-5,6-dihydro-[1,2,4]triazo-
lo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00075##
[0227] Example 8
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00076##
[0228] Example 9
(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]triazo-
lo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00077##
[0229] Example 10
(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00078##
[0230] Example 11
(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]p-
yrazin-7(8H)-yl)methanone
##STR00079##
[0231] Example 12
(R)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
-a]pyrazin-7(8H)-yl)methanone
##STR00080##
[0233] Example 12 was isolated following chiral SFC separation of
Example 11 on a CHIRALCEL OD-H 5 .mu.m 250.times.20mm column with
mobile phase consisting of 70% CO.sub.2, 30% MeOH. Example 12 was
the first eluting peak under these conditions. MS (ESI): mass
calcd. for C.sub.19H.sub.16Cl.sub.2N.sub.4O, 386.1; m/z found,
386.10 [M+H].sup.+.
Example 13
(S)-(2,3-dichlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-
-a]pyrazin-7(8H)-yl)methanone
##STR00081##
[0235] Example 13 was isolated following chiral SFC separation of
Example 11 on a CHIRALCEL OD-H 5 .mu.m 250.times.20mm column with
mobile phase consisting of 70% CO.sub.2, 30% MeOH. Example 13 was
the second eluting peak under these conditions. MS (ESI): mass
calcd. for C.sub.19H.sub.16Cl.sub.2N.sub.4O, 386.1; m/z found,
386.10 [M+H].sup.+.
Example 14
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00082##
[0237] Example 14 was isolated following chiral SFC separation of
Example 10 on a CHIRALCEL OD-H 5 .mu.m 250.times.20mm column with
mobile phase consisting of 75% CO.sub.2, 25% MeOH. Example 14 was
the first eluting peak under these conditions. MS (ESI): mass
calcd. for C.sub.20H.sub.16ClF.sub.3N.sub.4O, 420.1; m/z found,
420.10 [M+H].sup.+.
Example 15
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00083##
[0239] Example 15 was isolated following chiral SFC separation of
Example 10 on a CHIRALCEL OD-H 5 .mu.m 250.times.20mm column with
mobile phase consisting of 75% CO.sub.2, 25% MeOH. Example 14 was
the second eluting peak under these conditions. MS (ESI): mass
calcd. for C.sub.20H.sub.16ClF.sub.3N.sub.4O, 420.1; m/z found,
420.10 [M+H].sup.+.
Example 16
(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-methyl-3-(pyridin-2-yl)-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00084##
[0241] The title compound was prepared in a manner analogous to
Example 1 substituting 3-chloro-2-(trifluoromethyl)isonicotinic
acid for 2,3-dichlorobenzoic acid in Example 1, step d. MS (ESI)
mass calcd. C.sub.18H.sub.14ClF.sub.3N.sub.6O, 422.09; m/z found
423.1 [M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.76-8.52 (m, 2H),
8.37-8.30 (m, 1H), 7.89-7.81 (m, 1H), 7.54-7.46 (m, 1H), 7.41-7.30
(m, 1H), 6.23-6.15 (m, 1H), 5.25-4.91 (m, 2H), 4.43-4.08 (m, 1H),
3.80-3.36 (m, 2H), 1.85-1.61 (m, 3H).
Example 17
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00085##
[0243] The title compound was prepared in a manner analogous to
Example 1 substituting 2-chloro-3-(trifluoromethyl)benzoic acid for
2,3-dichlorobenzoic acid in Example 1, step d. MS (ESI) mass calcd.
C.sub.19H.sub.15ClF.sub.3N.sub.5O, 421.09; m/z found 422.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.67-8.51 (m, 1H), 8.37-8.29
(m, 1H), 7.88-7.79 (m, 2H), 7.59-7.30 (m, 3H), 6.25-6.18 (m, 1H),
5.21-4.93 (m, 2H), 4.42-4.11 (m, 1H), 3.76-3.33 (m, 1H), 1.85-1.58
(m, 3H).
Example 18
(R)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00086##
[0245] 1H NMR (400 MHz, DMSO) .delta. 7.91-7.34 (m, 7H), 5.72-5.39
(m, 1H), 4.76-3.70 (m, 4H), 1.27-0.96 (m, 3H). MS (ESI): mass
calcd. for C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z found,
405.1 [M+H].sup.+.
Example 19
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00087##
[0247] 1H NMR (400 MHz, DMSO) .delta. 7.93-7.31 (m, 7H), 5.72-5.35
(m, 1H), 4.80-3.75 (m, 4H), 1.23-0.96 (m, 3H). MS (ESI): mass
calcd. for C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z found,
405.1 [M+H].sup.+.
Example 20
(2-fluoro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyridin-2-yl)-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00088##
[0249] The title compound was prepared in a manner analogous to
Example 1 substituting 2-fluoro-3-(trifluoromethyl)benzoic acid for
2,3-dichlorobenzoic acid in Example 1, step d. MS (ESI) mass calcd.
C.sub.19H.sub.15F.sub.4N.sub.5O, 405.12; m/z found 406.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.68-8.54 (m, 1H), 8.37-8.29
(m, 1H), 7.87-7.52 (m, 3H), 7.43-7.31 (m, 2H), 6.19 (s, 1H),
5.22-4.96 (m, 2H), 4.43-4.17 (m, 1H), 3.85-3.37 (m, 1H), 1.86-1.65
(m, 3H).
Example 21
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00089##
[0250] Intermediate 21A: (S)-tert-butyl
(1-azidopropan-2-yl)carbamate
[0251] To a solution of Boc-L-alaninol (10.9 g, 61.7 mmol) in ether
(300 mL) at 0.degree. C. was added triethylamine (12.8 mL, 92.5
mmol) followed by methanesulonylchloride (4.8 mL, 61.7 mmol) and
the reaction mixture was stirred to 1 hour. Water was added and the
resulting reaction mixture was extracted with DCM. The organic
layers were combined, dried, concentrated and the resulting residue
was dissolved in DMF (100 mL). To the resulting solution was added
sodium azide (8.0 g, 123.4 mmol) and the reaction mixture was
heated to 70.degree. C. for 18 hours. The reaction mixture was
cooled to rt, water was added and the reaction mixture was
extracted with EtOAc. The organic layers were combined, washed with
brine, dried, concentrated and purified by flash column
chromatography (0-50% EtOAc in hexanes) to provide (S)-tert-butyl
(1-azidopropan-2-yl)carbamate (8.5 g). 1H NMR (400 MHz, DMSO)
.delta. 7.05-6.84 (d, J=8.0 Hz, 1H), 3.72-3.55 (m, 1H), 3.26-3.19
(d, J=6.1 Hz, 2H), 1.42-1.36 (s, 9H), 1.06-0.99 (d, J=6.8 Hz,
3H).
Intermediate 21B: (S)-tert-butyl
(1-(2-chloroacetamido)propan-2-yl)carbamate
[0252] To a solution of (S)-tert-butyl
(1-azidopropan-2-yl)carbamate (8.5g, 42.4 mmol) in EtOAc (300 mL)
was added 10% Pd/C (4.5g) and the reaction mixture was placed under
H.sub.2 atmosphere (60 psi) for 2 hours. The reaction mixture was
filtered through a pad of celite, concentrated and the resulting
residue was taken up in DCM (300 mL). The resulting solution was
cooled to -78.degree. C. and triethylamine (8.9 mL) was added
followed by chloroacetyl chloride (3.5 mL, 44.6 mmol). The reaction
mixture was stirred at -78.degree. C. for 20 minutes then warmed to
0.degree. C. where it was stirred for 1 hour. Water was added and
the resulting reaction mixture was extracted with DCM. The organic
layers were combined, washed with brine, dried, concentrated and
purified by flash column chromatography (0-100% EtOAc in hexanes)
to provide (S)-tert-butyl
(1-(2-chloroacetamido)propan-2-yl)carbamate (7.3 g). 1H NMR (400
MHz, DMSO) .delta. 8.28-8.10 (s, 1H), 6.79-6.60 (d, J=8.2 Hz, 1H),
4.16-3.97 (s, 2H), 3.64-3.48 (m, 1H), 3.13-2.99 (m, 2H), 1.44-1.30
(s, 9H), 1.05-0.91 (d, J=6.7 Hz, 3H).
Intermediate 21C: (S)-tert-butyl
2-methyl-5-oxopiperazine-1-carboxylate
[0253] (S)-Tert-butyl (1-(2-chloroacetamido)propan-2-yl)carbamate
(7.3 g, 29.1 mmol) was dissolved in trifluoroacetic acid (20 mL)
and stirred at rt for 15 minutes. The reaction mixture was
concentrated and the resulting residue was dissolved in THF (100
mL). To the resulting solution was added K.sub.2CO.sub.3 (20.1 g,
145.6 mmol) and the reaction mixture refluxed for 20 hours. The
reaction mixture was cooled to 60.degree. C. and catalytic DMAP was
added followed by BOC-anhydride (12.5 mL, 58.2 mmol). The reaction
mixture was stirred for 12 hours, water was added and the resulting
reaction mixture was extracted with EtOAc. The organic layers were
combined, dried, concentrated and purified by flash column
chromatography (0-50% iPrOH in EtOAc) to provide (S)-tert-butyl
2-methyl-5-oxopiperazine-1-carboxylate (4.6g). 1H NMR (400 MHz,
DMSO) .delta. 8.08-7.90 (s, 1H), 4.31-4.09 (s, 1H), 4.00-3.87 (d,
J=17.9 Hz, 1H), 3.63-3.51 (d, J=17.8 Hz, 1H), 3.37-3.33 (m, 1H),
3.04-2.93 (ddd, J=12.7, 4.9, 2.5 Hz, 1H), 1.46-1.35 (s, 9H),
1.15-1.06 (d, J=6.7 Hz, 3H).
Intermediate 21D: (S)-tert-butyl
3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(-
8H)-carboxylate
[0254] To a solution of (S)-tert-butyl
2-methyl-5-oxopiperazine-1-carboxylate (1.3 g, 6.1 mmol) in DCM (31
mL) was added trimethyloxonium tetrafluoroborate (1.0 g, 6.8 mmol)
and the reaction mixture was stirred at rt for 6 hours.
4-fluorobenzohydrazide (1.2 g, 8.0 mmol) was added and the reaction
mixture was allowed to stir at rt overnight. The reaction mixture
was concentrated via gentle N.sub.2 stream and the resulting
residue was dissolved in dioxane (15 mL). To the resulting solution
was added saturated aqueous sodium bicarbonate (15 mL) and the
reaction mixture was refluxed for 12 hours. The reaction mixture
was cooled to rt, diluted with EtOAc, washed with water, dried,
concentrated and purified by flash column chromatography (0-10%
MeOH in DCM) to provide (S)-tert-butyl
3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(-
8H)-carboxylate (1.2 g). MS (ESI) mass calcd.
C.sub.17H.sub.21FN.sub.4O.sub.2, 332.4; m/z found, 333.2
[M+H].sup.+.
Intermediate 21 E:
(S)-3-(4-fluorophenyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]-
pyrazine
[0255] (S)-Tert-butyl
3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(-
8H)-carboxylate (0.2 g, 0.6 mmol) was dissolved in trifluoroacetic
acid and stirred at rt for 20 minutes. The reaction mixture was
concentrated and the resulting residue was used without further
purification.
[0256] To a solution of
(S)-3-(4-fluorophenyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]-
pyrazine (90 mg, 0.387 mmol) and
2-chloro-3-(trifluoromethyl)benzoyl chloride (109 mg, 0.452 mmol)
in DCM (10 mL) was added triethylamine (0.2 mL, 1.5 mmol) and the
reaction mixture was stirred at rt for 1 hour. Water was added and
the resulting reaction mixture was extracted with DCM. The organic
layers were combined, dried, concentrated and purified by flash
column chromatography (0-70% iPrOH in EtOAc) to provide
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-di-
hydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (70 mg). 1H
NMR (400 MHz, DMSO) .delta. 7.93-7.31 (m, 7H), 5.72-5.35 (m, 1H),
4.80-3.75 (m, 4H), 1.23-0.96 (m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found,
439.1[M+H].sup.+.
Example 22
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00090##
[0258]
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in an
analogous fashion to that described for
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2,3-dichlorobenzoyl chloride was used in place of
2-chloro-3-(trifluoromethyl)benzoyl chloride and
pyrazine-2-carbohydrazide was used in place of
4-fluorobenzohydrazide. 1H NMR (400 MHz, DMSO) .delta. 9.54-8.01
(m, 3H), 7.90-7.25 (m, 3H), 5.73-5.46 (m, 1H), 4.90-4.02 (m, 3H),
3.85-3.57 (m, 1H), 1.34-0.98 (m, 3H). MS (ESI): mass calcd. for
C.sub.17H.sub.14Cl.sub.2N.sub.6O, 388.1; m/z found, 390.1
[M+H].sup.+.
Example 23
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(pyrazin-2-yl)-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00091##
[0260]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(pyrazin-2-yl)--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous manor to
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
pyrazine-2-carbohydrazide was used in place of
4-fluorobenzohydrazide. 1H NMR (400 MHz, DMSO) .delta. 9.51-8.68
(m, 3H), 8.22-7.45 (m, 3H), 5.75-5.24 (m, 1H), 4.91-4.05 (m, 3H),
3.92-3.57 (m, 1H), 1.37-0.98 (m, 3H). MS (ESI): mass calcd. for
C.sub.18H.sub.14ClF.sub.3N.sub.6O, 422.1; m/z found, 423.1
[M+H].sup.+.
Example 24
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-6-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00092##
[0261] Intermediate 23A: (S)-tert-butyl
6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate
##STR00093##
[0262] Step A: (S)-tert-butyl
6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate
[0263] (S)-tert-butyl
6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate
was prepared as described for Intermediate 21D, substituting formic
acid hydrazide for 4-fluorobenzohydrazide. MS (ESI) mass calcd.
C.sub.11H.sub.17BrN.sub.4O.sub.2, 316.05; m/z found, 317.1
[M+H].sup.+.
Intermediate 23B: (S)-tert-butyl
3-bromo-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxy-
late
##STR00094##
[0264] Step B: (S)-tert-butyl
3-bromo-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxy-
late
[0265] To a solution of (S)-tert-butyl
6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate
(75 mg, 0.32 mmol) in chloroform (3 mL) was added
N-bromosuccinimide (61 mg, 0.35 mmol) and sodium bicarbonate (53
mg, 0.63 mmol). The solution was allowed to stir overnight at rt
then saturated sodium bicarbonate (2 mL was added). The layers were
separated and the water layer was extracted two times more with
methylene chloride. The organic layers were combined, dried over
anhydrous MgSO.sub.4, filtered and concentrated. The residue was
purified by HPLC (Agilent prep system, Waters XBridge C18 5 .mu.m
50.times.100 mm column, 5-99% MeCN/20 nM NH.sub.4OH over 18 min at
80 mL/min) to provide the product (45 mg, 45%). MS (ESI) mass
calcd. C.sub.11H.sub.17BrN.sub.4O.sub.2, 316.05; m/z found, 317.1
[M+H].sup.+.
Intermediate 23C: (S)-tert-butyl
3-(furan-2-yl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)--
carboxylate
##STR00095##
[0266] Step C: (S)-tert-butyl
3-(furan-2-yl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)--
carboxylate
[0267] To a solution of (S)-tert-butyl
3-bromo-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxy-
late (44 mg, 0.14 mmol) in 1,4-dioxane (1 mL) was added
2-furanylboronic acid (47 mg, 0.42 mmol),
[1,1'-BIS(DIPHENYLPHOSPHINO)FERROCENE]DICHLOROPALLADIUM(II) (15 mg,
0.021 mmol), 1,1'-BIS(DIPHENYLPHOSPHINO)FERROCENE (5 mg, 0.008
mmol), and potassium phosphate (88 mg, 0.42 mmol). The flask was
flushed with nitrogen, sealed and heated to 100.degree. C.
overnight. The reaction was allowed to cool and then filtered
through celite. The filtrate was concentrated and the residue was
purified by silica gel chromatography (30-100% ethyl
acetate/hexanes) to provide the product (30 mg, 71%). MS (ESI) mass
calcd. C.sub.15H.sub.20N.sub.4O.sub.3, 304.15; m/z found, 305.2
[M+H].sup.+.
Example 24
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-6-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00096##
[0268] Step D:
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0269] To a solution of (S)-tert-butyl
3-(furan-2-yl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)--
carboxylate (30 mg, 0.1 mmol) in CH.sub.2Cl.sub.2 (1 mL) was added
TFA (0.15 mL, 2.0 mmol). The reaction was allowed to stir at rt for
3 h and then evaporated in vacuo. The residue was dissolved in
CH.sub.2Cl.sub.2 (5 mL) and cooled to 0.degree. C. after which was
added triethylamine (0.092 mL, 0.66 mmol) and (64 mg, 0.26 mmol).
The ice bath was removed and warmed to rt followed by the addition
of water (5 mL). The layers were separated and the water layer was
extracted with CH.sub.2Cl.sub.2 two times more. The organic layers
were combined, dried with MgSO4 and purified by prep HPLC (Agilent
prep system, Waters XBridge C18 5 .mu.m 50.times.100 mm column,
5-99% MeCN/20 nM NH.sub.4OH over 18 min at 80 mL/min) to provide
the product (42 mg, 77%). MS (ESI): mass calcd. for
C.sub.18H.sub.14ClF.sub.3N.sub.4O.sub.2, 410.1; m/z found, 411.1
[M+H].sup.+.
Example 25
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-fluorophenyl)-8-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00097##
[0270] Example 25
Step a: tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate
[0271] To a heterogeneous mixture of Lawesson's reagent (2.11 g,
5.06 mmol) in toluene (18 mL) was added the product of Example 1,
step a (1.01 g, 4.73 mmol). The mixture was heated at 80.degree. C.
for 1 h and then concentrated in vacuo. The residue was dissolved
in DCM and filter loaded on SiO.sub.2 column eluting with EtOAc/Hex
to afford the desired product as an orange solid (1.12 g, 100%). MS
(ESI) mass calcd. C.sub.10H.sub.18N.sub.2O.sub.2S, 230.11; m/z
found 231.1 [M+H].sup.+. 1H NMR (500 MHz, CDCl3): 8.39 (s, 1H),
5.27-4.84 (m, 1H), 4.30-3.92 (m, 1H), 3.53-3.42 (m, 1H), 3.39-3.30
(m, 1H), 3.30-3.05 (m, 1H), 1.62 (d, J=7.0 Hz, 3H), 1.48 (s,
9H).
Example 25
Step b: tert-butyl
3-(4-fluorophenyl)-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(-
8H)-carboxylate
[0272] The product of Example 25, step a (230 mg, 1.00 mmol) and
4-fluorobenzhydrazide (241 mg, 1.50 mmol) were added to a round
bottom flask followed by n-BuOH (4 mL). The mixture was heated at
140.degree. C. for 48 h. The mixture was concentrated in vacuo and
taken on to the next step without further purification. MS (ESI)
mass calcd. C.sub.17H.sub.21FN.sub.4O.sub.2, 332.16; m/z found
333.2 [M+H].sup.+.
Example 25
Step c:
3-(4-fluorophenyl)-8-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-
-a]pyrazine
[0273] To the product of Example 25, step b (332 mg, 1.00 mmol) in
DCM (5 mL) was added TFA (2 mL). After stirring 2 h, the reaction
was complete and concentrated in vacuo. The TFA salt was loaded on
SiO.sub.2 column eluting with 2 M NH.sub.3 in MeOH/DCM over 1 h to
afford the desired compound as a pale yellow solid. MS (ESI) mass
calcd. C.sub.12H.sub.13FN.sub.4, 232.11; m/z found 233.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 7.71-7.65 (m, 2H), 7.23-7.16
(m, 2H), 4.35-4.29 (m, 1H), 4.08-3.97 (m, 2H), 3.45-3.38 (m, 1H),
3.19-3.11 (m, 1H), 1.69 (d, J=6.7 Hz, 3H).
Example 25
Step d:
7-{[2-Chloro-3-(trifluoromethyl)phenyl]carbonyl}-3-(4-fluorophenyl-
)-8-methyl-5,6,7,8-tetrahydro[1,2,4]triazolo[4,3-a]pyrazine
[0274] To a solution of the product of Example 25, step c (189 mg,
0.82 mmol) in DCM (8 mL) was added TEA (0.14 mL, 0.98 mmol)
followed by 2-chloro-3-(trifluoromethyl)benzoyl chloride (208 mg,
0.86 mmol) in one portion. The reaction was stirred overnight and
then loaded directly on a SiO.sub.2 column eluting with IPA/EtOAc
to afford the title compound as a colorless solid (324 mg, 91%). MS
(ESI) mass calcd. C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.09; m/z
found 439.1 [M+H].sup.+. 1H NMR (500 MHz, CDCl3) d 7.86-7.80 (m,
1H), 7.74-7.63 (m, 2H), 7.60-7.40 (m, 2H), 7.26-7.16 (m, 2H),
6.23-6.15 (m, 1H), 5.20-4.97 (m, 1H), 4.34-3.95 (m, 2H), 3.73-3.25
(m, 1H), 1.86-1.53 (m, 3H).
Example 26
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00098##
[0275] Intermediate 26A: (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate
[0276] To a solution of (S)-tert-butyl
2-methyl-5-oxopiperazine-1-carboxylate (1.2 g, 5.6 mmol) in THF 20
mL) was added Lawesson's Reagent (2.6 g, 6.2 mmol) and the reaction
mixture was heated to 80.degree. C. for 1 hour. The reaction
mixture was concentrated and purified by flash column
chrmotatography (0-50% EtOAc in hexanes) to provide (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate with minor residual
impurities. 1H NMR (400 MHz, DMSO) .delta. 10.64-10.46 (s, 1H),
4.48-4.31 (d, J=18.9 Hz, 1H), 4.31-4.12 (s, 1H), 4.11-3.93 (m, 1H),
3.90-3.74 (m, 1H), 3.20-3.06 (m, 1H), 1.48-1.35 (s, 9H), 1.09-1.02
(d, J=6.6 Hz, 3H).
Intermediate 26B: (S)-tert-butyl
6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]-
pyrazine-7(8H)-carboxylate
[0277] To a solution of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate (350 mg, 1.52 mmol) in
n-butanol (3 mL) was added 4-(trifluoromethyl)benzohydrazide (479
mg, 2.28 mmol) and the reaction mixture was heated to 140.degree.
C. for 48 hours. The reaction mixture was cooled to rt and diluted
with methanol (10 ml). BOC-anhydride (0.65 mL, 3.04 mmol) was added
and the reaction mixture was stirred for 5 hours. The reaction
mixture was diluted with EtOAc, washed with water, dried,
concentrated and purified by flash column chromatography (0-100%
EtOAc in hexanes) to provide (S)-tert-butyl
6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]-
pyrazine-7(8H)-carboxylate (375 mg).
Intermediate 26C:
(S)-6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydro-[1,2,4]tria-
zolo[4,3-a]pyrazine
[0278] (S)-tert-butyl
6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]-
pyrazine-7(8H)-carboxylate was dissolved in trifluoroacetic acid
and stirred at rt for 10 minutes. The reaction mixture was
concentrated to provide
(S)-6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydro-[1,-
2,4]triazolo[4,3-a]pyrazine which was used without further
purification.
[0279] To a solution of
(S)-6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6,7,8-tetrahydro-[1,2,4]tria-
zolo[4,3-a]pyrazine (72 mg, 0.26 mmol) in DCM (5 mL) was added
2,3-dichlorobenzoyl chloride (107 mg, 0.51 mmol) and triethylamine
(0.18 mL, 1.28 mmol) and the reaction mixture was stirred for 4
hours at rt. Water was added and the reaction mixture was extracted
with DCM. The organic layers were combined, dried, concentrated and
purified by hplc. 1H NMR (400 MHz, DMSO) .delta. 8.18-7.38 (m, 7H),
5.78-5.17 (m, 1H), 4.81-3.81 (m, 4H), 1.25-0.96 (m, 3H). MS (ESI):
mass calcd. for C.sub.20H.sub.15Cl.sub.2F.sub.3N.sub.4O, 454.1; m/z
found, 455.1 [M+H].sup.+.
Example 27
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(4-(trifluoromethyl)ph-
enyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00099##
[0281]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(4-(trifluorome-
thyl)phenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (400 MHz, DMSO) .delta.
8.12-7.59 (m, 7H), 5.76-5.24 (m, 1H), 4.79-3.80 (m, 4H), 1.36-0.75
(m, 3H). MS (ESI): mass calcd. for
C.sub.21H.sub.15ClF.sub.6N.sub.4O, 488.1; m/z found, 489.1
[M+H].sup.+.
Example 28
(S)-(2,3-dichlorophenyl)(3-(3-fluoro-4-(trifluoromethyl)phenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00100##
[0283]
(S)-(2,3-dichlorophenyl)(3-(3-fluoro-4-(trifluoromethyl)phenyl)-6-m-
ethyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in analogous fashion
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
3-fluoro-4-(trifluoromethyl)benzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (400 MHz, DMSO) .delta.
8.07-7.37 (m, 6H), 5.74-5.19 (m, 1H), 4.85-3.87 (m, 4H), 1.26-0.96
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.14Cl.sub.2F.sub.4N.sub.4O, 472.0; m/z found, 473.1
[M+H].sup.+.
Example 29
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluoro-4-(trifluoromethyl)ph-
enyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanon-
e
##STR00101##
[0285]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluoro-4-(trifluorome-
thyl)phenyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)m-
ethanone was generated in an analogous fashion to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanonewherein
3-fluoro-4-(trifluoromethyl)benzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (400 MHz, DMSO) .delta.
8.15-7.62 (m, 6H), 5.73-5.26 (m, 1H), 4.83-3.86 (m, 4H), 1.30-0.97
(m, 3H). MS (ESI): mass calcd. for
C.sub.21H.sub.14ClF.sub.7N.sub.4O, 506.1; m/z found, 507.1
[M+H].sup.+.
Example 30
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(3,4,5-trifluorophenyl-
)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00102##
[0287]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(3,4,5-trifluor-
ophenyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in analogous fashion to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanonewherein
3,4,5-trifluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (400 MHz, DMSO) .delta.
8.08-7.59 (m, 5H), 5.67-5.21 (m, 1H), 4.79-3.84 (m, 4H), 1.23-0.94
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.13ClF.sub.6N.sub.4O, 474.1; m/z found, 475.1
[M+H].sup.+.
Example 31
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00103##
[0289]
(S)-(2,3-dichlorophenyl)(6-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in an
analogous manor to that describe for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
picolinohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (400 MHz, DMSO) .delta.
8.83-7.41 (m, 7H), 5.68-5.24 (m, 1H), 5.03-3.91 (m, 4H), 1.43-0.96
(m, 3H). MS (ESI): mass calcd. for
C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.1; m/z found,
388.1[M+H].sup.+.
Example 33
(S)-(2,3-dichlorophenyl)(3-(4-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00104##
[0290] Example 34
(2,3-dichlorophenyl)(5-methyl-3-(pyridin-2-yl)-5,6-dihydro-[1,2,4]triazolo-
[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00105##
[0291] Example 35
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(4-(trifluoromethyl)phenyl-
)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00106##
[0293] The title compound was prepared in a manner analogous to
Example 25 substituting 4-(trifluoromethyl)benzhydrazide for
4-fluorobenzhydrazide in Example 25, step b. MS (ESI) mass calcd.
C.sub.21H.sub.15ClF.sub.6N.sub.4O, 488.08; m/z found 489.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 7.89-7.74 (m, 5H), 7.60-7.41
(m, 2H), 6.24-6.18 (m, 1H), 5.21-5.00 (m, 1H), 4.36-4.16 (m, 1H),
4.08-4.00 (m, 1H), 3.73-3.64 (m, 1H), 3.56-3.28 (m, 1H), 1.86-1.61
(m, 3H).
Example 36
(2-chloro-3-(trifluoromethyl)phenyl)(8-methyl-3-(pyrazin-2-yl)-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00107##
[0295] The title compound was prepared in a manner analogous to
Example 25 substituting pyrazine-2-carbohydrazide for
4-fluorobenzhydrazide in Example 25, step b. MS (ESI) mass calcd.
C.sub.18H.sub.14ClF.sub.3N.sub.6O, 422.09; m/z found 423.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 9.63-9.55 (m, 1H), 8.68-8.48
(m, 2H), 7.85-7.77 (m, 1H), 7.60-7.36 (m, 2H), 6.27-6.21 (m, 1H),
5.21-4.83 (m, 2H), 4.43-4.09 (m, 1H), 3.78-3.35 (m, 1H), 1.86-1.57
(m, 3H).
Example 37
(2-chloro-3-(trifluoromethyl)phenyl)(3-(4-chlorophenyl)-8-methyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00108##
[0297] The title compound was prepared in a manner analogous to
Example 25 substituting 4-chlorobenzhydrazide for
4-fluorobenzhydrazide in Example 25, step b. MS (ESI) mass calcd.
C.sub.20H.sub.15Cl.sub.2F.sub.3N.sub.4O, 454.06; m/z found 455.1
[M+H].sup.+. 1H NMR (500 MHz, CDCl3): 7.86-7.81 (m, 1H), 7.69-7.45
(m, 6H), 6.21-6.16 (m, 1H), 5.19-4.98 (m, 1H), 4.31-3.97 (m, 2H),
3.72-3.26 (m, 1H), 1.85-1.60 (m, 3H).
Example 38
(S)-(2,3-dichlorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00109##
[0299]
(S)-(2,3-dichlorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in an
analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (400 MHz, DMSO) .delta.
7.90-7.32 (m, 7H), 5.65-5.20 (m, 1H), 4.85-3.62 (m, 4H), 1.30-0.91
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z found, 405.1
[M+H].sup.+.
Example 39
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2-fluorophenyl)-6-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00110##
[0301]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2-fluorophenyl)-6-methy-
l-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous manor to
(S(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (400 MHz, DMSO) .delta.
8.10-7.25 (m, 7H), 5.68-5.19 (m, 1H), 4.86-3.58 (m, 4H), 1.28-0.86
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found, 439.1
[M+H].sup.+.
Example 40
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00111##
[0303]
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2-fluorophenyl)-6-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous fashion to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(400 MHz, DMSO) .delta. 7.98-7.28 (m, 7H), 5.69-5.01 (m, 1H),
4.78-3.54 (m, 4H), 1.27-0.90 (m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found,
423.1[M+H].sup.+.
Example 41
(S)-(2,3-dichlorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00112##
[0305]
(S)-(2,3-dichlorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in an
analogous manor to
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-3-(4-(trifluoromethyl)p-
henyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
wherein 3-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazidec. 1H NMR (500 MHz, DMSO) .delta.
7.94-7.28 (m, 7H), 5.74-5.19 (m, 1H), 4.82-3.76 (m, 4H), 1.30-0.84
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z found, 405.1
[M+H].sup.+.
Example 42
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluorophenyl)-6-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00113##
[0307]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3-fluorophenyl)-6-methy-
l-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
3-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (400 MHz, DMSO) .delta.
8.14-7.28 (m, 7H), 5.73-5.19 (m, 1H), 4.90-3.80 (m, 4H), 1.42-0.90
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found, 439.1
[M+H].sup.+.
Example 43
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00114##
[0309]
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3-fluorophenyl)-6-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
3-fluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (400 MHz, DMSO) .delta.
7.96-7.23 (m, 6H), 5.72-5.18 (m, 1H), 4.78-3.75 (m, 4H), 1.30-0.89
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 423.1
[M+H].sup.+.
Example 44
(S)-(2,3-dichlorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dihydro-[1,2,-
4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00115##
[0311]
(S)-(2,3-dichlorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in
a manor analogous to that described for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2,3-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (600 MHz, DMSO) .delta.
7.85-7.34 (m, 6H), 5.71-5.20 (m, 1H), 4.83-3.66 (m, 4H), 1.30-0.90
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 423.1
[M+H].sup.+.
Example 45
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,3-difluorophenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00116##
[0313]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,3-difluorophenyl)-6-m-
ethyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
2,3-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (600 MHz, DMSO) .delta.
8.13-7.20 (m, 6H), 5.73-5.18 (m, 1H), 4.85-3.53 (m, 4H), 1.35-0.87
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.14ClF.sub.5N.sub.4O, 456.1; m/z found, 457.1
[M+H].sup.+.
Example 46
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00117##
[0315]
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,3-difluorophenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2,3-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(600 MHz, DMSO) .delta. 7.81-7.36 (m, 5H), 5.70-5.16 (m, 1H),
4.76-3.49 (m, 4H), 1.25-0.88 (m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.0; m/z found, 441.1
[M+H].sup.+.
Example 47
(S)-(2,3-dichlorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dihydro-[1,2,-
4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00118##
[0317]
(S)-(2,3-dichlorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in
a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
3,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (600 MHz, DMSO) .delta.
7.98-7.33 (m, 6H), 5.72-5.23 (m, 1H), 4.78-3.73 (m, 4H), 1.29-0.94
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 423.1
[M+H].sup.+.
Example 48
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3,4-difluorophenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00119##
[0319]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(3,4-difluorophenyl)-6-m-
ethyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in an analogous manor to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
3,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (600 MHz, DMSO) .delta.
8.11-7.48 (m, 6H), 5.75-5.23 (m, 1H), 4.80-3.81 (m, 4H), 1.29-0.96
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.14ClF.sub.5N.sub.4O, 456.1; m/z found, 457.1
[M+H].sup.+.
Example 49
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00120##
[0321]
(S)-(2,3-dichloro-4-fluorophenyl)(3-(3,4-difluorophenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in a manor analogous to that described for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
3,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(600 MHz, DMSO) .delta. 8.05-7.39 (m, 5H), 5.73-5.18 (m, 1H),
4.80-3.77 (m, 4H), 1.38-0.91 (m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.0; m/z found, 441.1
[M+H].sup.+.
Example 50
(S)-(2,3-dichlorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dihydro-[1,2,-
4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00121##
[0323]
(S)-(2,3-dichlorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was generated in
a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
2,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (600 MHz, DMSO) .delta.
7.88-7.26 (m, 6H), 5.71-5.17 (m, 1H), 4.79-3.53 (m, 4H), 1.35-0.87
(m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 423.1
[M+H].sup.+.
Example 51
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,4-difluorophenyl)-6-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00122##
[0325]
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(2,4-difluorophenyl)-6-m-
ethyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
was generated in a manor analogous to that described for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
2,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (600 MHz, DMSO) .delta.
8.12-7.23 (m, 6H), 5.68-5.20 (m, 1H), 4.81-3.58 (m, 4H), 1.27-0.89
(m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.14ClF.sub.5N.sub.4O, 456.1; m/z found, 457.1
[M+H].sup.+.
Example 52
(S)-(2,3-dichloro-4-fluorophenyl)(3-(2,4-difluorophenyl)-6-methyl-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00123##
[0327]
(S)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(3-(2,4-difluoroph-
enyl)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanon-
e was generated in a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
2,4-difluorobenzohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(600 MHz, DMSO) .delta. 7.88-7.22 (m, 5H), 5.66-5.19 (m, 1H),
4.74-3.60 (m, 4H), 1.26-0.84 (m, 3H). MS (ESI): mass calcd. for
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.0; m/z found, 441.1
[M+H].sup.+.
Example 53
(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00124##
[0328] Intermediate 53A: tert-butyl
(2-(2-bromo-2-phenylacetamido)ethyl)carbamate
##STR00125##
[0329] Step A: tert-butyl
(2-(2-bromo-2-phenylacetamido)ethyl)carbamate
[0330] A solution of tert-butyl N-(2-aminoethyl)carbamate (10 g,
59.29 mmol) in 40 mL of DCM was cooled to -78.degree. C.
Triethylamine (16.48 mL, 118.59 mmol) and 2-bromo-2-phenylacetyl
chloride (13.85 g, 59.29 mmol) were subsequently added and the
reaction mixture was stirred for 20 minutes then warmed to
0.degree. C. and stirred for 1 hour. The reaction mixture was
quenched with water and then extracted three times with DCM. The
combined organic layers were washed with brine, dried with MgSO4
and concentrated under reduced pressure. The resulting residue was
purified via silica gel chromatography (0-50% ethyl
acetate/hexanes) to provide the desired product (15.09 g, 71%) as a
white solid. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.51-7.28
(m, 5H), 5.43-5.31 (m, 1H), 4.89 (s, 2H), 3.58-3.16 (m, 4H),
1.56-1.32 (m, 9H). MS (ESI) mass calcd.
C.sub.15H.sub.21BrN.sub.2O.sub.3, 357.2; m/z found, 358.2
[M+H].sup.+.
Intermediate 53B: N-(2-aminoethyl)-2-bromo-2-phenylacetamide
##STR00126##
[0331] Step B: N-(2-aminoethyl)-2-bromo-2-phenylacetamide
[0332] To a solution of tert-butyl
(2-(2-bromo-2-phenylacetamido)ethyl)carbamate (7.8 g, 21.75 mmol)
in 30 mL of DCM was added Trifluoroacetic acid (16.6 mL, 217.49
mmol). The resulting reaction mixture was allowed to stir at room
temperature overnight. The reaction mixture was concentrated into a
brown residue under reduced pressure and then washed with conc.
NaHCO3 solution and extracted three times with DCM. The combined
organic layers were dried using MgSO4, filtered and concentrated to
provide the desired product (10.54 g, 99%). MS (ESI) mass calcd.
C.sub.10H.sub.13BrN.sub.2O, 257.2; m/z found, 258.2
[M+H].sup.+.
Intermediate 53C: tert-butyl
3-oxo-2-phenylpiperazine-1-carboxylate
##STR00127##
[0333] Step C: tert-butyl
3-oxo-2-phenylpiperazine-1-carboxylate
[0334] To a solution of N-(2-aminoethyl)-2-bromo-2-phenylacetamide
(19.28 g, 43.74 mmol) in 430 mL of THF was added anhydrous K2CO3
(60.46 g, 437.46 mmol). The resulting reaction mixture was refluxed
at 65.degree. C. overnight. Di-tert-butyldicarbonate (19.28 g,
87.49 mmol) was subsequently added and the reaction mixture was
refluxed at 65.degree. C. for an additional 5 hours. The resulting
reaction mixture was cooled to room temperature and diluted with
ethyl acetate then washed with water. The organic layer was
partitioned, dried with MgSO4, filtered and concentrated into a
residue. The resulting residue was purified via silica gel
chromatography (0-30% Ethyl acetate/hexanes) to provide the desired
product (9.54 g, 79%) as a white solid. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.42-7.27 (m, 5H), 6.02-5.71 (m, 1H), 4.15-4.06
(m, 1H), 3.80-3.69 (m, 1H), 3.69-3.61 (m, 1H), 3.42-3.30 (m, 1H),
1.51 (s, 9H). MS (ESI) mass calcd. C.sub.15H.sub.20N.sub.2O.sub.3,
276.3; m/z found, 277.2 [M+H].sup.+.
Intermediate 53D: tert-butyl
2-phenyl-3-thioxopiperazine-1-carboxylate
##STR00128##
[0335] Step D: tert-butyl
2-phenyl-3-thioxopiperazine-1-carboxylate
[0336] To a mixture of Lawesson's reagent (4.15 g, 9.95 mmol) in
125 mL of toluene was added tert-butyl
3-oxo-2-phenylpiperazine-1-carboxylate (2.5 g, 9.05 mmol) in a 5 mL
solution of toluene. The resulting reaction mixture was heated at
110.degree. C. for 3 hours in a sealed tube. The reaction was
worked up with 10% NaOH solution and extracted three times with
ethyl acetate. The combined organic layers were dried with MgSO4,
filtered and concentrated into a residue. The resulting residue was
purified via silica gel chromatography (0-50% ethyl
acetate/hexanes) to provide the desired product (613.8 mg, 23%) as
a crystalline orange solid. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 9.67 (s, 1H), 7.51-7.41 (m, 2H), 7.37-7.27 (m, 3H), 6.13
(s, 1H), 4.08-3.77 (m, 1H), 3.53-3.38 (m, 1H), 3.38-3.26 (m, 2H),
1.50 (s, 9H). MS (ESI) mass calcd. C.sub.15H.sub.20N.sub.2S0.sub.2,
292.4; m/z found, 293.2 [M+H].sup.+.
Intermediate 53E: tert-butyl
3-(methylthio)-2-phenyl-5,6-dihydropyrazine-1(2H)-carboxylate
##STR00129##
[0337] Step E: tert-butyl
3-(methylthio)-2-phenyl-5,6-dihydropyrazine-1(2H)-carboxylate
[0338] To a stirred solution of tert-butyl
2-phenyl-3-thioxopiperazine-1-carboxylate (390 mg, 1.334 mmol)) in
3 ml of acetonitrile was added iodomethane (227 mg, 1.601 mmol).
The resulting reaction mixture was stirred at room temperature
overnight and then concentrated under reduced pressure to provide
the desired product (407 mg, 99%). .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.51-7.41 (m, 3H), 7.40-7.31 (m, 2H), 6.17 (s,
1H), 4.25-4.08 (m, 2H), 4.06-3.90 (m, 1H), 3.50-3.37 (m, 1H), 3.08
(s, 3H), 1.48 (s, 9H). MS (ESI) mass calcd.
C.sub.16H.sub.22N.sub.2SO.sub.2, 306.4; m/z found, 307.2
[M+H].sup.+.
Intermediate 53F: tert-butyl
3-methyl-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carbox-
ylate
##STR00130##
[0339] Step F: tert-butyl
3-methyl-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carbox-
ylate
[0340] To a round-bottom flask was added tert-butyl
3-(methylthio)-2-phenyl-5,6-dihydropyrazine-1(2H)-carboxylate (606
mg, 1.978 mmol), acetic hydrazide (1.48 g, 19.76 mmol) followed by
10 mL of n-butanol. The resulting reaction mixture was heated to
155.degree. and stirred for 3 hours. The reaction mixture was
cooled to room temperature and di-tert-butyl dicarbonate (436 mg,
1.978 mmol) was added. The reaction mixture was subsequently
stirred for 1 hour at room temperature then isolated and
concentrated down into a brown residue which was purified via
silica gel chromatography (0-10% 2M NH3/MeOH in DCM) to produce the
desired product (390 mg, 63%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.40-7.20 (m, 5H), 1.54-1.48 (m, 9H), 6.67 (s, 1H), 4.45
(s, 1H), 3.98-3.77 (m, 2H), 3.32-3.16 (m, 1H), 2.44 (s, 3H). MS
(ESI) mass calcd. C.sub.17H.sub.22N.sub.4O.sub.2, 315.3; m/z found,
316.2 [M+H].sup.+.
Intermediate 53G:
3-methyl-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine
##STR00131##
[0341] Step G:
3-methyl-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine
[0342] To a solution of
3-methyl-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carbox-
ylate (390 mg, 1.241 mmol)) in 5 mL of DCM was added
Trifluoroacetic acid (0.390 mL, 5.096 mmol). The resulting reaction
mixture was allowed to stir at room temperature overnight. The
reaction mixture was concentrated into a brown residue under
reduced pressure and then washed with conc. NaHCO3 solution and
extracted three times with DCM. The combined organic layers were
dried using MgSO4, filtered and concentrated to provide the desired
product (120 mg, 45%). MS (ESI) mass calcd.
C.sub.12H.sub.14N.sub.4, 214.2; m/z found, 215.2 [M+H].sup.+.
Example 53
(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,2,4]-
triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00132##
[0343] Step H:
(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,2,4-
]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0344] To a solution of
3-methyl-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine
(120 mg, 0.560 mmol) in 5 mL of DCM was added triethylamine (0.234
mL, 1.68 mmol). The resulting reaction mixture was stirred for 5
min at room temperature and then cooled to 0.degree. C.
2-chloro-3-(trifluoromethyl)benzoyl chloride (272 mg, 1.120 mmol)
was subsequently added and the reaction was stirred at 0.degree. C.
for 20 min. The reaction was quenched with water and warmed to room
temperature then extracted three times with DCM. The combined
organic layers were dried using MgSO4 and concentrated into a
residue, which was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 .mu.m 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) to provide the racemic product
(157 mg, 67%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.87-7.70
(m, 1H), 7.60-7.29 (m, 7H), 6.21-5.93; 5.21-5.01 (m, 1H), 4.16-3.30
(m, 4H), 2.51-2.45 (m, 3H). MS (ESI): mass calcd. for
C.sub.20H.sub.16ClF.sub.3N.sub.4O, 420.1; m/z found, 421.0
[M+H].sup.+.
Example 54
(S)-(2,3-dichlorophenyl)(3-(6-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00133##
[0346] To a solution of
(S)-3-(6-fluoro-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,-
3-a]pyrazine (110 mg, 0.41 mmol) (prepared as described in Example
54, Intermediate 54B, replacing 4-methoxy-pyridine-2-carboxylic
acid hydrazide with 6-fluoro-pyridine-2-carboxylic acid hydrazide
in step for intermediate 54A) and triethylamine (0.283 mL, 2.04
mmol) in CH.sub.2Cl.sub.2 (3 mL) was added 2,3-dichlorobenzoyl
chloride (128.1 mg, 0.61 mmol) at 0.degree. C. The reaction mixture
was slowly warmed to room temperature and stirred for 1 h. The
reaction mixture was quenched with water and extracted with
CH.sub.2Cl.sub.2. The organic layers were separated, dried
(Na.sub.2SO.sub.4), filtered and the solvent concentrated in vacuo.
The crude compound was purified by column chromatography (silica,
MeOH in EtOAC 0:100 to 10:90), the desired fractiones were
collected, the solvent evaporated to give
(S)-(2,3-dichlorophenyl)(3-(6-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (100 mg, 60%) as a
white foam. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 1.22 (d,
J=6.9 Hz, 0.90H), 1.36-1.41 (m, 1.05H), 1.43 (d, J=6.9 Hz, 1.05H),
4.09-4.27 (m, 0.80H), 4.42-4.51 (m, 0.70H), 4.55-4.84 (m, 1.70H),
4.98 (d, J=13.9 Hz, 0.30H), 5.04 (d, J=13.9 Hz, 0.50H), 5.54-5.63
(m, 0.50H), 5.82 (d, J=18.5 Hz, 0.30H), 5.95 (d, J=18.2 Hz, 0.20H),
7.00 (ddd, J=15.4, 8.2, 2.5 Hz, 1H), 7.17-7.39 (m, 2H), 7.54-7.61
(m, 1H), 7.91-8.00 (m, 1H), 8.20-8.31 (m, 1H). MS (ESI): mass
calcd. For C.sub.18H.sub.14Cl.sub.2FN.sub.5O, 405.1; m/z found,
406.04 [M+H].sup.+, [.alpha.]=+84.4.degree. (589 nm, c 0.55 w/v %,
DMF, 20.degree. C.
Example 55
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00134##
[0347] Intermediate 55A: (S)-tert-butyl
3-(4-methoxy-2-pyridyl)-6-methyl-6,8-dihydro-5H-[1,2,4]triazolo[4,3-a]pyr-
azine-7-carboxylate
[0348] To a solution of (S)-tert-butyl
2-methyl-5-thioxo-piperazine-1-carboxylate (Intermediate 26A, 0.5
g, 2.17 mmol) in ethanol (5 mL) was added
4-methoxy-pyridine-2-carboxylic acid hydrazide (435 mg, 2.61 mmol).
The reaction mixture was heated at 150.degree. C. in a sealed tube
for 12 hours in a Q-TUBE. The solvent was then evaporated and the
crude product purified by column chromatography (silica, MeOH in
EtOAc 0:100 to 10:90). The desired fractions were collected and the
solvent was evaporated to afford (S)-tert-butyl
3-(4-methoxy-2-pyridyl)-6-methyl-6,8-dihydro-5H-[1,2,4]triazolo[4,3-a]pyr-
azine-7-carboxylate as an off-white solid that was used for the
next reaction step without further purification.
Intermediate 55B:
(S)-3-(4-methoxy-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4-
,3-a]pyrazine
[0349] Trifluoroacetic acid (2.5 mL, 32.67 mmol) was added to a
mixture of (S)-tert-butyl
3-(4-methoxy-2-pyridyl)-6-methyl-6,8-dihydro-5H-[1,2,4]triazolo[4,3-a]pyr-
azine-7-carboxylate (457 mg, 1.32 mmol) in CH.sub.2Cl.sub.2 (2.5
mL). The solution was stirred for 15 min at room temperature and
then the mixture was basified with aq. sat NaHCO.sub.3 and
extracted with CH.sub.2Cl.sub.2. The organic layers were separated,
dried (Na.sub.2SO.sub.4), filtered and the solvent concentrated in
vacuo to yield
(S)-3-(4-methoxy-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]tria-
zolo[4,3-a]pyrazine (121 mg, 37.3%) as an off-white solid.
Example 55
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0350] 2,3-Dichlorobenzoyl chloride (103.3 mg, 0.49 mmol) was added
to a stirred solution of
(S)-3-(4-methoxy-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4-
,3-a]pyrazine (121 mg, 0.49 mmol) and triethylamine (0.171 mL, 1.23
mmol) in CH.sub.2Cl.sub.2 under nitrogen at 0.degree. C. The
reaction mixture was slowly warmed to room temperature and stirred
for 1 h. The reaction mixture was quenched with water and extracted
with CH.sub.2Cl.sub.2 (1.5 mL). The organic layers were separated,
dried (Na.sub.2SO.sub.4), filtered and the solvent concentrated in
vacuo. The crude product was purified by column chromatography
(silica, MeOH in EtOAc 0:100 to 10:90) and the desired fractions
were collected and the solvent evaporated. The residue was further
purified by column chromatography (silica, CH3CN in
CH.sub.2Cl.sub.2 0/100 to 100/0). The desired fractions were
collected and the solvent evaporated. The residue was triturated
with diisopropyl ether to yield
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (81 mg, 39.3%) as
an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
1.19 (d, J=6.9 Hz, 0.90H), 1.37 (d, J=6.9 Hz, 0.60H), 1.38 (d,
J=6.9 Hz, 0.30H), 1.41 (d, J=7.2 Hz, 1.20H), 3.93 (s, 0.90H), 3.94
(s, 2.10H), 4.05-4.17 (m, 0.50H), 4.21 (dd, J=13.5, 4.5 Hz, 0.20H),
4.38-4.49 (m, 0.80H), 4.54-4.75 (m, 1.10H), 4.77 (d, J=17.1 Hz,
0.40H), 4.89 (d, J=13.6 Hz, 0.20H), 5.02-5.19 (m, 0.80H), 5.49-5.60
(m, 0.50H), 5.80 (d, J=18.3 Hz, 0.30H), 5.94 (d, J=18.3 Hz, 0.20H),
6.87 (ddd, J=12.5, 5.8, 2.5 Hz, 1H), 7.17-7.39 (m, 2H), 7.54-7.60
(m, 1H), 7.83-7.91 (m, 1H), 8.31-8.37 (m, 0.40H), 8.43 (d, J=5.8
Hz, 0.60H). MS (ESI): mass calcd. for
C.sub.19H.sub.17Cl.sub.2N.sub.5O.sub.2, 417.1; m/z found,
418.3[M+H].sup.+. [.alpha.]=+62.4.degree. (589 nm, c 0.42 w/v %,
DMF, 20.degree. C.).
Example 56
(2-chlorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyraz-
in-7(8H)-yl)methanone
##STR00135##
[0351] Example 57
(3,4-difluoro-2-methylphenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazol-
o[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00136##
[0352] Example 58
(2-chloro-4-fluorophenyl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4,-
3-a]pyrazin-7(8H)-yl)methanone
##STR00137##
[0353] Example 59
(2,3-dichloropyridin-4-yl)(6-methyl-3-phenyl-5,6-dihydro-[1,2,4]triazolo[4-
,3-a]pyrazin-7(8H)-yl)methanone
##STR00138##
[0354] Example 60
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-
(2,3-dichlorophenyl)methanone
##STR00139##
[0356]
(R,S)-(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyra-
zin-7(8H)-yl)(2,3-dichlorophenyl)methanone was generated in a manor
analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate and
cyclohexanecarbohydrazide was used in in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (600 MHz, DMSO) .delta.
7.92-7.36 (m, 3H), 6.02-5.81 (m, 1H), 4.23-3.53 (m, 3H), 2.95-2.75
(m, 1H), 2.06-1.16 (m, 14H). MS (ESI): mass calcd. for
C.sub.19H.sub.22Cl.sub.2N.sub.4O, 392.1; m/z found, 393.2
[M+H].sup.+.
Example 61
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclohexyl-8-methyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00140##
[0358]
(R,S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclohexyl-8-methyl-5,-
6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate and
cyclohexanecarbohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (600 MHz, DMSO) .delta.
8.10-7.57 (m, 3H), 5.98-5.81 (m, 1H), 4.35-3.42 (m, 3H), 3.01-2.80
(m, 1H), 2.02-1.19 (m, 14H). MS (ESI): mass calcd. for
C.sub.20H.sub.22ClF.sub.3N.sub.4O, 426.1; m/z found, 427.2
[M+H].sup.+.
Example 62
(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-
(2,3-dichloro-4-fluorophenyl)methanone
##STR00141##
[0360]
(R,S)-(3-cyclohexyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyra-
zin-7(8H)-yl)(2,3-dichloro-4-fluorophenyl)methanone was generated
in a manor analogous to that described for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate,
cyclohexanecarbohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(600 MHz, DMSO) .delta. 7.98-7.40 (m, 2H), 5.88-5.69 (d, J=6.7 Hz,
1H), 4.06-3.48 (m, 4H), 1.96-1.07 (m, 14H). MS (ESI): mass calcd.
for C.sub.19H.sub.21Cl.sub.2FN.sub.4O, 410.1; m/z found,411.2
[M+H].sup.+.
Example 63
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl-
)(2,3-dichlorophenyl)methanone
##STR00142##
[0362]
(R,S)-(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyr-
azin-7(8H)-yl)(2,3-dichlorophenyl)methanone was generated in a
manor analogous to that described for
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone wherein
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate and
cyclopropanecarbohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide. 1H NMR (600 MHz, DMSO) .delta.
7.89-7.31 (m, 3H), 5.94-5.79 (m, 1H), 4.93-2.96 (m, 4H), 2.15-1.95
(m, 1H), 1.48-0.66 (m, 7H).
Example 64
(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl-
)(2,3-dichloro-4-fluorophenyl)methanone
##STR00143##
[0364]
(R,S)-(3-cyclopropyl-8-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyr-
azin-7(8H)-yl)(2,3-dichloro-4-fluorophenyl)methanone was generated
in a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate,
cyclopropanecarbohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and 2,3-dichloro-4-fluorobenzoyl
chloride was used in place of 2,3-dichlorobenzoyl chloride. 1H NMR
(600 MHz, CDCl3) .delta. 12.48-12.11 (m, 2H), 10.61-10.45 (d, J=6.8
Hz, 1H), 9.53-8.35 (m, 4H), 6.81-6.64 (m, 1H), 6.44-5.46 (m,
7H).
Example 65
(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00144##
[0366]
(R,S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-methyl-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone was
generated in a manor analogous to
(S)-(2,3-dichlorophenyl)(6-methyl-3-(4-(trifluoromethyl)phenyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone where in
(R,S)-tert-butyl 2-methyl-3-thioxopiperazine-1-carboxylate was used
in place of (S)-tert-butyl
2-methyl-5-thioxopiperazine-1-carboxylate,
cyclopropanecarbohydrazide was used in place of
4-(trifluoromethyl)benzohydrazide and
2-chloro-3-(trifluoromethyl)benzoyl chloride was used in place of
2,3-dichlorobenzoyl chloride. 1H NMR (600 MHz, DMSO) .delta.
8.08-7.54 (m, 3H), 5.94-5.74 (m, 1H), 4.97-2.99 (m, 4H), 2.16-1.91
(m, 1H), 1.69-0.81 (m, 7H).
Example 66
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00145##
[0368] The desired product was prepared in an analogous manner to
example 53. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 65% CO.sub.2, 35% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.73 (m, 1H),
7.58-7.28 (m, 7H), 6.16-5.99; 5.21-5.05 (m, 1H), 4.16-3.31 (m, 4H),
2.53-2.40 (m, 3H). MS (ESI) mass calcd. for
C.sub.20H.sub.16ClF.sub.3N.sub.4O, 420.1; m/z found, 421.0
[M+H].sup.+.
Example 67
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-phenyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00146##
[0370] The desired product was prepared in an analogous manner to
example 53. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 65% CO.sub.2, 35% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.74 (m, 1H),
7.58-7.28 (m, 7H), 6.15-5.99; 5.19-5.05 (m, 1H), 4.17-3.31 (m, 4H),
2.53-2.43 (m, 3H). MS (ESI) mass calcd.
C.sub.20H.sub.16ClF.sub.3N.sub.4O, 420.1; m/z found, 421.0
[M+H].sup.+.
Example 68
(S)-(2,3-dichlorophenyl)(3-(4-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00147##
[0372] To a solution of
(S)-3-(4-iodopyridin-2-yl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,-
3-a]pyrazine (30 mg, 0.088 mmol) (generated in a manor analogous to
Intermediate 21E wherein 4-iodopicolinohydrazide was used in place
of 4-fluorobenzhydrazide) in CH.sub.2Cl.sub.2 (1 mL) was added
triethylamine (0.04 mL, 0.3 mmol) and 2,3-dichlorobenzoyl chloride
(24 mg, 0.11 mmol). The reaction was allowed to stir at rt for 30
min and then evaporated in vacuo. The residue was chromatographed
(SiO.sub.2, 0-10% 2 N NH.sub.3in MeOH)/CH.sub.2Cl.sub.2). After
concentration in vacuo the resulting residue was dissolved in dry
THF (0.2 mL) and tetrabutylammonium fluoride THF solution (0.041
mL, 0.041 mmol) was added. The reaction was heated to 120.degree.
C. for 5 min in the microwave. An additional 0.041 mmol TBAF was
added and the reaction was heated in the microwave to 150.degree.
C. for 5 min, followed by 160.degree. C. for 30 min and 165.degree.
C. for 40 min, and finally 160.degree. C. for 90 min. The reaction
was purified by prep HPLC (C18 XSelect 19.times.100 5 .mu.m, Mobile
phase Gradient from 80% 0.1% NH.sub.4CO.sub.3H/NH.sub.4OH pH 9
solution in Water, 20% CH.sub.3CN to 0% 0.1%
NH.sub.4CO.sub.3H/NH.sub.4OH pH 9 solution in Water, 100%
CH.sub.3CN) 1.6 mg, 9.6%). MS (ESI) mass calcd.
C.sub.18H.sub.14Cl.sub.2FN.sub.5O, 405.1; m/z found, 405.0.
Example 69
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00148##
[0374] 2,3-Dichlorobenzoyl chloride (94.3 mg, 0.45 mmol) was added
to a stirred solution of
(S)-3-(5-fluoro-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,-
3-a]pyrazine (70 mg, 0.30 mmol) (prepared as described in Example
54, Intermediate 54B, replacing 4-methoxy-pyridine-2-carboxylic
acid hydrazide with 5-fluoro-pyridine-2-carboxylic acid hydrazide
in step for intermediate 54A) and triethylamine (0.21 mL, 1.50
mmol) in CH.sub.2Cl.sub.2 (2 mL) under nitrogen at 0.degree. C. The
reaction mixture was slowly warmed to room temperature and stirred
for 30 additional min. The reaction mixture was quenched with water
and extracted with CH.sub.2Cl.sub.2. The organic layers were
separated, dried (Na.sub.2SO.sub.4), filtered and the solvent
concentrated in vacuo. The crude product was purified by column
chromatography (silica, MeOH in EtOAc 0:100 to 10:90), the desired
fractions were collected and the solvent evaporated in vacuo to
yield the desired compound with some impurities. This was purified
by RP HPLC on (C18 Sunfire 30.times.100 5 .mu.m). Mobile phase
(Gradient from 50% 0.1% NH.sub.4CO.sub.2CH.sub.3 solution in Water,
40% CH.sub.3CN to 40% 0.1% NH.sub.4CO.sub.2CH.sub.3 solution in
Water, 50% CH.sub.3CN), yielding
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (31.3 mg,
24.4%).
[0375] MS (ESI) mass calcd. C.sub.18H.sub.14Cl.sub.2FN.sub.5O,
405.1; m/z found, 406.0 [M+H].sup.+. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. ppm 1.20 (d, J=6.6 Hz, 0.75H), 1.38 (d, J=6.9
Hz, 0.75H), 1.38 (d, J=6.9 Hz, 0.45H), 1.42 (d, J=7.2 Hz, 1.05H),
4.07-4.18 (m, 0.45H), 4.21 (dd, J=13.6, 4.6 Hz, 0.15H), 4.38-4.49
(m, 0.80H), 4.53-4.83 (m, 1.80H), 4.92-5.06 (m, 0.85H), 5.52-5.61
(m, 0.55H), 5.81 (d, J=18.5 Hz, 0.25H), 5.94 (d, J=18.2 Hz, 0.15H),
7.17-7.39 (m, 2H), 7.54-7.60 (m, 2H), 8.34-8.44 (m, 1.30H), 8.50
(d, J=2.6 Hz, 0.70H).
Example 70
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-fluoropyridin-2-yl)-6-methyl-
-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00149##
[0377] 2-Chloro-3-(trifluoromethyl)benzoyl chloride (159.4 mg, 0.66
mmol) was added to a stirred solution of
(S)-3-(5-fluoro-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,-
3-a]pyrazine (102 mg, 0.44 mmol) and triethylamine (0.30 mL, 2.19
mmol) in CH.sub.2Cl.sub.2 (3 mL) at 0.degree. C. The reaction
mixture was slowly warmed to room temperature and stirred for 90
min. The reaction mixture was quenched with water and extracted
with CH.sub.2Cl.sub.2. The organic layers were separated, dried
(Na.sub.2SO.sub.4), filtered and the solvent concentrated in vacuo.
The crude product was purified by column chromatography (silica,
MeOH in EtOAc 0/100 to 10/90), the desired fractions were collected
and the solvent evaporated in vacuo. The product containing some
impurities was purified by RP HPLC on (C18 Sunfire 30.times.100 5
.mu.m). Mobile phase (Gradient from 60% 0.1%
NH.sub.4CO.sub.2CH.sub.3 solution in Water, 40% CH.sub.3CN to 40%
0.1% NH.sub.4CO.sub.2CH.sub.3 solution in Water, 60% CH.sub.3CN),
yielding
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-fluoropyridin-2-yl)-6-methy-
l-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(0.27 g, 44.2%). MS (ESI) mass calcd.
C.sub.19H.sub.14ClF.sub.4N.sub.5O, 439.1; m/z found, 440.0
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 1.22 (d,
J=6.9 Hz, 0.75H), 1.39 (d, J=7.2 Hz, 0.45H), 1.40 (d, J=6.7 Hz,
0.60H), 1.43 (d, J=7.2 Hz, 1.20H), 4.04-4.14 (m, 0.40H), 4.21 (dd,
J=13.6, 4.6 Hz, 0.15H), 4.39-4.49 (m, 0.85H), 4.56-4.85 (m, 1.80H),
4.93-5.09 (m, 0.85H), 5.53-5.63 (m, 0.55H), 5.82 (d, J=18.3 Hz,
0.25H), 5.96 (d, J=18.3 Hz, 0.15H), 7.45-7.61 (m, 3H), 7.79-7.86
(m, 1H), 8.33-8.44 (m, 1.40H), 8.51 (d, J=2.8 Hz, 0.60H).
[.alpha.]=+60.6.degree. (589 nm, c 0.5 w/v %, DMF, 20.degree.
C.).
Example 71
((S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyridin-2-yl)-6-meth-
yl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00150##
[0379] 2-Chloro-3-(trifluoromethyl)benzoyl chloride (127.8 mg,
0.526 mmol) was added to a stirred solution of
(S)-3-(5-methoxy-2-pyridyl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4-
,3-a]pyrazine (86 mg, 0.35 mmol) (prepared as described in Example
54, Intermediate 54B, replacing 4-methoxy-pyridine-2-carboxylic
acid hydrazide with 5-methoxy-pyridine-2-carboxylic acid hydrazide
in step for intermediate 54A) and triethylamine (0.24 mL, 1.75
mmol) in CH.sub.2Cl.sub.2 (3 mL) under nitrogen at 0.degree. C. The
reaction mixture was slowly warmed to room temperature and stirred
for 30 min. The reaction mixture was quenched with water and
extracted with CH.sub.2Cl.sub.2. The organic layers were separated,
dried (Na.sub.2SO.sub.4), filtered and the solvent concentrated in
vacuo. The crude product was purified twice by column
chromatography (silica, MeOH in EtOAc 0:100 to 10:90), the desired
fractions were collected and the solvent evaporated in vacuo. Final
purification was performed by RP HPLC on (C18 Sunfire 30.times.100
5 .mu.m). Mobile phase (Gradient from 60% 0.1%
NH.sub.4CO.sub.2CH.sub.3 solution in Water, 40% CH.sub.3CN to 40%
0.1% NH.sub.4CO.sub.2CH.sub.3 solution in Water, 60% CH.sub.3CN),
yielding
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyridin-2-y-
l)-6-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
(35.9 mg, 22.6%). MS (ESI) mass calcd.
C.sub.20H.sub.17ClF.sub.3N.sub.5O.sub.2, 451.1; m/z found,
452.1[M+H].sup.+. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm
1.20 (d, J=6.9 Hz, 0.75H), 1.38 (dd, J=6.9, 2.0 Hz, 1.20H), 1.43
(d, J=7.2 Hz, 1.05H), 3.90-3.92 (m, 1.30H), 3.93 (s, 1.70H),
4.01-4.12 (m, 0.40H), 4.19 (dd, J=13.6, 4.6 Hz, 0.15 H), 4.37-4.47
(m, 0.85H), 4.54-4.90 (m, 1.85H), 4.95-5.11 (m, 0.85H), 5.51-5.61
(m, 0.50H), 5.79 (d, J=18.2 Hz, 0.25H), 5.94 (d, J=18.2 Hz, 0.15H),
7.30-7.38 (m, 1H), 7.45-7.60 (m, 2H), 7.78-7.85 (m, 1H), 8.20-8.35
(m, 2H).
Example 72
(S)-(2,3-dichlorophenyl)(3-(5-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00151##
[0381] The desired product was prepared in an analogous manner to
Example 71, using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride. Further purification
was not required. MS (ESI) mass calcd.
C.sub.19H.sub.17Cl.sub.2N.sub.5O.sub.2, 417.1; m/z found, 418.0
[M+H].sup.+. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. ppm 1.19 (d,
J=6.9 Hz, 0.75H), 1.37 (d, J=6.6 Hz, 0.75H), 1.37 (d, J=6.9 Hz,
0.45H), 1.41 (d, J=6.9 Hz, 1.05H), 3.89-3.92 (m, 1.20H), 3.93 (s,
1.80H), 4.04-4.16 (m, 0.40H), 4.19 (dd, J=13.4, 4.8 Hz, 0.15H),
4.35-4.48 (m, 0.85H), 4.52-4.83 (m, 1.85H), 4.93-5.09 (m, 0.85H),
5.48-5.60 (m, 0.50H), 5.78 (d, J=18.2 Hz, 0.25H), 5.92 (d, J=18.2
Hz, 0.15H), 7.15-7.39 (m, 3H), 7.52-7.60 (m, 1H), 8.19-8.36 (m,
2H).
Example 73
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00152##
[0383] 2,3-Dichlorobenzoyl chloride (60 mg, 0.29 mmol) was added to
a stirred solution of
(S)-3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazol-
o[4,3-a]pyrazine (67 mg, 0.29 mmol) (prepared as described in
Example 54, Intermediate 54B, replacing
4-methoxy-pyridine-2-carboxylic acid hydrazide with
5-fluoropyrimidine-2-carboxylic acid hydrazide in step for
intermediate 54A) and triethylamine (0.20 mL, 1.43 mmol) in
CH.sub.2Cl.sub.2 (15 mL) under nitrogen at 0.degree. C. The
reaction mixture was slowly warmed to room temperature and stirred
for 1 h. The reaction mixture was quenched with water and extracted
with CH.sub.2Cl.sub.2. The organic layers were separated, dried
(Na.sub.2SO.sub.4), filtered and the solvent concentrated in vacuo.
The crude product was purified by column chromatography (silica,
MeOH in EtOAc 0:100 to 10:90), and the desired fractions were
collected and the solvent evaporated in vacuo to yield
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (45 mg, 38.5%). MS
(ESI) mass calcd. C.sub.17H.sub.13Cl.sub.2FN.sub.6O, 406.1; m/z
found, 406.9 [M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 1.21 (d, J=6.7 Hz, 0.75H), 1.38 (d, J=6.7 Hz, 0.60H), 1.39 (d,
J=7.2 Hz, 0.60H), 1.43 (d, J=7.2 Hz, 1.05H), 4.10-4.27 (m, 0.60H),
4.38-4.98 (m, 3.35H), 5.55-5.65 (m, 0.60H), 5.85 (d, J=18.5 Hz,
0.25H), 5.97 (d, J=18.5 Hz, 0.20H), 7.17-7.41 (m, 2H), 7.55-7.62
(m, 1H), 8.74 (s, 0.45H), 8.74 (s, 0.45H), 8.78 (br s, 1H), M.P.
258.9.degree. C. [.alpha.]=+56.8.degree. (589 nm, c 0.48 w/v %,
DMF, 20.degree. C.).
Example 74
(S)-(2,3-dichlorophenyl)(3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00153##
[0385] The desired product was prepared in an analogous manner to
Example 73, using 2-chloro-3-(trifluoromethyl)benzoyl chloride
instead of 2,3-dichlorobenzoyl chloride. MS (ESI) mass calcd.
C.sub.18H.sub.13ClF.sub.4N.sub.6O, 440.1; m/z found, 441.0
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 1.22 (d,
J=6.9 Hz, 0.75H), 1.40 (dd, J=6.9, 1.2 Hz, 1.05H), 1.44 (d, J=6.9
Hz, 1.20H), 4.06-4.19 (m, 0.55H), 4.23 (dd, J=13.5, 4.5 Hz, 0.15H),
4.39-4.53 (m, 0.80H), 4.59-5.01 (m, 2.50H), 5.56-5.67 (m, 0.60H),
5.86 (d, J=18.7 Hz, 0.25H), 5.99 (d, J=18.3 Hz, 0.15H), 7.46-7.60
(m, 2H), 7.80-7.87 (m, 1H), 8.73 (s, 0.30H), 8.75 (s, 0.50H), 8.78
(s, 1.20H). M.P. >300.degree. C. [.alpha.]=+51.0.degree. (589
nm, c 0.44 w/v %, DMF, 20.degree. C.).
Example 75
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5-
,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00154##
[0387] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step) and was purified via basic HPLC
(Agilent prep system, Waters XBridge C18 5 um 50.times.100 mm
column, 5-95% MeCN/20 nM NH.sub.4OH over 22 min at 80 mL/min) to
provide the racemic product (6.8 mg, 25%). .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.87-7.75 (m, 1H), 7.61-7.29 (m, 7H),
6.25-6.08; 5.22-5.09 (m, 1H), 4.39-3.35 (m, 4H). MS (ESI) mass
calcd. C.sub.20H.sub.13ClF.sub.6N.sub.4O, 474.1; m/z found, 475.1
[M+H].sup.+.
Example 76
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00155##
[0389] The desired product was prepared in an analogous manner to
example 53 (using cyclopropanecarbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.71 (m,
1H), 7.60-7.29 (m, 7H), 6.15-5.99; 5.20-5.02 (m, 1H), 4.25-3.30 (m,
4H), 1.78-1.68 (m, 1H), 1.28-1.16 (m, 2H), 1.16-0.99 (m, 2H). MS
(ESI) mass calcd. C.sub.22H.sub.18ClF.sub.3N.sub.4O, 446.1; m/z
found, 447.1 [M+H].sup.+.
Example 77
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-phenyl-5,6-dihydr-
o-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00156##
[0391] The desired product was prepared in an analogous manner to
example 53 (using cyclopropanecarbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.88-7.68 (m,
1H), 7.60-7.29 (m, 7H), 6.19-5.93; 5.21-5.03 (m, 1H), 4.26-3.31 (m,
4H), 1.79-1.67 (m, 1H), 1.28-1.16 (m, 2H), 1.16-1.02 (m, 2H). MS
(ESI) mass calcd. C.sub.22H.sub.18ClF.sub.3N.sub.4O, 446.1; m/z
found, 447.0 [M+H].sup.+.
Example 78
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(5-methoxypyrimidin-2-yl)-6-met-
hyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00157##
[0393] The desired product was prepared in an analogous manner to
Example 74, replacing
(S)-3-(5-fluoropyrimidin-2-yl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazol-
o[4,3-a]pyrazine with
(S)-3-(5-methoxypyrimidin-2-yl)-6-methyl-5,6,7,8-tetrahydro-[1,2,4]triazo-
lo[4,3-a]pyrazine. MS (ESI) mass calcd.
C.sub.19H.sub.16ClF.sub.3N.sub.6O.sub.2, 452.1; m/z found, 453.0
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 1.21 (d,
J=6.9 Hz, 0.75H), 1.39 (d, J=7.4 Hz, 0.45H), 1.39 (d, J=6.7 Hz,
0.60H), 1.43 (d, J=7.2 Hz, 1.20H), 3.99 (s, 0.60H), 4.00 (s,
0.75H), 4.02 (s, 1.65H), 4.04-4.16 (m, 0.55H), 4.20 (dd, J=13.8,
4.5 Hz, 0.15H), 4.37-4.50 (m, 0.80H), 4.57-5.03 (m, 2.50H),
5.54-5.64 (m, 0.60H), 5.83 (d, J=18.5 Hz, 0.25H), 5.97 (d, J=18.3
Hz, 0.15H), 7.45-7.60 (m, 2H), 7.78-7.88 (m, 1H), 8.50 (s, 0.30H),
8.52 (s, 0.50H), 8.55 (s, 1.20H). M.P. 129.4.degree. C.
Example 79
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-di-
hydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00158##
[0395] The desired product was prepared in an analogous manner to
example 53 (using oxazole-2-carbohydrazide instead of acetic
hydrazide in Step F.) .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.86-7.71 (m, 1H), 7.63-7.59 (m, 1H), 7.57-7.28 (m, 7H), 7.21-7.13
(m, 1H), 6.65-6.54 (m, 1H), 6.26-6.00; 5.17-5.04 (m, 1H), 4.61-3.36
(m, 4H). MS (ESI) mass calcd.
C.sub.23H.sub.16ClF.sub.3N.sub.4O.sub.2, 472.1; m/z found, 473.1
[M+H].sup.+.
Example 80
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(1-hydroxyethyl)-8-phenyl-5,-
6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00159##
[0397] The desired product was prepared in an analogous manner to
example 53 (using 2-hydroxypropanehydrazide instead of acetic
hydrazide in Step F.) .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.83-7.74 (m, 1H), 7.59-7.29 (m, 7H), 6.16-5.99; 5.15-4.97 (m, 2H),
4.47-2.76 (m, 5H), 1.79-1.65 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.18ClF.sub.3N.sub.4O.sub.2, 450.1; m/z found, 451.2
[M+H].sup.+.
Example 81
(R)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8-
H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
##STR00160##
[0399] The desired product was prepared in an analogous manner to
example 53 (using pivalohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 80% CO.sub.2, 20% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.71 (m, 1H),
7.60-7.28 (m, 7H), 6.20-6.01; 5.13-4.91 (m, 1H), 4.36-3.31 (m, 4H),
1.53-1.46 (m, 9H). MS (ESI) mass calcd.
C.sub.23H.sub.22ClF.sub.3N.sub.4O, 462.1; m/z found, 463.1
[M+H].sup.+.
Example 82
(S)-(3-(tert-butyl)-8-phenyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8-
H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
##STR00161##
[0401] The desired product was prepared in an analogous manner to
example 53 (using pivalohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 80% CO.sub.2, 20% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.72 (m, 1H),
7.58-7.28 (m, 7H), 6.20-6.01; 5.12-4.96 (m, 1H), 4.34-3.29 (m, 4H),
1.53-1.45 (m, 9H). MS (ESI) mass calcd.
C.sub.23H.sub.22ClF.sub.3N.sub.4O, 462.1; m/z found, 463.1
[M+H].sup.+.
Example 83
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00162##
[0403] The desired product was prepared in an analogous manner to
example 53 (using oxazole-2-carbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 68% CO.sub.2, 32% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.85-7.74 (m,
1H), 7.65-7.30 (m, 8H), 7.21-7.13 (m, 1H), 6.65-6.52 (m, 1H),
6.23-6.04; 5.19-5.05 (m, 1H), 4.61-3.34 (m, 4H). MS (ESI) mass
calcd. C.sub.23H.sub.16ClF.sub.3N.sub.4O.sub.2, 472.1; m/z found,
473.1 [M+H].sup.+.
Example 84
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(furan-2-yl)-8-phenyl-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00163##
[0405] The desired product was prepared in an analogous manner to
example 53 (using oxazole-2-carbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 68% CO.sub.2, 32% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.85-7.75 (m,
1H), 7.64-7.28 (m, 8H), 7.21-7.14 (m, 1H), 6.64-6.57 (m, 1H),
6.20-6.08; 5.21-5.01 (m, 1H), 4.64-3.35 (m, 4H). MS (ESI) mass
calcd. C.sub.23H.sub.16ClF.sub.3N.sub.4O.sub.2, 472.1; m/z found,
473.0 [M+H].sup.+.
Example 85
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00164##
[0407] To a solution of 3-methyl-8-(pyridin-2-yl)-5,6,7,
8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine (200 mg, 0.766 mmol) in
5 mL of DCM was added triethylamine (0.639 mL, 4.596 mmol). The
resulting reaction mixture was stirred for 5 min at room
temperature and then cooled to 0.degree. C.
2-chloro-3-(trifluoromethyl)benzoyl chloride (372 mg, 1.532 mmol)
was subsequently added and the reaction was stirred at 0.degree. C.
for 20 min. The reaction was quenched with water and warmed to room
temperature then extracted three times with DCM. The combined
organic layers were dried using MgSO4 and concentrated into a
residue, which was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 um 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) to provide the racemic product
(114 mg, 35.2%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.68-8.39 (m, 1H), 7.97-7.12 (m, 6H), 5.98-5.90; 5.23-5.09 (m, 1H),
4.47-3.43 (m, 4H), 2.51-2.39 (m, 3H). MS (ESI) mass calcd.
C.sub.19H.sub.15ClF.sub.3N.sub.5O, 421.1; m/z found, 422.0
[M+H].sup.+.
Example 86
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00165##
[0409] The desired product was prepared in an analogous manner to
example 53 (using propionohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 70% CO.sub.2, 30% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.73 (m, 1H),
7.58-7.28 (m, 7H), 6.20-6.01; 5.19-5.02 (m, 1H), 4.17-3.30 (m, 4H),
2.89-2.64 (m, 2H), 1.49-1.37 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.18ClF.sub.3N.sub.4O, 434.1; m/z found, 435.4
[M+H].sup.+.
Example 87
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-ethyl-8-phenyl-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00166##
[0411] The desired product was prepared in an analogous manner to
example 53 (using propionohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 70% CO.sub.2, 30% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87-7.71 (m, 1H),
7.60-7.27 (m, 7H), 6.20-6.01; 5.19-5.01 (m, 1H), 4.16-3.28 (m, 4H),
2.88-2.67 (m, 2H), 1.48-1.39 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.18ClF.sub.3N.sub.4O, 434.1; m/z found, 435.2
[M+H].sup.+.
Example 88
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00167##
[0413] The desired product was prepared in an analogous manner to
example 53 (using isobutyrohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 70% CO.sub.2, 30% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.74 (m, 1H),
7.58-7.28 (m, 7H), 6.19-6.01; 5.17-5.00 (m, 1H), 4.21-3.30 (m, 4H),
3.08-2.92 (m, 1H), 1.52-1.44 (m, 3H), 1.44-1.37 (m, 3H). MS (ESI)
mass calcd. C.sub.22H.sub.20ClF.sub.3N.sub.4O, 448.1; m/z found,
449.3 [M+H].sup.+.
Example 89
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-isopropyl-8-phenyl-5,6-dihydro--
[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00168##
[0415] The desired product was prepared in an analogous manner to
example 53 (using isobutyrohydrazide instead of acetic hydrazide in
Step F.) The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 70% CO.sub.2, 30% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.84-7.73 (m, 1H),
7.58-7.27 (m, 7H),6.19-6.01; 5.19-5.02 (m, 1H), 4.20-3.28 (m, 4H),
3.09-2.92 (m, 1H), 1.51-1.44 (m, 3H), 1.43-1.37 (m, 3H). MS (ESI)
mass calcd. C.sub.22H.sub.20ClF.sub.3N.sub.4O, 448.1; m/z found,
449.4 [M+H].sup.+.
Example 90
(.+-.)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,-
4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00169##
[0417] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and 2,3-dichlorobenzoyl chloride
instead of 2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E)
and was purified via basic HPLC (Agilent prep system, Waters
XBridge C18 5 .mu.m 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) to provide the racemic product
(100 mg, 55%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.61-7.52
(m,12H), 7.52-7.17 (m, 7H), 6.34-6.11; 5.20-5.05 (m, 1H), 4.40-3.32
(m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.1; m/z found, 441.1
[M+H].sup.+.
Example 91
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00170##
[0419] The desired product was prepared in an analogous manner to
example 85. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 60% CO.sub.2, 40% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.63-8.44 (m, 1H),
7.97-7.13 (m, 6H), 5.99-5.91; 5.26-5.10 (m, 1H), 4.50-3.52 (m, 4H),
2.51-2.43 (m, 3H). MS (ESI) mass calcd.
C.sub.19H.sub.15ClF.sub.3N.sub.5O, 421.1; m/z found, 421.8
[M+H].sup.+.
Example 92
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dih-
ydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00171##
[0421] The desired product was prepared in an analogous manner to
example 85. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 60% CO.sub.2, 40% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.63-8.44 (m, 1H),
7.97-7.14 (m, 6H), 5.99-5.94; 5.24-5.10 (m, 1H), 4.49-3.53 (m, 4H),
2.52-2.41 (m, 3H). MS (ESI) mass calcd.
C.sub.19H.sub.15ClF.sub.3N.sub.5O, 421.1; m/z found, 421.8
[M+H].sup.+.
Example 93
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00172##
[0423] The desired product was prepared in an analogous manner to
example 53 (using cyclobutanecarbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.83-7.74 (m,
1H), 7.56-7.28 (m, 7H), 6.18-5.97; 5.13-4.98 (m, 1H), 4.08-3.27 (m,
5H), 2.66-2.49 (m, 2H), 2.50-2.34 (m, 2H), 2.21-2.00 (m, 2H). MS
(ESI) mass calcd. C.sub.23H.sub.20ClF.sub.3N.sub.4O, 460.1; m/z
found, 461.1 [M+H].sup.+.
Example 94
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclobutyl-8-phenyl-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00173##
[0425] The desired product was prepared in an analogous manner to
example 53 (using cyclobutanecarbohydrazide instead of acetic
hydrazide in Step F.) The racemic product was separated via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.83-7.73 (m,
1H), 7.56-7.28 (m, 7H), 6.16-5.98; 5.13-4.98 (m, 1H), 4.08-3.24 (m,
5H), 2.68-2.49 (m, 2H), 2.49-2.34 (m, 2H), 2.23-2.00 (m, 2H). MS
(ESI) mass calcd. C.sub.23H.sub.20ClF.sub.3N.sub.4O, 460.1; m/z
found, 461.1 [M+H].sup.+.
[0426] Examples 95 and 96 were prepared as described in Example
108, substituting trifluoracetic anhydride for difluoroacetic
anhydride in Step C and 2-chloro-4-fluoro-3-trifluoromethyl benzoic
acid for 2-chloro-3-(trifluoromethyl)benzoic acid in Step E. The
racemic mixture was separated by prep HPLC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20mm), Mobile phase: 80% CO.sub.2,
20% EtOH) to provide the (R) and (S) enantiomers.
Example 95
(R*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluorome-
thyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00174##
[0428] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.60-7.13 (m, 7H),
6.21-6.23; 6.10-6.04; 5.20-5.07 (m, 1H), 4.38-4.15; 4.02-3.88;
3.78-3.38 (m, 4H). MS (ESI) mass calcd.
C.sub.20H.sub.12ClF.sub.7N.sub.4O, 492.1; m/z found, 492.0.
Example 96
(S*)-(2-chloro-4-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluorome-
thyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00175##
[0430] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.60-7.13 (m, 7H),
6.21-6.23; 6.10-6.04; 5.20-5.07 (m, 1H), 4.38-4.15; 4.02-3.88;
3.78-3.38 (m, 4H). MS (ESI) mass calcd.
C.sub.20H.sub.12ClF.sub.7N.sub.4O, 492.1; m/z found, 492.0.
Example 97
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00176##
[0432] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C) and was separated via chrial
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 80% CO.sub.2, 20% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.77 (m,
1H), 7.60-7.32 (m, 7H), 6.26-6.08; 5.23-5.09 (m, 1H), 4.40-3.36 (m,
4H). MS (ESI) mass calcd. C.sub.20H.sub.13ClF.sub.6N.sub.4O, 474.1;
m/z found, 474.7 [M+H].sup.+.
Example 98
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00177##
[0434] The desired product was separated from example 97 via chiral
SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm),
(Mobile phase: 80% CO.sub.2, 20% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.76 (m,
1H), 7.60-7.31 (m, 7H), 6.26-6.07; 5.23-5.09 (m, 1H), 4.41-3.36 (m,
4H). MS (ESI) mass calcd. C.sub.20H.sub.13ClF.sub.6N.sub.4O, 474.1;
m/z found, 474.8 [M+H].sup.+.
[0435] Examples 99 and 100 were prepared as described in Example
108, substituting trifluoracetic anhydride for difluoroacetic
anhydride in Step C and 3-chloro-2-(trifluoromethyl)isonicotinic
acid for 2-chloro-3-(trifluoromethyl)benzoic acid in Step E. The
racemic mixture was separated by prep HPLC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20mm), Mobile phase: 80% CO.sub.2,
20% EtOH) to provide the (R) and (S) enantiomers.
Example 99
(R*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-(trifluoromethy-
l)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00178##
[0437] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.78-8.59 (m, 1H),
7.61-7.20 (m, 6H), 6.20-6.12; 6.04-5.92; 5.21-5.06 (m, 1H),
4.42-4.16; 4.07-3.93; 3.81-3.41 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.12ClF.sub.6N.sub.5O, 475.1; m/z found, 475.9
[M+H].sup.+.
Example 100
(S*)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(8-phenyl-3-(trifluoromethy-
l)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00179##
[0439] .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.78-8.59 (m, 1H),
7.61-7.20 (m, 6H), 6.20-6.12; 6.04-5.92; 5.21-5.06 (m, 1H),
4.42-4.16; 4.07-3.93; 3.81-3.41 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.12ClF.sub.6N.sub.5O, 475.1; m/z found, 475.9
[M+H].sup.+.
Example 101
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,-
6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00180##
[0441] To a solution of
8-(4-fluorophenyl)-3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine (120 mg, 0.435 mmol) in 5 mL of DCM was added triethylamine
(0.363 mL, 2.610 mmol). The resulting reaction mixture was stirred
for 5 min at room temperature and then cooled to 0.degree. C.
2-chloro-3-(trifluoromethyl)benzoyl chloride (211 mg, 0.870 mmol)
was subsequently added and the reaction was stirred at 0.degree. C.
for 20 min. The reaction was quenched with water and warmed to room
temperature then extracted three times with DCM. The combined
organic layers were dried using MgSO4 and concentrated into a
residue, which was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 um 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) to provide the racemic product
(124 mg, 65%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.85-7.75
(m, 1H), 7.61-7.33 (m, 4H), 7.10-6.99 (m, 2H), 6.12-5.92; 5.19-5.04
(m, 1H), 4.14-3.27 (m, 4H), 2.51-2.41 (m, 3H). MS (ESI) mass calcd.
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found, 439.2
[M+H].sup.+.
Example 102
(R)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00181##
[0443] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and 2,3-dichlorobenzoyl chloride
instead of 2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E)
and was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H
5 .mu.m 250.times.20 mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH)
yielding the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.61-7.52 (m, 1H), 7.52-7.19 (m, 7H), 6.34-6.11; 5.19-5.08
(m, 1H), 4.40-3.32 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.0; m/z found, 440.8
[M+H].sup.+.
Example 103
(S)-(2,3-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00182##
[0445] The desired product was separated from example 102 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.61-7.53 (m,
1H), 7.53-7.19 (m, 7H), 6.33-6.12; 5.19-5.08 (m, 1H), 4.40-3.31 (m,
4H). MS (ESI) mass calcd. C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O,
440.0; m/z found, 440.8 [M+H].sup.+.
Example 104
(R)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00183##
[0447] The desired product was prepared in an analogous manner to
example 101 (using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride.) The racemic product
was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5
.mu.m 250.times.20 mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH)
yielding the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.60-7.51 (m, 1H), 7.47-7.15 (m, 4H), 7.10-6.98 (m, 2H),
6.20-5.98; 5.20-5.02 (m, 1H), 4.14-3.23 (m, 4H), 2.53-2.40 (m, 3H).
MS (ESI) mass calcd. C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z
found, 404.8 [M+H].sup.-.
Example 105
(S)-(2,3-dichlorophenyl)(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00184##
[0449] The desired product was prepared in an analogous manner to
example 101 (using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride.) The racemic product
was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5
.mu.m 250.times.20 mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH)
yielding the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.60-7.50 (m, 1H), 7.47-7.16 (m, 4H), 7.10-6.98 (m, 2H),
6.20-5.99; 5.18-5.03 (m, 1H), 4.16-3.21 (m, 4H), 2.53-2.44 (m, 3H).
MS (ESI) mass calcd. C.sub.19H.sub.15Cl.sub.2FN.sub.4O, 404.1; m/z
found, 404.7 [M+H].sup.-.
Example 106
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-fluorophenyl)--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00185##
[0451] The desired product was prepared in an analogous manner to
example 101 (using
3-cyclopropyl-8-(4-fluorophenyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a-
]pyrazine instead of
8-(4-fluorophenyl)-3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine.) The racemic product was separated via chiral SFC (Stationary
phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 75%
CO.sub.2, 25% iPrOH) yielding the desired product. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.86-7.75 (m, 1H), 7.59-7.27 (m, 4H),
7.11-6.98 (m, 2H), 6.13-5.92; 5.20-5.05 (m, 1H), 4.25-3.27 (m, 4H),
1.79-1.65 (m, 1H), 1.29-1.17 (m, 2H), 1.16-1.04 (m, 2H). MS (ESI)
mass calcd. C.sub.22H.sub.17ClF.sub.4N.sub.4O, 464.1; m/z found,
464.8 [M+H].sup.+.
Example 107
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-cyclopropyl-8-(4-fluorophenyl)--
5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00186##
[0453] The desired product was prepared in an analogous manner to
example 101 (using
3-cyclopropyl-8-(4-fluorophenyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a-
]pyrazine instead of
8-(4-fluorophenyl)-3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine.) The racemic product was separated via chiral SFC (Stationary
phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 75%
CO.sub.2, 25% iPrOH) yielding the desired product. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.86-7.75 (m, 1H), 7.60-7.27 (m, 4H),
7.11-6.98 (m, 2H), 6.12-5.92; 5.22-5.05 (m, 1H), 4.25-3.25 (m, 4H),
1.81-1.65 (m, 1H), 1.33-1.17 (m, 2H), 1.17-1.02 (m, 2H). MS (ESI)
mass calcd. C.sub.22H.sub.17ClF.sub.4N.sub.4O, 464.1; m/z found,
464.8 [M+H].sup.+.
Example 108
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00187##
[0454] Intermediate 108A: 2-chloro-3-phenylpyrazine
##STR00188##
[0455] Step A: 2-chloro-3-phenylpyrazine
[0456] To a solution of 2,3-dichloropyrazine (1.50 g, 10.07 mmol)
and phenylboronic acid (1.23 g, 10.07 mmol) in 35 mL of DME was
added Na2CO3 (1.07 g, 10.07 mmol) in 15 mL of water. N2 gas was
bubbled through the reaction mixture for 15 min then the flask was
equipped with a condenser and purged with N2 for another 15 in
before adding tetrakis(triphenylphosphine)palladium (581.75 mg,
0.503 mmol). The resulting reaction mixture was heated to reflux
(85.degree. C.) and allowed to stir overnight. The reaction was
cooled to rt and diluted with 80 mL of water then extracted three
times with DCM. The combined organic extracts were dried with
MgSO4, filtered and concentrated under reduced pressure. The
resulting yellow residue was purified via silica gel chromatography
(0-30% ethyl acetate/hexanes) to provide the desired product (1.39
g, 72%) as a white solid. MS (ESI) mass calcd.
C.sub.10H.sub.7ClN.sub.2, 190.63; m/z found, 191.0 [M+H].sup.+.
Intermediate 108B: 2-hydrazinyl-3-phenylpyrazine
##STR00189##
[0457] Step B: 2-hydrazinyl-3-phenylpyrazine
[0458] A neat suspension of 2-chloro-3-phenylpyrazine (1.39 g, 7.23
mmol) in hydrazine monohydrate (3.6 mL, 72.78 mmol) was placed in
microwave vial and irradiates at 120.degree. C. for 1 hour. The
resulting reaction mixture was cooled down to rt and diluted with
30 mL of water and then extracted three times with 30 mL of DCM.
The combined organic extracts were dried using MgSO4 and
concentrated under reduced pressure to provide the desired product
(1.21 g, 89%). MS (ESI) mass calcd. C.sub.10H.sub.10N.sub.4, 186.2;
m/z found, 187.2 [M+H].sup.+.
Intermediate 108C:
3-(difluoromethyl)-8-phenyl-[1,2,4]triazolo[4,3-a]pyrazine
##STR00190##
[0459] Step C:
3-(difluoromethyl)-8-phenyl-[1,2,4]triazolo[4,3-a]pyrazine
[0460] A neat residue of 2-hydrazinyl-3-phenylpyrazine (665 mg,
3.571 mmol) was cooled to 0.degree. C. and Difluoroacetic anhydride
(4.44 mL, 35.71 mmol) was added drop-wise. The resulting reaction
mixture was allowed to stir at room temperature for 2 hours then
concentrated into a brown residue under reduced pressure. The brown
residue was suspended in 4 mL of polyphosphoric acid to form a
gelatinous mixture, which was heated to 140.degree. C. and stirred
overnight The reaction mixture was then neutralized to pH 7 with
NaOH pellets and ice water. The resulting aqueous solution was
extracted three times with ethyl acetate. The combined organic
extracts were dried with MgSO4 and concentrated into a brown
residue which was purified via silica gel chromatography (0-50%
ethyl acetate/hexanes) to provide the desired product (500 mg,
57%). .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.84-8.77 (m, 2H),
8.20 (d, J=4.6 Hz, 1H), 8.12 (d, J=4.6 Hz, 1H), 7.60-7.53 (m, 3H),
7.45-7.22 (m, 1H). MS (ESI) mass calcd.
C.sub.12H.sub.8F.sub.2N.sub.4, 246.2; m/z found, 274.1
[M+H].sup.+.
Intermediate 108D:
3-(difluoromethyl)-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine
##STR00191##
[0461] Step D:
3-(difluoromethyl)-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine
[0462] To a round-bottom flask containing a solution of
3-(difluoromethyl)-8-phenyl-[1,2,4]triazolo[4,3-a]pyrazine (500 mg,
2.031 mmol) in 5 mL ethanol was added 10% palladium on carbon (wet
Degussa powder) (108 mg, 0.102 mmol). The resulting reaction vessel
was purged with N2 gas and fitted with a hydrogen balloon (1 atm),
then the reaction mixture was stirred at rt overnight. The reaction
mixture was then filtered through a pad of celite and concentrated
under reduced pressure to provide the desired product (470 mg,
92%). MS (ESI) mass calcd. C.sub.12H12F.sub.2N.sub.4, 250.2; m/z
found, 251.1 [M+H].sup.+.
Example 108
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00192##
[0463] Step E:
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0464] To a solution of
3-(difluoromethyl)-8-phenyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyra-
zine (150 mg, 0.600 mmol) in 5 mL of DCM was added triethylamine
(0.25 mL, 1.798 mmol). The resulting reaction mixture was stirred
for 5 min at room temperature and then cooled to 0.degree. C.
2-chloro-3-(trifluoromethyl)benzoyl chloride (291 mg, 1.200 mmol)
was subsequently added and the reaction was stirred at 0.degree. C.
for 20 min. The reaction was quenched with water and warmed to room
temperature then extracted three times with DCM. The combined
organic layers were dried using MgSO4 and concentrated into a
residue, which was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 .mu.m 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) to provide the racemic product
(132 mg, 48.2%). The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 70% CO.sub.2, 30% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87-7.77 (m, 1H),
7.61-7.32 (m, 7H), 7.14-6.84 (m, 1H), 6.29-6.08; 5.17-5.10 (m, 1H),
4.43-3.27 (m, 4H). MS (ESI) mass calcd.
C.sub.20H.sub.14ClF.sub.5N.sub.4O, 456.1; m/z found, 456.8
[M+H].sup.+.
Example 109
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(3-(difluoromethyl)-8-phenyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00193##
[0466] The desired product was separated from example 108 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87-7.76 (m,
1H), 7.60-7.30 (m, 7H), 7.14-6.84 (m, 1H), 6:30-6.09; 5.17-5.07 (m,
1H), 4.44-3.31 (m, 4H). MS (ESI) mass calcd.
C.sub.20H.sub.14ClF.sub.5N.sub.4O, 456.1; m/z found, 456.8
[M+H].sup.+.
Example 110
(R)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00194##
[0468] The desired product was prepared in an analogous manner to
example 108 (using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E) and was
separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m
250.times.20 mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding
the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.61-7.51 (m, 1H), 7.49-7.18 (m, 7H), 7.14-6.84 (m, 1H), 6.30-6.09;
5.15-5.08 (m, 1H), 4.44-3.28 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 422.8
[M+H].sup.+.
Example 111
(S)-(2,3-dichlorophenyl)(3-(difluoromethyl)-8-phenyl-5,6-dihydro-[1,2,4]tr-
iazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00195##
[0470] The desired product was separated from example 110 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.61-7.51 (m,
1H), 7.50-7.18 (m, 7H), 7.14-6.84 (m, 1H), 6:30-6.09; 5.17-5.07 (m,
1H), 4.44-3.31 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.14Cl.sub.2F.sub.2N.sub.4O, 422.1; m/z found, 422.8
[M+H].sup.+.
Example 112
(R)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00196##
[0472] The desired product was prepared in an analogous manner to
example 85 (using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride.) The racemic product
was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5
.mu.m 250.times.20 mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH)
yielding the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.62-8.43 (m, 1H), 7.96-7.09 (m, 6H), 6.10-5.94; 5.20-5.07
(m, 1H), 4.45-3.57 (m, 4H), 2.52-2.39 (m, 3H). MS (ESI) mass calcd.
C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.1; m/z found, 387.7
[M+H].sup.+.
Example 113
(S)-(2,3-dichlorophenyl)(3-methyl-8-(pyridin-2-yl)-5,6-dihydro-[1,2,4]tria-
zolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00197##
[0474] The desired product was prepared in an analogous manner to
example 85 (using 2,3-dichlorobenzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride.) The racemic product
was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5
.mu.m 250.times.20 mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH)
yielding the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.61-8.42 (m, 1H), 7.94-7.08 (m, 6H), 6.11-5.90; 5.22-5.07
(m, 1H), 4.46-3.57 (m, 4H), 2.54-2.41 (m, 3H). MS (ESI) mass calcd.
C.sub.18H.sub.15Cl.sub.2N.sub.5O, 387.1; m/z found, 387.7
[M+H].sup.+.
Example 114
(R)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00198##
[0476] The desired product was prepared in an analogous manner to
example 101. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 80% CO.sub.2, 20% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.85-7.75 (m, 1H),
7.57-7.27 (m, 4H), 7.11-6.99 (m, 2H), 6.13-5.93; 5.20-5.07 (m, 1H),
4.15-3.27 (m, 4H), 2.52-2.43 (m, 3H). MS (ESI) mass calcd.
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found, 438.7
[M+H].sup.+.
Example 115
(S)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(4-fluorophenyl)-3-methyl-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00199##
[0478] The desired product was prepared in an analogous manner to
example 101. The racemic product was separated via chiral SFC
(Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile
phase: 80% CO.sub.2, 20% iPrOH) yielding the desired product.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86-7.76 (m, 1H),
7.57-7.27 (m, 4H), 7.11-6.99 (m, 2H), 6.12-5.94; 5.20-5.04 (m, 1H),
4.15-3.26 (m, 4H), 2.53-2.41 (m, 3H). MS (ESI) mass calcd.
C.sub.20H.sub.15ClF.sub.4N.sub.4O, 438.1; m/z found, 438.7
[M+H].sup.-.
Example 116
(R)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone
##STR00200##
[0480] The desired product was prepared in an analogous manner to
example 101 (using 2-methyl-3-(trifluoromethyl)benzoyl chloride
instead of 82-chloro-3-(trifluoromethyl)benzoyl chloride.) The
racemic product was separated via chiral SFC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 75%
CO.sub.2, 25% iPrOH) yielding the desired product. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.76-7.66 (m, 1H), 7.47-7.27 (m, 4H),
7.11-6.96 (m, 2H), 6.21-5.93; 5.21-5.12 (m, 1H), 4.13-3.28 (m, 4H),
2.53-2.18 (m, 6H). MS (ESI) mass calcd.
C.sub.21H.sub.18F.sub.4N.sub.4O, 418.1; m/z found, 418.8
[M+H].sup.+.
Example 117
(S)-(8-(4-fluorophenyl)-3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-
-7(8H)-yl)(2-methyl-3-(trifluoromethyl)phenyl)methanone
##STR00201##
[0482] The desired product was prepared in an analogous manner to
example 101 (using 2-methyl-3-(trifluoromethyl)benzoyl chloride
instead of 82-chloro-3-(trifluoromethyl)benzoyl chloride.) The
racemic product was separated via chiral SFC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 75%
CO.sub.2, 25% iPrOH) yielding the desired product. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.77-7.65 (m, 1H), 7.47-7.27 (m, 4H),
7.13-6.98 (m, 2H), 6.21-5.92; 5.20-5.14 (m, 1H), 4.15-3.29 (m, 4H),
2.55-2.16 (m, 6H). MS (ESI) mass calcd.
C.sub.21H.sub.18F.sub.4N.sub.4O, 418.1; m/z found, 418.8
[M+H].sup.+.
Example 118
(R)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00202##
[0484] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and 2-chloro-4-fluorobenzoyl
chloride instead of 2-chloro-3-(trifluoromethyl)benzoyl chloride in
Step E) and was separated via chiral SFC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 70%
CO.sub.2, 30% iPrOH) yielding the desired product. .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.54-7.48 (m, 1H), 7.46-7.02 (m, 7H),
6.17-6.09; 5.18-5.10 (m, 1H), 4.38-3.29 (m, 4H). MS (ESI) mass
calcd. C.sub.19H.sub.13ClF.sub.4N.sub.4O, 424.1; m/z found, 424.7
[M+H].sup.+.
Example 119
(S)-(2-chloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,-
2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00203##
[0486] The desired product was separated from example 118 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.55-7.46 (m,
1H), 7.45-7.01 (m, 7H), 6.17-6.09; 5.18-5.10 (m, 1H), 4.36-3.33 (m,
4H). MS (ESI) mass calcd. C.sub.19H.sub.13ClF.sub.4N.sub.4O, 424.1;
m/z found, 424.7 [M+H].sup.+.
Example 120
(R)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00204##
[0488] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and 2,4-dichlorobenzoyl chloride
instead of 2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E)
and was separated via chiral SFC (Stationary phase: CHIRALPAK AD-H
5 .mu.m 250.times.20 mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH)
yielding the desired product. .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.53-7.47 (m, 1H), 7.47-7.30 (m, 7H), 6.15-6.09; 5.17-5.09
(m, 1H), 4.37-3.30 (m, 4H). MS (ESI) mass calcd.
C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O, 440.0; m/z found, 440.7
[M+H].sup.+.
Example 121
(S)-(2,4-dichlorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00205##
[0490] The desired product was separated from example 120 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 70% CO.sub.2, 30% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.54-7.47 (m,
1H), 7.47-7.29 (m, 7H), 6.15-6.09; 5.17-5.08 (m, 1H), 4.37-3.31 (m,
4H). MS (ESI) mass calcd. C.sub.19H.sub.13Cl.sub.2F.sub.3N.sub.4O,
440.0; m/z found, 440.7 [M+H].sup.+.
Example 122
(R)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00206##
[0492] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and
2-methyl-3-(trifluoromethyl)benzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E) and was
separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m
250.times.20 mm), (Mobile phase: 80% CO.sub.2, 20% iPrOH) yielding
the desired product. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.77-7.68 (m, 1H), 7.60-7.27 (m, 7H), 6.31-6.06; 5.25-5.15 (m, 1H),
4.43-3.40 (m, 4H), 2.51-2.18 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.16F.sub.6N.sub.4O, 454.1; m/z found, 454.8
[M+H].sup.+.
Example 123
(S)-(2-methyl-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00207##
[0494] The desired product was separated from example 122 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 80% CO.sub.2, 20% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.78-7.67 (m,
1H), 7.60-7.28 (m, 7H), 6.33-6.06; 5.26-5.14 (m, 1H), 4.43-3.37 (m,
4H), 2.51-2.16 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.16F.sub.6N.sub.4O, 454.1; m/z found, 454.8
[M+H].sup.+.
Example 124
(R)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00208##
[0496] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and 2,3-dichloro-4-fluorobenzoyl
chloride instead of 2-chloro-3-(trifluoromethyl)benzoyl chloride in
Step E) and was separated via chiral SFC (Stationary phase:
CHIRALPAK AD-H 5 .mu.m 250.times.20 mm), (Mobile phase: 75%
CO.sub.2, 25% iPrOH) yielding the desired product. .sup.1H NMR (500
MHz, CDCl.sub.3) .delta. 7.51-7.47 (m, 1H), 7.43-7.13 (m, 6H),
6.33-6.06; 5.17-5.09 (m, 1H), 4.37-3.34 (m, 4H). MS (ESI) mass
calcd. C.sub.19H.sub.12Cl.sub.2F.sub.4N.sub.4O, 458.0; m/z found,
458.7 [M+H].sup.+.
Example 125
(S)-(2,3-dichloro-4-fluorophenyl)(8-phenyl-3-(trifluoromethyl)-5,6-dihydro-
-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00209##
[0498] The desired product was separated from example 124 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 75% CO.sub.2, 25% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.46 (m,
1H), 7.44-7.11 (m, 6H), 6.33-6.08; 5.17-5.09 (m, 1H), 4.39-3.32 (m,
4H). MS (ESI) mass calcd. C.sub.19H.sub.12Cl.sub.2F.sub.4N.sub.4O,
458.0; m/z found, 458.7 [M+H].sup.+.
Example 126
(.+-.)-(8-(1H-pyrazol-5-yl)-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazol-
o[4,3-a]pyrazin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
##STR00210##
[0500] The desired product was prepared in an analogous manner to
example 108 (using 1-(2-tetrahydropyranyl)-1H-pyrazole-5-boronic
acid pinacol ester instead of phenylboronic acid in Step A and
trifluoracetic anhydride instead of difluoroacetic anhydride in
Step C) and was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 .mu.m 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) yielding the desired product.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.87-7.74 (m, 1H),
7.61-7.41 (m, 3H), 12.01-10.82 (m, 1H), 7.41-7.37; 6.50-6.14 (m,
2H), 6.13-6.03; 5.16-5.04 (m, 1H), 4.48-3.50 (m, 4H). MS (ESI) mass
calcd. C.sub.17H.sub.11ClF.sub.6N.sub.6O, 464.1; m/z found, 465.1
[M+H].sup.+.
Example 127
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(8-(pyridin-3-yl)-3-(trifluorom-
ethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00211##
[0502] The desired product was prepared in an analogous manner to
example 108 (using 1-(2-tetrahydropyranyl)-1H-pyrazole-5-boronic
acid pinacol ester instead of phenylboronic acid in Step A and
trifluoracetic anhydride instead of difluoroacetic anhydride in
Step C) and was purified via basic HPLC (Agilent prep system,
Waters XBridge C18 5 .mu.m 50.times.100 mm column, 5-95% MeCN/20 nM
NH.sub.4OH over 22 min at 80 mL/min) yielding the desired product.
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.68-8.54 (m, 2H),
7.92-7.29 (m, 4H), 6.20-6.11; 5.28-5.14 (m, 1H), 4.43-3.30 (m, 4H).
MS (ESI) mass calcd. C.sub.19H.sub.12ClF.sub.6N.sub.5O, 475.1; m/z
found, 476.1 [M+H].sup.+.
Example 128
(R)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00212##
[0504] The desired product was prepared in an analogous manner to
example 108 (using trifluoracetic anhydride instead of
difluoroacetic anhydride in Step C and
2-fluoro-3-(trifluoromethyl)benzoyl chloride instead of
2-chloro-3-(trifluoromethyl)benzoyl chloride in Step E) and was
separated via chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m
250.times.20 mm), (Mobile phase: 85% CO.sub.2, 15% iPrOH) yielding
the desired product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.83-7.67 (m, 1H), 7.60-7.29 (m, 7H), 6.29-6.21; 5.19-5.09 (m, 1H),
4.38-3.38 (m, 4H). MS (ESI) mass calcd.
C.sub.20H.sub.13F.sub.7N.sub.4O, 458.1; m/z found, 458.7
[M+H].sup.+.
Example 129
(S)-(2-fluoro-3-(trifluoromethyl)phenyl)(8-phenyl-3-(trifluoromethyl)-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00213##
[0506] The desired product was separated from example 128 via
chiral SFC (Stationary phase: CHIRALPAK AD-H 5 .mu.m 250.times.20
mm), (Mobile phase: 85% CO.sub.2, 15% iPrOH) yielding the desired
product. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.82-7.67 (m,
1H), 7.62-7.28 (m, 7H), 6.30-6.21; 5.19-5.09 (m, 1H), 4.39-3.35 (m,
4H). MS (ESI) mass calcd. C.sub.20H.sub.13F.sub.7N.sub.4O, 458.1;
m/z found, 458.7 [M+H].sup.+.
Example 130
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-8-phenyl-3-(trifluoro-
methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00214##
[0507] Intermediate 130A:
6-methyl-8-phenyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[-
4,3-a]pyrazine
##STR00215##
[0508] Step A:
6-methyl-8-phenyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[-
4,3-a]pyrazine
[0509] The desired product was prepared in an analogous manner to
example 108 (using 2,3-dichloro-5-methylpyrazine instead of
2,3-dichloropyrazine in Step A and trifluoracetic anhydride instead
of difluoroacetic anhydride in Step C) yielding the desired product
(502 mg, 46%). MS (ESI) mass calcd. C.sub.13H.sub.13F3N.sub.4,
282.2; m/z found, 283.2 [M+H].sup.+.
Example 130
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-8-phenyl-3-(trifluoro-
methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
##STR00216##
[0510] Step 130B:
(.+-.)-(2-chloro-3-(trifluoromethyl)phenyl)(6-methyl-8-phenyl-3-(trifluor-
omethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone
[0511] In a flask purged with N2 was added
6-methyl-8-phenyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[-
4,3-a]pyrazine (54mg, 0.191 mmol) followed by 1 mL of THF. The
resulting solution was cooled to -78.degree. C. and n-butylLithium
(2.0M in cyclohexane) (115 uL, 0.23 mmol) was added. The solution
was left to stir at -78.degree. C. for 15 minutes then a 1 mL
solution of 2-chloro-3-(trifluoromethyl)benzoyl chloride (70 mg,
0.287 mmol) in THF was added drop-wise. Upon complete addition, the
reaction mixture was left to stir at -78.degree. C. for 30 minutes
then quenched with addition of 3 mL of water. The resulting
reaction mixture was extracted three times with DCM and the
combined organic layers were dried with MgSO4 then concentrated
under reduced pressure. The resulting residue was purified via
basic HPLC (Agilent prep system, Waters XBridge C18 5 um
50.times.100 mm column, 5-95% MeCN/20 nM NH.sub.4OH over 22 min at
80 mL/min) yielding the desired product. .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.74-7.21 (m, 8H), 5.65-5.25; (m, 1H),
4.23-3.12 (m, 3H), 1.41-1.34 (m, 3H). MS (ESI) mass calcd.
C.sub.21H.sub.15ClF.sub.6N.sub.4O, 488.1; m/z found,
[M+H].sup.+.
Example 131
(.+-.)-Benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazi-
n-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
##STR00217##
[0512] Intermediate 131A:
3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine
7-oxide
##STR00218##
[0513] Step A:
3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine
7-oxide
[0514] To a stirring mixture of
3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine
(5.00 g, 26.0 mmol) and sodium tungstate dihydrate (0.343 g, 1.04
mmol) in water (5 mL) at 0.degree. C. was added 30% hydrogen
peroxide soln (6.1 mL) dropwise over 15 min. The reaction was
warmed to rt, kept at rt for 5 min, then cooled again over an ice
bath. After 20 min sodium bisulfate (1g) was added portionwise
followed by CH.sub.2Cl.sub.2 (300 mL), MeOH (30 mL) and NaCl to
saturate the mixture. After stirring for 16 h the solids were
allowed to settle and the liquids were decanted away. The solids
were washed with 10% MeOH/CH.sub.2Cl.sub.2. The organics were
combined, dried with Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash chromatography (SiO.sub.2,
0-10% (10% 2N NH.sub.3/MeOH)/DCM) to obtain the product as a yellow
oil (5.36 g, 26%). .sup.1H NMR (500 MHz, CH.sub.3OD) .delta.
8.16-8.13 (m, 1H), 4.71-4.64 (m, 2H), 4.52-4.44 (m, 2H). MS (ESI)
mass calcd. C.sub.6H.sub.5F.sub.3N.sub.4O, 206.04; m/z found, 206.9
[M+H].sup.+.
Intermediate 131B:
8-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(-
8H)-ol
##STR00219##
[0515] Step B:
8-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(-
8H)-ol
[0516] To a solution of
3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine
7-oxide (626 mg, 3.04 mmol) in THF (5 mL) at 0.degree. C. was added
benzylmagnesium chloride in THF (3.8 mL, 7.6 mmol). After stirring
for 30 min at 0.degree. C. saturated ammonium chloride was added.
The layers were separated and the water layer was extracted two
times more with methylene chloride. The organic layers were
combined, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated. The residue was purified by flash chromatography
(SiO.sub.2, 0-10% (10% 2N NH.sub.3/MeOH)/DCM) to obtain the product
as a yellow oil (750 mg, 83%). MS (ESI) mass calcd.
C.sub.13H.sub.13F.sub.3N.sub.4O, 298.10; m/z found, 299.7
[M+H].sup.-.
Intermediate 131C:
8-benzyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyr-
azine
##STR00220##
[0517] Step C:
8-benzyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyr-
azine
[0518] To a solution of
(8-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7-
(8H)-ol (750 mg, 2.52 mmol) in acetic acid (5 mL) and water (5 mL)
was added zinc dust (839 mg, 12.6 mmol). The reaction was stirred
at 60 oC for 30 min followed by the addition of more zinc (800 mg).
The reaction was allowed to stir at 40 oC for 72 h after which time
it was filtered over celite. The filtrate was evaporated in vacuo
followed by the addition of CH.sub.2Cl.sub.2 and aq NaHCO.sub.3
(sat). The layers were separated and the water layer was extracted
two times more with methylene chloride. The organic layers were
combined, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated. The residue was purified by flash chromatography
(SiO.sub.2, 0-10% (10% 2N NH.sub.3/MeOH)/DCM) to obtain the title
compound (413 mg, 58%). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.36-7.19 (m, 5H), 4.39 (dd, J=9.5, 3.8 Hz, 1H), 4.22-4.13 (m, 1H),
4.10-4.00 (m, 1H), 3.61-3.53 (dd, J=14.0, 3.7 Hz, 1H), 3.12-2.97
(m, 2H). MS (ESI) mass calcd. C.sub.13H.sub.13F.sub.3N.sub.4,
282.11; m/z found, 283.6 [M+H].sup.+.
##STR00221##
Step D:
(8-benzyl-3-(trifluoromethyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]p-
yrazin-7(8H)-yl)(2-chloro-3-(trifluoromethyl)phenyl)methanone
[0519] To a solution of
8-benzyl-3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyr-
azine (93 mg, 0.32 mmol) in CH.sub.2Cl.sub.2 (1 mL) was added
triethylamine (0.1 mL, 0.7 mmol) and
2-chloro-3-(trifluoromethyl)benzoyl chloride. The reaction was
allowed to stir at rt for 30 min and then evaporated in vacuo. The
residue was dissolved MeOH and purified by prep HPLC to afford the
title compound as a white solid (95 mg, 59%). MS (ESI): mass calcd.
for C.sub.21H.sub.15ClF.sub.6N.sub.4O, 488.08; m/z found, 489.1
[M+H].sup.+.
Example 132
(S)-(2,3-dichlorophenyl)(3-(4-hydroxypyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (Precursor for the
Radiolabeling of
(S)-(2,3-dichlorophenyl)(3-(44.sup.11C]methoxypyridin-2-yl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanon
##STR00222##
[0521] Iodotrimethylsilane (71.45 .mu.L, 0.502 mmol) was added to a
solution of
(S)-(2,3-dichlorophenyl)(3-(4-methoxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (70 mg, 0.167 mmol)
in CH.sub.3CN (1 mL). The mixture was stirred at 150.degree. C. for
6 min under microwave irradation. MeOH (0.1mL) was added and the
solvents were evaporated in vacuo. The crude was purified by column
chromatography (silica, MeOH in EtOAc 0:100 to 10:90), the desired
fractions were collected and the solvent evaporated in vacuo. The
residue was diluted into a mixture of water and CH.sub.2Cl.sub.2.
The organic layer was separated, dried (Na.sub.2SO.sub.4), filtered
and the solvent was evaporated in vacuo. Finally, the compound was
triturated with diisopropyl ether to afford
(S)-(2,3-dichlorophenyl)(3-(4-hydroxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (41.4 mg,
61.2%).
Radiosynthesis, Biodistribution and Radiometabolite Analysis
[0522] HPLC analysis was performed on a LaChrom Elite HPLC system
(Hitachi, Armstadt, Germany) connected to a UV spectrometer set at
220 nm. For the analysis of radiolabeled compounds, the HPLC
eluate, after passage through the UV detector, was led over a
shielded 3-inch NaI(Tl) scintillation detector connected to a
multichannel analyser (Gabi box, Raytest, Straubenhardt, Germany).
The output signal was recorded and analysed using a GINA Star data
acquisition system (Raytest, Straubenhardt, Germany). Radioactivity
in samples of biodistribution studies, cell uptake experiments and
radiometabolite analysis was quantified using an automated gamma
counter equipped with a 3-inch NaI(Tl) well crystal coupled to a
multichannel analyser (Wallac 2480 Wizard, Wallac, Turku, Finland).
Results were corrected for background radiation, physical decay and
counter dead time.
[0523] Animals were housed in individually ventilated cages in a
thermoregulated (.about.22.degree. C.), humidity-controlled
facility under a 12 h/12 h light/dark cycle with access to food and
water ad libitum. All animal experiments were conducted according
to the Belgian code of practice for the care and use of animals,
after approval from the KU Leuven University Ethics Committee for
Animals.
Radiosynthesis of
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.11C]methoxypyridin-2-yl)-6-methyl-5,6-
-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (Example
[.sup.11C]55)
##STR00223##
[0525] Carbon-11 was produced via a [.sup.14N(p,.alpha.).sup.11C]
nuclear reaction. The target gas, which was a mixture of N.sub.2
(95%) and H.sub.2 (5%), was irradiated using 18-MeV protons at a
beam current of 25 .mu.A. The irradiation was done for about 30 min
to yield [.sup.11C]methane ([.sup.11C]CH.sub.4). The
[.sup.11C]CH.sub.4 was then transferred to a home-built
recirculation synthesis module and trapped on a Porapak column that
was immersed in liquid nitrogen. After flushing with helium, the
condensed [.sup.11C]CH.sub.4 was converted to the gaseous phase by
bringing the Porapak loop to room temperature. This
[.sup.11C]CH.sub.4 was then reacted with vaporous I.sub.2 at
650.degree. C. to convert it to [.sup.11C]methyl iodide
([.sup.11C]MeI). Subsequently, the [.sup.11C]MeI was passed over a
silver triflate column (6 mm.times.50 mm) at 180.degree. C. The
resulting [.sup.11C]methyl-triflate ([.sup.11C]MeOTf) was bubbled
with a flow of helium through a solution of the precursor
(S)-(2,3-dichlorophenyl)(3-(4-hydroxypyridin-2-yl)-6-methyl-5,6-dihydro-[-
1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (0.2 mg) and
Cs.sub.2CO.sub.3 (1-3 mg) in anhydrous DMF (0.2 mL). When the
amount of radioactivity in the reaction vial had stabilized, the
reaction mixture was left at room temperature for 3 min. The crude
mixture was diluted with water (0.6 mL) and injected onto an HPLC
system (XBridge C.sub.18, 5 .mu.m, 4.6 mm.times.150 mm; Waters)
eluted with a mixture of 0.05 M NaOAc (pH 5.5) and EtOH (60:40 v/v)
at a flow rate of 1 mL/min. UV detection of the HPLC eluate was
performed at 254 nm. The radiolabeled product was collected after
11 min. The collected peak corresponding to the desired
radioligand, (Example [.sup.11C]55), was then diluted with saline
(Mini Plasco.RTM., Braun, Melsungen, Germany) to obtain a final
EtOH concentration of 10% and the solution was sterile filtered
through 0.22 .mu.m membrane filter (Millex.RTM.-GV, Millipore).
[0526] Chemical and radiochemical purity of Example [.sup.11C]55)
formulation was analyzed on an analytical HPLC system consisting of
an XBridge C.sub.18 column (3.5 .mu.m, 3.0 mm.times.100 mm, Waters)
eluted with a mixture of 0.05 M NaOAc (pH 5.5) and CH.sub.3CN
(70:30 v/v) at a flow rate of 0.8 mL/min. UV detection was
performed at 220 nm. The crude radiolabeling mixture was purified
using semi-preparative RP-HPLC affording
(S)-(2,3-dichlorophenyl)(3-(4-[11C]methoxypyridin-2-yl)-6-methy-
l-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone in
good radiochemical yields (40-60%, relative to starting
radioactivity of [.sup.11C]MeOTf, non-decay corrected, n=12), with
a radiochemical purity of >98% and an average specific
radioactivity of 233.+-.99 GBq/.mu.mol at end of synthesis (EOS)
(n=12). The identity of the radiotracer was confirmed by co-elution
with the non-radioactive analogue after co-injection onto an
analytical HPLC system
Radiosynthesis of
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.18F]fluoropyridin-2-yl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (Example
[.sup.18F]68)
##STR00224##
[0528] Fluorine-18 was produced via a [.sup.18O(p,n).sup.18F]
nuclear reaction in a Cyclone 18/9 cyclotron (Ion Beam
Applications, Louvain-la-Neuve, Belgium). After irradiation,
[.sup.18F]F.sup.- was trapped on a SepPak Light Accell plus QMA
anion exchange cartridge (Waters) and eluted with a kryptofix 222
14 mg/K.sub.2CO.sub.3 1.2 mg dissolved in 750 .mu.l
CH.sub.3CN/H.sub.2O mixture (95:5 v/v) into the reaction vial. The
solvent was evaporated under a stream of helium at 80.degree. C. by
applying microwave heating (Resonance instruments 521, Skokie Ill.
USA) with a power of 50 W and further dried by azeotropic
distillation of traces of water using CH.sub.3CN (1 mL in four
fractions) at the above applied microwave settings. Finally, the
residue was dried under a stream of helium at 50 W until complete
dryness.
[0529] A solution of the precursor
(S)-(2,3-dichlorophenyl)(3-(4-chloropyridin-2-yl)-6-methyl-5,6-dihydro-[1-
,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone (1.5 mg), which was
prepared in an analogous manner to Example 68, in DMSO (0.5 mL) was
added to the dried [.sup.18F]F.sup.-/K.sub.2CO.sub.3/kryptofix
residue and the mixture was heated using microwave irradiation at
50 W and (temperature setting 170.degree. C.) for 3 min. Next, the
crude mixture was diluted with a mixture of EtOH/NaOAc 0.025M pH
5.5 (17/83 v/v; 0.5 mL) and injected onto the HPLC system. The HPLC
system consisted of an XBridge column (C.sub.18, 5 .mu.m, 4.6
mm.times.150 mm; Waters) that was eluted with a mixture of
EtOH/NaOAc 0.025M pH 5.5 (35/65 v/v) at a flow rate of 1 mL/min. UV
detection of the HPLC eluate was performed at 254 nm. The
radiolabeled product
(S)-(2,3-dichlorophenyl)(3-(4-[.sup.18F]fluoropyridin-2-yl)-6-methyl-5,6--
dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methanone, (Example
[.sup.18F]68), was collected after a total synthesis time of 45 min
with an average radiochemical yield of 15% and a specific activity
of 22 GBq/.mu.mol.
[0530] Radiochemical purity and identity was assayed using an HPLC
system consisting of an XBridge column (C18, 3.mu., 3.0.times.100
mm; Waters) eluted with NaOAc 0.05M pH 5.5/CH.sub.3CN (70:30) at a
flow rate of 0.8 ml/min. The radioligand had a retention time of
7.7 min and had a radiochemical purity >98%.
[0531] Biodistribution Studies
[0532] The biodistribution studies were performed in healthy female
Wistar rats (body weight, 185-220 g) at 2, 30 and 60 min after
tracer injection (n=3/time point). Rats were anesthetized with
isoflurane (2.5% in oxygen at a flow rate of 1 L/min) and injected
with the radioligand via a tail vein. The animals were sacrificed
by decapitation at the indicated time points. Blood and major
organs were collected in tared tubes and weighed. The radioactivity
in the dissected organs and blood was counted using an automated
gamma counter. For calculation of total blood radioactivity, blood
mass was assumed to be 7% of the body mass.
Biodistribution Study of [.sup.11C]55
[0533] Table 2 presents the percentage injected dose (% ID) in the
different organs and body fluids at 2, 30 and 60 min after
radiotracer injection. 10.0% of the injected dose was detected in
the blood at 2 min after injection, which cleared to 5.3% at 60 min
after tracer injection. The total initial brain uptake of
[.sup.11C]55 was 0.6% at 2 min after injection and this cleared to
0.4% at 60 min after tracer injection. At 60 min after tracer
injection, 43.8% of the injected dose was retained in the liver and
the intestines. Urinary excretion of the radiotracer was minimal,
with 2.9% ID in urine and kidneys at 60 min after injection.
[0534] Table 3 shows the standardized uptake values (SUV) for
different brain regions and blood. At 2 min after tracer injection,
the radioactivity concentration in the cerebellum was highest of
all brain regions. Clear wash-out was observed between 2 and 30 min
after tracer injection for all brain regions with relative wash-out
ratios higher than 1.3 (2 min-to-30 min). The radioactivity
concentration at 30 and 60 min after tracer injection was
comparable for all studied brain regions, and also for total brain
and the blood at the three studied time points.
TABLE-US-00003 TABLE 2 Biodistribution of [.sup.11C]55 in normal
rats at 2, 30 and 60 minutes after tracer injection. % ID.sup.a 2
min 30 min 60 min urine 0.1 .+-. 0.1 0.9 .+-. 0.0 1.1 .+-. 0.5
kidneys 4.5 .+-. 0.8 1.9 .+-. 0.1 1.8 .+-. 0.2 liver 34.8 .+-. 2.1
19.4 .+-. 0.5 17.1 .+-. 0.3 spleen + pancreas 1.4 .+-. 0.4 0.6 .+-.
0.1 0.6 .+-. 0.1 lungs 2.1 .+-. 0.3 1.0 .+-. 0.1 1.1 .+-. 0.4 heart
1.3 .+-. 0.2 0.4 .+-. 0.0 0.4 .+-. 0.1 intestines 14.0 .+-. 1.8
23.8 .+-. 1.1 26.7 .+-. 9.1 stomach 1.7 .+-. 0.3 2.1 .+-. 1.0 5.1
.+-. 3.4 cerebrum 0.5 .+-. 0.0 0.3 .+-. 0.0 0.3 .+-. 0.1 cerebellum
0.1 .+-. 0.0 0.1 .+-. 0.0 0.1 .+-. 0.0 blood 10.0 .+-. 2.4 5.2 .+-.
0.6 5.3 .+-. 0.6 carcass 35.1 .+-. 4.1 46.9 .+-. 0.5 43.2 .+-. 5.3
.sup.aPercentage of injected dose calculated as counts per minute
(cpm) in organ/total cpm recovered. Data are expressed as mean .+-.
SD; n = 3 per time point.
TABLE-US-00004 TABLE 3 [.sup.11C]55 concentration in the different
rat brain regions and blood at 2, 30 and 60 minutes after tracer
injection. SUV.sup.a 2 min 30 min 60 min striatum 0.78 .+-. 0.0
0.45 .+-. 0.0 0.50 .+-. 0.1 hippocampus 0.74 .+-. 0.0 0.43 .+-. 0.0
0.52 .+-. 0.1 cortex 0.80 .+-. 0.2 0.58 .+-. 0.1 0.65 .+-. 0.1 rest
of cerebrum 0.86 .+-. 0.0 0.46 .+-. 0.0 0.53 .+-. 0.1 whole
cerebrum 0.83 .+-. 0.0 0.46 .+-. 0.0 0.54 .+-. 0.1 cerebellum 1.02
.+-. 0.1 0.53 .+-. 0.0 0.59 .+-. 0.1 blood 1.42 .+-. 0.3 0.74 .+-.
0.1 0.75 .+-. 0.1 .sup.aCalculated as (radioactivity in cpm in
organ/weight of organ in grams)/(total cpm recovered/body weight
rat in grams). Data are expressed as mean .+-. SD; n = 3 per time
point.
Rat Plasma Radiometabolite Analysis of [.sup.11C]55
[0535] Radiometabolites of [.sup.11C]55 in plasma of normal female
Wistar rats (n=2) were quantified at 30 min after tracer injection.
The Chromolith C.sub.18 column was eluted with gradient mixtures of
0.05 M NaOAc (pH 5.5) (A) and CH.sub.3CN (B) (0-4 min: isocratic 0%
B and flow rate of 0.5 mL/min; 4-14 min: linear gradient 0% B to
90% B and flow rate of 1 mL/min; and 14-17 min: isocratic 90% B and
flow rate of 1mL/min). UV detection was done at 220 nm. The
reconstructed radiochromatogram demonstrated two peaks,
corresponding to intact [.sup.11C]55 eluting at 10 min and a polar
radiometabolite eluting at 2 min (chromatograms not shown). 30 min
after radiotracer injection, 70.+-.6% of the recovered
radioactivity in the plasma was in the form of intact tracer and
30.+-.6% was in the form of polar radiometabolite(s). The fraction
of more lipophilic radiometabolites eluting after the intact tracer
was negligible (<1.5%).
Perfused Rat Brain Radiometabolite Analysis of [.sup.11C]55
[0536] Radiometabolites of [.sup.11C]55 in perfused cerebrum and
cerebellum of normal female Wistar rats (n=2) were quantified at 30
min after tracer injection. Homogenates were analysed using an
analytical XBridge column (C.sub.18, 5 .mu.m, 3.times.100 mm;
Waters) eluted with a mixture of 0.05 M sodium acetate (pH 5.5) and
CH.sub.3CN (65:35 v/v) at a flow rate of 0.8 mL/min. UV detection
was performed at 220 nm. The reconstructed radiochromatograms from
perfused rat cerebellum and cerebrum HPLC analysis at 30 min post
tracer injection showed only one radioactive peak corresponding to
intact [.sup.11C]55 eluting at 9 min (chromatograms not shown).
Both the fraction of more polar and more lipophilic
radiometabolites were negligible (<2%).
[0537] The studies using [.sup.18F]68, were performed in a manner
analogus manner to those performed with[.sup.11C]55 and the results
of those experiments are shown in Tables 4 and 5.
Biodistribution Study of [.sup.18F]68
TABLE-US-00005 [0538] TABLE 4 Biodistribution of [.sup.18F]68 in
normal rats at 2, 30 and 60 minutes after tracer injection. %
ID.sup.a 2 min 30 min 60 min urine 0.1 .+-. 0.0 8.4 .+-. 1.8 8.4
.+-. 1.8 kidneys 3.3 .+-. 0.5 1.1 .+-. 0.2 0.9 .+-. 0.1 liver 34.6
.+-. 2.4 7.5 .+-. 0.6 3.61 .+-. 0.3 spleen + pancreas 1.3 .+-. 0.2
0.3 .+-. 0.1 0.2 .+-. 0.0 lungs 1.6 .+-. 0.1 0.7 .+-. 0.1 0.6 .+-.
0.2 heart 0.9 .+-. 0.1 0.2 .+-. 0.0 0.1 .+-. 0.0 intestines 10.6
.+-. 2.1 20.9 .+-. 6.0 11.2 .+-. 0.6 stomach 1.5 .+-. 0.5 1.6 .+-.
0.9 10.7 .+-. 3.3 cerebrum 0.6 .+-. 0.0 0.1 .+-. 0.0 0.1 .+-. 0.0
cerebellum 0.1 .+-. 0.0 0.0 .+-. 0.0 0.0 .+-. 0.0 blood 6.8 .+-.
0.6 4.3 .+-. 0.3 2.0 .+-. 0.5 carcass 40.8 .+-. 0.8 56.6 .+-. 2.8
60.4 .+-. 2.3 bone.sup.b 8.8 .+-. 0.8 44.5 .+-. 3.8 64.5 .+-. 5.9
.sup.aPercentage of injected dose calculated as counts per minute
(cpm) in organ/total cpm recovered. Data are expressed as mean .+-.
SD; n = 3 per time point. .sup.bcalculated to estimated total bone
tissue (% ID/g bone * body mass * 0.12)
TABLE-US-00006 TABLE 5 [.sup.18F]68 concentration in the different
rat brain regions and blood at 2, 30 and 60 minutes after tracer
injection. SUV.sup.a 2 min 30 min 60 min striatum 1.19 .+-. 0.13
0.24 .+-. 0.05 0.19 .+-. 0.03 hippocampus 1.11 .+-. 0.10 0.23 .+-.
0.04 0.19 .+-. 0.03 cortex 1.27 .+-. 0.13 0.94 .+-. 0.29 0.69 .+-.
0.17 rest of cerebrum 1.20 .+-. 0.11 0.31 .+-. 0.06 0.25 .+-. 0.04
whole cerebrum 1.20 .+-. 0.11 0.31 .+-. 0.06 0.25 .+-. 0.04
cerebellum 1.21 .+-. 0.22 0.33 .+-. 0.07 0.38 .+-. 0.08 blood 0.97
.+-. 0.08 0.61 .+-. 0.04 0.29 .+-. 0.07 .sup.aCalculated as
(radioactivity in cpm in organ/weight of organ in grams)/(total cpm
recovered/body weight rat in grams). Data are expressed as mean
.+-. SD; n = 3 per time point.
Pharmacological Examples
[0539] The in vitro affinity of the compounds of the invention for
the rat and human P2X7 receptor was determined using a human
peripheral blood mononuclear cells (PBMCs), a human whole blood
assay, a Ca.sup.2+ flux and radioligand binding assay in
recombinant human P2X7 cells and recombinant rat P2X7 cells. In
Table 6, when the data cell has been left blank, it is intended to
mean that the compound was not tested in that assay. The data
represented in Tables 2 may represent a value from a single
determination or when the experiment was run more than once, the
data represent averages from between 2-12 runs.
[0540] P2X7 Antagonism in Human Peripheral Blood Mononuclear Cells
(PBMCs) and Human Whole Blood.
[0541] Human blood was collected using a blood donor program. PBMCs
were isolated from blood using a Ficoll density gradient technique.
Briefly, blood was laid on Ficoll solution and centrifuged at RT
for 20 minutes at 2000 rpm. The buffy layer (between red blood
cells and plasma) was carefully collected by aspiration, washed
with PBS and centrifuged again at 1500 rpm for 15 minutes. The
resulting cell pellet was washed and plated on 96 well-plates for
experiments. For the Human Whole Blood experiments, 150 .mu.l of
human blood was platted on 96 well-plates. Lipopolysaccharide (LPS)
(30 ng/ml) was added to each well and incubated for 1 hour. Test
compounds were then added and incubated for 30 minutes. The P2X7
agonist, 2'(3')-O-(4-benzoylbenzoyl) adenosine 5' -triphosphate
(Bz-ATP) was then added at a final concentration of 0.5 mM (PBMC)
or 1 mM (blood). Cells were incubated for an additional 1.5 hours.
At that point, supernatant was collected and stored for IL-1.beta.
assay using manufacturer's protocol for enzyme-linked immunosorbent
assay (ELISA). Data was expressed as percent control, where control
is defined as the difference in IL-1.beta. release in LPS+Bz-ATP
samples and LPS only samples. Data was plotted as response (%
control) versus concentration to generate IC.sub.50 values. In
Tables 2, this data is represented by PBMC 1 .mu.M (% control) and
PBMC 10 .mu.M (% control) and human whole blood IC.sub.50 (.mu.M).
Data are analyzed and graphed on Graphpad Prism 5. For analysis,
each concentration point is averaged from triplicate values and the
averaged values are plotted on Graphpad Prism. The IC.sub.50 for
each compound is then uploaded into 3DX.
P2X7 Antagonism in Recombinant Human P2X7 Cells or Recombinant Rat
P2X7 Cells: (a) Ca.sup.2+ Flux and (b) Radioligand Binding
[0542] (a) Ca.sup.2+ flux: 1321N1 cells expressing the recombinant
human or rat P2X7 channel was cultured in HyQ
DME/(HyClone/Dulbecco's Modified Eagle Medium) high glucose
supplemented with 10% Fetal Bovine Serum (FBS) and appropriate
selection marker. Cells were seeded at a density of 25000
cells/well (96-well clear bottom black walled plates) in 100 .mu.l
volume/well. On the day of the experiment, cell plates were washed
with assay buffer, containing (in mM): 130 NaCl, 2 KCl, 1
CaCl.sub.2, 1 MgCl.sub.2, 10 HEPES, 5 glucose; pH 7.40 and 300 mOs.
After the wash, cells were loaded with the Calcium-4 dye (Molecular
Device) and incubated in the dark for 60 minutes. Test compounds
were prepared at 250.times. the test concentration in neat DMSO.
Intermediate 96-well compound plates were prepared by transferring
1.2 .mu.L of the compound into 300 .mu.L of assay buffer. A further
3.times. dilution occurred when transferring 50 .mu.L/well of the
compound plate to 100 .mu.L/well in the cell plate. Cells were
incubated with test compounds and dye for 30 minutes. Calcium dye
fluorescence was monitored in FLIPR as the cells were challenged by
adding 50 .mu.L/well of BzATP (final concentration is 250 .mu.M
BzATP (human and rat)). The fluorescence change was measured 180
seconds after adding the agonist. Peak fluorescence was plotted as
a function of BzATP concentration using Origin 7 software and the
resultant IC.sub.50 is shown in Tables 2 under the column headings
FLIPR (human) IC.sub.50 (.mu.M) and FLIPR (rat) IC.sub.50
(.mu.M).
[0543] (b) Radioligand binding: human or rat P2X7-1321N1 cells were
collected and frozen @-80.degree. C. On the day of the experiment,
cell membrane preparations were made according to standard
published methods. The total assay volume was 100 .mu.l:10 .mu.l
compound (10.times.)+(b) 40 .mu.l tracer (2.5.times.)+50 .mu.l
membrane (2.times.). The tracer used for the assay was tritiated
A-804598. The compound can be prepared as described in the
literature. (Donnelly-Roberts, D. Neuropharmacology 2008, 56 (1),
223-229.) Compounds, tracer and membranes were incubated for 1 hour
@4.degree. C. The assay was terminated by filtration (GF/B filters
pre-soaked with 0.3% PEI) and washed with washing buffer (Tris-HCl
50 mM). The IC.sub.50 generated in the binding assay was corrected
for tracer concentration and affinity of the tracer to derive at
the affinity (K.sub.i) of the test compounds. The data are
presented in Table 6 under the headings: P2X7 human K.sub.i (.mu.M)
and P2X7 rat K.sub.i (.mu.M). Data are analyzed and graphed on
Graphpad Prism 5. For analysis, each concentration point is
averaged from triplicate values and the averaged values are plotted
on Graphpad Prism.
TABLE-US-00007 TABLE 6* P2X7 activity of the compounds of Formula
(I) in a panel of in-vitro assays FLIPR FLIPR Human PBMC PBMC P2X7
P2X7 (human) (rat) whole blood 1 .mu.M 10 .mu.M human rat IC.sub.50
IC.sub.50 IC.sub.50 Ex # (% control) (% control) K.sub.i (.mu.M)
K.sub.i (.mu.M) (.mu.M) (.mu.M) (.mu.M) 1 -11.8 nt 0.0550 nt 0.0065
1.2070 nt 2 15.0 7.1 0.0214 0.0093 0.0013 0.8153 0.009 3 100.4 nt
nt 1.7783 >10 >10 nt 4 98.2 nt nt nt nt nt nt 5 88.7 nt nt nt
nt nt nt 6 100.4 nt nt 0.5012 1.2589 15.8489 nt 7 99.9 nt nt 0.3162
1.9953 10.0000 nt 8 13.9 nt 0.0427 nt 0.0286 0.1600 nt 9 16.1 nt
0.0437 nt 0.0179 1.0233 nt 10 6.5 nt 0.0955 nt 0.0152 0.0198 nt 11
1.4 nt 0.0468 nt 0.0070 1.3772 nt 12 nt 15.0 0.0158 0.0045 0.0035
0.1023 nt 13 nt 14.9 1.5849 0.3162 0.2972 >10 nt 14 nt 8.9
0.0316 0.0050 0.0109 0.0240 nt 15 nt 49.4 3.1623 0.5012 8.5114
>10 nt 16 nt 14.4 0.1000 nt 0.0195 0.0177 nt 17 nt 3.0 0.0794 nt
0.0060 0.0650 nt 18 nt 102.4 nt nt 1.4656 2.2387 nt 19 nt 7.1
0.0251 0.0079 0.0064 0.0062 0.182 20 nt 10.6 0.0398 nt 0.0105
0.0512 nt 21 nt 7.0 0.0282 nt 0.0102 0.0091 0.035 22 nt 2.9 0.0200
nt 0.0100 0.0050 0.016 23 nt 6.2 0.0631 nt 0.0146 0.0092 0.006 24
nt 8.9 0.0219 nt 0.0068 0.0047 0.025 25 nt 4.7 0.0372 nt 0.0100
7.8886 nt 26 nt 28.0 nt nt 4.1305 3.9537 nt 27 nt 20.4 nt nt 1.6634
2.9648 nt 28 nt 19.8 nt nt 7.4817 3.7757 nt 29 nt -0.5 nt nt 3.5481
2.9717 nt 30 nt -9.6 0.0501 nt 0.0838 0.9268 1.585 31 nt -11.8
0.0079 nt 0.0067 0.0753 0.016 33 nt 5.9 0.3162 nt 0.3681 2.3496 nt
34 nt 18.1 nt nt 12.8529 >10 nt 35 nt 19.9 nt nt >10 >10
nt 36 nt 8.1 0.1259 nt 0.0553 0.0111 nt 37 nt 4.8 0.2512 nt 0.1718
4.5920 nt 38 nt 25.6 nt 0.0562 9.9312 7.6736 nt 39 nt 22.0 nt nt
2.4322 9.4406 nt 40 nt 39.8 nt nt 9.0365 14.7571 nt 41 nt 33.5 nt
nt >10 10.6905 nt 42 nt 28.3 nt 0.0186 2.1257 9.2257 nt 43 nt
31.7 nt 0.1334 9.7051 >10 nt 44 nt 8.4 nt nt 3.9174 1.4588 nt 45
nt 11.7 nt nt 1.8155 0.4083 nt 46 nt 4.4 nt nt 2.5410 2.4210 nt 47
nt 36.0 nt nt 10.3753 1.6069 nt 48 nt 31.2 nt nt 1.6634 0.6368 nt
49 nt 20.4 nt nt 4.1115 2.7290 nt 50 nt 41.0 nt nt 10.0000 1.6827
nt 51 nt 17.5 nt nt 1.9320 0.5781 nt 52 nt 40.5 nt nt 7.0469 2.0893
nt 53 nt 6.8 0.0158 0.0398 0.0020 0.0628 0.079 54 nt 0.8 0.0141
0.0100 0.0197 1.8239 nt 55 nt -1.3 0.0063 0.0010 0.0126 0.0200 nt
56 nt 102.0 nt nt nt nt nt 57 nt 83.5 nt nt nt nt nt 58 nt 93.9 nt
nt nt nt nt 59 nt 69.8 nt nt nt nt nt 60 nt 54.0 nt nt nt nt nt 61
nt 29.3 nt nt 0.3396 >10 nt 62 nt 83.0 nt nt nt nt nt 63 nt 81.5
nt nt nt nt nt 64 nt 81.2 nt nt nt nt nt 65 nt 23.9 nt nt >10
>10 nt 66 nt -0.5 0.0079 0.0020 0.0027 0.0269 nt 67 nt -3.7
0.3162 nt 0.4831 2.0464 nt 68 nt 7.3 0.0063 0.0016 0.0052 0.0066 nt
69 nt 5.6 0.0032 0.0025 0.0031 0.3350 nt 70 nt 10.8 0.0126 0.0040
0.0069 0.0077 nt 71 nt 16.1 nt nt 1.2218 >10 nt 72 nt 25.9 nt
0.1778 8.5901 >10 nt 73 nt 52.6 nt nt 0.0068 0.1072 nt 74 nt
39.9 nt nt 0.0049 0.0101 nt 75 nt 23.8 0.0288 0.0251 0.0529 0.5416
nt 76 nt 22.6 0.0100 0.0050 0.0067 0.0168 nt 77 nt 14.8 nt nt
0.5888 1.1092 nt 78 nt 32.3 nt nt 0.0143 6.6069 nt 79 nt -0.9
0.1000 nt 0.0151 0.0071 nt 80 nt 10.7 0.0200 nt 0.0563 0.3954 nt 81
nt 10.1 0.0501 0.0089 0.0113 0.0270 nt 82 nt 1.6 nt nt 0.9594
1.8493 nt 83 nt 17.6 nt nt 1.7906 0.9977 nt 84 nt 32.6 0.0200 nt
0.0025 0.0031 nt 85 nt 11.0 0.0398 nt 0.0301 1.0186 nt 86 nt 12.3
0.0126 0.0100 0.0023 0.0585 nt 87 nt 12.3 nt nt 3.8107 9.9312 nt 88
7.4 0.0200 0.0251 0.0102 0.1662 89 nt 10.3 nt nt 0.9795 >10 nt
90 nt 2.3 0.0200 nt 0.0119 0.9638 nt 91 nt 1.9 0.0316 nt 0.0075
2.1316 nt 92 nt 37.1 nt nt 0.7551 17.6604 nt 93 nt 7.9 0.0158
0.0063 0.0081 0.0178 nt 94 nt -3.8 nt nt 0.4335 2.1577 nt 95 nt 8.9
0.0316 nt 0.0097 0.2113 nt 96 nt 43.6 nt nt 4.8865 >10 nt 97 nt
-1.7 0.0126 0.0047 0.0078 0.0428 nt 98 nt 44.1 nt nt 5.5081 >10
nt 99 nt 4.9 0.0174 0.0063 0.0091 0.0348 nt 100 nt 102.0 nt nt nt
nt nt 101 nt 12.3 0.0316 nt 0.0278 5.0583 nt 102 nt -20.1 0.0100
0.0295 0.0105 0.4883 nt 103 nt 15.0 nt nt 1.3964 >10 nt 104 nt
59.1 nt nt 4.4771 >10 nt 105 nt -16.6 0.0219 nt 0.0986 9.9541 nt
106 nt 26.7 nt nt >10 >10 nt 107 nt -25.8 0.0100 nt 0.0128
0.1782 nt 108 nt -4.2 0.0058 0.0100 0.0086 0.0816 nt 109 nt 42.7 nt
nt >10 >10 nt 110 nt -2.2 0.0079 0.0166 0.0034 0.4406 nt 111
nt 67.2 nt nt >10 >10 nt 112 nt 6.8 0.0447 nt 0.1340 2.8249
nt 113 nt 54.0 nt nt 3.8107 >10 nt 114 nt 15.6 nt nt 4.0832
>10 nt 115 nt -9.9 0.0100 nt 0.0080 0.6227 nt 116 nt 59.5 nt nt
nt nt nt 117 nt 2.5 0.0200 nt 0.0640 7.0307 nt 118 nt -22.9 nt nt
0.9205 19.6336 nt 119 nt 87.3 nt nt nt nt nt 120 nt -16.0 0.0501 nt
0.4764 >10 nt 121 nt 79.6 nt nt nt nt nt 122 nt -5.4 0.0100 nt
0.0098 0.2618 nt 123 nt 88.6 nt nt nt nt nt 124 nt -28.8 0.0079 nt
0.0106 0.2884 nt 125 nt -14.1 nt nt 11.7219 >10 nt 126 nt 2.1 nt
nt 1.5488 >10 nt 127 nt 10.4 0.1259 nt 0.2553 28.2488 nt 128 nt
2.0 0.0501 nt 0.0685 0.2911 nt 129 nt 114.8 nt nt nt nt nt 130 nt
12.4 nt nt >10 >10 nt 131 nt nt nt nt nt nt nt *means not
tested
* * * * *