U.S. patent application number 15/119639 was filed with the patent office on 2017-03-09 for 2-amino-3,5,5-trifluoro-3,4,5,6-tetrahydropyridines as bace1 inhibitors for treatment of alzheimer's disease.
This patent application is currently assigned to H. Lundbeck A/S. The applicant listed for this patent is H. Lundbeck A/S. Invention is credited to Thomas Jensen, Karsten Juhl, Mauro Marigo, Lena Tagmose.
Application Number | 20170066741 15/119639 |
Document ID | / |
Family ID | 59009438 |
Filed Date | 2017-03-09 |
United States Patent
Application |
20170066741 |
Kind Code |
A1 |
Juhl; Karsten ; et
al. |
March 9, 2017 |
2-Amino-3,5,5-Trifluoro-3,4,5,6-Tetrahydropyridines as BACE1
Inhibitors for Treatment of Alzheimer's Disease
Abstract
The present invention is directed to novel inhibitors of the
BACE1 enzyme. Separate aspects of the invention are directed to
pharmaceutical compositions comprising said compounds and uses of
the compounds to treat disorders for which the reduction of A.beta.
deposits is beneficial such as Alzheimer's disease.
Inventors: |
Juhl; Karsten; (Greve,
DK) ; Marigo; Mauro; (Skovlunde, DK) ;
Tagmose; Lena; (Lyngby, DK) ; Jensen; Thomas;
(Copenhagen O, DK) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
H. Lundbeck A/S |
Valby |
|
DK |
|
|
Assignee: |
H. Lundbeck A/S
Valby
DK
|
Family ID: |
59009438 |
Appl. No.: |
15/119639 |
Filed: |
February 17, 2015 |
PCT Filed: |
February 17, 2015 |
PCT NO: |
PCT/EP2015/053327 |
371 Date: |
August 17, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 413/12 20130101;
C07D 211/72 20130101; C07D 413/14 20130101; C07D 405/12 20130101;
C07D 227/10 20130101; C07D 417/12 20130101; A61P 25/28 20180101;
C07D 401/12 20130101 |
International
Class: |
C07D 401/12 20060101
C07D401/12; C07D 211/72 20060101 C07D211/72; C07D 405/12 20060101
C07D405/12; C07D 413/12 20060101 C07D413/12; C07D 417/12 20060101
C07D417/12 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 19, 2014 |
DK |
PA 201400086 |
Jul 1, 2014 |
DK |
PA 201400349 |
Claims
1. A compound of formula I: ##STR00073## wherein: Ar is selected
from the group consisting of phenyl, pyridyl, pyrimidyl, pyrazinyl,
imidazolyl, pyrazolyl, 1,2,4-triazolyl, thiophenyl, thiazolyl,
oxazolyl, isoxazolyl, 1,3,4-thiadiazolyl, isothiazolyl,
1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl, furazanyl and
1,2,4-thiadiazolyl and wherein Ar is optionally substituted with
one or more halogen, CN, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 fluoroalkyl or
C.sub.1-C.sub.6 alkoxy; R.sup.1 is C.sub.1-C.sub.3 alkyl or
C.sub.1-C.sub.3 fluoroalkyl; R.sup.2 is hydrogen, halogen,
C.sub.1-C.sub.3 fluoroalkyl or C.sub.1-C.sub.3 alkyl; and R.sup.3
is C.sub.1-C.sub.3 alkyl; or a pharmaceutically acceptable salt
thereof.
2. The compound of claim 1, wherein the compound is of formula Ia:
##STR00074## or a pharmaceutically acceptable salt thereof.
3. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein R.sup.1 is CH.sub.3.
4. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein R.sup.2 is F or H.
5. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein R.sup.3 is CH.sub.3.
6. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein Ar is optionally substituted with one or more F,
C.sub.1, CN, C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 fluoroalkyl or
C.sub.1-C.sub.3 alkoxy.
7. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein the stereochemistry of said compound is (2R,5S),
or a.
8. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein the compound is selected from the group consisting
of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
and (7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrah-
ydropyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide.
9. The compound or pharmaceutically acceptable salt thereof of
claim 1, wherein the compound is selected from the group consisting
of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
(7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydro-
pyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (8)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (9)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(10)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide; (11)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide; (12)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide; (13)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(14)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(15)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(16)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide;
(17)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (18)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; (19)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxy-3-methylpyrazine-2-carboxamide;
(20)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,3,4-oxadiazole-2-carboxamide;
(21)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide;
(22)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-methyl-1H-1,2,4-triazole-3-carboxamide;
(23) N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,
5-tetrahydropyridin-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole--
3-carboxamide; (24)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,
5-tetrahydropyridin-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-ca-
rboxamide; (25)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,
5-tetrahydropyridin-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide-
; (26) N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,
5-tetrahydropyridin-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxami-
de; (27) N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,
5-tetrahydropyridin-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide;
(28)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide;
(29)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(30)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(31)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(32)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (33)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (34)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (35)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,2,4-oxadiazole-3-carboxamide;
(36)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (37)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(38)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1,4-dimethyl-1H-imidazole-2-carboxamide;
(39)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1-methyl-1H-pyrazole-3-carboxamide;
(40)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methylisoxazole-5-carboxamide; (41)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methylfuran-2-carboxamide; (42)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-5-carboxamide; (43)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; (44)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-chlorobenzamide; (45)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethoxy)picolinamide; (46)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-fluoro-5-methoxypicolinamide; (47)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2,5-dimethyloxazole-4-carboxamide; (48)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; and (49)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoro-3-methylpicolinamide.
10. The compound or pharmaceutically acceptable salt thereof of
claim 9, wherein the compound is selected from the group consisting
of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; and (5)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide.
11. A pharmaceutical composition comprising the compound or a
pharmaceutically acceptable salt thereof of claim 1 and a
pharmaceutically acceptable carrier.
12-15. (canceled)
16. The pharmaceutical composition of claim 11, wherein the
compound is of formula Ia: ##STR00075##
17. The pharmaceutical composition of claim 11, wherein the
compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
(7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydro-
pyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (8)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (9)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(10)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide; (11)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide; (12)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide; (13)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(14)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(15)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(16)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide;
(17)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (18)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; (19)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxy-3-methylpyrazine-2-carboxamide;
(20)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,3,4-oxadiazole-2-carboxamide;
(21)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide;
(22)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-methyl-1H-1,2,4-triazole-3-carboxamide;
(23)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
(24)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide;
(25)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide;
(26)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide; (27)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide; (28)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide; (29)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(30)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(31)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(32)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (33)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (34)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (35)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,2,4-oxadiazole-3-carboxamide;
(36)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (37)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(38)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1,4-dimethyl-1H-imidazole-2-carboxamide;
(39)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1-methyl-1H-pyrazole-3-carboxamide;
(40)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methylisoxazole-5-carboxamide; (41)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methylfuran-2-carboxamide; (42)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-5-carboxamide; (43)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; (44)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-chlorobenzamide; (45)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethoxy)picolinamide; (46)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-fluoro-5-methoxypicolinamide; (47)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2,5-dimethyloxazole-4-carboxamide; (48)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; and (49)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoro-3-methylpicolinamide.
18. The pharmaceutical composition of claim 17, wherein the
compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
and (7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrah-
ydropyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide.
19. The pharmaceutical composition of claim 17, wherein the
compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; and (5)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide.
20. A method of treating Alzheimer's disease comprising
administering a therapeutically effective amount of the compound or
pharmaceutically acceptable salt thereof of claim 1.
21. The method of treating Alzheimer's disease of claim 20, wherein
said compound is of formula Ia: ##STR00076##
22. The method of treating Alzheimer's disease of claim 20, wherein
said compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
(7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydro-
pyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (8)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (9)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(10)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide; (11)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide; (12)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide; (13)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(14)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(15)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(16)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide;
(17)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (18)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; (19)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxy-3-methylpyrazine-2-carboxamide;
(20)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,3,4-oxadiazole-2-carboxamide;
(21)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide;
(22)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-methyl-1H-1,2,4-triazole-3-carboxamide;
(23)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
(24)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide;
(25)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide;
(26)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide; (27)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide; (28)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide; (29)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide;
(30)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide;
(31)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide;
(32)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (33)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (34)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide; (35)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methyl-1,2,4-oxadiazole-3-carboxamide;
(36)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-thiazole-2-carboxamide; (37)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide;
(38)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1,4-dimethyl-1H-imidazole-2-carboxamide;
(39)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydr-
opyridin-2-yl)-4-fluorophenyl)-1-methyl-1H-pyrazole-3-carboxamide;
(40)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-methylisoxazole-5-carboxamide; (41)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methylfuran-2-carboxamide; (42)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2-methyloxazole-5-carboxamide; (43)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; (44)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-4-chlorobenzamide; (45)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethoxy)picolinamide; (46)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-3-fluoro-5-methoxypicolinamide; (47)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-2,5-dimethyloxazole-4-carboxamide; (48)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide; and (49)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoro-3-methylpicolinamide.
23. The method of treating Alzheimer's disease of claim 22, wherein
said compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-chloropicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyanopicolinamide; (5)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (6)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide;
and (7)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrah-
ydropyridin-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide.
24. The method of treating Alzheimer's disease of claim 22, wherein
said compound is selected from the group consisting of: (1)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide; (2)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide; (3)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-methoxypicolinamide; (4)
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide; and (5)
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide.
Description
FIELD OF THE INVENTION
[0001] The present invention provides compounds which act as BACE1
inhibitors. Separate aspects of the invention are directed to
pharmaceutical compositions comprising said compounds and uses of
the compounds to treat Alzheimer's disease.
BACKGROUND ART
[0002] Dementia is a clinical syndrome characterized by deficits in
multiple areas of cognition that cannot be explained by normal
aging, a noticeable decline in function, and an absence of
delirium. In addition, neuropsychiatric symptoms and focal
neurological findings are usually present. Dementia is further
classified based on etiology Alzheimer's disease (AD) is the most
common cause of dementia, followed by mixed AD and vascular
dementia, vascular dementia, Lewy body dementia (DLB), and
fronto-temporal dementia.
[0003] .beta.-Amyloid deposits and neurofibrillary tangles are
considered to be major pathologic characterizations associated with
AD which is characterized by the loss of memory, cognition,
reasoning, judgment, and orientation. Also affected, as the disease
progresses, are motor, sensory and linguistic abilities until
global impairment of multiple cognitive functions occurs.
.beta.-Amyloid deposits are predominantly an aggregate of A.beta.
peptide, which in turn is a product of the proteolysis of amyloid
precursor protein (APP) as part of the .beta.-amyloidogenic
pathway, A.beta. peptide results from the cleavage of APP at the
C-terminals by one or more .gamma.-secretases and at the N-terminus
by .beta.-secretase enzyme (BACE1) also known as aspartyl protease
2. BACE1 activity is correlated directly to the generation of
A.beta. peptide from APP.
[0004] Studies indicate that the inhibition of BACE1 impedes the
production of A.beta. peptide. Further, BACE1 co-localizes with its
substrate APP in Golgi and endocytic compartments (Willem M, et al.
Semin. Cell Dev. Biol, 2009, 20, 175-182). Knock-out studies in
mice have demonstrated the absence of amyloid peptide formation
while the animals are healthy and fertile (Ohno M, et al.
Neurobiol. Dis., 2007, 26, 134-145). Genetic ablation of BACE1 in
APP-overexpressing mice has demonstrated absence of plaque
formation, and the reversal of cognitive deficits (Ohno M, et al.
Neuron; 2004, 41, 27-33). BACE1 levels are elevated in the brains
of sporadic AD patients (Hampel and Shen, Scand. J. Clin. Lab.
Invest. 2009, 69, 8-12).
[0005] These convergent findings indicate that the inhibition of
BACE1 may be a therapeutic target for the treatment of AD as well
as disorders for which the reduction of A.beta. deposits is
beneficial.
[0006] At the 2012 Alzheimer's Association International Conference
in Vancouver, several drug developers announced their BACE1
inhibitors in clinical trials. Eli Lilly scientists reported
preclinical research on LY2886721 which had entered a Phase II
study. Merck presented a series of posters detailing the Phase I
studies of its BACE inhibitor, MK-8931, and announced the start of
separate Phase III studies which will test the compound for two
years in people with prodromal Alzheimer's disease. Numerous patent
applications directed to BACE1 inhibitors have been published over
the past several years.
[0007] AstraZeneca announced the discovery of AZD3839, a potent and
selected BACE1 inhibitor clinical candidate for the treatment of AD
(Jeppsson, F., et al. JBC, 2012, 287, 41245-41257) in October 2012.
The effort which led to the discovery of AZD3839 was further
described in Ginman, T., et al. Journal of Medicinal Chemistry,
2013, 56, 4181-4205. The Ginman publication describes the issues
which were overcome in connection with the discovery and
identification of AZD3839. These issues related to poor blood brain
barrier penetration and P-glycoprotein mediated efflux of the
compounds resulting in lack of brain exposure.
[0008] The Ginman manuscript hypothesized that the differences
would largely be due to the core structures and Structure Activity
Relationship data was provided wherein the in vitro properties on
the reported compounds were given into four tables according to
core sub-types. In table 4, a series of amidine containing
compounds are described that were considered interesting from an
activity perspective. However, the data suggests that the amidine
containing core did not exhibit a favourable blood brain barrier
permeability profile.
[0009] Researchers from Hoffmann-La Roche and Siena Biotech also
reported the discovery of amidine containing compounds (Woltering,
T. J., et al. Bioorg. Med. Chem. Lett. 2013, 23, 4239-4243). These
compounds (compounds 17 and 18 in the paper) were found not to have
any in vivo effect (lack of A.beta.40 reduction in brain in wild
type mice).
SUMMARY OF THE INVENTION
[0010] Contrary to the teachings of Ginman, et al. and Woltering,
T. J., et al., the inventors have discovered a series of amidine
compounds which are brain penetrant and are thus able to inhibit
BACE1 in the brain after peripheral dose of said compounds.
Accordingly, the present invention relates to novel compounds
having BACE1 inhibitory activity, to their preparation, to their
medical use and to medicaments comprising them.
[0011] An objective of the present invention is to provide
compounds that inhibit BACE1. Accordingly, the present invention
relates to compounds of Formula I.
##STR00001##
[0012] Ar is selected from the group consisting of phenyl, pyridyl,
pyrimidyl, pyrazinyl, imidazolyl, pyrazolyl, 1,2,4-triazolyl,
thiophenyl, thiazolyl, oxazolyl, isoxazolyl, 1,3,4-thiadiazolyl,
isothiazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl, furazanyl and
1,2,4-thiadiazolyl and where the Ar is optionally substituted with
one or more halogen, CN, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 fluoroalkyl or
C.sub.1-C.sub.6 alkoxy;
[0013] R.sup.1 is C.sub.1-C.sub.3 alkyl or C.sub.1-C.sub.3
fluoroalkyl;
[0014] R.sup.2 is hydrogen, halogen, C.sub.1-C.sub.3 fluoroalkyl or
C.sub.1-C.sub.3 alkyl; and
[0015] R.sup.3 is C.sub.1-C.sub.3 alkyl; or a pharmaceutically
acceptable salt thereof.
[0016] The present invention further provides a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula I and a pharmaceutically acceptable
carrier.
[0017] The present invention provides a method of treating a
subject suffering from Alzheimer's disease comprising administering
to the subject a therapeutically effective amount of a compound of
Formula I.
[0018] The present invention is directed to the use of a compound
as defined in Formula I for the manufacture of a medicament for
treating Alzheimer's disease.
[0019] Further embodiments of the invention are provided
immediately below:
[0020] In one embodiment, the compound is of formula Ia
##STR00002##
or a pharmaceutically acceptable salt thereof.
[0021] In one embodiment, R.sup.1 is CH.sub.3.
[0022] In one embodiment, R.sup.2 is F or H.
[0023] In one embodiment, R.sup.3 is CH.sub.3.
[0024] In one embodiment, Ar is optionally substituted with one or
more F, Cl, CN, C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 fluoroalkyl
or C.sub.1-C.sub.3 alkoxy.
[0025] In one embodiment, the stereochemistry is (2R,5S).
[0026] In one embodiment, Ar is optionally substituted phenyl.
[0027] In one embodiment, Ar is optionally substituted pyridyl.
[0028] In one embodiment, Ar is optionally substituted
pyrimidyl.
[0029] In one embodiment, Ar is optionally substituted
pyrazinyl.
[0030] In one embodiment, Ar is optionally substituted
imidazolyl.
[0031] In one embodiment, Ar is optionally substituted
pyrazolyl.
[0032] In one embodiment, Ar is optionally substituted
1,2,4-triazolyl.
[0033] In one embodiment, Ar is optionally substituted
thiophenyl.
[0034] In one embodiment, Ar is optionally substituted
oxazolyl.
[0035] In one embodiment, Ar is optionally substituted
isoxazolyl.
[0036] In one embodiment, Ar is optionally substituted
1,3,4-thiadiazolyl.
[0037] In one embodiment, Ar is optionally substituted
thiazolyl.
[0038] In one embodiment, Ar is optionally substituted
isothiazolyl.
[0039] In one embodiment, Ar is optionally substituted
1,3,4-oxadiazolyl.
[0040] In one embodiment, Ar is optionally substituted
1,2,4-oxadiazolyl.
[0041] In one embodiment, Ar is optionally substituted
furazanyl.
[0042] In one embodiment, Ar is optionally substituted
1,2,4-thiadiazolyl.
[0043] In separate embodiments of the invention, the compound is
selected from one of the exemplified compounds disclosed in the
Experimental Section.
[0044] A separate embodiment is directed to a pharmaceutical
composition comprising the compound or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable
carrier.
[0045] Another embodiment is directed to a method of treating
Alzheimer's disease comprising administering a therapeutically
effective amount of the compound.
[0046] Yet another embodiment is directed to a use of a compound
for the manufacture of a medicament for treating Alzheimer's
disease.
[0047] One embodiment is a compound for use in therapy.
[0048] Yet another embodiment is directed to a compound for use in
the treatment of Alzheimer's disease. Included also in this
invention are isotopically labelled compounds, which are identical
to those claimed in formula I, where one or more atoms are replaced
by an atom having an atomic mass or mass number different from the
atomic mass or mass number usually found in nature. Examples of
isotopes that can be incorporated into compounds of the invention
include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur,
fluorine, chlorine and iodine, such as .sup.2H, .sup.3H, .sup.13C,
.sup.11C, .sup.14C, .sup.15N, .sup.18O, .sup.17O, .sup.35S,
.sup.18F, .sup.36Cl and .sup.125I, respectively. Compounds of the
invention, prodrugs thereof, and pharmaceutically acceptable salts
of said compounds or said prodrugs which contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of this invention. Certain isotopically labelled compounds of the
invention, such as those with radioactive isotopes incorporated
including but not limited to .sup.3H and .sup.14C, are useful in
drug and/or substrate tissue distribution assays.
BRIEF DESCRIPTION OF THE DRAWINGS
[0049] FIG. 1 shows the two molecules in the asymmetric unit as
found in the crystal structure. The X-ray method used cannot
distinguish between hydrogen (.sup.1H) and deuterium (D or
.sup.2H). Hence, the deuterium atoms in the d3-methoxy group are
depicted as hydrogen.
DETAILED DESCRIPTION OF THE INVENTION
[0050] The present invention is based on the discovery that the
compounds of Formula I are inhibitors of BACE1, and as such, are
useful for the treatment of related disorders. Certain aspects of
the invention are explained in greater detail below but this
description is not intended to be a detailed catalog of all the
different ways in which the invention may be implemented, or all
the features that may be added to the present invention. Hence, the
following specification is intended to illustrate some embodiments
of the invention, and not to exhaustively specify all permutations,
combinations and variations thereof.
[0051] As used herein, the term "C.sub.1-C.sub.6 alkyl" refers to a
straight chained or branched saturated hydrocarbon having from one
to six carbon atoms, inclusive. Examples of such substituents
include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl,
1-butyl, 2-butyl, 2-methyl-2-propyl, 2-methyl-1-propyl, n-pentyl
and n-hexyl. Similarly, the term "straight chained or branched
C.sub.1-C.sub.3 alkyl" refers to a saturated hydrocarbon having
from one to three carbon atoms, inclusive. Examples of such
substituents include, but are not limited to, methyl, ethyl and
n-propyl.
[0052] Likewise, the term "C.sub.1-C.sub.6 alkoxy" refers to a
straight chained or branched saturated alkoxy group having from one
to six carbon atoms, inclusive, with the open valency on the
oxygen. Examples of such substituents include, but are not limited
to, methoxy, ethoxy, n-butoxy, t-butoxy and n-hexyloxy. The
"C.sub.1-C.sub.6 alkoxy" is optionally substituted with one or more
fluorine atoms.
[0053] As used herein, the term "C.sub.1-C.sub.6 fluoroalkyl"
refers to a straight chained or branched saturated hydrocarbon
having from one to six carbon atoms inclusive substituted with one
or more fluorine atoms. Examples of such substituents include, but
are not limited to, trifluoromethyl, pentafluoroethyl,
1-fluoroethyl, monofluoromethyl, difluoromethyl, 1,2-difluoroethyl
and 3,4 difluorohexyl. Similarly, the term "straight chained or
branched C.sub.1-C.sub.3 fluoroalkyl" refers to a saturated
hydrocarbon having from one to three carbon atoms, inclusive,
substituted with one or more fluorine atoms per carbon atom.
[0054] The term "halogen" refers to fluorine, chlorine, bromine and
iodine.
[0055] The term "C.sub.2-6-alkenyl" refers to a branched or
unbranched alkenyl group having from two to six carbon atoms and
one double bond, including but not limited to ethenyl, propenyl,
and butenyl.
[0056] The term "C.sub.2-6-alkynyl" shall mean a branched or
unbranched alkynyl group having from two to six carbon atoms and
one triple bond, including but not limited to ethynyl, propynyl and
butynyl.
[0057] Throughout the description and the claims, each compound
number corresponds to the number of the experiment in which the
method of manufacture is disclosed. Compounds 4 and 18 have
resynthesized using a modified method of manufacture as disclosed
in examples 4a and 18a.
[0058] As used herein, the phrase "effective amount" when applied
to a compound of the invention, is intended to denote an amount
sufficient to cause an intended biological effect. The phrase
"therapeutically effective amount" when applied to a compound of
the invention is intended to denote an amount of the compound that
is sufficient to ameliorate, palliate, stabilize, reverse, slow or
delay the progression of a disorder or disease state, or of a
symptom of the disorder or disease. In an embodiment, the method of
the present invention provides for administration of combinations
of compounds. In such instances, the "effective amount" is the
amount of the combination sufficient to cause the intended
biological effect.
[0059] The term "treatment" or "treating" as used herein means
ameliorating or reversing the progress or severity of a disease or
disorder, or ameliorating or reversing one or more symptoms or side
effects of such disease or disorder. "Treatment" or "treating", as
used herein, also means to inhibit or block, as in retard, arrest,
restrain, impede or obstruct, the progress of a system, condition
or state of a disease or disorder. For purposes of this invention,
"treatment" or "treating" further means an approach for obtaining
beneficial or desired clinical results, where "beneficial or
desired clinical results" include, without limitation, alleviation
of a symptom, diminishment of the extent of a disorder or disease,
stabilized (i.e., not worsening) disease or disorder state, delay
or slowing of a disease or disorder state, amelioration or
palliation of a disease or disorder state, and remission of a
disease or disorder, whether partial or total, detectable or
undetectable.
[0060] The present invention also provides a method of treating a
disease or disorder, the method comprises administering a
therapeutically effective amount of at least one compound of the
present invention or a pharmaceutically acceptable salt thereof to
a mammal in need thereof, wherein the disease or disorder is a
neurodegenerative or cognitive disease or disorder.
[0061] In an embodiment of the invention, the neurodegenerative or
cognitive disease or disorder is Alzheimer's disease, mild
cognitive impairment, Trisomy 21 (Down Syndrome), cerebral amyloid
angiopathy, Hereditary Cerebral Hemorrhage with Amyloidosis of the
Dutch-Type (HCHWA-DT), degenerative dementia, amyotrophic lateral
sclerosis, traumatic brain injury or stroke.
[0062] In an embodiment, the disease or disorder is a peripheral
amyloidosis, such as amyloid neuropathy or pancreatitis.
[0063] In an embodiment, the disease or disorder is peripheral
nerve damage.
[0064] The present invention provides a method of treating
Alzheimer's disease in a patient comprising administering to a
patient in need of such treatment a therapeutically effective
amount of at least one compound of formula I.
[0065] The present invention further provides a method of
inhibiting BACE1 in a patient comprising administering to a patient
in need thereof a therapeutically effective amount of at least one
compound of formula I.
[0066] The present invention also provides a method of inhibiting
.beta.-secretase mediated cleavage of amyloid precursor protein
comprising administering to a patient in need of such treatment a
therapeutically effective amount of at least one compound of
formula I.
[0067] In further embodiments, the present invention provides the
use of a compound of formula I for the manufacture of a medicament
for the treatment of Alzheimer's disease. The present invention
also provides the use of a compound of formula I for the
manufacture of a medicament for the inhibition of BACE1. The
present invention further provides the use of a compound of formula
I for the manufacture of a medicament for the inhibition of
production or accumulation of A.beta. peptide.
[0068] The invention also provides a compound of formula I for use
in therapy of a patient, for example, in the treatment of
Alzheimer's disease or to slow the progression of a patient's mild
cognitive impairment to Alzheimer's disease.
[0069] In a further embodiment, the invention provides a
pharmaceutical formulation adapted for any of the above treatments
and uses.
[0070] In some embodiments, the mammal of the method of the
invention is a human.
[0071] In other embodiments, the patient of the method of the
invention is a human patient.
[0072] In some embodiments of the invention, at least one symptom
of the neurodegenerative or cognitive disease or disorder is
treated.
Pharmaceutically Acceptable Salts
[0073] The present invention also comprises salts of the present
compounds, typically, pharmaceutically acceptable salts. Such salts
include pharmaceutically acceptable acid addition salts. Acid
addition salts include salts of inorganic acids as well as organic
acids.
[0074] Representative examples of suitable inorganic acids include
hydrochloric, hydrobromic, hydroiodic, phosphoric, sulfuric,
sulfamic, nitric acids and the like. Representative examples of
suitable organic acids include formic, acetic, trichloroacetic,
trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric,
glycolic, itaconic, lactic, methanesulfonic, maleic, malic,
malonic, mandelic, oxalic, picric, pyruvic, salicylic, succinic,
methane sulfonic, ethanesulfonic, tartaric, ascorbic, pamoic,
bismethylene salicylic, ethanedisulfonic, gluconic, citraconic,
aspartic, stearic, palmitic, EDTA, glycolic, p-aminobenzoic,
glutamic, benzenesulfonic, p-toluenesulfonic acids, theophylline
acetic acids, as well as the 8-halotheophyllines (for example,
8-bromotheophylline and the like). Further examples of
pharmaceutically acceptable inorganic or organic acid addition
salts include the pharmaceutically acceptable salts listed in S. M.
Berge, et al., J. Pharm. Sci., 1977, 66, 2.
[0075] Furthermore, the compounds of this invention may exist in
unsolvated as well as in solvated forms with pharmaceutically
acceptable solvents such as water, ethanol and the like.
[0076] The compounds of the present invention may have one or more
asymmetric centres and it is intended that any optical isomers
(i.e. enantiomers or diastereomers), as separated, pure or
partially purified optical isomers, and any mixtures thereof
including racemic mixtures, i.e. a mixture of stereoisomeres, are
included within the scope of the invention.
[0077] In this context is understood that when specifying the
enantiomeric form, then the compound is in enantiomeric excess,
e.g. essentially in a pure form. Accordingly, one embodiment of the
invention relates to a compound of the invention having an
enantiomeric excess of at least 60%, at least 70%, at least 80%, at
least 85%, at least 90%, at least 96%, preferably at least 98%.
[0078] Racemic forms may be resolved into the optical antipodes by
known methods, for example, by separation of diastereomeric salts
thereof with an optically active acid, and liberating the optically
active amine compound by treatment with a base. Separation of such
diastereomeric salts can be achieved, e.g. by fractional
crystallization. The optically active acids suitable for this
purpose may include, but are not limited to d- or l-tartaric,
mandelic or camphorsulfonic acids. Another method for resolving
racemates into the optical antipodes is based upon chromatography
on an optically active matrix. The compounds of the present
invention may also be resolved by the formation and chromatographic
separation of diastereomeric derivatives from chiral derivatizing
reagents, such as, chiral alkylating or acylating reagents,
followed by cleavage of the chiral auxiliary. Any of the above
methods may be applied either to resolve the optical antipodes of
the compounds of the invention per se or to resolve the optical
antipodes of synthetic intermediates, which can then be converted
by methods described herein into the optically resolved final
products which are the compounds of the invention.
[0079] Additional methods for the resolution of optical isomers,
known to those skilled in the art, may be used. Such methods
include those discussed by J. Jaques, A. Collet and S. Wilen in
Enantiomers, Racemates, and Resolutions, John Wiley and Sons, New
York, 1981. Optically active compounds can also be prepared from
optically active starting materials.
Pharmaceutical Compositions
[0080] The present invention further provides a pharmaceutical
composition comprising a therapeutically effective amount of a
compound of Formula I and a pharmaceutically acceptable carrier.
The present invention also provides a pharmaceutical composition
comprising a therapeutically effective amount of one of the
specific compounds disclosed in the description, such as in the
experimental section and a pharmaceutically acceptable carrier.
[0081] The compounds of the invention may be administered alone or
in combination with pharmaceutically acceptable carriers or
excipients, in either single or multiple doses. The pharmaceutical
compositions according to the invention may be formulated with
pharmaceutically acceptable carriers or diluents as well as any
other known adjuvants and excipients in accordance with
conventional techniques such as those disclosed in Remington: The
Science and Practice of Pharmacy, 21.sup.th Edition, Troy, Ed.,
Lippincott Williams &Wilkins, Baltimore, Md., USA.
[0082] Pharmaceutical compositions for oral administration include
solid dosage forms such as capsules, tablets, dragees, pills,
lozenges, powders and granules. Where appropriate, the compositions
may be prepared with coatings such as enteric coatings or they may
be formulated so as to provide controlled release of the active
ingredient such as sustained or prolonged release according to
methods well known in the art. Liquid dosage forms for oral
administration include solutions, emulsions, suspensions, syrups
and elixirs. Pharmaceutical compositions for parenteral
administration include sterile aqueous and nonaqueous injectable
solutions, dispersions, suspensions or emulsions as well as sterile
powders to be reconstituted in sterile injectable solutions or
dispersions prior to use. Other suitable administration forms
include, but are not limited to, suppositories, sprays, ointments,
creams, gels, inhalants, dermal patches and implants.
[0083] Typical oral dosages range from about 0.01 to about 100
mg/kg body weight per day.
[0084] The compounds of this invention are generally utilized as
the free substance or as a pharmaceutically acceptable salt
thereof. One example is an acid addition salt of a compound having
the utility of a free base. When a compound of Formula I contains a
free base such salts are prepared in a conventional manner by
treating a solution or suspension of a free base of Formula I with
a molar equivalent of a pharmaceutically acceptable acid.
Representative examples of suitable organic and inorganic acids are
described above.
[0085] Suitable pharmaceutical carriers include inert solid
diluents or fillers, sterile aqueous solutions and various organic
solvents. Examples of solid carriers include lactose, terra alba,
sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia,
magnesium stearate, stearic acid and lower alkyl ethers of
cellulose. Examples of liquid carriers include, but are not limited
to, syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty
acid amines, polyoxyethylene and water. Similarly, the carrier or
diluent may include any sustained release material known in the
art, such as glyceryl monostearate or glyceryl distearate, alone or
mixed with a wax. The pharmaceutical compositions formed by
combining the compounds of Formula I and a pharmaceutically
acceptable carrier are then readily administered in a variety of
dosage forms suitable for the disclosed routes of administration.
The formulations may conveniently be presented in unit dosage form
by methods known in the art of pharmacy.
[0086] If a solid carrier is used for oral administration, the
preparation may be tabletted, placed in a hard gelatin capsule in
powder or pellet form or it may be in the form of a troche or
lozenge. The amount of solid carrier will vary widely but will
range from about 25 mg to about 1 g per dosage unit. If a liquid
carrier is used, the preparation may be in the form of a syrup,
emulsion, soft gelatin capsule or sterile injectable liquid such as
an aqueous or non-aqueous liquid suspension or solution.
Experimental Section
[0087] The compounds of the present invention of the general
formula I, wherein R.sup.1, R.sup.2, R.sup.3 and Ar are as defined
above, can be prepared by the methods outlined in the following
reaction schemes 1-3 and in the examples. In the described methods,
it is possible to make use of variants or modifications, which are
themselves known to chemists skilled in the art or could be
apparent to the person of ordinary skill in this art. Furthermore,
other methods for preparing compounds of the invention will be
readily apparent to the person skilled in the art in light of the
following reaction schemes and examples.
[0088] For example, Schemes 1-2 describe the use of selective
protecting groups during the synthesis of the compounds of the
invention. One skilled in the art would be able to select the
appropriate protecting group for a particular reaction. Moreover,
it may be necessary to incorporate protection and deprotection
strategies for substituents such as amino, amido, carbaldehyde and
hydroxyl groups in the synthetic methods described below to
synthesize the compounds of Formula I. Methods for protection and
deprotection of such groups are well known in the art, and may be
found in T. Green, et al., Protective Groups in Organic Synthesis,
1991, 2.sup.nd Edition, John Wiley & Sons, New York.
[0089] For compounds, which can exist as a mixture or equilibrium
between two or more tautomers, only one tautomer is represented in
the schemes, although it may not be the most stable tautomer. For
compounds, which can exist in enantiomeric, stereoisomeric or
geometric isomeric forms their geometric configuration is
specified; otherwise the structure represents a mixture of
stereoisomers.
Analytical LC-MS Data were Obtained Using the Following
Methods.
Method A:
[0090] LC-MS were run on Waters Acquity UPLC-MS consisting of
Waters Acquity including column manager, binary solvent manager,
sample organizer, PDA detector (operating at 254 nm), ELS detector,
and TQ-MS equipped with APPI-source operating in positive ion
mode.
[0091] LC-conditions: The column was Acquity UPLC BEH C18 1.7
.mu.m; 2.1.times.50 mm operating at 60.degree. C. with 1.2 ml/min
of a binary gradient consisting of water+0.05% trifluoroacetic acid
(A) and acetonitrile+5% water+0.05% trifluoroacetic acid. Gradient:
0.00 min: 10% B; 1.00 min: 100% B; 1.01 min: 10% B; 1.15 min: 10%
B. Total run time: 1.15 min.
Method B:
[0092] LC-MS were run on Waters Aquity UPLC-MS consisting of Waters
Aquity including column manager, binary solvent manager, sample
organizer, PDA detector (operating at 254 nM), ELS detector, and
SQ-MS equipped with APPI-source operating in positive ion mode.
[0093] LC-conditions: The column was Acquity UPLC BEH C18 1.7
.mu.m; 2.1.times.150 mm operating at 60.degree. C. with 0.6 ml/min
of a binary gradient consisting of water+0.05% trifluoroacetic acid
(A) and acetonitrile+5% water+0.03% trifluoroacetic acid. Gradient:
0.00 min: 10% B; 3.00 min: 99.9% B; 3.01 min: 10% B; 3.60 min: 10%
B. Total run time: 3.60 min.
[0094] .sup.1H NMR spectra were recorded at 600 MHz on a Bruker
Avance AV-III-600 instrument. Chemical shift values are expressed
in ppm-values relative. The following abbreviations are used for
multiplicity of NMR signals: s=singlet, d=doublet, t=triplet,
q=quartet, dd=double doublet, ddd=double double doublet, dt=double
triplet, br=broad, and m=multiplet.
[0095] As an example and wherein R.sup.2 is fluorine in the ortho
position of the phenyl ring, compounds of the general formulae XVIa
and XVIb may be prepared as shown in Scheme 1.
##STR00003## ##STR00004## ##STR00005##
where R.sup.1 and R.sup.3 are as defined under formula I, R.sup.4
and R.sup.5 are an alkyl group such as methyl or ethyl and R.sup.6
and R.sup.7 are independently selected amine protection groups such
as a tert-butoxy carbonyl group.
[0096] Compounds of the general formula IV (Scheme 1) may be
prepared by reacting compounds of the general formula II with a
sulfinamide such as III in the presence of a Lewis acid/drying
agent such as titanium tetraethoxide. Treatment of compounds of the
general formula IV with compounds of the general formula V such as
ethyl bromodifluoroacetate in the presence of Zn powder or in the
presence of diethyl zinc and tris(triphenylphosphine)rhodium(I)
chloride gives compounds of the general formula VI. Compounds of
the general formula VII are obtained from compounds of the general
formula VI by treatment with a reducing agent such as
diisobutylaluminium hydride. In some cases compound VII might be in
the hydrate form or an oligomeric form thereof. Treatment of
compounds of the general formula VII with conditions such as ethyl
2-(diethoxyphosphoryl)-2-fluoroacetate in the presence of lithium
chloride and a base such as N,N-diisopropylethylamine gives
compounds of the general formula VIII. Compounds of the general
formula IX are obtained by hydrogenation of compounds of the
general formula VIII in the presence of a catalyst such as
palladium on carbon. Compounds of the general formula X are
obtained by treatment of compounds of the general formula IX with
an acid such as hydrochloric acid in methanol followed by treatment
with potassium carbonate in methanol. Compounds of the general
formula XI are obtained by treatment of compounds of the general
formula X with di-tert-butyl dicarbonate in the presence of a
catalytic amount of DMAP (N,N-dimethyl-4-amino-pyridine). Compounds
of the general formula XII are obtained by treatment of compounds
of the general formula XI with a base such as lithium
hexamethyldisilazide follow by alkylation with a alkylhalide.
Deprotection of compounds of the general formula XII gives
compounds of the general formula XIII which can be nitrated using
nitric acid to give compounds of the general formula XIV. Reduction
of the nitro group of compounds of the general formula XIV followed
by protection of the formed aniline moiety gives compounds of the
general formula XX. Treatment of compounds of the general formula
XV with a reagent such as Lawesson's reagent
(2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide)
followed by chromatographic separation gives compounds of the
general formulae XVIa and XVIb.
[0097] Compounds of the general formula I may be prepared as shown
in Scheme 2.
##STR00006##
where R.sup.1, R.sup.2, R.sup.3 and Ar are as defined under formula
I and R.sup.7 is an amine protection groups such as a tert-butoxy
carbonyl group.
[0098] Compounds of the general formula XVII (Scheme 2) can be
obtained by deprotection of compounds of the general formula XVI.
Compounds of the general formula XX may be prepared by reacting
compounds of the general formula XVII with a carboxylic acid
chloride of general formula XVIII or by reaction with a carboxylic
acid of general formula XIX using procedures known to chemists
skilled in the art. Treatment of compounds of the general formula
XX with ammonia gives compounds of the general formula I. In some
cases, the addition of an oxidizing reagent such as tert-butyl
hydroperoxide might be necessary to facilitate the reaction.
[0099] Compounds of the general formula I may be prepared as shown
in Scheme 3.
##STR00007##
where R.sup.1, R.sup.2, R.sup.3 and Ar are as defined under formula
I.
[0100] Compounds of the general formula XXI (Scheme 3) can be
obtained by treatment of compounds of the general formula XVII with
ammonia. Compounds of the general formula I may be prepared by
reacting compounds of the general formula XXI with a carboxylic
acid chloride of general formula XVIII or by reaction with a
carboxylic acid of general formula XIX using procedures known to
chemists skilled in the art.
Preparation of Intermediates
Intermediate:
(R)--N-(1-(2-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide
##STR00008##
[0102] 1-(2-Fluorophenyl)ethanone (15 g, 109 mmol) and
(R)-2-methylpropane-2-sulfinamide (15.79 g, 130 mmol) were placed
in a round bottom flask fitted with a reflux condenser.
Tetrahydrofuran (90 ml) (dried over 4 .ANG. MS) was added followed
by Ti(OEt).sub.4 (49.5 g, 217 mmol) and the resulting yellow
solution was stirred at gentle reflux overnight. The reaction was
allowed to cool to room temperature and concentrated under reduced
pressure. The residue was dissolved in ethyl acetate (200 mL) and
brine (100 mL) was added dropwise while stirring vigorously. The
addition of brine resulted in immediate formation of copious
amounts of a white precipitate. After 10 min stirring at room
temperature the suspension was filtered through a plug of celite
using ethyl acetate for elution. The filtrate was transferred to a
separation funnel, the layers were separated, and the organic layer
was washed with brine (150 mL). The organic layer was dried over
MgSO.sub.4, filtered and concentrated under reduced pressure and
was purified using a CombiFlash system (330 g SiO.sub.2, gradient
elution; heptanes:ethyl acetate 100:0.fwdarw.70:30) to afford
(R)--N-(1-(2-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide
(20.6 g, 70.8% yield) .sup.1H NMR (600 MHz, CDCl.sub.3) .delta.
7.69 (t, J=7.1 Hz, 1H), 7.43 (dd, J=13.0, 6.0 Hz, 1H), 7.18 (td,
J=7.7, 1.0 Hz, 1H), 7.11 (ddd, J=11.3, 8.3, 0.8 Hz, 1H), 2.78 (d,
J=3.4 Hz, 3H), 1.32 (s, 9H).
Intermediate: (R)-ethyl
3-((R)-1,1-dimethylethylsulfinamido)-2,2-difluoro-3-(2-fluorophenyl)-buta-
noate
##STR00009##
[0104] Tris(triphenylphosphine)rhodium(I) chloride (1.50 g, 1.62
mmol) was placed in a dry round bottom flask. The flask was
evacuated and filled with argon (.times.3).
(R)--N-(1-(2-Fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide
(15.6 g, 64.6 mmol) was dissolved in tetrahydrofuran (265 ml)
(dried over 4 .ANG. MS) and added to the reaction flask followed by
ethyl bromodifluoroacetate (26.2 g, 16.6 ml, 129 mmol). The dark
red/orange reaction mixture was cooled to 0.degree. C. using an
ice/water bath. Diethyl zinc (126 ml, 126 mmol, 1 M in hexane) was
added in a dropwise manner. Upon complete addition the reaction was
stirred at 0.degree. C. for an additional 1 h, the cooling was
removed and the reaction was stirred at room temperature overnight.
The reaction was diluted with ethyl acetate (250 mL) and quenched
with saturated aqueous NaHCO.sub.3 (100 mL). The resulting
suspension was filtered through a plug of celite, the phases were
separated, and the organic layer was dried over MgSO.sub.4,
filtered, and concentrated under reduced pressure. The crude
material was purified using a CombiFlash system (330 g SiO.sub.2,
gradient elution; heptanes:ethyl acetate 100:0.fwdarw.60:40) to
afford (R)-ethyl
3-((R)-1,1-dimethyl-ethylsulfinamido)-2,2-difluoro-3-(2-fluorop-
henyl)butanoate (14.1 g, 59.7% yield). The stereochemistry was
assigned based on literature precedence (WO2012110459) .sup.1H NMR
(600 MHz, CDCl.sub.3) .delta. 7.46 (t, J=8.0 Hz, 1H), 7.41-7.35 (m,
1H), 7.19-7.14 (m, 1H), 7.07 (dd, J=13.0, 8.2 Hz, 1H), 4.65 (d,
J=2.6 Hz, 1H), 4.25 (q, J=7.2 Hz, 2H), 2.07 (s, 3H), 1.30-1.21 (m,
12H).
Intermediate: (R)-ethyl
5-((R)-1,1-dimethylethylsulfinamido)-2,4,4-trifluoro-5-(2-fluorophenyl)-h-
ex-2-enoate
##STR00010##
[0106] (R)-Ethyl
3-((R)-1,1-dimethylethylsulfinamido)-2,2-difluoro-3-(2-fluorophenyl)butan-
oate (7.6 g, 20.8 mmol) was dissolved in toluene (100 ml) (dried
over 4 .ANG. MS) and transferred to a dry round bottom flask. The
solution was cooled to -78.degree. C. using a dry ice/acetone bath.
DIBAL-H (41.6 ml, 41.6 mmol, 1 M in toluene) was added in a
dropwise manner using a syringe pump (addition rate 1 mL/min) Upon
complete addition the reaction was stirred at -78.degree. C. for an
additional 1 h 20 min. The reaction was quenched at -78.degree. C.
by addition of 10 mL of ethyl acetate followed by addition of 150
mL of a saturated aqueous solution of sodium potassium tartrate.
Upon complete addition the cooling was removed, the reaction
allowed to warm to room temperature and stirred at this temperature
for 1 h. The mixture was diluted with ethyl acetate (200 mL) and
filtered through a plug of celite using ethyl acetate for elution.
The filtrate was transferred to a separation funnel and the organic
layer was isolated. The aqueous phase was extracted with ethyl
acetate (2.times.100 mL), the combined organics were washed with
brine (100 mL), dried over MgSO.sub.4, filtered, and concentrated
under reduced pressure to afford an intermediate. The intermediate
was used immediately in the subsequent step without further
purification. Lithium chloride (2.20 g, 52.0 mmol) was placed in a
round bottom flask, dried under vacuum with heating and allowed to
cool to room temperature under vacuum. Acetonitrile (87 mL) was
added followed by ethyl 2-(diethoxyphosphoryl)-2-fluoroacetate
(5.79 g, 23.9 mmol). The solution was cooled to 0.degree. C. using
an ice/water bath and N,N-diisopropylethylamine (4.03 g, 5.5 ml,
31.2 mmol) was added. After 10 min stirring at this temperature a
solution of the intermediate mentioned above in acetonitrile (33
ml) was added. Upon complete addition the cooling was removed and
the reaction was stirred overnight at room temperature. The
reaction mixture was concentrated to approximately 50 mL (under
vacuum), ethyl acetate (250 mL), water (50 mL) and saturated
aqueous NH.sub.4Cl (50 mL) were added. The phases were separated
and the aqueous layer was extracted with ethyl acetate (2.times.100
mL). The combined organics were dried over MgSO.sub.4, filtered,
and concentrated under reduced pressure. The crude material was
purified using a CombiFlash system (220 g SiO.sub.2, gradient
elution; heptanes:ethyl acetate 100:0.fwdarw.60:40) to afford
(R)-ethyl
5-((R)-1,1-dimethylethylsulfinamido)-2,4,4-trifluoro-5-(2-fluorophenyl)he-
x-2-enoate (5.1 g, 60% yield). .sup.1H NMR (600 MHz, CDCl.sub.3)
.delta. 7.45 (tt, J=4.3, 2.2 Hz, 1H), 7.40-7.33 (m, 1H), 7.16 (tt,
J=5.4, 2.7 Hz, 1H), 7.09 (ddd, J=13.2, 8.2, 1.2 Hz, 1H), 6.00 (dt,
J=20.3, 14.5 Hz, 1H), 4.98 (d, J=4.1 Hz, 1H), 4.30 (q, J=7.1 Hz,
2H), 2.03 (s, 3H), 1.32 (t, J=7.2 Hz, 3H), 1.24 (s, 9H).
Intermediate:
(6R)-3,5,5-trifluoro-6-(2-fluorophenyl)-6-methylpiperidin-2-one
##STR00011##
[0108] (R)-Ethyl
5-((R)-1,1-dimethylethylsulfinamido)-2,4,4-trifluoro-5-(2-fluorophenyl)he-
x-2-enoate (5.1 g, 12.5 mmol) was dissolved in ethyl acetate (200
mmol) and placed in a Parr-flask. Palladium on carbon (2.65 g, 2.49
mmol, 10%) was added and the Parr-flask was placed in a Parr-shaker
(H.sub.z-pressure=2.8 bar initially). After 16 h in the Parr shaker
at room temperature the reaction mixture was filtered through a
plug of celite using ethyl acetate for elution. The filtrate was
concentrated under reduced pressure. This material was dissolved in
ethyl acetate (200 ml, 3494 mmol) and the reaction mixture was
split equally into two Parr-flasks. Palladium on carbon (2.65 g,
2.49 mmol, 10%) was split in two equal portions and added to the
two Parr-flasks. The flasks were placed in two different
Parr-shakers (Hz-pressure=2.8 bar initially) and run in parallel.
After 16 h in the Parr-shaker at room temperature the two
suspensions were combined and filtered through a plug of celite
using ethyl acetate for elution. The filtrate was concentrated
under reduced pressure. The material thus obtained was dissolved in
methanol (330 ml). HCl (4.7 ml, 19 mmol, 4 M in 1,4-dioxane) was
added and the reaction was stirred at room temperature for 1 h 30
min. K.sub.2CO.sub.3 (5.16 g, 37.4 mmol) was added and the reaction
was stirred at room temperature for another 1 h 30 min. The
reaction was concentrated to dryness under reduced pressure and the
residue was partitioned between water (200 mL) and ethyl acetate
(250 mL). The phases were separated and the aqueous layer was
extracted with ethyl acetate (2.times.100 mL). The combined
organics were washed with brine, dried over MgSO.sub.4, filtered,
and concentrated under reduced pressure. The crude material was
purified using a CombiFlash system (120 g SiO.sub.2, gradient
elution; heptanes:ethyl acetate 100:0.fwdarw.45:55) to afford
(6R)-3,5,5-trifluoro-6-(2-fluorophenyl)-6-methylpiperidin-2-one
(1.86 g, 57.2% yield) as a semi-solid/foam (1:1 mixture of
diastereomers) LC-MS (m/z) 262.2 (MH.sup.+) t.sub.R=0.57 minutes
(Method B).
Intermediate: (2R)-tert-butyl
3,3,5-trifluoro-2-(2-fluorophenyl)-2-methyl-6-oxopiperidine-1-carboxylate
##STR00012##
[0110]
(6R)-3,5,5-Trifluoro-6-(2-fluorophenyl)-6-methylpiperidin-2-one
(2.42 g, 9.26 mmol) (1:1 mixture of diastereomers) was placed in a
round bottom flask. DMAP (0.283 g, 2.316 mmol) was added followed
by a solution of di-tert-butyl dicarbonate (6.07 g, 27.8 mmol) in
tetrahydrofuran (170 ml). The solution was stirred at room
temperature for 1 h. The reaction was diluted with ethyl acetate
(200 mL) and washed with a solution of water (50 mL) and saturated
aqueous NH.sub.4Cl (50 mL). The phases were separated and the
aqueous layer was extracted with ethyl acetate (2.times.100 mL).
The combined organics were washed with brine (50 mL), dried over
MgSO.sub.4, filtered, and concentrated under reduced pressure. The
crude material was purified using a CombiFlash system (120 g
SiO.sub.2, gradient elution; heptanes:ethyl acetate
100:0.fwdarw.55:45) to afford (2R)-tert-butyl
3,3,5-trifluoro-2-(2-fluorophenyl)-2-methyl-6-oxopiperidine-1-carboxylate
(2.60 g, 78% yield) (1:1 mixture of diastereomers).
Intermediate:
(6R)-3,5,5-trifluoro-6-(2-fluorophenyl)-3,6-dimethylpiperidin-2-one
##STR00013##
[0112] For the first step, two reactions were run in parallel under
identical conditions with total amounts as described below.
[0113] (2R)-Tert-butyl
3,3,5-trifluoro-2-(2-fluorophenyl)-2-methyl-6-oxopiperidine-1-carboxylate
(1.30 g, 3.60 mmol) (1:1 mixture of diastereomers) was dissolved in
tetrahydrofuran (36 mL) (dried over 4 .ANG. MS) and added to a dry
round bottom flask. The solution was cooled to -78.degree. C. using
a dry ice/acetone bath. LiHMDS (lithium hexamethyldisilazide) (4.50
mL, 4.50 mmol, 1.0 M in tetrahydrofuran) was added in a dropwise
manner and the resulting solution was stirred at -78.degree. C. for
1 h. Methyl iodide (2.55 g, 1.13 mL, 18 mmol) was added in a
dropwise manner and the solution was stirred at -78.degree. C. for
45 min then the cooling was removed and the solution was stirred
for another 15 min at room temperature. The solution was re-cooled
to -78.degree. C. and quenched with saturated aqueous NH.sub.4Cl
(25 mL). The cooling bath was removed and the reaction was allowed
to warm to room temperature. The two reaction mixtures were
combined and ethyl acetate (200 mL) and water (50 mL) were added.
The phases were separated and the aqueous layer was extracted with
ethyl acetate (2.times.100 mL). The combined organics were washed
with brine, dried over MgSO.sub.4, filtered, and concentrated under
reduced pressure. All of the thus obtained material was dissolved
in 1,2-dichloroethane (85 ml) and the solution was cooled to
0.degree. C. using an ice/water bath. TFA (21 ml, 273 mmol) was
added and the cooling bath was allowed to slowly expire overnight,
with stirring of the reaction mixture. The reaction was diluted
with toluene (50 mL) and concentrated to approximately 25 mL under
vacuum. The residue was diluted with ethyl acetate (150 mL) and
washed with saturated aqueous NaHCO.sub.3 (50 mL). The phases were
separated and the aqueous layer was extracted with ethyl acetate
(2.times.75 mL). The combined organics were washed with brine,
dried over MgSO.sub.4, filtered, and concentrated under reduced
pressure. The crude material was purified using a CombiFlash system
(80 g SiO.sub.2, gradient elution; heptanes:ethyl acetate
100:0.fwdarw.50:50) to afford
(6R)-3,5,5-trifluoro-6-(2-fluorophenyl)-3,6-dimethylpiperidin-2-one
(1.86 g, 6.76 mmol, 94% yield) (1:1.8 mixture of diastereomers)
LC-MS (m/z) 276.2 (MH.sup.+) t.sub.R=0.62 minutes (Method B).
Intermediate:
(6R)-3,5,5-trifluoro-6-(2-fluoro-5-nitrophenyl)-3,6-dimethylpiperidin-2-o-
ne
##STR00014##
[0115]
(6R)-3,5,5-Trifluoro-6-(2-fluorophenyl)-3,6-dimethylpiperidin-2-one
(1.86 g, 6.76 mmol) (1:1.8 mixture of diastereomers) was suspended
in trifluoroacteic acid (11.5 ml, 149 mmol). The mixture was cooled
to 0.degree. C. and concentrated H.sub.2SO.sub.4 (2.86 ml, 52.0
mmol, 97%) was added. Finally, fuming HNO.sub.3 (0.33 ml, 7.4 mmol)
was added in a dropwise manner and the reaction was stirred at
0.degree. C. for 10 min. The reaction mixture was poured onto 150 g
ice and basified to pH >11 using 5 M NaOH. The resulting
suspension was extracted with ethyl acetate (250 mL). The phases
were separated and the aqueous layer was extracted with ethyl
acetate (2.times.100 mL). The combined organics were washed with a
solution of saturated aqueous NH.sub.4Cl (50 mL) and water (50 mL),
dried over MgSO.sub.4, filtered, and concentrated under reduced
pressure to afford
(6R)-3,5,5-trifluoro-6-(2-fluoro-5-nitrophenyl)-3,6-dimethylpiperidin-2-o-
ne (2.08 g, 6.50 mmol, 96% yield) (1:1.8 mixture of diastereomers)
LC-MS (m/z) 321.1 (MH.sup.+) t.sub.R=0.62 minutes (Method B).
Intermediate:
(6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidin-2-o-
ne
##STR00015##
[0117]
(6R)-3,5,5-Trifluoro-6-(2-fluoro-5-nitrophenyl)-3,6-dimethylpiperid-
in-2-one (2.08 g, 6.50 mmol) (1:1.8 mixture of diastereomers) was
dissolved in methanol (28 ml). Ammonium formate (2.05 g, 32.5 mmol)
was added followed by portionwise addition of palladium on carbon
(1.38 g, 1.30 mmol, 10%). The reaction is slightly exothermic and
the reaction mixture was briefly immersed into an ice/water bath to
control the temperature increase. After the initial temperature
increase had settled, the reaction was stirred at room temperature
for another 10 min. The reaction was filtered through a plug of
celite using methanol for elution. The filtrate was concentrated
under reduced pressure and partitioned between ethyl acetate (100
mL) and saturated aqueous NaHCO.sub.3 (50 mL). The phases were
separated and the aqueous layer was extracted with ethyl acetate
(2.times.50 mL). The combined organics were dried over MgSO.sub.4,
filtered and concentrated under reduced pressure to afford
(6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpip-
eridin-2-one (1.73 g, 5.96 mmol, 92% yield) (1:1.8 mixture of
diastereomers). LC-MS Major: (m/z) 291.0 (MH.sup.+-tert-butyl)
t.sub.R=0.40 minutes (Method B); Minor: (m/z) 291.0 (MH.sup.+)
t.sub.R=0.41 minutes (Method B). The crude material was used in the
next reaction step without further purification.
Intermediate: tert-butyl
(4-fluoro-3-((2R)-3,3,5-trifluoro-2,5-dimethyl-6-oxopiperidin-2-yl)-pheny-
l)carbamate
##STR00016##
[0119]
(6R)-6-(5-Amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperid-
in-2-one (1.73 g, 5.96 mmol) (1:1.8 mixture of diastereomers) was
placed in a round bottom flask and a solution of di-tert-butyl
dicarbonate (1.56 g, 7.15 mmol) in tetrahydrofuran (25 ml) (dried
over 4 .ANG. MS) was added. The solution was heated to 50.degree.
C. and stirred at this temperature overnight. The reaction was
concentrated under reduced pressure and the crude material was
purified using a CombiFlash system (80 g SiO.sub.2, gradient
elution; heptanes: ethyl acetate 100:0.fwdarw.50:50) to afford
tert-butyl
(4-fluoro-3-((2R)-3,3,5-tri-fluoro-2,5-dimethyl-6-oxopiperidin-2-yl)pheny-
l)carbamate (2.1 g, 5.38 mmol, 90% yield) (1:1.7 mixture of
diastereomers) LC-MS Major: (m/z) 335.0 (MH.sup.+-tert-butyl)
t.sub.R=0.74 minutes (Method B); Minor: (m/z) 391.2 (MH.sup.+)
t.sub.R=0.76 minutes (Method B).
Intermediate: tert-butyl
(4-fluoro-3-((2R,5S)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
-phenyl)carbamate and tert-butyl
(4-fluoro-3-((2R,5R)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
phenyl)carbamate
##STR00017##
[0121] Tert-butyl
(4-fluoro-3-((2R)-3,3,5-trifluoro-2,5-dimethyl-6-oxopiperidin-2-yl)phenyl-
)carbamate (2.1 g, 5.38 mmol) (1:1.7 mixture of diastereomers) was
placed in a round bottom flask and dissolved in toluene (60 ml)
(dried over 4 .ANG. MS). Argon was bubbled through the reaction for
10 min followed by addition of Lawesson's reagent
(2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane
2,4-disulfide) (2.18 g, 5.38 mmol). The reaction was carefully
evacuated and backfilled with argon (.times.3). The suspension was
heated to 80.degree. C. The reaction was stirred at this
temperature for 3 h 30 min. The reaction was allowed to cool to
room temperature and concentrated under reduced pressure. The crude
material was suspended in CHCl.sub.3 and filtered. The filtrate was
concentrated under reduced pressure and the crude material was
purified using a CombiFlash system (120 g SiO.sub.2, gradient
elution; heptanes:ethyl acetate 100:0.fwdarw.80:20) to afford
tert-butyl
(4-fluoro-3-((2R,5S)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
phenyl)carbamate (1.15 g, 52.6% yield) (fast eluting isomer) LC-MS
(m/z) 407.4 (MH.sup.+) t.sub.R=0.83 minutes (Method B) and
tert-butyl
(4-fluoro-3-((2R,5R)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
phenyl)carbamate (0.816 g (60% purity), 22.4% yield) (slow eluting
isomer) LC-MS (m/z) 407.4 (MH.sup.+) t.sub.R=0.82 minutes (Method
B).
Intermediate:
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione
##STR00018##
[0123] tert-butyl
(4-fluoro-3-((2R,5S)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
phenyl)carbamate (1.15 g, 2.83 mmol) was dissolved in
dichloromethane (13 ml). The solution was cooled to 0.degree. C.
and TFA (6.5 ml, 84 mmol) was added. The solution was stirred at
0.degree. C. for 1 h 20 min. The reaction was diluted with toluene
(25 mL) and concentrated to approx 10 mL under reduced pressure.
The residue was diluted with ethyl acetate (50 mL) and washed with
saturated aqueous NaHCO.sub.3 (25 mL). The phases were separated
and the aqueous layer was extracted with ethyl acetate (2.times.25
mL). The combined organics were dried over MgSO.sub.4, filtered,
and concentrated under reduced pressure to afford
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione (548 mg (70% purity), 63.2% yield). The crude product was
used in the next reaction step without further purification.
[0124] LC-MS (m/z) 307.2 (MH.sup.+) t.sub.R=0.49 minutes (Method B)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 7.93 (bs, 1H), 6.93-6.88
(m, 1H), 6.66-6.61 (m, 1H), 6.59-6.55 (m, 1H), 2.62-2.57 (m, 2H),
1.90 (s, 3H), 1.86 (d, J=22.4 Hz, 3H)
[.alpha.].sup.20,.sub.D=211.degree. (589 nm, c=0.1 g/100 mL,
MeOH
Intermediate:
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione
##STR00019##
[0126] tert-butyl
(4-fluoro-3-((2R,5R)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopiperidin-2-yl)-
phenyl)carbamate (816 mg, 2.01 mmol) was dissolved in
dichloromethane (9.2 mL). The solution was cooled to 0.degree. C.
and TFA (4.6 mL, 59.5 mmol) was added. The solution was stirred at
0.degree. C. for 1 h 20 min. The reaction was diluted with toluene
(15 mL) and concentrated to approx. 10 mL under reduced pressure.
The residue was diluted with ethyl acetate (50 mL) and washed with
saturated aqueous NaHCO.sub.3 (25 mL). The phases were separated
and the aqueous layer was extracted with ethyl acetate (2.times.25
mL). The combined organics were dried over MgSO.sub.4, filtered,
and concentrated under reduced pressure to afford
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione (587 mg, (50% purity), 47.7% yield). The material was
used in the next reaction step without further purification.
[0127] LC-MS (m/z) 307.0 (MH.sup.+) t.sub.R=0.47 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 8.18 (bs, 1H), 6.92 (dd,
J=12.0, 8.7 Hz, 1H), 6.69-6.63 (m, 1H), 6.52-6.45 (m, 1H),
2.78-2.66 (m, 1H), 2.56-2.43 (m, 1H), 1.91 (s, 3H), 1.79 (dd,
J=20.3, 12.5 Hz, 3H).
Intermediate: 5-(Methoxy-d.sub.3) picolinic acid
##STR00020##
[0129] Methyl 5-hydroxypicolinate (2.88 g, 18.81 mmol) was
dissolved in DMF (108 mL). Potassium carbonate (7.20 g, 52.1 mmol)
was added and the suspension was stirred for 45 minutes at room
temperature. Methyl-d.sub.3-iodide (3.27 g, 1.40 ml, 22.6 mmol) was
added. The reaction mixture was stirred at room temperature for 2
hours.
[0130] Water and ethyl acetate were added. The mixture was
extracted three times with ethyl acetate. The combined organic
phases were washed with brine, dried over MgSO.sub.4, filtered and
concentrated in vacuo.
[0131] The product was chromatographed on silicagel to obtain
methyl 5-(methoxy-d.sub.3) picolinate (2.17 g, 68% yield).
[0132] Methyl 5-(methoxy-d.sub.3) picolinate (0.58 g, 3.41 mmol)
was dissolved in water (4 ml) and 1,4-dioxane (12 mL). LiOH (0.20
g, 8.5 mmol) was added and the reaction mixture was stirred at room
temperature for 2 hours. The reaction mixture was acidified to pH 2
with 6M HCl (aq). The reaction mixture was concentrated in vacuo
followed by azetropic removal of residual water with two portions
of toluene to give 5-(methoxy-d.sub.3) picolinic acid. Used in next
step without further purification. LC-MS (m/z) 157.1 (MH.sup.+)
t.sub.R=0.19 minutes (Method A)
PREPARATION OF THE COMPOUNDS OF THE INVENTION
Example 1
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-fluoropicolinamide
##STR00021##
[0134] 5-fluoropicolinic acid (269 mg, 1.906 mmol) and
1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo-[4,5-b]pyridinium
3-oxid hexafluorophosphate (HATU) (797 mg, 2.10 mmol) were placed
in a round bottom flask, dissolved in DMF (5.2 mL), and stirred at
room temperature for 5 min.
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione (292 mg, 0.953 mmol) was added followed by
N,N-diisopropylethylamine (830 .mu.l, 4.77 mmol) and the reaction
was stirred at room temperature for 5 min. The reaction was diluted
with ethyl acetate (50 mL) and washed with a mixture of water (25
mL) and saturated aqueous NH.sub.4Cl (25 mL). The phases were
separated and the aqueous layer was extracted with ethyl acetate
(2.times.50 mL). The combined organics were dried over MgSO.sub.4,
filtered, and concentrated under reduced pressure. The intermediate
5-fluoro-N-(4-fluoro-3-((2R,5S)-3,3,5-trifluoro-2,5-dimethyl-6-thioxopipe-
ridin-2-yl)phenyl)picolinamide was purified using a CombiFlash
system (40 g SiO.sub.2, gradient elution; heptanes:ethyl acetate
100:0.fwdarw.60:40). The intermediate (225 mg, 0.523 mmol) was
split in two equal portions and placed in two separate reaction
vials. Ammonia (14.6 mL, 102 mmol, 7 M in methanol) was also split
in two equal portions and added to the two vials. The vials were
capped and heated to 65.degree. C. using an oil bath. After 6 h
stirring at this temperature the reactions were allowed to cool to
room temperature, the mixtures were combined and concentrated under
reduced pressure. The crude material was subjected to silica-gel
chromatography (eluent; heptane:ethyl acetate=50:50.fwdarw.0:100)
to afford
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-fluoropicolinamide (124 mg, 57%
yield).
[0135] LC-MS (m/z) 413.2 (MH+); t.sub.R=0.54 (Method A) .sup.1H NMR
(600 MHz, CDCl.sub.3) .delta. 9.78 (br s, 1H), 8.45 (d, J=2.7 Hz,
1H), 8.33 (dd, J=8.7, 4.6 Hz, 1H), 7.89 (ddd, J=8.8, 3.8, 2.9 Hz,
1H), 7.59 (ddd, J=8.6, 8.0, 2.8 Hz, 1H), 7.57 (dd, J=6.8, 2.7 Hz,
1H), 7.09 (dd, J=11.8, 8.8 Hz, 1H), 4.74 (br s, 2H), 2.60-2.37 (m,
2H), 1.79 (t, J=2.8 Hz, 3H), 1.76 (d, J=23.6 Hz, 3H).
[0136] The following compounds were prepared in a way similar to
example 1:
Example 2
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-fluoropicolinamide
##STR00022##
[0138] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-fluoropicolinic acid
[0139] LC-MS (m/z) 413.1 (MH.sup.+) t.sub.R=0.55 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 9.78 (br s, 1H), 8.45 (d,
J=2.8 Hz, 1H), 8.34-8.31 (m, 1H), 7.84 (ddd, J=8.8, 3.8, 2.9 Hz,
1H), 7.61-7.57 (m, 2H), 7.08 (dd, J=11.6, 8.8 Hz, 1H), 4.69 (br s,
2H), 2.75-2.62 (m, 1H), 2.47-2.40 (m, 1H), 1.82 (s, 3H), 1.75 (d,
J=23.9 Hz, 3H)
Example 3
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-chloropicolinamide
##STR00023##
[0141] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-chloropicolinic acid
[0142] LC-MS (m/z) 429.2 (MH.sup.+) t.sub.R=0.57 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 9.81 (br s, 1H), 8.56
(dd, J=2.4, 0.7 Hz, 1H), 8.24 (dd, J=8.4, 0.7 Hz, 1H), 7.91-7.85
(m, 2H), 7.59 (dd, J=6.9, 2.7 Hz, 1H), 7.09 (dd, J=11.8, 8.8 Hz,
1H), 2.61-2.35 (m, 2H), 1.80 (t, J=2.8 Hz, 3H), 1.76 (d, J=23.6 Hz,
3H)
Example 4
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-cyanopicolinamide
##STR00024##
[0144] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-cyanopicolinic acid
[0145] LC-MS (m/z) 420.0 (MH.sup.+) t.sub.R=1.79 minutes (Method B)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.82 (br s, 1H), 9.20
(dd, J=2.0, 0.8 Hz, 1H), 8.58 (dd, J=8.1, 2.0 Hz, 1H), 8.28 (dd,
J=8.2, 0.7 Hz, 1H), 7.94 (dd, J=7.2, 2.7 Hz, 1H), 7.89-7.83 (m,
1H), 7.15 (dd, J=11.9, 8.8 Hz, 1H), 6.22 (br s, 2H), 2.74-2.59 (m,
1H), 2.49-2.38 (m, 1H), 1.67 (d, J=22.7 Hz, 3H), 1.62 (s, 3H)
Example 5
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide
##STR00025##
[0147] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxypyrazine-2-carboxylic acid
[0148] LC-MS (m/z) 426.3 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 9.50 (br s, 1H), 9.00 (d,
J=1.3 Hz, 1H), 8.13 (d, J=1.3 Hz, 1H), 7.88 (ddd, J=8.8, 3.8, 2.9
Hz, 1H), 7.58 (dd, J=6.8, 2.7 Hz, 1H), 7.08 (dd, J=11.8, 8.8 Hz,
1H), 4.80 (br s, 2H), 4.06 (s, 3H), 2.59-2.35 (m, 2H), 1.79 (t,
J=2.8 Hz, 3H), 1.75 (d, J=23.6 Hz, 3H)
Example 6
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide
##STR00026##
[0150] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 1-(difluoromethyl)-1H-pyrazole-3-carb oxylic acid
[0151] LC-MS (m/z) 434.2 (MH.sup.+) t.sub.R=0.49 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 8.66 (br s, 1H), 7.86 (d,
J=2.7 Hz, 1H), 7.78 (ddd, J=8.8, 3.7, 2.9 Hz, 1H), 7.58 (dd, J=6.8,
2.7 Hz, 1H), 7.18 (t, J=60.3 Hz, 1H), 7.07 (dd, J=11.7, 8.8 Hz,
1H), 7.03 (d, J=2.7 Hz, 1H), 4.57 (br s, 2H), 2.59-2.34 (m, 2H),
1.80 (t, J=2.8 Hz, 3H), 1.74 (d, J=23.6 Hz, 3H)
Example 7
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-2-methyloxazole-4-carboxamide
##STR00027##
[0153] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2-methyloxazole-4-carboxylic acid
[0154] LC-MS (m/z) 399 (MH.sup.+) t.sub.R=0.46 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.12 (br s, 1H), 8.62
(s, 1H), 7.82 (dd, J=7.1, 2.5 Hz, 1H), 7.75 (dt, J=8.5, 3.2 Hz,
1H), 7.09 (dd, J=11.9, 8.8 Hz, 1H), 6.19 (br s, 2H), 2.71-2.57 (m,
1H), 2.51 (s, 3H), 2.47-2.35 (m, 1H), 1.66 (d, J=22.8 Hz, 3H), 1.61
(s, 3H)
Example 8
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)thiazole-2-carboxamide
##STR00028##
[0156] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and thiazole-2-carboxylic acid
[0157] LC-MS (m/z) 401 (MH.sup.+) t.sub.R=0.49 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.83 (br s, 1H), 8.13
(d, J=3.1 Hz, 1H), 8.10 (d, J=3.1 Hz, 1H), 7.97 (dd, J=7.2, 2.7 Hz,
1H), 7.78 (dt, J=8.7, 3.4 Hz, 1H), 7.14 (dd, J=11.9, 8.8 Hz, 1H),
6.22 (br s, 2H), 2.73-2.59 (m, 1H), 2.49-2.37 (m, 1H), 1.68 (d,
J=22.7 Hz, 3H), 1.63 (s, 3H)
Example 9
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide
##STR00029##
[0159] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-bromo-1-methyl-1H-imidazole-2-carb oxylic acid
[0160] LC-MS (m/z) 475.9 (MH.sup.+) t.sub.R=0.55 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.42 (br s, 1H), 7.83
(dd, J=7.2, 2.7 Hz, 1H), 7.76-7.72 (m, 1H), 7.63 (s, 1H), 7.09 (dd,
J=11.9, 8.8 Hz, 1H), 6.20 (br s, 2H), 3.96 (s, 3H), 2.72-2.58 (m,
1H), 2.49-2.36 (m, 1H), 1.67 (d, J=22.7 Hz, 3H), 1.61 (s, 3H)
Example 10
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-4-methylthiazole-2-carboxamide
##STR00030##
[0162] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-methylthiazole-2-carboxylic acid
[0163] LC-MS (m/z) 415 (MH.sup.+) t.sub.R=0.53 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.74 (br s, 1H), 7.95
(dd, J=7.2, 2.7 Hz, 1H), 7.78 (dt, J=8.7, 3.4 Hz, 1H), 7.68 (d,
J=0.8 Hz, 1H), 7.12 (dd, J=11.9, 8.8 Hz, 1H), 6.22 (br s, 2H),
2.72-2.59 (m, 1H), 2.50 (s, 3H), 2.49-2.36 (m, 1H), 1.67 (d, J=22.8
Hz, 3H), 1.62 (s, 3H)
Example 11
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-5-(trifluoromethyl)picolinamide
##STR00031##
[0165] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(trifluoromethyl)picolinic acid
[0166] LC-MS (m/z) 463 (MH.sup.+) t.sub.R=0.61 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.81 (br s, 1H),
9.13-9.11 (m, 1H), 8.49 (dd, J=8.3, 2.1 Hz, 1H), 8.34 (d, J=8.2 Hz,
1H), 7.95 (dd, J=7.1, 2.7 Hz, 1H), 7.90 (dt, J=8.7, 3.4 Hz, 1H),
7.16 (dd, J=11.9, 8.8 Hz, 1H), 6.25 (br s, 2H), 2.74-2.61 (m, 1H),
2.51-2.38 (m, 1H), 1.69 (d, J=22.7 Hz, 3H), 1.64 (s, 3H)
Example 12
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-5-methoxypyrimidine-2-carboxamide
##STR00032##
[0168] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxypyrimidine-2-carboxylic acid (Prepared as
described in Scott, Jack D. et al. PCT Int. Appl. 2011044181)
[0169] LC-MS (m/z) 426 (MH.sup.+) t.sub.R=0.45 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.60 (br s, 1H), 8.72
(s, 2H), 7.90-7.86 (m, 1H), 7.83 (dd, J=7.1, 2.7 Hz, 1H), 7.14 (dd,
J=11.9, 8.8 Hz, 1H), 6.23 (br s, 2H), 4.02 (s, 3H), 2.74-2.59 (m,
1H), 2.49-2.37 (m, 1H), 1.67 (d, J=22.7 Hz, 3H), 1.63 (s, 3H)
Example 13
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-2-(difluoromethyl)oxazole-4-carboxamide
##STR00033##
[0171] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2-(difluoromethyl)oxazole-4-carboxylic acid
[0172] LC-MS (m/z) 435 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.42 (br s, 1H), 9.01
(s, 1H), 7.81 (dd, J=7.1, 2.7 Hz, 1H), 7.76 (dt, J=8.5, 3.3 Hz,
1H), 7.33 (t, J=51.9 Hz, 1H), 7.12 (dd, J=11.9, 8.8 Hz, 1H), 6.20
(br s, 2H), 2.74-2.58 (m, 1H), 2.48-2.36 (m, 1H), 1.67 (d, J=22.7
Hz, 3H), 1.62 (s, 3H)
Example 14
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-4-(fluoromethyl)oxazole-2-carboxamide
##STR00034##
[0174] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-(fluoromethyl)oxazole-2-carboxylic acid
[0175] LC-MS (m/z) 417 (MH.sup.+) t.sub.R=0.46 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.04 (br s, 1H), 8.56
(d, J=5.0 Hz, 1H), 7.88 (dd, J=7.2, 2.7 Hz, 1H), 7.80-7.75 (m, 1H),
7.14 (dd, J=11.9, 8.8 Hz, 1H), 6.21 (br s, 2H), 5.43 (d, J=48.0 Hz,
2H), 2.74-2.58 (m, 1H), 2.48-2.35 (m, 1H), 1.67 (d, J=22.8 Hz, 3H),
1.62 (s, 3H)
Example 15
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-4-(fluoromethyl)thiazole-2-carboxamide
##STR00035##
[0177] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-(fluoromethyl)thiazole-2-carboxylic acid
[0178] LC-MS (m/z) 433 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.90 (br s, 1H), 8.25
(d, J=3.2 Hz, 1H), 7.94 (dt, J=11.2, 5.6 Hz, 1H), 7.81-7.73 (m,
1H), 7.13 (dd, J=11.9, 8.8 Hz, 1H), 6.21 (br s, 2H), 5.57 (d,
J=47.7 Hz, 2H), 2.72-2.57 (m, 1H), 2.48-2.36 (m, 1H), 1.67 (d,
J=22.8 Hz, 3H), 1.62 (s, 3H)
Example 16
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluoro-phenyl)-5-(difluoromethyl)pyrazine-2-carboxamide
##STR00036##
[0180] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(difluoromethyl)pyrazine-2-carboxylic acid
[0181] LC-MS (m/z) 446 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 9.63 (br s, 1H), 9.53 (s,
1H), 8.92 (s, 1H), 7.90-7.86 (m, 1H), 7.63 (dd, J=6.8, 2.7 Hz, 1H),
7.11 (dd, J=11.6, 8.8 Hz, 1H), 6.80 (t, J=54.5 Hz, 1H), 4.76 (br s,
2H), 2.64-2.34 (m, 2H), 1.80 (t, J=2.7 Hz, 3H), 1.77 (d, J=23.5 Hz,
3H)
Example 17
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methoxypicolinamide
##STR00037##
[0183] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxypicolinic acid
[0184] LC-MS (m/z) 425 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.42 (br s, 1H), 8.39
(dd, J=2.9, 0.4 Hz, 1H), 8.13-8.11 (m, 1H), 7.91-7.85 (m, 2H), 7.61
(dd, J=8.8, 2.9 Hz, 1H), 7.16-7.09 (m, 1H), 6.27 (br s, 2H), 3.93
(s, J=2.9 Hz, 3H), 2.73-2.59 (m, 1H), 2.49-2.38 (m, 1H), 1.68 (d,
J=22.7 Hz, 3H), 1.63 (s, 3H)
Example 18
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide
##STR00038##
[0186] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-cyano-3-methylpicolinic acid (Prepared as described
in Badiger, Sangamesh et al. PCT Int. Appl., 2012095469)
[0187] LC-MS (m/z) 434 (MH.sup.+) t.sub.R=0.53 minutes (Method
A).
Example 19
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methoxy-3-methylpyrazine-2-carb
oxamide
##STR00039##
[0189] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxy-3-methylpyrazine-2-carboxylic acid
(Prepared as described in Yoshizawa, Kazuhiro et al. PCT Int.
Appl., 2013162065)
[0190] LC-MS (m/z) 440.1 (MH.sup.+) t.sub.R=0.58 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.42 (s, 1H), 8.23 (s,
J=0.5 Hz, 1H), 7.88-7.83 (m, 1H), 7.76 (dd, J=7.1, 2.7 Hz, 1H),
7.12 (dd, J=11.9, 8.8 Hz, 1H), 6.23 (s, 2H), 3.99 (s, 3H), 2.75 (s,
3H), 2.73-2.58 (m, 1H), 2.49-2.38 (m, 1H), 1.67 (d, J=22.7 Hz, 3H),
1.63 (s, 3H)
Example 20
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methyl-1,3,4-oxadiazole-2-carboxamide
##STR00040##
[0192] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methyl-1,3,4-oxadiazole-2-carboxylic acid
[0193] LC-MS (m/z) 400.1 (MH.sup.+) t.sub.R=0.42 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.51 (d, J=37.6 Hz,
1H), 11.40 (s, 1H), 10.01 (s, 1H), 9.89 (s, 1H), 8.06-8.01 (m, 1H),
7.97 (dd, J=7.3, 2.5 Hz, 1H), 7.35 (dd, J=12.3, 9.0 Hz, 1H),
3.18-3.04 (m, 1H), 2.93-2.80 (m, 1H), 2.64 (s, 3H), 1.94-1.89 (m,
6H)
Example 21
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-3-methyl-1,2,4-oxadiazole-5-carboxamide
##STR00041##
[0195] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 3-methyl-1,2,4-oxadiazole-5-carboxylic acid
[0196] LC-MS (m/z) 400 (MH.sup.+) t.sub.R=0.46 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.54 (s, 1H), 11.40
(s, 1H), 10.01 (s, 1H), 9.89 (s, 1H), 8.06-8.00 (m, 1H), 7.97 (dd,
J=7.3, 2.5 Hz, 1H), 7.34 (dd, J=12.3, 9.0 Hz, 1H), 3.19-3.04 (m,
1H), 2.92-2.79 (m, 1H), 2.64 (s, 3H), 1.92 (m, 6H)
Example 22
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-1-methyl-1H-1,2,4-triazole-3-carboxamide
##STR00042##
[0198] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 1-methyl-1H-1,2,4-triazole-3-carboxylic acid
[0199] LC-MS (m/z) 399 (MH.sup.+) t.sub.R=0.39 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.40 (s, 1H), 8.68 (s,
1H), 7.84 (dd, J=7.2, 2.7 Hz, 1H), 7.82-7.76 (m, 1H), 7.12 (dd,
J=11.9, 8.8 Hz, 1H), 6.50-6.03 (s, 2H), 3.98 (s, 3H), 2.75-2.60 (m,
1H), 2.49-2.35 (m, 1H), 1.68 (d, J=22.8 Hz, 3H), 1.62 (s, 3H)
Example 23
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-1-(difluoromethyl)-1H-pyrazole-3-carboxamide
##STR00043##
[0201] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 1-(difluoromethyl)-1H-pyrazole-3-carb oxylic acid
[0202] LC-MS (m/z) 434 (MH.sup.+) t.sub.R=0.5 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.49 (s, 1H), 8.41 (t,
J=3.4 Hz, 1H), 7.92 (t, J=58.7 Hz, 1H), 7.80-7.64 (m, 2H), 7.13
(dd, J=11.9, 8.7 Hz, 1H), 7.01 (d, J=2.7 Hz, 1H), 6.32 (s, 2H),
2.63-2.45 (d, J=23.4 Hz, 1H), 2.28-2.11 (m, 1H), 1.69 (d, J=23.3
Hz, 3H), 1.67 (s, 3H)
Example 24
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-(difluoromethyl)pyrazine-2-carboxamide
##STR00044##
[0204] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(difluoromethyl)pyrazine-2-carboxylic acid
[0205] LC-MS (m/z) 446 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.03 (s, 1H), 9.37 (t,
J=2.4 Hz, 1H), 9.09 (s, 1H), 7.89-7.80 (m, 2H), 7.26 (t, J=53.9 Hz,
1H), 7.18 (dd, J=11.9, 8.7 Hz, 1H), 6.34 (s, 2H), 2.62-2.51 (m,
1H), 2.29-2.14 (m, 1H), 1.71 (d, J=23.2 Hz, 3H), 1.68 (s, 3H)
Example 25
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methoxypyrazine-2-carboxamide
##STR00045##
[0207] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxypyrazine-2-carboxylic acid
[0208] LC-MS (m/z) 426 (MH.sup.+) t.sub.R=0.5 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.63 (s, 1H), 8.88 (d,
J=1.3 Hz, 1H), 8.41 (d, J=1.3 Hz, 1H), 7.82 (dd, J=7.2, 2.6 Hz,
1H), 7.78 (dt, J=8.6, 3.4 Hz, 1H), 7.14 (dd, J=11.9, 8.8 Hz, 1H),
6.31 (s, 2H), 4.02 (s, 3H), 2.60-2.49 (m, 1H), 2.28-2.13 (m, 1H),
1.70 (d, J=23.0 Hz, 3H), 1.67 (s, 3H)
Example 26
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methoxypyrimidine-2-carboxamide
##STR00046##
[0210] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methoxypyrimidine-2-carboxylic acid
[0211] LC-MS (m/z) 426 (MH.sup.+) t.sub.R=0.45 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.65 (s, 1H), 10.99
(s, 1H), 10.20 (s, 1H), 10.08 (s, 1H), 8.74 (s, 2H), 8.13 (m, 1H),
8.05 (d, J=7.1 Hz, 1H), 7.34 (t, J=10.6 Hz, 1H), 4.04 (s, 3H),
3.13-2.99 (m, 1H), 2.76-2.60 (m, 1H), 1.97-1.86 (m, 6H)
Example 27
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-4-methylthiazole-2-carboxamide
##STR00047##
[0213] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-methylthiazole-2-carboxylic acid
[0214] LC-MS (m/z) 415 (MH.sup.+) t.sub.R=0.54 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.89 (s, 1H),
7.83-7.76 (m, 2H), 7.69 (d, J=0.9 Hz, 1H), 7.15 (dd, J=11.9, 8.8
Hz, 1H), 6.40 (s, 2H), 2.59-2.52 (m, 1H), 2.50 (s, 3H), 2.19 (m,
1H), 1.74-1.67 (m, 6H)
Example 28
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-(trifluoromethyl)picolinamide
##STR00048##
[0216] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(trifluoromethyl)picolinic acid
[0217] LC-MS (m/z) 463 (MH.sup.+) t.sub.R=0.61 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.96 (s, 1H),
9.18-9.09 (m, 1H), 8.48 (dd, J=8.3, 2.1 Hz, 1H), 8.33 (d, J=8.2 Hz,
1H), 7.89-7.81 (m, 2H), 7.17 (dd, J=11.9, 8.7 Hz, 1H), 6.58-6.29
(m, 2H), 2.62-2.52 (m, 1H), 2.31-2.16 (m, 1H), 1.77-1.66 (m,
6H)
Example 29
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-2-(difluoromethyl)oxazole-4-carboxamide
##STR00049##
[0219] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2-(difluoromethyl)oxazole-4-carboxylic acid
[0220] LC-MS (m/z) 435 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.52 (s, 1H), 9.03 (s,
1H), 7.73 (dd, J=7.2, 2.6 Hz, 1H), 7.72-7.68 (m, 1H), 7.33 (t,
J=51.9 Hz, 1H), 7.14 (dd, J=11.9, 8.8 Hz, 1H), 6.31 (s, 2H),
2.58-2.48 (m, 1H), 2.25-2.11 (m, 1H), 1.72-1.64 (m, 6H)
Example 30
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-4-(fluoromethyl)oxazole-2-carboxamide
##STR00050##
[0222] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-(fluoromethyl)oxazole-2-carboxylic acid
[0223] LC-MS (m/z) 417 (MH.sup.+) t.sub.R=0.47 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.15 (s, 1H), 8.57 (d,
J=5.0 Hz, 1H), 7.81-7.71 (m, 2H), 7.16 (dd, J=11.9, 8.7 Hz, 1H),
6.34 (s, 2H), 5.43 (d, J=48.0 Hz, 2H), 2.58-2.48 (m, 1H), 2.23-2.09
(m, 1H), 1.73-1.64 (m, 6H)
Example 31
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-4-(fluoromethyl)thiazole-2-carboxamide
##STR00051##
[0225] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-(fluoromethyl)thiazole-2-carboxylic acid
[0226] LC-MS (m/z) 433 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.03 (s, 1H), 8.25 (d,
J=3.2 Hz, 1H), 7.85-7.75 (m, 2H), 7.16 (dd, J=11.9, 8.7 Hz, 1H),
6.35 (dd, J=22.8, 15.9 Hz, 1H), 5.58 (d, J=47.7 Hz, 2H), 2.61-2.52
(m, 1H), 2.19 (m, 1H), 1.76-1.64 (m, 6H)
Example 32
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-cyanopicolinamide
##STR00052##
[0228] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-cyanopicolinic acid
[0229] LC-MS (m/z) 420 (MH.sup.+) t.sub.R=0.52 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.96 (s, 1H), 9.20
(dd, J=2.0, 0.8 Hz, 1H), 8.57 (dd, J=8.2, 2.1 Hz, 1H), 8.31-8.26
(m, 1H), 7.87-7.78 (m, 2H), 7.16 (dd, J=11.9, 8.7 Hz, 1H), 6.34 (s,
2H), 2.62-2.52 (m, 1H), 2.28-2.13 (m, 1H), 1.73-1.65 (m, 6H)
Example 33
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-chloropicolinamide
##STR00053##
[0231] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-chloropicolinic acid
[0232] LC-MS (m/z) 429 (MH.sup.+) t.sub.R=0.55 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.80 (s, 1H),
8.80-8.76 (m, 1H), 8.18 (dd, J=8.4, 2.4 Hz, 1H), 8.15 (d, J=8.4 Hz,
1H), 7.85-7.77 (m, 2H), 7.15 (dd, J=11.9, 8.7 Hz, 1H), 6.31 (d,
J=25.7 Hz, 2H), 2.58-252 (m, 1H), 2.28-2.15 (m, 1H), 1.75-1.66 (m,
6H)
Example 34
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-2-methyloxazole-4-carboxamide
##STR00054##
[0234] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2-methyloxazole-4-carboxylic acid
[0235] LC-MS (m/z) 399 (MH.sup.+) t.sub.R=0.46 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.29 (br s, 1H), 7.79
(m, 1H), 7.72 (m, 1H), 7.59 (s, 1H), 7.13 (dd, J=11.7, 9.0 Hz, 1H),
6.40 (br s, 2H), 2.55 (m, 1H), 2.44 (s, 3H), 2.20 (m, 1H), 1.71 (d,
J=23.4 Hz, 3H), 1.69 (s, 3H)
Example 35
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methyl-1,2,4-oxadiazole-3-carboxamide
##STR00055##
[0237] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methyl-1,2,4-oxadiazole-3-carboxylic acid
[0238] LC-MS (m/z) 400 (MH.sup.+) t.sub.R=0.43 minutes (Method
A)
Example 36
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)thiazole-2-carb oxamide
##STR00056##
[0240] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and thiazole-2-carboxylic acid
[0241] LC-MS (m/z) 401 (MH.sup.+) t.sub.R=0.47 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.97 (br s, 1H), 8.14
(d, J=3.1 Hz, 1H), 8.10 (d, J=3.1 Hz, 1H), 7.79 (m, 2H), 7.15 (dd,
J=11.9, 8.8 Hz, 1H), 6.33 (br s, 2H), 2.55 (m, 1H), 2.18 (m, 1H),
1.70 (d, J=23.0 Hz, 3H), 1.67 (s, 3H)
Example 37
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-4-bromo-1-methyl-1H-imidazole-2-carboxamide
##STR00057##
[0243] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-bromo-1-methyl-1H-imidazole-2-carboxylic acid
[0244] LC-MS (m/z) 476 (MH.sup.+) t.sub.R=0.53 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.59 (br s, 1H), 7.74
(dt, J=8.7, 3.3 Hz, 1H), 7.69 (dd, J=7.2, 2.6 Hz, 1H), 7.63 (s,
1H), 7.11 (dd, J=11.9, 8.8 Hz, 1H), 6.32 (br s, 2H), 3.95 (s, 3H),
2.55 (m, 1H), 2.18 (m, 1H), 1.69 (d, J=23.1 Hz, 3H), 1.66 (s,
3H)
Example 38
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-1,4-dimethyl-1H-imidazole-2-carboxamide
##STR00058##
[0246] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 1,4-dimethyl-1H-imidazole-2-carboxylic acid
[0247] LC-MS (m/z) 412.3 (MH.sup.+) t.sub.R=0.46 minutes (Method B)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.39 (s, 1H),
7.74-7.71 (m, 1H), 7.68 (dd, J=7.2, 2.6 Hz, 1H), 7.14 (s, J=15.0
Hz, 1H), 7.12-7.07 (m, 1H), 6.30 (s, 2H), 3.91 (s, 3H), 2.58-2.52
(m, 1H), 2.26-2.15 (m, 1H), 2.17 (s, 3H), 1.69 (d, J=23.0 Hz, 3H),
1.66 (s, 3H).
Example 39
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-1-methyl-1H-pyrazole-3-carboxamide
##STR00059##
[0249] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 1-methyl-1H-pyrazole-3-carb oxylic acid
[0250] LC-MS (m/z) 398 (MH.sup.+) t.sub.R=0.42 minutes (Method B)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 9.93 (br s, 1H), 8.31
(s, 1H), 8.00 (t, J=2.3 Hz, 1H), 7.63 (m, 1H), 7.61 (m, 3H), 6.31
(br s, 2H), 2.56 (m, 1H), 2.21 (m, 1H), 1.69 (d, J=23.3 Hz, 3H),
1.67 (s, 3H)
Example 40
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-3-methylisoxazole-5-carboxamide
##STR00060##
[0252] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 3-methylisoxazole-5-carboxylic acid
[0253] LC-MS (m/z) 399.1 (MH.sup.+) t.sub.R=0.47 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.84 (br s, 1H), 7.73
(m, 1H), 7.70 (dd, J=7.1, 2.7 Hz, 1H), 7.16 (dd, J=11.9, 8.8 Hz,
1H), 7.11 (s, 1H), 6.35 (br s, 2H), 2.55 (m, 1H), 2.34 (s, 3H),
2.20 (m, 1H), 1.68 (d, J=22.6 Hz, 3H), 1.67 (s, 3H)
Example 41
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-methylfuran-2-carboxamide
##STR00061##
[0255] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-methylfuran-2-carb oxylic acid
[0256] LC-MS (m/z) 398.3 (MH.sup.+) t.sub.R=0.52 minutes (Method B)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.16 (br s, 1H), 7.67
(dd, J=7.2, 2.7 Hz, 1H), 7.63 (ddd, J=8.7, 3.8, 2.9 Hz, 1H), 7.23
(m, 1H), 7.12 (dd, J=11.9, 8.8 Hz, 1H), 6.32 (dd, J=3.4, 1.0 Hz,
1H), 6.31 (br s, 2H), 2.55 (m, 1H), 2.38 (s, 3H), 2.19 (m, 1H),
1.69 (d, J=23.1 Hz, 3H), 1.66 (s, 3H)
Example 42
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-2-methyloxazole-5-carboxamide
trifluoroacetic acid salt
##STR00062##
[0258] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2-methyloxazole-5-carboxylic acid
[0259] LC-MS (m/z) 399 (MH.sup.+) t.sub.R=0.43 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 11.64 (s, 1H), 10.62
(s, 1H), 10.17 (s, 1H), 10.07 (s, 1H), 7.90-7.85 (m, 3H), 7.36-7.31
(m, 1H), 3.14-3.02 (m, 1H), 2.77-2.62 (m, 1H), 2.54 (s, 3H), 1.92
(s, 3H), 1.89 (d, J=23.0 Hz, 4H)
Example 43
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-(methoxy-d.sub.3)picolinamide
##STR00063##
[0261] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(methoxy-d.sub.3)picolinic acid
[0262] LC-MS (m/z) 428.2 (MH.sup.+) t.sub.R=0.54 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.58 (s, 1H), 8.39 (d,
J=2.9 Hz, 1H), 8.12 (d, J=8.7 Hz, 1H), 7.84-7.81 (m, 1H), 7.78 (d,
J=8.1 Hz, 1H), 7.60 (dd, J=8.7, 2.9 Hz, 1H), 7.13 (dd, J=11.8, 8.9
Hz, 1H), 6.32 (s, 2H), 2.59-2.45 (m, 1H), 2.36-2.10 (m, 1H), 1.71
(d, J=23.0 Hz, 3H), 1.67 (s, 3H)
Example 44
N-(3-((2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-4-chlorobenzamide
##STR00064##
[0264] Prepared from
(3R,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 4-chlorobenzoic acid
[0265] LC-MS (m/z) 428.1 (MH.sup.+) t.sub.R=0.58 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.46 (s, 1H), 7.96 (d,
J=8.4 Hz, 2H), 7.76-7.65 (m, 2H), 7.60 (d, J=8.4 Hz, 2H), 7.15 (dd,
J=11.7, 8.9 Hz, 1H), 6.37 (s, 2H), 2.61-2.49 (m, 1H), 2.29-2.14 (m,
1H), 1.70 (d, J=22.2 Hz, 3H), 1.68 (s, 3H)
Example 45
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-(difluoromethoxy)picolinamide
##STR00065##
[0267] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(difluoromethoxy)picolinic acid
[0268] LC-MS (m/z) 461.1 (MH.sup.+) t.sub.R=0.55 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.59 (s, 1H), 8.62 (d,
J=2.8 Hz, 1H), 8.22 (dd, J=8.7, 0.5 Hz, 1H), 7.93-7.89 (m, 2H),
7.89-7.85 (m, 1H), 7.50 (t, J=72.9 Hz, 1H), 7.14 (dd, J=11.9, 8.8
Hz, 1H), 6.23 (s, 2H), 2.74-2.59 (m, 1H), 2.55 (s, 3H), 2.49-2.38
(m, 1H), 1.68 (d, J=22.7 Hz, 3H), 1.63 (s, 3H)
Example 46
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-3-fluoro-5-methoxypicolinamide
##STR00066##
[0270] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 3-fluoro-5-methoxypicolinic acid
[0271] LC-MS (m/z) 443.1 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.40 (s, 1H), 8.28
(dd, J=2.3, 0.8 Hz, 1H), 7.84-7.81 (m, 1H), 7.78 (dd, J=7.2, 2.7
Hz, 1H), 7.62 (dd, J=12.7, 2.4 Hz, 1H), 7.12 (dd, J=12.0, 8.8 Hz,
1H), 6.22 (s, 2H), 3.94 (s, 3H), 2.72-2.59 (m, 1H), 2.49-2.36 (m,
1H), 1.67 (d, J=22.7 Hz, 3H), 1.62 (s, 3H)
Example 47
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4 fluorophenyl)-2,5-dimethyloxazole-4-carboxamide
##STR00067##
[0273] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 2,5-dimethyloxazole-4-carboxylic acid
[0274] LC-MS (m/z) 413.1 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 9.90 (s, 1H), 7.80 (dd,
J=7.2, 2.7 Hz, 1H), 7.75 (ddd, J=8.7, 3.8, 2.9 Hz, 1H), 7.08 (dd,
J=12.0, 8.8 Hz, 1H), 6.21 (s, 2H), 2.71-2.60 (m, 1H), 2.57 (s, 3H),
2.47-2.40 (m, 1H), 2.45 (s, 3H), 1.67 (d, J=22.8 Hz, 3H), 1.61 (s,
3H)
Example 48
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-5-(methoxy-d3)picolinamide
##STR00068##
[0276] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-(methoxy-d.sub.3)picolinic acid
[0277] LC-MS (m/z) 428.1 (MH.sup.+) t.sub.R=0.51 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.41 (s, 1H), 8.39
(dd, J=2.9, 0.5 Hz, 1H), 8.12 (dd, J=8.7, 0.5 Hz, 1H), 7.88-7.85
(m, 2H), 7.61 (dd, J=8.7, 2.9 Hz, 1H), 7.15-7.09 (m, 1H), 6.22 (s,
2H), 2.72-2.59 (m, 1H), 2.48-2.38 (m, 1H), 1.67 (d, J=22.7 Hz, 3H),
1.62 (s, 3H)
Example 49
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-fluoro-3-methylpicolinamide
##STR00069##
[0279] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-fluoro-3-methylpicolinic acid
[0280] LC-MS (m/z) 427.1 (MH.sup.+) t.sub.R=0.54 minutes (Method A)
.sup.1H NMR (600 MHz, DMSO-d.sub.6) .delta. 10.51 (s, 1H), 8.53
(dd, J=2.7, 0.4 Hz, 1H), 7.89-7.84 (m, 1H), 7.81 (ddd, J=9.8, 2.7,
0.6 Hz, 1H), 7.75 (dd, J=7.2, 2.7 Hz, 1H), 7.12 (dd, J=12.0, 8.8
Hz, 1H), 6.22 (s, 2H), 2.75-2.61 (m, 1H), 2.58 (s, 3H), 2.49-2.37
(m, 1H), 1.67 (d, J=22.7 Hz, 3H), 1.62 (s, 3H)
Example 18a
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide
##STR00070##
[0282]
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpipe-
ridine-2-thione (750 mg, 2.44 mmol) was dissolved in 7M ammonia in
methanol (36 ml, 252 mmol). The reaction mixture was stirred in a
sealed vial at 60.degree. C. overnight. The reaction mixture was
allowed to cool to room temperature and concentrated under reduced
pressure to afford
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethyl-3,4,5,6-t-
etrahydropyridin-2-amine (708 mg, 2.448 mmol, 100% yield) as a pale
yellow solid that was used in the next reaction without further
purification.
[0283] 5-Cyano-3-methylpicolinic acid (232 mg, 1.432 mmol)
(Prepared as described in Badiger, Sangamesh et al. PCT Int. Appl.,
2012095469) was placed in a round bottom flask and dissolved in DMF
(7 mL). HATU (669 mg, 1.760 mmol) was added and the reaction was
stirred at room temperature for 5 min, N,N-diisopropylethylamine
(0.7 mL, 4.1 mmol) was added. The reaction mixture was cooled to
0.degree. C. and added dropwise to a solution of
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethyl-3,4,5,6-t-
etrahydropyridin-2-amine (470 mg, 1.63 mmol) in DMF (7 mL) at
0.degree. C. The reaction mixture was stirred at 0.degree. C. for
30 min then at 30 min at room temperature. The reaction mixture was
diluted with ethyl acetate (50 mL) and washed with water. The
phases were separated and the aqueous layer was extracted with
ethyl acetate (2.times.100 mL). The combined organic phases were
dried over MgSO.sub.4, filtered, and concentrated in vacuo. The
crude material was purified using a RediSep Automated flash system
on 80 g silica gel (eluent: ethyl acetate/heptane). The product was
further purified by the following procedure: The product was
dissolved in ethyl acetate (50 mL) and washed with a solution of
saturated aqueous NaHCO.sub.3/water (1/1). The organic phase was
washed total of 15 times (using 10 mL each time). The organic phase
was dried over MgSO.sub.4, filtered, and evaporated to give
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyri-
din-2-yl)-4-fluorophenyl)-5-cyano-3-methylpicolinamide (153 mg, 26%
yield).
[0284] LC-MS (m/z) 434 (MH.sup.+) t.sub.R=0.53 minutes (Method
A).
[0285] .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 9.97 (s, 1H), 8.72
(dd, J=1.9, 0.6 Hz, 1H), 7.99-7.90 (m, 2H), 7.44 (dd, J=6.8, 2.8
Hz, 1H), 7.09 (dd, J=11.7, 8.8 Hz, 1H), 4.71 (s, 2H), 2.86 (s, 3H),
2.48 (m, 2H), 1.81-1.74 (m, 6H).
[0286] The following compound was prepared in a way similar to
example 18a:
Example 4a
N-(3-((2R,5S)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyrid-
in-2-yl)-4-fluorophenyl)-5-cyanopicolinamide
##STR00071##
[0288] Prepared from
(3S,6R)-6-(5-amino-2-fluorophenyl)-3,5,5-trifluoro-3,6-dimethylpiperidine-
-2-thione and 5-cyanopicolinic acid
[0289] LC-MS (m/z) 420.0 (MH.sup.+) t.sub.R=1.79 minutes (Method
B)
Stereochemistry
[0290] Crystals were obtained by recrystallization of compound 48
from a mixture of heptane and ethyl acetate. The structure of
compound 48 was elucidated by X-ray crystallography of said
crystals. The two molecules in the asymmetric unit as found in the
X-ray structure of compound 48 are shown in FIG. 1 and shows that
the stereoconfiguration is (2R,5S).
[0291] The absolute configurations of the exemplified compounds of
the present invention can thus be rationalized. All examples were
synthesized from the intermediates XVIa or XVIb with
R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy carbonyl
##STR00072##
[0292] The relative and absolute stereochemistry of intermediate
XVIa (R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy carbonyl) has
been assigned as (2R,5S) based on the X-ray structure of example
48. The absolute stereochemistry of the 2-position in intermediate
XVIa (R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy carbonyl) was
assigned as (2R) based on the absolute configuration of
intermediate XV (R.sup.1=methyl and R.sup.4=ethyl) for which the
absolute configuration was assigned based on literature precedence
(WO2012110459). The two ways of assigning the stereochemistry at
the 5-position are in agreement.
[0293] The absolute stereochemistry of intermediate XVIb
(R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy carbonyl) was based
on the absolute configuration of intermediate XV (R.sup.1=methyl
and R.sup.4=ethyl) for which the absolute configuration was
assigned based on literature precedence. The stereochemistry at the
5-position of the
6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyridine
substructure is opposite to the stereochemistry at that position of
intermediate XVIa (R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy
carbonyl), hence, the stereochemistry of intermediate XVIb is
(2R,5S).
[0294] The stereochemistry of the exemplified compounds containing
the
(2R,5R)-6-amino-3,3,5-trifluoro-2,5-dimethyl-2,3,4,5-tetrahydropyridine
substructure, e.g. example 2, is based on the stereochemistry of
intermediate XVIb (R.sup.1=R.sup.3=methyl and R.sup.7=tert-butoxy
carbonyl).
Pharmacological Testing
BACE1 Binding Assay
[0295] The binding assay was performed as SPA-based assay using a
biotinylated form of human BACE1 recombinantly expressed and
subsequently purified from Freestyle HEK293 cells. The binding
assay was run in a 50 mM sodium acetate buffer, pH 4.5 containing
50 mM NaCl and 0.03% Tween-20 in white clear bottom 384 plates
(Corning #3653). 10 nM (final concentration) radioligand
([.sup.3H]--N-((1
S,2R)-1-benzyl-3-cyclopropylamino-2-hydroxy-propyl)-5-(methanesulfonyl-me-
thyl-amino)-N--((R)-1-phenyl-ethyl)-isophthalamide) (TRQ11569
purchased from GE Healthcare) was mixed with test compound at a
given concentration, 6 nM (final concentration) human BACE1 and 25
.mu.g Streptavidin coated PVT core SPA beads (RPNQ0007, GE
Healthcare Life Sciences) in a total volume of 40 .mu.l. Several
concentrations of each test compound were tested in the assay for
IC.sub.50 determination. The plates were incubated for one hour at
room temperature and counted in a Wallac Trilux counter. Total and
non-specific binding were determined using buffer and 1 .mu.M
(final concentration) of the high affinity BACE1 reference
inhibitor
(S)-6-[3-chloro-5-(5-prop-1-ynyl-pyridin-3-yl)-thiophen-2-yl]-2-imino-3,6-
-dimethyl-tetrahydro-pyrimidin-4-one, respectively. For each test
compound, a IC.sub.50 value (the concentration mediating 50%
inhibition of the specific binding of the radioligand) was
determined from concentration-response curve and used to calculate
the from the equation K.sub.i=IC.sub.50/(1+L/K.sub.d), where L and
K.sub.d are the final concentration of the radioligand used in the
assay and the dissociation constant of the radioligand,
respectively. The K.sub.d of the radioligand was determined from
saturation binding experiments.
TABLE-US-00001 TABLE 1 binding affinity of selected compounds
Compound BACE1 No Ki (nM) 1 83 2 210 3 12 4 11 5 18 6 19 7 33 8 66
9 100 10 44 11 63 12 29 13 28 14 37 15 46 16 58 17 11 18 4.3 19 32
20 1600 21 58 22 110 23 71 24 420 25 130 26 130 27 140 28 170 29 92
30 260 31 92 32 51 33 62 34 210 35 680 36 390 37 420 38 1000 39
4800 40 8200 41 520 42 1500 43 91 44 740 45 14 46 14 47 65 48 14 49
60
BACE1 Efficacy Assay
[0296] The efficacy assay was performed as a FRET-based assay using
a commercially available BACE1 kit (Life Technologies, P2985). 2
.mu.l test compound at 10 .mu.M (final concentration) and 15 .mu.l
BACE1 enzyme from the kit (final concentration 3 nM) were
preincubated for 15 minutes at room temperature before addition of
15 ml of substrate from the kit (250 nM final concentration) and
incubated for additional 90 minutes at room temperature. The assay
plate was subsequently read in a Pherastar (Ex540/Em590). The
enzyme activity observed in presence of test compound were
normalized to the enzyme activity observed in presence of buffer
and 10 .mu.M (final concentration) of the high affinity BACE1
reference inhibitor
(S)-6-[3-Chloro-5-(5-prop-1-ynyl-pyridin-3-yl)-thiophen-2-yl]-2-
-imino-3,6-dimethyl-tetra-hydropyrimidin-4-one, respectively. The
efficacy of the test compounds was evaluated at 10 .mu.M (final
concentration) and defined as the percent inhibition of the enzyme
activity using the equation % inhibition=100%-normalized enzyme
activity in percent.
TABLE-US-00002 TABLE 2 BACE1 activity of selected compounds BACE1
inhibition Compound at 10 .mu.M No (%) 1 104 2 100 3 102 4 99 5 96
6 100 7 100 8 104 9 103 10 102 11 99 12 104 13 99 14 102 15 101 16
105 17 101 18 102 19 98 20 88 21 102 22 92 23 102 24 98 25 102 26
102 27 98 28 102 29 101 30 103 31 102 32 101 33 103 34 102 35 100
36 100 37 104 38 98 39 74 40 69 41 93 42 84 43 94 44 92 45 103 46
101 47 102 48 99 49 98
Assessment of A.beta. Levels in Rat Brain and Plasma Following
BACE1 Inhibition.
Animals.
[0297] All rat care and experimental procedures were approved by
Lundbeck Veterinary Staff, according to Danish legislature. The
rats were maintained in a barrier facility with a 12/12-h
light/dark cycle and ad libitum food and water access.
Treatment of Naive Rats.
[0298] Young adult Male Sprague Dawley rats of approximately 250 g
weight were purchased from Charles River and received 0-30 mg/kg of
vehicle (10% HP betaCD+1M MeSO.sub.4, pH 2.5) or test compounds
(dissolved in vehicle) only by oral gavage (p.o). The compounds are
dosed at a volume of 5 ml/kg. Cohorts of 5-10 animals were
established for each treatment condition.
[0299] The animals undergoing treatment were closely monitored by
veterinary staff for any signs of toxicity. Monitoring parameters
included body weight, physical appearance, changes in coat
appearance, occurrence of unprovoked behavior, and blunted or
exaggerated responses to external stimuli.
Tissue Collection.
[0300] At T=180 minutes after initial dosing the animals were
stunned and decapitated with a guillotine. Trunk-blood was sampled
in EDTA coated tubes after decapitation of the animal. The blood
was centrifuged at 2200 G at 4.degree. C. for 15 minutes and the
plasma was collected and frozen at -80.degree. C. The blood was
aliquoted for A.beta. ELISA and DMPK analysis Immediately following
sacrifice, the brain was extracted and split into 2 halves. The
right hemibrains were snap frozen on dry ice and stored at
-80.degree. C. The left half was dissected; with the front
forebrain taken for A.beta. ELISA and the remainder used for DMPK
analysis. These samples were also snap frozen on dry ice and stored
at -80.degree. C. until use for analysis.
Tissue Processing.
[0301] The cortex samples were thawed slightly on wet ice before
they were homogenized with a small volume dispersing instrument
(T10 basic ULTRA-TURRAX.RTM.) which was set at speed 5 for
approximately 5-7 sec. The tissue was processed in a 10 times
volume of the weight, for example 100 mg of tissue was homogenized
in 1000 .mu.L of Homogenization buffer. Homogenization buffer: 50
ml Milli Q water+50 nM NaCl+0.2% Diethylamin (DEA)+1 tablet of
Complete Protease inhibitor cocktail+1 nM 4-(2-aminoethyl)
benzenesulfonyl fluoride hydrochloride irreversible serine protease
inhibitor (AEBSF).
[0302] After homogenization 450 .mu.L aliquots of the samples are
collected into a 1.5 ml Eppendorf tube and placed on wet ice, 0.5%
NP-40 (50 ul) was added to all samples and then they were incubated
on ice for 30 min. After which all samples were sonicated using an
Ultrasonic homogenizer with 20 kHz homogeneous sound (SONOPLUS
HD2070, Bandelin Electronic) 10 pulse set at 12-13% power to
extract all the A.beta. species. The samples were then centrifuged
(Ole Dich 157 MPRF Micro centrifuge) at 20000 G for 20 minutes at
4.degree. C. After centrifugation 285 .mu.L of the supernatant was
pipetted into 600 .mu.L microtubes tubes and neutralized with 15
.mu.L of 1M Tris-HCL buffer.
ELISA Protocol.
[0303] WAKO 294-62501 Human/Rat Abeta amyloid (40) kit was used for
all ELISA analyses. 30 .mu.L plasma samples or 30 .mu.L of the
cortex supernatants generated as described above were placed in 600
.mu.L microtubes tubes on wet ice. To this 30 .mu.L of 8M Urea
(AppliChem A1049, 9025) are added to generate a 2-fold dilution.
Both plasma and cortex supernatants are incubated on ice for 30
min.
[0304] Standard rows were prepared from the standard peptide stock
provided in the kit and standard diluent containing 1.6M Urea (200
.mu.L 8M Urea+800 .mu.L of standard diluent) and 0.8M Urea (400
.mu.L 8M Urea+3600 .mu.L Standard diluent). A serial 2-fold
dilution of A.beta.40 from 100 pmol/ml to 0 pmol/L was prepared for
the assay.
[0305] After incubation with urea, all samples were further diluted
by addition of 5 times standard diluent from the Kit. This was done
by adding 240 .mu.L Standard Diluent to 60 .mu.L sample/urea
mixture, which was then mixed well. 100 .mu.L of each diluted
sample was pipetted into designated wells of the ELISA plate in
duplicates. The plate was then covered and incubated overnight at
4.degree. C. The following day, the ELISA kit was brought to room
temperature before use. The incubated plate was washed 5 times with
the 20.times. washing solution diluted in Milli Q water. 100 .mu.L
HRP-conjugate was applied to each well, and the plate was covered
and incubates at 4.degree. C. for 1 hr. The wash was repeated again
for 5 times. 100 .mu.L 3,3',5,5'-Tetramethylbenzidine (TMB)
solution was applied to each well and the plate was covered and
incubated in the dark at room temperature for 30 minutes. 100 .mu.L
STOP-solution was next applied to each well, and the plate was read
at 450 nm wavelength in a spectrophotometer (Labsystems Multiscan
Ascent) within 30 min of adding the STOP-solution to the wells.
[0306] Concentration of A.beta. in the samples was determined based
on a standard curve generated from standards containing known
concentrations of synthetic A.beta.40. Those skilled in the art
will appreciate that diethylamine (DEA) and urea extractions will
release soluble A.beta., and insoluble A.beta. respectively. Since
the ELISA kit is validated and widely used, it is accepted that as
long as the treatment conditions and assay conditions are the same
for each compound tested, then the assay should yield consistent
robust data for the compounds tested and produce minimal
discrepancies.
Data Analysis
[0307] To determine the concentration of A.beta.40 in the samples,
the interpolated values of the samples loaded on plates are
multiplied by 20 to account for the dilutions made when the volumes
of DEA, urea and neutralization solution were added up. Values are
calculated as percentage change in A.beta.40 compared to vehicle
treated animals.
[0308] Compounds 1, 5, 17, 18, and 24 were administered at doses of
10 or 30 mg/kg p.o. and brain and plasma samples were collected at
3 hours post dose and the following exposures and reductions in
A.beta.40 levels were measured as described above.
TABLE-US-00003 TABLE 3 Results for compound 1 Dose Exp Brain/Plasma
A.beta.40 (mg/kg) (ng/g) ratio reduction (%) Brain Rat 30 2188 0.62
56 Plasma Rat 3545 41
TABLE-US-00004 TABLE 4 Results for compound 5 Dose Exp Brain/Plasma
A.beta.40 (mg/kg) (ng/g) ratio reduction (%) Brain Rat 10 174 1.3
34 Plasma Rat 137 34 Brain Rat 30 954 1.5 61 Plasma Rat 632 39
TABLE-US-00005 TABLE 5 Results for example 17 Dose Exp Brain/Plasma
A.beta.40 (mg/kg) (ng/g) ratio reduction (%) Brain Rat 30 1223 1.48
63 Plasma Rat 828 49
TABLE-US-00006 TABLE 6 Results for example 18 Dose Exp Brain/Plasma
A.beta.40 (mg/kg) (ng/g) ratio reduction (%) Brain Rat 10 412 0.53
66 Plasma Rat 778 54 Brain Rat 30 1606 0.54 61 Plasma Rat 3000
50
TABLE-US-00007 TABLE 7 Results for example 24 Dose Exp Brain/Plasma
A.beta.40 (mg/kg) (ng/g) ratio reduction (%) Brain Rat 10 134 1.39
15 Plasma Rat 96.2 28 Brain Rat 30 809 1.20 49 Plasma Rat 673
61
[0309] As shown in tables 3-7, compounds of the present invention
are able to penetrate the blood brain barrier and are efficacious
in lowering A.beta.40 levels in the brain of animals.
* * * * *