U.S. patent application number 12/430420 was filed with the patent office on 2009-08-20 for soluble guanylate cyclase activators.
Invention is credited to Amy R. Bittner, Jiang Chang, Ronald M. Kim, J. W. Mirc, Emma R. Parmee, Christopher Joseph Sinz, Qiang Tan.
Application Number | 20090209556 12/430420 |
Document ID | / |
Family ID | 40070857 |
Filed Date | 2009-08-20 |
United States Patent
Application |
20090209556 |
Kind Code |
A1 |
Bittner; Amy R. ; et
al. |
August 20, 2009 |
SOLUBLE GUANYLATE CYCLASE ACTIVATORS
Abstract
A compound having the structure ##STR00001## useful for
treatment or prevention of cardiovascular diseases, endothelial
dysfunction, diastolic dysfunction, atherosclerosis, hypertension,
angina pectoris, thromboses, restenoses, myocardial infarction,
strokes, cardiac insufficiency, pulmonary hypertonia, erectile
dysfunction, asthma bronchiale, chronic kidney insufficiency,
diabetes, or cirrhosis of the liver in a human or animal
patient.
Inventors: |
Bittner; Amy R.; (Princeton,
NJ) ; Sinz; Christopher Joseph; (Cranford, NJ)
; Chang; Jiang; (Westfield, NJ) ; Kim; Ronald
M.; (Summit, NJ) ; Mirc; J. W.; (Union City,
NJ) ; Parmee; Emma R.; (Schotch Plains, NJ) ;
Tan; Qiang; (Westfield, NJ) |
Correspondence
Address: |
MERCK AND CO., INC
P O BOX 2000
RAHWAY
NJ
07065-0907
US
|
Family ID: |
40070857 |
Appl. No.: |
12/430420 |
Filed: |
April 27, 2009 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
PCT/US2008/010321 |
Sep 2, 2008 |
|
|
|
12430420 |
|
|
|
|
60967827 |
Sep 6, 2007 |
|
|
|
Current U.S.
Class: |
514/255.05 ;
514/333; 514/341; 544/405; 546/256; 546/275.4 |
Current CPC
Class: |
C07D 403/04 20130101;
A61P 15/10 20180101; C07D 403/14 20130101; A61P 3/06 20180101; A61P
9/10 20180101; A61P 3/10 20180101; A61P 1/16 20180101; A61P 7/10
20180101; C07D 405/14 20130101; A61P 11/06 20180101; A61P 13/12
20180101; A61P 9/08 20180101; C07D 401/14 20130101; A61P 9/04
20180101; C07D 417/14 20130101; A61P 9/00 20180101; C07D 417/04
20130101; A61P 43/00 20180101; C07D 409/14 20130101; C07D 413/14
20130101; A61P 9/12 20180101; C07D 401/04 20130101; A61P 7/02
20180101 |
Class at
Publication: |
514/255.05 ;
514/333; 514/341; 544/405; 546/256; 546/275.4 |
International
Class: |
A61K 31/497 20060101
A61K031/497; A61K 31/4545 20060101 A61K031/4545; A61K 31/4439
20060101 A61K031/4439; C07D 403/04 20060101 C07D403/04; C07D 401/14
20060101 C07D401/14; C07D 401/04 20060101 C07D401/04; A61P 9/00
20060101 A61P009/00 |
Claims
1. A compound of formula I, or a pharmaceutically acceptable salt
or hydrate thereof: ##STR00373## wherein Z.sup.1 is selected from
the group consisting of CH and N; A is a ring selected from the
group consisting of ##STR00374## D.sup.1 is CH, CR.sup.4 or N;
R.sup.7 is selected from the group consisting of 1) hydrogen, 2)
C.sub.1-6 alkyl wherein the alkyl group may be unsubstituted or
substituted with 1-3 fluorine atoms and unsubstituted or
monosubstituted with OC.sub.1-3 alkyl, 3) C.sub.3-6 cycloalkyl
wherein the cycloalkyl group may be unsubstituted or substituted
with 1-3 fluorine atoms and unsubstituted or monosubstituted with
OC.sub.1-3 alkyl, and 4) phenyl, wherein the phenyl group is
unsubstituted or substituted with C.sub.1-4 alkyl, --OC.sub.1-4
alkyl, halogen, CN, NO.sub.2, and S(O).sub.0-2C.sub.1-4 alkyl,
wherein C.sub.1-4 alkyl and --OC.sub.1-4 alkyl are unsubstituted or
substituted with 1-3 flourine atoms; L.sup.1 is selected from the
group consisting of O, S, C(R.sup.12).sub.2; and CF.sub.2; L.sup.2
is selected from the group consisting of (CH.sub.2).sub.2-4,
--C(R.sup.12).sub.2, --CF.sub.2--O, and S, provided that when
L.sup.1 is O or S, L.sup.2 is not O or S; R.sup.12 is independently
selected from the group consisting of hydrogen and C.sub.1-3 alkyl,
wherein C.sub.1-3 alkyl is unsubstituted or substituted with 1-3
flourine atoms; E is a ring selected from the group consisting of
1) a 6-10 membered aryl ring, 2) a 5-10 membered heteroaryl ring
having 1, 2 or 3 heteroatoms independently selected from the group
consisting of 0, 1, 2, and 3 N atoms, 0 or 1 O atoms, and 0 or 1 S
atoms, 3) a C.sub.3-8 cycloalkyl ring; wherein aryl, heteroaryl,
and C.sub.3-8 cycloalkyl are unsubstituted or monosubstituted with
R.sup.5, and unsubstituted, monosubstituted or independently
disubstituted with R.sup.8; R.sup.4, in each instance in which it
occurs, is independently selected from the group consisting of
halogen, C.sub.1-6 alkyl, wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.1-6 alkyl, wherein the alkyl group may be unsubstituted
or substituted with 1-3 fluorine atoms, C.sub.3-8 cycloalkyl,
unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.3-8 cycloalkyl, unsubstituted or substituted with 1-3
fluorine atoms, CN, and NO.sub.2; R.sup.5, in each instance in
which it occurs, is independently selected from the group
consisting of 1) R.sup.6, 2) --OR.sup.6, 3) C.sub.1-6 alkyl which
may be unsubstituted or substituted with 1-3 fluorine atoms, and
unsubstituted or monosubstituted with a group independently
selected from C.sub.3-6 cycloalkyl, --O--C.sub.1-4 alkyl, OH,
.dbd.O, S(O).sub.0-2C.sub.1-4 alkyl, --OR.sup.6 and R.sup.6, 4)
C.sub.1-6 alkenyl which may be unsubstituted or substituted with
1-3 fluorine atoms and unsubstituted or monosubstituted with a
group independently selected from --O--C.sub.1-4 alkyl, OH, .dbd.O,
S(O).sub.0-2C.sub.1-4 alkyl, --OR.sup.6 and R.sup.6, 5)
O--C.sub.1-6 alkyl wherein the alkyl group may be unsubstituted or
substituted with 1-3 fluorine atoms, and unsubstituted or
monosubstituted with a group independently selected from C.sub.3-6
cycloalkyl and R.sup.6, 6) --S--C.sub.1-6 alkyl, 7) a C.sub.3-8
cycloalkyl ring which is unsubstituted or mono, di- or
tri-substituted with groups independently selected from fluoro and
C1-4 alkyl, and unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl wherein the alkyl group
may be unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.1-4 alkyl, OH, .dbd.O, S(O).sub.0-2C.sub.1-4 alkyl,
--OR.sup.6, R.sup.6, and NR.sup.9R.sup.10, 8) a C.sub.5-8
cycloalkenyl ring which is unsubstituted or mono, di- or
tri-substituted with a group independently selected from fluoro and
C.sub.1-4 alkyl, and unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl, wherein the alkyl
group may be unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.1-4 alkyl, OH, .dbd.O, S(O).sub.0-2C.sub.1-4 alkyl, and
R.sup.6, 9) a 5- to 6 membered heterocyclyl ring having 1 or 2
heteroatoms selected from the group consisting of N, O and S, and
which is unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl wherein the alkyl group
may be unsubstituted or substituted with 1-3 fluorine atoms,
--OC.sub.1-4 alkyl, and .dbd.O, and 10) halogen; R.sup.6 is
selected from the group consisting of 1) a phenyl ring which is
unsubstituted, monosubstituted or disubstituted with a group
independently selected from the group consisting of halogen, OH,
CN, C.sub.1-4 alkyl wherein the alkyl group may be unsubstituted or
substituted with 1-3 fluorine atoms, OC.sub.1-4 alkyl wherein the
alkyl group may be unsubstituted or substituted with 1-3 fluorine
atoms, NO.sub.2, S(O).sub.0-2C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
O--C.sub.2-4 alkenyl, NR.sup.9R.sup.10, and COOH, and 2) a 5-6
membered heteroaryl ring containing 1-2 heteroatoms which are
independently selected from N, O and S, wherein the heteroaryl ring
is unsubstituted, monosubstituted or disubstituted with a group
independently selected from: halogen, OH, CN, C.sub.1-4 alkyl
wherein the alkyl group may be unsubstituted or substituted with
1-3 fluorine atoms, OC.sub.1-4 alkyl wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms, NO.sub.2,
S(O).sub.0-2C.sub.1-6 alkyl, S(O).sub.0-2 aryl, C.sub.2-6 alkenyl,
OC.sub.2-6 alkenyl, NR.sup.9R.sup.10, and COOH; R.sup.8 is selected
from the group consisting of C.sub.1-4 alkyl wherein the alkyl
group may be unsubstituted or substituted with 1-3 fluorine atoms,
C.sub.2-4 alkenyl, halogen, C.sub.3-6 cycloalkyl, wherein the
cycloalkyl group may be unsubstituted or substituted with 1-3
fluorine atoms OC.sub.1-4 alkyl wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms, O--C.sub.2-4
alkenyl, NO.sub.2, S(O).sub.0-2C.sub.1-4 alkyl, and CN; R.sup.9 and
R.sup.10 are independently selected from the group consisting of
hydrogen and C.sub.1-6 alkyl; and R.sup.11 is selected from the
group consisting of hydrogen and C.sub.1-6 alkyl.
2. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein ##STR00375## is selected from the group consisting
of ##STR00376## wherein E.sup.1 is CH or N.
3. A compound of claim 2, or a pharmaceutically acceptable salt
thereof, wherein ##STR00377## is selected from the group consisting
of ##STR00378##
4. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Z.sup.1 is CH.
5. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.7 is selected from the group consisting of
CH.sub.3, CF.sub.3 and CF.sub.2H.
6. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein L.sup.1 is selected from a group consisting of O
and S.
7. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein L.sup.2 is selected from the group consisting of
CH.sub.2, CH(CH.sub.3), O, CH.sub.2CH.sub.2, CF.sub.2 and
CH.sub.2CH.sub.2CH.sub.2, provided that when L.sup.2 is O L.sup.1
is not O.
8. A compound of claim 7, or a pharmaceutically acceptable salt
thereof, wherein L.sup.2 is selected from the group consisting of
CH.sub.2 and CF.sub.2.
9. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.4 is selected from the group consisting of
Cl, F, Br, CH.sub.3, cyclopropyl, NO.sub.2, and CF.sub.3.
10. A compound of claim 9, or a pharmaceutically acceptable salt
thereof, wherein R.sup.4 is selected from the group consisting of
Cl and CH.sub.3.
11. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.6 is a phenyl ring which is unsubstituted or
mono, di- or tri-substituted with a group independently selected
from the group consisting of Cl, F, --CH.sub.3,
--C(CH.sub.3).sub.3, CF.sub.3, --OCF.sub.3, --OCH.sub.3,
--OCH(CH.sub.3).sub.2 and COOH.
12. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.5 is selected from the group consisting of
1) R.sup.6, 2) a C.sub.3-6 cycloalkyl ring which is unsubstituted
or mono, di- or tri-substituted with a group independently selected
from phenyl, F, CF.sub.3, CH.sub.3, OH, and .dbd.O, 3) a pyridinyl
ring, wherein the point of attachment to the pyridinyl ring is a
carbon atom, and wherein the pyridinyl ring is unsubstituted or
mono-substituted with CF.sub.3, 4) --CH.sub.2-L.sup.3-R.sup.6,
wherein L.sup.3 is --CH.sub.2-- or --O--, 5) --OR.sup.6, 6)
--OCH.sub.2R.sup.6, 7) ##STR00379## 8) --CF.sub.3 9) Cl, F, or Br,
10) --CH.sub.3, 11) OCH.sub.3, 12) OCF.sub.3, 13)
--CH.dbd.CHR.sup.6, and 14) --SCH.sub.2CH.sub.3.
13. A compound of claim 12, or a pharmaceutically acceptable salt
thereof, wherein R.sup.5 is selected from the group consisting of
Cl, F, Br, --CH.sub.3, --C(CH.sub.3).sub.3, OCH.sub.3, OCF.sub.3,
--SCH.sub.2CH.sub.3, ##STR00380## ##STR00381## ##STR00382##
14. A compound of claim 13, or a pharmaceutically acceptable salt
thereof, wherein R.sup.5 is selected from the group consisting of
--CH.sub.3, - ##STR00383##
15. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.8 is selected from the group consisting of
CH.sub.3, Cl, F, cyclopropyl, and CF.sub.3.
16. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein A is a ring selected from the group consisting of
##STR00384##
17. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.11 is hydrogen.
18. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, selected from the group consisting of
1-[6-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluorome-
thyl)-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-(6-{2-[(4-{2-[4-(trifluoromethyl)phenyl]ethyl}benzy-
l)-oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid,
1-{6-[2-({4-[(1S,2S)-2-Phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[2-({4-[(1R,2R)-2-Phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(4-Chlorophenoxy)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-{6-[2-({4-[4-(trifluoromethyl)phenoxy]benzyl}oxy)ph-
enyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-(6-{2-[(4-{[4-(trifluoromethyl)phenoxy]methyl}benzy-
l)-oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Methyl-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(4-Oxocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(4,4-Difluorocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(trans-4-Methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5--
(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(cis-4-Methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(t-
rifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{[4-(trans)-4-Methoxycyclohexyl)-2-methylbenzyl]oxy}phenyl)pyridi-
n-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-{6-[2-({4-[6-(trifluoromethyl)pyridin-3-yl]benzyl}--
oxy)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid,
1-(6-{2-[(2,4-Dimethylbenzyl)oxy]-3-methylphenyl}pyridin-2-yl)-5-(trifluo-
romethyl)-1H-pyrazole-4-carboxylic acid, Ethyl
1-{6-[5-chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}-oxy)phe-
nyl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate,
Ethyl
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}-oxy)pheny-
l]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate,
5-(Trifluoromethyl)-1-[4-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)-1,3-thiazol-2-yl]-1H-pyrazole-4-carboxylic acid,
1-[2-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyrimidin-4-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid,
1-{4-Methyl-6-[5-methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl-
}-oxy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid,
5-(Trifluoromethyl)-1-[6-(2-{2-[4'-(trifluoromethyl)biphenyl-4-yl]e-
thyl}phenyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-(2'-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-2,-
3'-bipyridin-6-yl)-1H-pyrazole-4-carboxylic acid,
1-(5'-Methyl-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-2,3-
'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid,
1-(5'-Chloro-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-2,3-
'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid,
1-[2'-{[3-Methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-5'-(trifluoro-
methyl)-2,3'-bipyridin-6-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid,
5-(Trifluoromethyl)-1-{6-[2-({[4'-(trifluoromethyl)biphenyl-4-yl]ox-
y}methyl)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid,
5-(Trifluoromethyl)-1-(6-{2-[({5-[4-(trifluoromethyl)phenyl]pyridin-2-yl}-
oxy)methyl]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Methyl-2-({[4'-(trifluoromethyl)biphenyl-4-yl]methyl}thio)phenyl]-
pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-[6-(2-{Difluoro[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}phenyl)pyridi-
n-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[2-(Difluoro
{4-[trans-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)phenyl]pyridin-2-y-
l}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[2-(difluoro
{4-[cis-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)phenyl]pyridin-2-yl}-
-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-(6-{2-[{2-Ethyl-4-[4
(trifluoromethyl)cyclohexyl]phenyl}(difluoro)methoxy]phenyl}pyridin-2-yl)-
-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[2-(Difluoro
{[4'-(trifluoromethyl)biphenyl-4-yl]oxy}methyl)phenyl]pyridin-2-yl}-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid, and
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)pyridin-2-yl]-1H-1,2,3-triazole-4-carboxylic acid.
19. A compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.1 is H, Z.sup.1 is CH, R.sup.7 is CF.sub.3
or CF.sub.2H, D.sup.1 is CH, L.sup.1 is O, L.sup.2 is CH.sub.2 or
CF.sub.2, ring A is ##STR00385## R.sup.5 is ##STR00386##
20. A compound of claim 18, or a pharmaceutically acceptable salt
thereof, selected from the group consisting of
1-{6-[5-Methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Methyl-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
and
1-{6-[5-Chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid.
21. A composition useful for the treatment or prevention of
cardiovascular diseases, endothelial dysfunction, diastolic
dysfunction, atherosclerosis, hypertension, angina pectoris,
thromboses, restenoses, myocardial infarction, strokes, cardiac
insufficiency, pulmonary hypertonia, erectile dysfunction, asthma
bronchiale, chronic kidney insufficiency, diabetes or cirrhosis of
the liver, said composition comprising an amount efficacious for
said treatment or prevention of a compound of formula I as claimed
in claim 1, a stereoisomeric form thereof, or a physiologically
acceptable salt thereof, or a mixture of two or more of any of the
foregoing, and one or more pharmaceutically acceptable
carriers.
22. A method for activating soluble guanylate cyclase, said method
comprising the step of administering an amount efficacious
therefore of a compound of formula I as claimed in claim 1, a
stereoisomeric form thereof, a physiologically acceptable salt
thereof, or a mixture of any two or more of the foregoing.
23. A composition comprising a compound of claim 1 and a compound
selected from the group consisting of an angiotensin converting
enzyme inhibitor, an angiotensin II receptor antagonist, a neutral
endopeptidase inhibitor, an aldosterone antagonist, a renin
inhibitor, an endothelin receptors antagonist, a vasodilator, a
calcium channel blocker, a potassium channel activators, a
diuretic, a sympatholitic, a beta-adrenergic blocking drug, an
alpha adrenergic blocking drug, a central alpha adrenergic agonist,
a peripheral vasodilator, a lipid lowering agent, and a metabolic
altering agents.
24. A method for treatment or prevention of cardiovascular
diseases, endothelial dysfunction, diastolic dysfunction,
atherosclerosis, hypertension, angina pectoris, thromboses,
restenoses, myocardial infarction, strokes, cardiac insufficiency,
pulmonary hypertonia, erectile dysfunction, asthma bronchiale,
chronic kidney insufficiency, diabetes, or cirrhosis of the liver
in a human or animal patient, said method comprising the step of
administering to the patient an amount efficacious for said
treatment or prevention of a compound of formula I as claimed in
claim 1, a stereoisomeric form thereof, a physiologically
acceptable salt thereof, or a mixture of any two or more of the
foregoing.
Description
BACKGROUND OF THE INVENTION
[0001] Cyclic GMP is an important intracellular messenger which
triggers a multitude of different effects via the modulation of
cGMP-dependent protein kinases, phosphodiesterases and ion
channels. Examples are the relaxation of smooth muscles, the
inhibition of thrombocyte activation and the inhibition of the
proliferation of smooth-muscle cells and of leukocyte adhesion.
cGMP is produced by particulate and soluble guanylate cyclases as a
response to a number of extracellular and intracellular stimuli. In
the case of the particulate guanylate cyclases, stimulation is
essentially effected by peptidic messengers, such as the atrial
natriuretic peptide or the cerebral natriuretic peptide. The
soluble guanylate cyclases ("sGC"), which are cytosolic
heterodimeric heme proteins, in contrast, are essentially regulated
by a family of low-molecular-weight factors which are formed
enzymatically. The most important stimulant is nitrogen monoxide
("NO") or a closely related species. The function of other factors
such as carbon monoxide or the hydroxyl radical is still largely
unclear. The binding of NO to the heme with formation of a
penta-coordinate heme-nitrosyl complex is proposed as the mechanism
of the activation by NO. The associated release of the histidine
which is bound in the basal state to the iron converts the enzyme
into the active conformation.
[0002] Active soluble guanylate cyclases are composed of an .alpha.
and .beta. subunit each. Several subunit subtypes have been
described which differ from one another with respect to sequence,
tissue-specific distribution and expression in different
development stages. The subtypes .alpha..sub.1 and .beta..sub.1 are
mainly expressed in brain and lung, while .beta..sub.2 is found in
particular in liver and kidney. The subtype .alpha..sub.2 was shown
to be present in human fetal brain. The subunits referred to as
.alpha..sub.3 and .beta..sub.3 were isolated from human brain and
are homologous to .alpha..sub.1 and .beta..sub.1. More recent works
indicate an .alpha..sub.2i subunit which contains an insert in the
catalytic domain. All subunits show great homologies in the region
of the catalytic domain. The enzymes presumably contain one heme
per heterodimer, which is bound via .beta..sub.1-Cys-78 and/or
.beta..sub.1-His-105 and is part of the regulatory center.
[0003] Under pathologic conditions, the formation of
guanylate-cyclase-activating factors can be reduced, or their
degradation may be promoted owing to the increased occurrence of
free radicals. The resulting reduced activation of the sGC leads,
via a weakening of the respective cGMP-mediated cellular response,
for example to an increase of the blood pressure, to platelet
activation or to increased cell proliferation and cell adhesion. As
a consequence, formation of endothelial dysfunction,
atherosclerosis, hypertension, stable or unstable angina pectoris,
thromboses, myocardial infarction, strokes or erectile dysfunction
results. Pharmacological stimulation of sGC offers a possibility to
normalize cGMP production and therefore makes possible the
treatment and/or prevention of such disorders.
[0004] For the pharmacological stimulation of the sGC, use has been
made of compounds whose activity is based on an intermediate NO
release, for example organic nitrates. The drawback of this
treatment is the development of tolerance and a reduction of
activity, and the higher dosage which is required because of
this.
[0005] Various sGC stimulators which do not act via NO release were
described by Vesely in a series of publications. However, the
compounds, most of which are hormones, plant hormones, vitamins or
natural compounds such as, for example, lizard poisons
predominantly only have weak effects on the cGMP formation in cell
lysates. D. L. Vesely, Eur. J. Clin. Invest., vol. 15, 1985, p.
258; D. L. Vesely, Biochem. Biophys. Res. Comm., vol. 88, 1979, p.
1244. A stimulation of heme-free guanylate cyclase by
protoporphyrin IX was demonstrated by Ignarro et al., Adv.
Pharmacol., vol. 26, 1994, p. 35. Pettibone et al., Eur. J.
Pharmacol., vol. 116, 1985 p. 307, described an antihypertensive
action of diphenyliodonium hexafluorophosphate and attributed this
to a stimulation of sGC. According to Yu et al., Brit. J.
Pharmacol, vol. 114, 1995, p. 1587, isoliquiritigenin, which has a
relaxing action on isolated rat aortas, also activates sGC. Ko et
al., Blood vol. 84, 1994, p. 4226, Yu et al., Biochem. J. vol. 306,
1995, p. 787, and Wu et al., Brit. J. Pharmacol vol. 116, 1995, p.
1973, demonstrated a sGC-stimulating activity of
1-benzyl-3-(5-hydroxymethyl-2-furyl)indazole and demonstrated an
antiproliferative and thrombocyte-inhibiting action. Pyrazoles and
fused pyrazoles which exhibit a sGC-stimulating activity are
described in European Patent Application No. 908,456 and German
Patent Application No. 19,744,027.
[0006] A series of 2-sulfonylaminobenzoic acid N-arylamides, the
N-aryl group of which carries a thio substituent, have been
mentioned in the literature. These compounds in which the N-aryl
group generally carries as further substituents groups which are
readily oxidizable such as, for example, two hydroxy groups being
in para position with respect to one another and which in this case
can be regarded as hydroquinone derivatives, are auxiliaries for
the preparation of photographic materials (see, for example,
Chemical Abstracts 119, 105757; 120, 41858; 123, 70224; or 126,
257007). British patent publication No. 876,526 (Chemical Abstracts
56, 15432e) discloses 3,5-dichloro-2-methylsulfonylaminobenzoic
acid N-(5-chloro-2-(4-chlorophenylmercapto)-phenyl)-amide which can
be used for the protection of wool against moths.
[0007] It has now been found that the compounds of the present
invention effect a strong activation of guanylate cyclase and are
therefore suitable for the therapy and prophylaxis of disorders
which are associated with a low cGMP level.
SUMMARY OF THE INVENTION
[0008] The present invention relates to compounds which activate
soluble guanylate cyclase which are valuable pharmaceutically
active compounds for the therapy and prophylaxis of diseases, for
example for cardiovascular diseases such as hypertension, angina
pectoris, diabetes, cardiac insufficiency, thromboses or
atherosclerosis. The compounds of the formula I are capable of
modulating the body's production of cyclic guanosine monophosphate
("cGMP") and are generally suitable for the therapy and prophylaxis
of diseases which are associated with a disturbed cGMP balance. The
invention furthermore relates to processes for preparing compounds
of the formula I, to their use for the therapy and prophylaxis of
the above-mentioned diseases and for preparing pharmaceuticals for
this purpose, and to pharmaceutical preparations which comprise
compounds of the formula I.
DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS
[0009] The invention concerns compounds of formula I which activate
soluble guanylate cyclase:
##STR00002## [0010] and pharmaceutically acceptable salts thereof,
wherein [0011] Z.sup.1 is selected from the group consisting of CH
and N; [0012] A is a ring selected from the group consisting of
[0012] ##STR00003## [0013] D.sup.1 is CH, CR.sup.4 or N; [0014]
R.sup.7 is selected from the group consisting of [0015] 1)
hydrogen, [0016] 2) C.sub.1-6 alkyl wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms and
unsubstituted or monosubstituted with OC.sub.1-3 alkyl, [0017] 3)
C.sub.3-6 cycloalkyl wherein the cycloalkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms and
unsubstituted or monosubstituted with OC.sub.1-3 alkyl, and [0018]
4) phenyl, wherein the phenyl group is unsubstituted or substituted
with C.sub.1-4 alkyl, --OC.sub.1-4 alkyl, halogen, CN, NO.sub.2,
and S(O).sub.0-2C.sub.1-4 alkyl, wherein C.sub.1-4 alkyl and
--OC.sub.1-4 alkyl are unsubstituted or substituted with 1-3
flourine atoms; [0019] L.sup.1 is selected from the group
consisting of O, S, C(R.sup.12).sub.2; and CF.sub.2; [0020] L.sup.2
is selected from the group consisting of (CH.sub.2).sub.2-4,
--C(R.sup.12).sub.2, --CF.sub.2--O, and S, provided that when
L.sup.1 is O or S, L.sup.2 is not O or S; [0021] R.sup.12 is
independently selected from the group consisting of hydrogen and
C.sub.1-3 alkyl, wherein C 1-3 alkyl is unsubstituted or
substituted with 1-3 flourine atoms; [0022] E is a ring selected
from the group consisting of [0023] 1) a 6-10 membered aryl ring,
[0024] 2) a 5-10 membered heteroaryl ring having 1, 2 or 3
heteroatoms independently selected from the group consisting of 0,
1, 2, and 3 N atoms, 0 or 1 O atoms, and 0 or 1 S atoms, [0025] 3)
a C.sub.3-8 cycloalkyl ring; [0026] wherein aryl, heteroaryl, and
C.sub.3-8 cycloalkyl are unsubstituted or monosubstituted with
R.sup.5, and unsubstituted, monosubstituted or independently
disubstituted with R.sup.8; [0027] R.sup.4, in each instance in
which it occurs, is independently selected from the group
consisting of halogen, [0028] C.sub.1-6 alkyl, wherein the alkyl
group may be unsubstituted or substituted with 1-3 fluorine atoms,
[0029] --O--C.sub.1-6 alkyl, wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms, [0030]
C.sub.3-8 cycloalkyl, unsubstituted or substituted with 1-3
fluorine atoms, [0031] --O--C.sub.3-8 cycloalkyl, unsubstituted or
substituted with 1-3 fluorine atoms, [0032] CN, and [0033]
NO.sub.2; [0034] R.sup.5, in each instance in which it occurs, is
independently selected from the group consisting of [0035] 1)
R.sup.6, [0036] 2) --OR.sup.6, [0037] 3) C.sub.1-6 alkyl which may
be unsubstituted or substituted with 1-3 fluorine atoms, and
unsubstituted or monosubstituted with a group independently
selected from C.sub.3-6 cycloalkyl, --O--C.sub.1-4 alkyl, OH,
.dbd.O, S(O).sub.0-2C.sub.1-4 alkyl, --OR.sup.6 and R.sup.6, [0038]
4) C.sub.1-6 alkenyl which may be unsubstituted or substituted with
1-3 fluorine atoms and unsubstituted or monosubstituted with a
group independently selected from --O--C.sub.1-4 alkyl, OH, .dbd.O,
S(O).sub.0-2C.sub.1-4 alkyl, --OR.sup.6 and R.sup.6, [0039] 5)
O--C.sub.1-6 alkyl wherein the alkyl group may be unsubstituted or
substituted with 1-3 fluorine atoms, and unsubstituted or
monosubstituted with a group independently selected from C.sub.3-6
cycloalkyl and R.sup.6, [0040] 6) --S--C.sub.1-6 alkyl, [0041] 7) a
C.sub.3-8 cycloalkyl ring which is unsubstituted or mono, di- or
tri-substituted with groups independently selected from fluoro and
C.sub.1-4 alkyl, and unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl wherein the alkyl group
may be unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.1-4 alkyl, OH, .dbd.O, S(O).sub.0-2C.sub.1-4 alkyl,
--OR.sup.6, R.sup.6, and NR.sup.9R.sup.10, [0042] 8) a C.sub.5-8
cycloalkenyl ring which is unsubstituted or mono, di- or
tri-substituted with a group independently selected from fluoro and
C.sub.1-4 alkyl, and unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl, wherein the alkyl
group may be unsubstituted or substituted with 1-3 fluorine atoms,
--O--C.sub.1-4 alkyl, OH, .dbd.O, S(O).sub.0-2C.sub.1-4 alkyl, and
R.sup.6, [0043] 9) a 5- to 6 membered heterocyclyl ring having 1 or
2 heteroatoms selected from the group consisting of N, O and S, and
which is unsubstituted or monosubstituted with a group
independently selected from C.sub.1-4 alkyl wherein the alkyl group
may be unsubstituted or substituted with 1-3 fluorine atoms,
--OC.sub.1-4 alkyl, and .dbd.O, and [0044] 10) halogen; [0045]
R.sup.6 is selected from the group consisting of [0046] 1) a phenyl
ring which is unsubstituted, monosubstituted or disubstituted with
a group independently selected from the group consisting of
halogen, OH, CN, C.sub.1-4 alkyl wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms, OC.sub.1-4
alkyl wherein the alkyl group may be unsubstituted or substituted
with 1-3 fluorine atoms, NO.sub.2, S(O).sub.0-2C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, O--C.sub.2-4 alkenyl, NR.sup.9R.sup.10, and
COOH, and [0047] 2) a 5-6 membered heteroaryl ring containing 1-2
heteroatoms which are independently selected from N, O and S,
wherein the heteroaryl ring is unsubstituted, monosubstituted or
disubstituted with a group independently selected from: halogen,
OH, CN, C.sub.1-4 alkyl wherein the alkyl group may be
unsubstituted or substituted with 1-3 fluorine atoms, OC.sub.1-4
alkyl wherein the alkyl group may be unsubstituted or substituted
with 1-3 fluorine atoms, NO.sub.2, S(O).sub.0-2C.sub.1-6 alkyl,
S(O).sub.0-2 aryl, C.sub.2-6 alkenyl, OC.sub.2-6 alkenyl,
NR.sup.9R.sup.10, and COOH; [0048] R.sup.8 is selected from the
group consisting of [0049] C.sub.1-4 alkyl wherein the alkyl group
may be unsubstituted or substituted with 1-3 fluorine atoms, [0050]
C.sub.2-4 alkenyl, [0051] halogen, [0052] C.sub.3-6 cycloalkyl,
wherein the cycloalkyl group may be unsubstituted or substituted
with 1-3 fluorine atoms [0053] OC.sub.1-4 alkyl wherein the alkyl
group may be unsubstituted or substituted with 1-3 fluorine atoms,
[0054] O--C.sub.2-4 alkenyl, [0055] NO.sub.2, [0056]
S(O).sub.0-2C.sub.1-4 alkyl, and [0057] CN; [0058] R.sup.9 and
R.sup.10 are independently selected from the group consisting of
hydrogen and C.sub.1-6 alkyl; and [0059] R.sup.11 is selected from
the group consisting of hydrogen and C.sub.1-6 alkyl.
[0060] In another embodiment, A is a ring selected from the group
consisting of
##STR00004##
and all other variables are as previously defined.
[0061] In another embodiment, R.sup.11 is hydrogen, and all other
variables are as previously defined.
[0062] In another embodiment,
##STR00005##
is selected from the group consisting of
##STR00006##
wherein E.sup.1 is CH or N, and all other variables are as
previously defined.
[0063] In another embodiment,
##STR00007##
is selected from the group consisting of
##STR00008##
and all other variables are as previously defined.
[0064] In another embodiment, Z.sup.1 is CH, and all other
variables are as previously defined.
[0065] In another embodiment, R.sup.7 is selected from the group
consisting of CH.sub.3, CF.sub.3 and CF.sub.2H, and all other
variables are as previously defined.
[0066] In another embodiment, L.sup.1 is selected from a group
consisting of O and S, and all other variables are as previously
defined.
[0067] In another embodiment, L.sup.2 is selected from the group
consisting of CH.sub.2, CH(CH.sub.3), O, CH.sub.2CH.sub.2, CF.sub.2
and CH.sub.2CH.sub.2CH.sub.2, provided that when L.sup.2 is O
L.sup.1 is not 0, and all other variables are as previously
defined
[0068] In another embodiment, L.sup.2 is selected from the group
consisting of CH.sub.2 and CF.sub.2, and all other variables are as
previously defined.
[0069] In another embodiment, R.sup.4 is selected from the group
consisting of Cl, F, Br, CH.sub.3, cyclopropyl, NO.sub.2, and
CF.sub.3, and all other variables are as previously defined.
[0070] In another embodiment, R.sup.4 is selected from the group
consisting of Cl and CH.sub.3, and all other variables are as
previously defined
[0071] In another embodiment, R.sup.6 is a phenyl ring which is
unsubstituted or mono, di- or tri-substituted with a group
independently selected from the group consisting of Cl, F,
--CH.sub.3, --C(CH.sub.3).sub.3, CF.sub.3, --OCF.sub.3,
--OCH.sub.3, --OCH(CH.sub.3).sub.2 and COOH, and all other
variables are as previously defined.
[0072] In another embodiment, R.sup.5 is selected from the group
consisting of [0073] 1) R.sup.6, [0074] 2) a C.sub.3-6 cycloalkyl
ring which is unsubstituted or mono, di- or tri-substituted with a
group independently selected from phenyl, F, CF.sub.3, CH.sub.3,
OH, and .dbd.O, [0075] 3) a pyridinyl ring, wherein the point of
attachment to the pyridinyl ring is a carbon atom, and wherein the
pyridinyl ring is unsubstituted or mono-substituted with CF.sub.3,
[0076] 4) --CH.sub.2-L.sup.3-R.sup.6, wherein L.sup.3 is
--CH.sub.2-- or --O--, [0077] 5) --OR.sup.6, [0078] 6)
--OCH.sub.2R.sup.6, [0079] 7)
[0079] ##STR00009## [0080] 8) --CF.sub.3, [0081] 9) Cl, F, or Br,
[0082] 10) --CH.sub.3, [0083] 11) OCH.sub.3, [0084] 12) OCF.sub.3,
[0085] 13) --CH.dbd.CHR.sup.6, and [0086] 14) --SCH.sub.2CH.sub.3,
and all other variables are as previously defined.
[0087] In another embodiment, R.sup.5 is selected from the group
consisting of Cl, F, Br, --CH.sub.3, --C(CH.sub.3).sub.3,
OCH.sub.3, OCF.sub.3, --SCH.sub.2CH.sub.3,
##STR00010## ##STR00011## ##STR00012##
and all other variables are as previously defined.
[0088] In another embodiment, R.sup.5 is selected from the group
consisting of --CH.sub.3, -
##STR00013##
and all other variables are as previously defined.
[0089] In another embodiment, R.sup.8 is selected from the group
consisting of CH.sub.3, Cl, F, cyclopropyl, and CF.sub.3.
[0090] In another embodiment, compounds of the invention are
selected from the group consisting of [0091]
1-[6-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluorome-
thyl)-1H-pyrazole-4-carboxylic acid, [0092]
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid, [0093]
5-(Trifluoromethyl)-1-(6-{2-[(4-{2-[4-(trifluoromethyl)phenyl]ethyl}benzy-
l)-oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid, [0094]
1-{6-[2-({4-[(1S,2S)-2-Phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0095]
1-{6-[2-({4-[(1R,2R)-2-Phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0096]
1-[6-(2-{[4-(4-Chlorophenoxy)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid, [0097]
5-(Trifluoromethyl)-1-{6-[2-({4-[4-(trifluoromethyl)phenoxy]benzyl}oxy)ph-
enyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid, [0098]
5-(Trifluoromethyl)-1-(6-{2-[(4-{[4-(trifluoromethyl)phenoxy]methyl}benzy-
l)-oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid, [0099]
1-{6-[5-Methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0100]
1-{6-[5-Methyl-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0101]
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0102]
1-{6-[5-Chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0103]
1-[6-(2-{[4-(4-Oxocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid, [0104]
1-[6-(2-{[4-(4,4-Difluorocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0105]
1-[6-(2-{[4-(trans-4-Methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5--
(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0106]
1-[6-(2-{[4-(cis-4-Methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(t-
rifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0107]
1-[6-(2-{[4-(trans)-4-Methoxycyclohexyl)-2-methylbenzyl]oxy}phenyl)pyridi-
n-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0108]
5-(Trifluoromethyl)-1-{6-[2-({4-[6-(trifluoromethyl)pyridin-3-yl]benzyl}--
oxy)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid, [0109]
1-(6-{2-[(2,4-Dimethylbenzyl)oxy]-3-methylphenyl}pyridin-2-yl)-5-(trifluo-
romethyl)-1H-pyrazole-4-carboxylic acid, [0110] Ethyl
1-{6-[5-chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}-oxy)phe-
nyl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate,
[0111] Ethyl
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}-oxy-
)phenyl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate,
[0112]
5-(Trifluoromethyl)-1-[4-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]me-
thoxy}phenyl)-1,3-thiazol-2-yl]-1H-pyrazole-4-carboxylic acid,
[0113]
1-[2-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyrimidin-4-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylic acid, [0114]
1-{4-Methyl-6-[5-methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl-
}-oxy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid, [0115]
5-(Trifluoromethyl)-1-[6-(2-{2-[4'-(trifluoromethyl)biphenyl-4-yl]ethyl}p-
henyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid, [0116]
5-(Trifluoromethyl)-1-(2'-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-2,-
3'-bipyridin-6-yl)-1H-pyrazole-4-carboxylic acid, [0117]
1-(5'-Methyl-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-2,3-
'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid, [0118]
1-(5'-Chloro-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]metho-
xy}-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid, [0119]
1-[2'-{[3-Methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-5'-(trifluoro-
methyl)-2,3'-bipyridin-6-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid, [0120]
5-(Trifluoromethyl)-1-{6-[2-({[4'-(trifluoromethyl)biphenyl-4-yl]oxy}meth-
yl)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid, [0121]
5-(Trifluoromethyl)-1-(6-{2-[({5-[4-(trifluoromethyl)phenyl]pyridin-2-yl}-
oxy)methyl]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid,
[0122]
1-{6-[5-Methyl-2-({[4'-(trifluoromethyl)biphenyl-4-yl]methyl}thio)phenyl]-
pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0123]
1-[6-(2-{Difluoro[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}phenyl)pyridi-
n-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0124]
1-{6-[2-(Difluoro
{4-[trans-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)phenyl]pyridin-2-y-
l}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0125]
1-{6-[2-(difluoro
{4-[cis-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)phenyl]pyridin-2-yl}-
-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0126]
1-(6-{2-[{2-Ethyl-4-[4
(trifluoromethyl)cyclohexyl]phenyl}(difluoro)methoxy]phenyl}pyridin-2-yl)-
-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid, [0127]
1-{6-[2-(Difluoro
{[4'-(trifluoromethyl)biphenyl-4-yl]oxy}methyl)phenyl]pyridin-2-yl}-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid, and [0128]
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)pyridin-2-yl]-1H-1,2,3-triazole-4-carboxylic acid and
pharmaceutically acceptable salts thereof.
[0129] In another embodiment, wherein R.sup.1 is H, Z.sup.1 is CH,
R.sup.7 is CF.sub.3 or CF.sub.2H, D.sup.1 is CH, L.sup.1 is O,
L.sup.2 is CH.sub.2 or CF.sub.2,
ring A is
##STR00014##
R.sup.5 is
##STR00015##
[0131] In another embodiment, compounds of the invention are
selected from the group consisting of [0132]
1-{6-[5-Methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0133]
1-{6-[5-Methyl-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
[0134]
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid,
and [0135]
1-{6-[5-Chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy-
)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid, and pharmaceutically acceptable salts thereof.
[0136] As used herein except where noted, "alkyl" is intended to
include both branched- and straight-chain saturated aliphatic
hydrocarbon groups having the specified number of carbon atoms. The
term "cycloalkyl" means carbocycles containing no heteroatoms.
Examples of cycloalkyl include cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, decahydronaphthyl and the
like. Commonly used abbreviations for alkyl groups are used
throughout the specification, e.g. methyl may be represented by
conventional abbreviations including "Me" or CH.sub.3 or a symbol
that is an extended bond without defined terminal group, e.g.
##STR00016##
ethyl may be represented by "Et" or CH.sub.2CH.sub.3, propyl may be
represented by "Pr" or CH.sub.2CH.sub.2CH.sub.3, butyl may be
represented by "Bu" or CH.sub.2CH.sub.2CH.sub.2CH.sub.3, etc.
"C.sub.1-6 alkyl" (or "C.sub.1-C.sub.6 alkyl") for example, means
linear or branched chain alkyl groups, including all isomers,
having the specified number of carbon atoms. C.sub.1-6 alkyl
includes all of the hexyl alkyl and pentyl alkyl isomers as well as
n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl.
"C.sub.1-4 alkyl" means n-, iso-, sec- and t-butyl, n- and
isopropyl, ethyl and methyl. If no number is specified, 1-10 carbon
atoms are intended for linear or branched alkyl groups. The phrase
"C.sub.1-6 alkyl, wherein the alkyl group may be unsubstituted or
substituted with 1-3 fluorine atoms" refers to alkyl groups having
0, 1, 2 or 3 fluorine atoms attached to one or more carbon atoms.
The group "CF.sub.3", for example, is a methyl group having three
fluorine atoms attached the same carbon atom.
[0137] "Alkenyl" unless otherwise indicated, means carbon chains
which contain at least one carbon-carbon double bond, and which may
be linear or branched or combinations thereof. Examples of alkenyl
include vinyl, allyl, isopropenyl, pentenyl, hexenyl, heptenyl,
1-propenyl, 2-butenyl, 2-methyl-2-butenyl, and the like. The term
"cycloalkenyl" means carbocycles containing no heteroatoms having
at least one carbon-carbon double bond.
[0138] "Aryl" unless otherwise indicated, means mono- and bicyclic
aromatic rings containing 6-12 carbon atoms. Examples of aryl
include phenyl, naphthyl, indenyl and the like. "Aryl" also
includes monocyclic rings fused to an aryl group. Examples include
tetrahydronaphthyl, indanyl and the like.
[0139] "Heteroaryl" unless otherwise indicated, means a mono- or
bicyclic aromatic ring or ring system containing at least one
heteroatom selected from O, S and N, with each ring containing 5 to
10 atoms. Examples include pyrrolyl, isoxazolyl, isothiazolyl,
pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl,
imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl,
pyrimidyl, pyridazinyl, pyrazinyl, and the like. Heteroaryl also
includes aromatic heterocyclic groups fused to heterocycles that
are non-aromatic or partially aromatic, and aromatic heterocyclic
groups fused to cycloalkyl rings. Heteroaryl also includes such
groups in charged form, e.g., pyridinium.
[0140] "Heterocyclyl", unless otherwise indicated, means a 5- or
6-membered monocyclic saturated ring containing at least one
heteroatom selected from N, S and 0, in which the point of
attachment may be carbon or nitrogen. Examples of "heterocyclyl"
include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl,
imidazolidinyl, 2,3-dihydrofuro(2,3-b)pyridyl, benzoxazinyl, and
the like. The term also includes partially unsaturated monocyclic
rings that are not aromatic, such as 2- or 4-pyridones attached
through the nitrogen or N-substituted-(1H,
3H)-pyrimidine-2,4-diones (N-substituted uracils). Heterocyclyl
moreover includes such moieties in charged form, e.g.,
piperidinium.
[0141] "Halogen" unless otherwise indicated, includes fluorine,
chlorine, bromine and iodine.
[0142] Unless expressly stated to the contrary, substitution by a
named substituent is permitted on any atom in a ring (e.g., aryl, a
heteroaryl ring, or a saturated heterocyclic ring) provided such
ring substitution is chemically allowed and results in a stable
compound. A "stable" compound is a compound which can be prepared
and isolated and whose structure and properties remain or can be
caused to remain essentially unchanged for a period of time
sufficient to allow use of the compound for the purposes described
herein (e.g., therapeutic or prophylactic administration to a
subject).
[0143] In the representation of rings that define variable A,
##STR00017##
the 1,3 substitution of each ring is oriented such that the carbon
atom of ring A numbered "1" is attached to the group
##STR00018##
and the carbon atom of ring A numbered "3" is attached to the
group
##STR00019##
[0144] In one embodiment of A, the same 1,3 substitution pattern is
followed:
##STR00020##
[0145] The present invention includes all stereoisomeric forms of
the compounds of the formula I. Centers of asymmetry that are
present in the compounds of formula I can all independently of one
another have S configuration or R configuration. The invention
includes all possible enantiomers and diastereomers and mixtures of
two or more stereoisomers, for example mixtures of enantiomers
and/or diastereomers, in all ratios. Thus, enantiomers are a
subject of the invention in enantiomerically pure form, both as
levorotatory and as dextrorotatory antipodes, in the form of
racemates and in the form of mixtures of the two enantiomers in all
ratios. In the case of a cis/trans isomerism the invention includes
both the cis form and the trans form as well as mixtures of these
forms in all ratios. The preparation of individual stereoisomers
can be carried out, if desired, by separation of a mixture by
customary methods, for example by chromatography or
crystallization, by the use of stereochemically uniform starting
materials for the synthesis or by stereoselective synthesis.
Optionally a derivatization can be carried out before a separation
of stereoisomers. The separation of a mixture of stereoisomers can
be carried out at the stage of the compounds of the formula I or at
the stage of an intermediate during the synthesis. The present
invention also includes all tautomeric forms of the compounds of
formula I.
[0146] If the compounds of the formula I contain one or more acidic
or basic groups the invention also includes the corresponding
physiologically or toxicologically acceptable salts, in particular
the pharmaceutically utilizable salts. Thus, the compounds of the
formula I which contain acidic groups can be present on these
groups and can be used according to the invention, for example, as
alkali metal salts, alkaline earth metal salts or as ammonium
salts. Examples of such salts are sodium salts, potassium salts,
calcium salts, magnesium salts or salts with ammonia or organic
amines such as, for example, ethylamine, ethanolamine,
triethanolamine or amino acids. Compounds of the formula I which
contain one or more basic groups, i.e. groups which can be
protonated, can be present and can be used according to the
invention in the form of their acid addition salts with inorganic
or organic acids, for example as salts with hydrogen chloride,
hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid,
methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic
acids, oxalic acid, acetic acid, tartaric acid, lactic acid,
salicylic acid, benzoic acid, formic acid, propionic acid, pivalic
acid, diethylacetic acid, malonic acid, succinic acid, pimelic
acid, fumaric acid, maleic acid, malic acid, sulfaminic acid,
phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic
acid, citric acid, adipic acid, etc. If the compounds of the
formula I simultaneously contain acidic and basic groups in the
molecule the invention also includes, in addition to the salt forms
mentioned, inner salts or betaines (zwitterions). Salts can be
obtained from the compounds of the formula I by customary methods
which are known to the person skilled in the art, for example by
combination with an organic or inorganic acid or base in a solvent
or dispersant, or by anion exchange or cation exchange from other
salts. The present invention also includes all salts of the
compounds of the formula I which, owing to low physiological
compatibility, are not directly suitable for use in pharmaceuticals
but which can be used, for example, as intermediates for chemical
reactions or for the preparation of physiologically acceptable
salts.
[0147] The present invention also relates to processes for the
preparation of the compounds of the formula I which are described
in the following and by which the compounds of the invention are
obtainable.
[0148] The compounds of the formula I according to the invention
effect an increase of the cGMP concentration via the activation of
the soluble guanylate cyclase (sGC), and they are therefore useful
agents for the therapy and prophylaxis of disorders which are
associated with a low or decreased cGMP level or which are caused
thereby, or for whose therapy or prophylaxis an increase of the
present cGMP level is desired. The activation of the sGC by the
compounds of the formula I can be examined, for example, in the
activity assay described below.
[0149] Disorders and pathological conditions which are associated
with a low cGMP level or in which an increase of the cGMP level is
desired and for whose therapy and prophylaxis it is possible to use
compounds of the formula I are, for example, cardiovascular
diseases, such as endothelial dysfunction, diastolic dysfunction,
atherosclerosis, hypertension, stable and unstable angina pectoris,
thromboses, restenoses, myocardial infarction, strokes, cardiac
insufficiency or pulmonary hypertonia, or, for example, erectile
dysfunction, asthma bronchiale, chronic kidney insufficiency and
diabetes. Compounds of the formula I can additionally be used in
the therapy of cirrhosis of the liver and also for improving a
restricted memory performance or ability to learn.
[0150] The compounds of the formula I and their physiologically
acceptable salts can be administered to animals, preferably to
mammals, and in particular to humans, as pharmaceuticals by
themselves, in mixtures with one another or in the form of
pharmaceutical preparations. A subject of the present invention
therefore also are the compounds of the formula I and their
physiologically acceptable salts for use as pharmaceuticals, their
use for activating soluble guanylate cyclase, for normalizing a
disturbed cGMP balance and in particular their use in the therapy
and prophylaxis of the abovementioned syndromes as well as their
use for preparing medicaments for these purposes.
[0151] Furthermore, a subject of the present invention are
pharmaceutical preparations (or pharmaceutical compositions) which
comprise as active component an effective dose of at least one
compound of the formula I and/or a physiologically acceptable salt
thereof and a customary pharmaceutically acceptable carrier, i.e.,
one or more pharmaceutically acceptable carrier substances and/or
additives. A subject of the present invention is also those
compounds of the formula I which were already known per se and
which are excluded by disclaimer from the above-defined compounds
of the formula I which are per se a subject of the present
invention, and their physiologically acceptable salt as activators
of soluble guanylate cyclase.
[0152] Thus, a subject of the invention are, for example, said
compound and its physiologically acceptable salts for use as a
pharmaceutical, pharmaceutical preparations which comprise as
active component an effective dose of said compound and/or a
physiologically acceptable salt thereof and a customary
pharmaceutically acceptable carrier, and the uses of said compound
and/or a physiologically acceptable salt thereof in the therapy or
prophylaxis of the abovementioned syndromes as well as their use
for preparing medicaments for these purposes.
[0153] The pharmaceuticals according to the invention can be
administered orally, for example in the form of pills, tablets,
lacquered tablets, sugar-coated tablets, granules, hard and soft
gelatin capsules, aqueous, alcoholic or oily solutions, syrups,
emulsions or suspensions, or rectally, for example in the form of
suppositories. Administration can also be carried out parenterally,
for example subcutaneously, intramuscularly or intravenously in the
form of solutions for injection or infusion. Other suitable
administration forms are, for example, percutaneous or topical
administration, for example in the form of ointments, tinctures,
sprays or transdermal therapeutic systems, or the inhalative
administration in the form of nasal sprays or aerosol mixtures, or,
for example, microcapsules, implants or rods. The preferred
administration form depends, for example, on the disease to be
treated and on its severity.
[0154] The amount of active compound of the formula I and/or its
physiologically acceptable salts in the pharmaceutical preparations
normally is from 0.2 to 200 mg, preferably from 1 to 200 mg, per
dose, but depending on the type of the pharmaceutical preparation
it can also be higher. The pharmaceutical preparations usually
comprise 0.5 to 90 percent by weight of the compounds of the
formula I and/or their physiologically acceptable salts. The
preparation of the pharmaceutical preparations can be carried out
in a manner known per se. For this purpose, one or more compounds
of the formula I and/or their physiologically acceptable salts,
together with one or more solid or liquid pharmaceutical carrier
substances and/or additives (or auxiliary substances) and, if
desired, in combination with other pharmaceutically active
compounds having therapeutic or prophylactic action, are brought
into a suitable administration form or dosage form which can then
be used as a pharmaceutical in human or veterinary medicine.
[0155] For the production of pills, tablets, sugar-coated tablets
and hard gelatin capsules it is possible to use, for example,
lactose, starch, for example maize starch, or starch derivatives,
talc, stearic acid or its salts, etc. Carriers for soft gelatin
capsules and suppositories are, for example, fats, waxes, semisolid
and liquid polyols, natural or hardened oils, etc. Suitable
carriers for the preparation of solutions, for example of solutions
for injection, or of emulsions or syrups are, for example, water,
physiologically sodium chloride solution, alcohols such as ethanol,
glycerol, polyols, sucrose, invert sugar, glucose, mannitol,
vegetable oils, etc. It is also possible to lyophilize the
compounds of the formula I and their physiologically acceptable
salts and to use the resulting lyophilisates, for example, for
preparing preparations for injection or infusion. Suitable carriers
for microcapsules, implants or rods are, for example, copolymers of
glycolic acid and lactic acid.
[0156] Besides the active compounds and carriers, the
pharmaceutical preparations can also contain customary additives,
for example fillers, disintegrants, binders, lubricants, wetting
agents, stabilizers, emulsifiers, dispersants, preservatives,
sweeteners, colorants, flavorings, aromatizers, thickeners,
diluents, buffer substances, solvents, solubilizers, agents for
achieving a depot effect, salts for altering the osmotic pressure,
coating agents or antioxidants.
[0157] The dosage of the active compound of the formula I to be
administered and/or of a physiologically acceptable salt thereof
depends on the individual case and is, as is customary, to be
adapted to the individual circumstances to achieve an optimum
effect. Thus, it depends on the nature and the severity of the
disorder to be treated, and also on the sex, age, weight and
individual responsiveness of the human or animal to be treated, on
the efficacy and duration of action of the compounds used, on
whether the therapy is acute or chronic or prophylactic, or on
whether other active compounds are administered in addition to
compounds of the formula I. In general, a daily dose of
approximately 0.01 to 100 mg/kg, preferably 0.01 to 10 mg/kg, in
particular 0.3 to 5 mg/kg (in each case mg per kg of bodyweight) is
appropriate for administration to an adult weighing approximately
75 kg in order to obtain the desired results. The daily dose can be
administered in a single dose or, in particular when larger amounts
are administered, be divided into several, for example two, three
or four individual doses. In some cases, depending on the
individual response, it may be necessary to deviate upwards or
downwards from the given daily dose.
[0158] The compounds of the formula I activate the soluble
guanylate cyclase. On account of this property, apart from use as
pharmaceutically active compounds in human medicine and veterinary
medicine, they can also be employed as a scientific tool or as aid
for biochemical investigations in which such an effect on guanylate
cyclase is intended, and also for diagnostic purposes, for example
in the in vitro diagnosis of cell samples or tissue samples. The
compounds of the formula I and salts thereof can furthermore be
employed, as already mentioned above, as intermediates for the
preparation of other pharmaceutically active compounds.
[0159] The above-mentioned compounds are also of use in combination
with other pharmacologically active compounds comprising
angiotensin converting enzyme inhibitors (e.g, alacepril,
benazepril, captopril, ceronapril, cilazapril, delapril, enalapril,
enalaprilat, fosinopril, imidapril, lisinopril, moveltipril,
perindopril, quinapril, ramipril, spirapril, temocapril, or
trandolapril), angiotensin II receptor antagonists (e.g., losratan,
valsartan, candesartan, olmesartan, telmesartan) neutral
endopeptidase inhibitors (e.g., thiorphan and phosphoramidon),
aldosterone antagonists, renin inhibitors (e.g. urea derivatives of
di- and tri-peptides (See U.S. Pat. No. 5,116,835), amino acids and
derivatives (U.S. Pat. Nos. 5,095,119 and 5,104,869), amino acid
chains linked by non-peptidic bonds (U.S. Pat. No. 5,114,937), di-
and tri-peptide derivatives (U.S. Pat. No. 5,106,835), peptidyl
amino diols (U.S. Pat. Nos. 5,063,208 and 4,845,079) and peptidyl
beta-aminoacyl aminodiol carbamates (U.S. Pat. No. 5,089,471);
also, a variety of other peptide analogs as disclosed in the
following U.S. Pat. Nos. 5,071,837; 5,064,965; 5,063,207;
5,036,054; 5,036,053; 5,034,512 and 4,894,437, and small molecule
renin inhibitors (including diol sulfonamides and sulfinyls (U.S.
Pat. No. 5,098,924), N-morpholino derivatives (U.S. Pat. No.
5,055,466), N-heterocyclic alcohols (U.S. Pat. No. 4,885,292) and
pyrolimidazolones (U.S. Pat. No. 5,075,451); also, pepstatin
derivatives (U.S. Pat. No. 4,980,283) and fluoro- and
chloro-derivatives of statone-containing peptides (U.S. Pat. No.
5,066,643), enalkrein, RO 42-5892, A 65317, CP 80794, ES 1005, ES
8891, SQ 34017, aliskiren
(2(S),4(S),5(S),7(S)-N-(2-carbamoyl-2-methylpropyl)-5-amino-4-hydroxy-2,7-
-diisopropyl-8-[4-methoxy-3-(3-methoxypropoxy)-phenyl]-octanamid
hemifumarate) SPP600, SPP630 and SPP635), endothelin receptor
antagonists, vasodilators, calcium channel blockers (e.g.,
amlodipine, nifedipine, veraparmil, diltiazem, gallopamil,
niludipine, nimodipins, nicardipine), potassium channel activators
(e.g., nicorandil, pinacidil, cromakalim, minoxidil, aprilkalim,
loprazolam), diuretics (e.g., hydrochlorothiazide), sympatholitics,
beta-adrenergic blocking drugs (e.g., propranolol, atenolol,
bisoprolol, carvedilol, metoprolol, or metoprolol tartate), alpha
adrenergic blocking drugs (e.g., doxazocin, prazocin or alpha
methyldopa) central alpha adrenergic agonists, peripheral
vasodilators (e.g. hydralazine), lipid lowering agents (e.g.,
simvastatin, lovastatin, ezetamibe, atorvastatin, pravastatin),
metabolic altering agents including insulin sensitizing agents and
related compounds (e.g., muraglitazar, glipizide, metformin,
rosiglitazone) or with other drugs beneficial for the prevention or
the treatment of the above-mentioned diseases including
nitroprusside and diazoxide.
[0160] The compounds of formula I can be synthesized in accordance
with the general schemes provided below where Z.sup.1, A, D.sup.1,
L.sup.1, L.sup.2, E, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8
and R.sup.11 are defined as above, taking into account the specific
examples that are provided. Throughout the synthetic schemes,
abbreviations are used with the following meanings unless otherwise
indicated:
TABLE-US-00001 aq, aq. = aqueous BuLi, n-BuLi = n-butyllithium Ar =
aryl DME = 1,2-dimethoxyethane Ac = acetate Bn = benzyl Bu = butyl,
t-Bu = fert-butyl CBZ, Cbz = Benzyloxycarbonyl cPr = cyclopropyl
conc, conc. = concentrated BOC, Boc = t-butyloxycarbonyl DAST =
(diethylamino)sulfur trifluoride DCM = dichloromethane dba =
dibenzylideneacetone; Pd.sub.2dba.sub.3 = DIEA =
diisopropylethylamine tris(dibenzylidineacetone)dipalladium DMAC,
DMA = dimethylacetamide DIAD = diisopropylazodicarboxylate DMSO =
dimethylsulfoxide DMAP = 4-dimethylaminopyridine Et = ethyl DMF =
N,N-dimethylformamide EtOAc = ethyl acetate dppf, DPPF = 1,1'- eq.
= equivalent(s) bis(diphenylphosphino)ferrocene HOAc = acetic acid
DIBAL, DIBAL-H = diisobutylaluminum hydride iPr = isopropyl ESI =
electrospray ionization h, hr = hour EtOH = ethanol IPA, i-PrOH =
isopropanol HPLC = High pressure liquid chromatography MeOH =
methanol LAH = Lithium aluminum hydride Me = methyl LCMS = liquid
chromatography - mass OMs, mesyl = methanesulfonyl spectroscopy NMP
= N-methylpyrrolidinone LHMDS = lithium bis(trimethylsilyl)amide
NMR = nuclear magnetic resonance min, min. = minute Ph = phenyl Py
= pyridyl Pr = propyl Pd/C = palladium on activated carbon THF =
tetrahydrofuran RT, rt = room temperature TBAI = tetrabutylammonium
iodide sat. = saturated TFA = Trifluoroacetic acid Tosyl =
toluenesulfonyl OTf, triflate = trifluoromethanesulfonate; triflic
= trifluoromethanesulfonic TLC = thin layer chromatography; prep
TLC = preparative thin layer chromatography Xantphos =
4,5-bis(diphenylphosphino)-9,9- dimethylxanthene
[0161] Where Z.sup.1=CH, such pyrazole acids and corresponding
esters may be obtained commercially, are known in the literature,
or may readily prepared by those skilled in the art. One such
procedure is shown in Scheme 1, involving reaction of an aryl or
heteroaryl hydrazine 1 with a .beta.-ketoester derivative 2 in
presence of a base such as Et.sub.3N and a solvent such as
acetonitrile at ambient or elevated temperatures to provide
pyrazole 3 (J. Comb. Chem. 2003, 5, 465; Heterocycles 1992, 34,
791).
##STR00021##
[0162] Such aryl and heteroaryl hydrazines 1 may be obtained
commercially, are known in the literature and may be prepared by a
variety of methods by those skilled in the art. One such synthetic
method for forming 2-hydrazinopyridines 1b is shown in Scheme 2,
involving reaction of a 2-chloropyridine derivative 4 with
hydrazine hydrate in refluxing ethanol. Another method also shown
in Scheme 2 involves reaction of 4 with
di-tert-butylhydrazine-1,2-dicarboxylate in presence of metal
catalyst such as Pd.sub.2 dba.sub.3, a ligand such as dppf and a
base such as Cs.sub.2CO.sub.3 in a solvent such as toluene at
elevated temperatures to provide the bis-Boc-hydrazinopyridine 5,
followed by deprotection in an acidic solution such as
dioxane/conc. HCl to provide 2-hydrazinopyridine 1b (Org. Lett.
2001, 3 (9), 1351-1354).
##STR00022##
[0163] Outlined in Scheme 3, thiazolylpyrazoles 8 may readily be
prepared by reaction of thiosemicarbazide with a .beta.-ketoester 2
in a solvent such as EtOH to form the intermediate
thioamidopyrazoline 6, followed by reaction with an
.alpha.-bromoketone such as 7 in a solvent such as EtOH at elevated
temperatures to provide the thiazolylpyrazole 8 (J. Comb. Chem.
2002, 4, 23).
##STR00023##
[0164] Where Z.sup.1=N, such triazoles 11 may be prepared as
outlined in Scheme 4, involving reaction of an aryl or heteroaryl
azide 9 with a .beta.-keto ester 10 in an appropriate solvent such
as MeCN in the presence of a base such as sodium ethoxide or
Et.sub.3N at elevated temperatures (J. Med. Chem. 1990, 33 (9),
2646; U.S. Pat. No. 4,474,599). Such azides 9 may be obtained
commercially, are known in the literature and may be obtained by
various methods by those skilled in the art. One such method is
also shown in Scheme 4, involving reaction of hydrazine 1 with
NaNO.sub.2 in an appropriate acidic solvent such as a mixed solvent
of diethyl ether and conc. HCl (U.S. Pat. No. 4,474,599).
##STR00024##
[0165] When desired, an appropriate phenyl or pyridyl ring may be
attached to ring A to provide compounds 14 using various approaches
by those skilled in the art. One such method is shown in Scheme 5,
involving a Suzuki cross coupling reaction between an appropriately
substituted intermediate 12 (Y=Cl, Br, I, OTf) and an aryl- or
pyridylboronic acid 13, utilizing a catalyst such as dichloro
bis(triphenylphosphine)palladium(II) and a base such as aqueous
sodium carbonate in an appropriate solvent such as acetonitrile,
often at elevated temperatures (Heterocycles, 2003, 60, 1891).
Conversely, 12 (Y=Cl, Br, I) can be converted to the boronate ester
15 by reaction with bis(pinacolato)diboron using a catalyst such as
Pd(dppf)Cl.sub.2 in presence of a base such as potassium acetate
and an appropriate solvent such as DMSO at elevated temperatures
(J. Org. Chem. 1995, 60, 7508), or employing a catalyst such as
bis(tricyclohexylphosphine)palladium(0) and a base such as sodium
carbonate in a solvent such as acetonitrile (Tetrahedron, 2001, 57,
9813). The resultant boronate ester can then be cross-coupled to an
appropriately substituted aryl or heteroaryl ring 16 (Y=Cl, Br, I,
OTf) using Suzuki coupling conditions, as described above, to
provide compound 14.
##STR00025##
[0166] The compounds may further be elaborated by methods known to
those skilled in the art. These manipulations may include, but are
not limited to, substitution, reduction, oxidation, alkylation,
acylation, and hydrolysis reactions. One such example for compounds
wherein L.sup.1.dbd.O is shown in Scheme 6, and involves alkylation
of a phenol or hydroxypyridine 17 (as will be known to those
skilled in the art, the hydroxypyridine can also exist in the
tautomeric pyridone form, but is shown as the hydroxypyridine
throughout for simplicity) with an alkyl or benzyl halide
Y-L.sup.2-E (Y=Cl, Br) in presence of a base such as
K.sub.2CO.sub.3 or Cs.sub.2CO.sub.3, typically in a polar solvent
such as DMF at ambient or slightly elevated temperatures to afford
ether 18. Such ethers 18 may also be formed using Mitsunobu
conditions, involving reaction of 17 with an alkyl or benzyl
alcohol E-L.sup.2-OH, typically in an aprotic solvent such as DCM
or THF, in presence of a phosphine such as triphenylphosphine and
an azodicarbonyl reagent such as diisopropyl azodicarboxylate
(Synthesis 1981, p. 1).
##STR00026##
[0167] Shown in Scheme 7, when D.sup.1=N, compounds 18b can also be
formed by reaction of an appropriately reactive intermediate, such
as, for example, fluoropyridine 19 with an alkyl or benzyl alcohol
HO-L.sup.2-E in the presence of a strong base such as NaOtBu in a
polar solvent such as DMF.
##STR00027##
[0168] In cases where the phenyl or pyridyl ring is appropriately
substituted, the compounds may be modified using cross coupling
conditions. One such example shown in Scheme 8, in which an aryl
triflate 20 is reacted with the corresponding alkyne ECCH in the
presence of copper (I) iodide,
dichorobis(triphenylphosphine)palladium(II), tetrabutylammonium
iodide and Et.sub.3N in acetonitrile at ambient temperature to
afford the alkyne 21 (Tetrahedron Lett. 2001, p. 5275). Reduction
of the triple bond by hydrogenation using a metal catalyst such as
PtO.sub.2 in a solvent such as EtOAc under a hydrogen atmosphere
provides ethylene derivative 22.
##STR00028##
[0169] Compounds wherein L.sup.1=S can also be obtained from
triflate 20 as shown in Scheme 9. Reaction with 4-methoxy
.alpha.-toluenethiol in presence of a metal catalyst such as
Pd.sub.2 dba.sub.3, a ligand such as Xantphos and a base such as
DIEA in an appropriate solvent such as dioxane at elevated
temperatures for 15 h provides the methoxybenzyl thioether 23
(Organic Letters 2004, 6 (24), 4587). Removal of the benzyl group
in an acidic solvent such as TFA in presence of a trapping agent
such as anisole provides the thiol 24, which is sometimes
accompanied by the corresponding disulfide dimer. Alkylation with
the desired alkyl or benzyl halide Y-L.sup.2-E (Y=Cl, Br) in
presence of a base such as or Cs.sub.2CO.sub.3, typically in a
polar solvent such as DMF, provides the thioether 25. In instances
where disulfide is present, addition of a reductant such as
NaBH.sub.4 to the reaction can aid in improving the product yield
by converting the disulfide to the thiol in situ.
##STR00029##
[0170] One method for obtaining compounds wherein L.sup.1=CH.sub.2
and L.sup.2=0 is shown in Scheme 10, involving the Suzuki
cross-coupling of an appropriately substituted compound 12 (Y=I,
Br, Cl, OTf) with an appropriate boronate such as 26 to provide the
hydroxymethyl analog 27. Such compounds can be further modified if
desired, for example, by reaction with aromatic and heteroaromatic
species E-OH using Mitsunobu coupling conditions as described above
(vide supra) to provide compounds 28.
##STR00030##
[0171] In some instances, further modification of the compounds
thus described may be desired. One such example is shown in Scheme
11, whereby treatment with of compound 17 with a chlorinating agent
such as benzyltrimethylammonium tetrachloroiodate in DCM at ambient
temperature for 24 h provides predominantly the para-chloro
derivative 29. Conversely, also shown in Scheme 11, treatment of 17
with iodine and silver sulfate in a solvent such as EtOH affords a
mixture of the para-iodo compound 30 and the ortho-iodo isomer 31.
If desired, the iodine may be further modified by a variety of
methods by those skilled in the art. These transformations include,
but are not limited to, cross-coupling reactions, cyanation
reactions, halogen exchange reactions and carbonylation reactions.
The phenol group may then be further modified as described above
(vide supra).
##STR00031##
[0172] In some instances further modification of ring E may be
desired. An example of such a transformation is depicted in Scheme
12, wherein ring E of compound 32 is an aryl or heteroaryl ring.
Such compounds may be alkylated with RY (R=alkyl, benzyl; Y=Br, Cl,
I, OMs, OTosyl) under basic conditions, or with ROH (R=alkyl,
benzyl) using Mitsunobu coupling conditions to provide ether 33, as
described previously (vide supra).
##STR00032##
[0173] The E ring may also be modified via cross coupling
reactions. For example, as shown in Scheme 13, when E is an
aromatic or heteroaromatic ring, compounds 34 (Y=Br, Cl, I, OTf)
may be coupled to alkyl, alkenyl, heteroaryl and aryl boronic acids
R.sup.5--B(OH).sub.2 utilizing Suzuki cross-coupling conditions to
provide products 35, as described above (vide supra). Aryl halides
may also be coupled with heteroatomic species such as phenols using
the procedures of Ullman (Org. Lett. 2002, p. 1623), involving
reaction in presence of a catalyst such as CuCl, a ligand such as
2,2,6,6-tetramethyl-3,5-heptane dione, and a base such as
Cs.sub.2CO.sub.3 in an appropriate solvent such as
N-methylpyrrolidinone at elevated temperatures to provide the aryl
ethers 36. Alternatively, compounds 34 (Y=Br, I) may be converted
to the corresponding boronate esters 37, then cross coupled to an
appropriate R.sup.5--Y (Y=Cl, Br, I, OTf) under Suzuki coupling
conditions, as described previously, to provide compounds 35 (vide
supra).
##STR00033##
[0174] As will be understood by those skilled in the art, the
compounds thus described may be further modified by a variety of
chemical reactions including, but not limited to, substitution,
reduction, oxidation, alkylation, acylation, cross-coupling and
hydrolysis reactions.
[0175] Shown in Scheme 14, when R.sup.11 is an alkyl group, such
pyrazole and triazole esters 38a, as well as synthetic
intermediates, may readily be converted to the corresponding
carboxylic acids using methods known to those skilled in the art.
For example, saponification of esters 38a may be achieved using a
base such as aqueous lithium- or sodium hydroxide in a polar
solvent such as tetrahydrofuran, dioxane, methanol, ethanol or a
mixture of similar solvents to provide the corresponding carboxylic
acids 38b. In addition, when R.sup.11 is a tert-butyl group, such
esters may be conveniently converted to the carboxylic acids 38b by
treatment with an acid such as trifluoroacetic acid, commonly as a
1:1 mixture with methylene chloride, for 0.5 to 8 h at ambient
temperature.
##STR00034##
[0176] As will be known to those skilled in the art, in all
schemes, the product I and all synthetic intermediates may be
purified from unwanted side products, reagents and solvents by
recrystallization, trituration, preparative thin layer
chromatography, flash chromatography on silica gel as described by
W. C. Still et al, J. Org. Chem. 1978, 43, 2923, or reverse-phase
HPLC. Compounds purified by HPLC may be isolated as the
corresponding salt.
[0177] Additionally, in some instances final compound I and
synthetic intermediates may be comprised of a mixture of cis and
trans isomers, enantiomers or diastereomers. As will be known to
those skilled in the art, such cis and trans isomers, enantiomers
and diastereomers may be separated by various methods including
crystallization, chromatography using a homochiral stationary phase
and, in the case of cis/trans isomers and diastereomers,
normal-phase and reverse-phase chromatography.
[0178] The following examples of compounds of the formula I and of
intermediates for their preparation illustrate the invention
without limiting it.
[0179] Chemical reactions were monitored by LCMS, and the purity
and identity of the reaction products were assayed by LCMS
(electrospray ionization) and NMR. .sup.1H NMR spectra are
internally referenced to residual protio solvent signals. Data for
.sup.1H NMR are reported with chemical shift (.delta. ppm),
multiplicity (s=singlet, d=doublet, t=triplet, q=quartet,
m=multiplet), coupling constant (Hz), and integration.
[0180] Preparative HPLC was performed on either a YMC-Pack Pro C18
column (150.times.20 mm i.d.) or a Kromasil 100-10C8 column
(100.times.30 mm i.d.) at an initial flow rate of 4 mL/min for 1.35
min, followed by 20 mL/min for 13.6 min. The gradients employed
during the faster part of the run are described, and all runs were
followed with 100% organic at 20 mL/min for 0.5 min.
[0181] Reactions with a sunlamp used a Fisher 120V, 3A lamp with a
250W bulb.
[0182] Flash chromatography on silica gel was performed using
pre-packed silica gel columns on Biotage Horizon or Biotage SP-1
instruments equipped with UV detectors.
[0183] The following examples are provided so that the invention
might be more fully understood. They should not be construed as
limiting the invention in any way.
Example 1
##STR00035##
[0184] Step A.
Ethyl-1-(6-chloropyridin-2-yl)-5-trifluoromethyl-1H-pyrazole-4-carboxylat-
e
[0185] To a solution 2-chloro-6-hydrazinopyridine (1.00 g, 6.97
mmol) and triethylamine (0.971 mL, 6.97 mmol) in acetonitrile (35
mL) was added ethyl
2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutyrate (1.36 mL, 6.97
mmol). After 20 min, the reaction mixture was placed in a
60.degree. C. oil bath. After 30 min, the reaction mixture was
allowed to cool to ambient temperature, then was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 30%
EtOAc in hexanes, then 30 to 100% EtOAc in hexanes) gave the title
compound: LCMS m/z 319.9 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.10 (s, 1H), 7.88 (t, J=7.5 Hz, 1H), 7.58 (d,
J=8.0 Hz, 1H), 7.47 (d, J=8.0 Hz, 1H), 4.38 (q, J=7.0 Hz, 2H), 1.38
(t, J=7.0 Hz, 3H).
Step B. Ethyl
1-[6-(2-hydroxylphenyl)pyridine-2-yl]-5-trifluoromethyl-1H-pyrazole-4-car-
boxylate
[0186] To a flask containing the title compound from the Example 1
Step A (500 mg, 1.56 mmol) were added 2-hydroxyphenylboronic acid
(237 mg, 1.72 mmol) and
trans-dichlorobis(triphenylphosphine)palladium (II) (112 mg, 0.16
mmol). Acetonitrile (4 mL) and sodium carbonate (3.9 mL, 1.0 M
aqueous, 3.9 mmol) were added, and the resulting mixture was
degassed via nitrogen sparge. The reaction mixture was stirred at
70.degree. C. for 3 h, then was allowed to cool to ambient
temperature and was poured into water. The mixture was extracted
with EtOAc, and the organic phase was concentrated in vacuo.
Purification by chromatography on silica gel (0 to 30% EtOAc in
hexanes, then 30 to 100% EtOAc in hexanes) provided the title
compound: LCMS m/z 378.5 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 12.02 (s, 1H), 8.18 (s, 1H), 8.09-8.04 (m, 2H),
7.82 (dd, J=8.0, 1.5 Hz, 1H), 7.50 (dd, J=7.5, 1.5 Hz, 1H),
7.38-7.34 (m, 1H), 7.06-7.03 (m, 1H), 6.99-6.95 (m, 1H), 4.40 (q,
J=7.0 Hz, 2H), 1.40 (t, J=7.0 Hz, 3H).
Step C. Ethyl
1-[6-(2-{[4-(2-phenylethyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluorome-
thyl)-1H-pyrazole-4-carboxylate
[0187] A vial was charged with the title compound from Example 1
Step B (36.0 mg, 0.095 mmol), cesium carbonate (62.2 mg, 0.191
mmol), and 4-chloromethyl dibenzyl (33.0 mg, 0.143 mmol). DMF (0.5
mL) was added, and the resulting suspension was stirred vigorously.
After 2 h, the reaction mixture was diluted with EtOAc and washed
with brine. The organic phase was separated and concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 25%
EtOAc in hexanes, then 25 to 100% EtOAc in hexanes) provided the
title compound: LCMS m/z 572.5 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.16 (d, J=8.0 Hz, 1H), 8.12 (s, 1H), 7.98 (dd,
J=7.5, 1.5 Hz, 1H), 7.86 (t, J=7.5 Hz, 1H), 7.53 (d, J=7.0 Hz, 1H),
7.39-7.36 (m, 1H), 7.29-7.26 (m, 4H), 7.22-7.17 (m, 5H), 7.11 (t,
J=7.5 Hz, 1H), 7.07 (d, J=8.5 Hz, 1H), 5.13 (s, 2H), 4.39 (q, J=7.0
Hz, 2H), 2.93 (app s, 4H), 1.39 (t, J=7.0 Hz, 3H).
Step D.
1-[6-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trif-
luoromethyl)-1H-pyrazole-4-carboxylic acid
[0188] To a solution of the title compound from Example 1 Step C
(27.0 mg, 0.048 mmol) in 1,4-dioxane (2 mL) was added lithium
hydroxide (1.0 mL, 2.0 M aqueous, 2.0 mmol), and the resulting
mixture was stirred at 60.degree. C. After 15 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (30 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 544.4 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.25 (s, 1H), 8.14-8.07 (m, 2H), 7.74 (d,
J=7.5 Hz, 1H), 7.69 (d, J=7.5 Hz, 1H), 7.42 (t, J=7.5 Hz, 1H), 7.32
(d, J=8.0 Hz, 1H), 7.27-7.14 (m, 8H), 7.09 (t, J=8.0 Hz, 1H), 5.19
(s, 2H), 2.86 (app s, 4H).
Example 2
##STR00036##
[0189] Step A. Ethyl
1-(6-{2-[(4-bromobenzyl)oxy]phenyl}pyridin-2-yl)-5-(trifluoromethyl)-1H-p-
yrazole-4-carboxylate
[0190] To a solution of the title compound from Example 1 Step B
(682 mg, 1.81 mmol) in DMF (10 mL) were added 4-bromobenzyl bromide
(678 mg, 2.71 mmol) and cesium carbonate (1.77 g, 5.42 mmol). After
1.5 h, the reaction mixture was poured into sat. aq. NH.sub.4Cl and
extracted with EtOAc. The organic phase was separated, dried over
sodium sulfate, filtered, and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 25% EtOAc in hexanes,
then 25 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 548.0 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.13 (s, 1H), 8.08 (d, J=8.0 Hz, 1H), 7.95 (dd, J=7.5, 1.5 Hz, 1H),
7.88 (t, J=8.0 Hz, 1H), 7.54 (d, J=8.0 Hz, 1H), 7.48 (d, J=8.0 Hz,
2H), 7.39-7.36 (m, 1H), 7.23 (d, J=8.0 Hz, 2H), 7.12 (t, J=7.5 Hz,
1H), 7.03 (d, J=7.5 Hz, 1H), 5.10 (s, 2H), 4.39 (q, J=7.0 Hz, 2H),
1.39 (t, J=7.0 Hz, 3H).
Step B. Ethyl
5-(trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}p-
henyl)pyridin-2-yl]-1H-pyrazole-4-carboxylate
[0191] To a flask containing the title compound from Example 2 Step
A (40.0 mg, 0.073 mmol) were added 4-trifluoromethylphenyl boronic
acid (21.0 mg, 0.110 mmol) and
dichlorobis(triphenylphosphine)palladium(II) (2.6 mg, 0.004 mmol).
Degassed acetonitrile (0.5 mL) and sodium carbonate (0.183 mL, 1.0
M aqueous, 0.183 mmol) were added, and the reaction mixture was
stirred at 70.degree. C. After 1.5 h, the reaction mixture was
allowed to cool to ambient temperature, then was filtered through a
short plug of silica gel, eluting with DCM. The Suzuki product was
used in the subsequent step without further purification: LCMS m/z
612.2 [M+H].sup.+.
Step C.
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]me-
thoxy}phenyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid
[0192] To a solution of the title compound from Example 2 Step B
(ca. 0.073 mmol) in 1,4-dioxane (0.5 mL) was added lithium
hydroxide (0.50 mL, 2.0 M in water, 1.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 2 h, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (50 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 584.1 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.30 (s, 1H), 8.18 (d, J=8.0 Hz, 1H), 8.14
(t, J=8.0 Hz, 1H), 7.88 (d, J=8.0 Hz, 2H), 7.80 (d, J=8.0 Hz, 2H),
7.75-7.71 (m, 4H), 7.56 (d, J=8.0 Hz, 2H), 7.46-7.43 (m, 1H), 7.30
(d, J=7.5 Hz, 1H), 7.11 (t, J=7.5 Hz, 1H), 5.32 (s, 2H).
Example 3
##STR00037##
[0193] Step A. Ethyl
5-(trifluoromethyl)-1-(6-{2-[(4-{(E)-2-[4-(trifluoromethyl)phenyl]vinyl}b-
enzyl)oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylate
[0194] To a vial containing the title compound from Example 2 Step
A (50.0 mg, 0.092 mmol) were added 2-(4-trifluoromethylphenyl)vinyl
boronic acid (29.6 mg, 0.137 mmol) and
trans-dichlorobis(triphenylphosphine) palladium (II) (6.4 mg, 0.009
mmol). Acetonitrile (0.400 mL) and sodium carbonate (0.229 mL, 1.0
M aqueous, 0.229 mmol) were added, and the resulting mixture was
degassed via nitrogen sparge. The reaction vial was capped and
placed in a pre-heated oil bath (70.degree. C.). After 18 h, the
reaction mixture was allowed to cool to ambient temperature and was
poured into water. The mixture was extracted with DCM, and the
organic phase was concentrated in vacuo. Purification by
chromatography on silica gel (0 to 30% EtOAc in hexanes, then 30 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 638.4
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.16 (d,
J=8.0 Hz, 1H), 8.13 (s, 1H), 7.97 (d, J=7.5 Hz, 1H), 7.88 (t, J=7.5
Hz, 1H), 7.61 (br s, 4H), 7.54 (d, J=8.0 Hz, 1H), 7.52 (d, J=8.5
Hz, 2H), 7.41-7.39 (m, 1H), 7.37 (d, J=7.5 Hz, 1H), 7.21-7.06 (m,
4H), 5.18 (s, 2H), 4.38 (q, J=7.0 Hz, 2H), 1.39 (t, J=7.0 Hz,
3H).
Step B.
5-(Trifluoromethyl)-1-(6-{2-[(4-{2-[4-(trifluoromethyl)phenyl]ethy-
l}benzyl)oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid
[0195] To a degassed solution of the title compound from Example 3
Step A (24.0 mg, 0.038 mmol) in EtOAc (2 mL) was added platinum(IV)
oxide (8.0 mg). The reaction mixture was fitted with a hydrogen
balloon attached to a 3-way adapter. The reaction flask was then
evacuated and back-filled with hydrogen. After this process was
repeated three times, the reaction mixture was placed under a
hydrogen atmosphere and was stirred vigorously. After 45 min, the
reaction mixture was filtered through Celite, rinsing with EtOAc.
The mixture was concentrated in vacuo and used without further
purification: LCMS m/z 640.6 [M+H].sup.+. To a solution of the
hydrogenation product in 1,4-dioxane (2 mL) was added lithium
hydroxide (1.0 mL, 2.0 M in water, 2.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 30 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (50 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 612.5 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.12 (d, J=8.0 Hz, 1H), 8.11 (s, 1H), 8.08
(t, J=8.0 Hz, 1H), 7.74 (dd, J=8.0, 1.5 Hz, 1H), 7.69 (d, J=7.5 Hz,
1H), 7.60 (d, J=8.5 Hz, 2H), 7.43 (d, J=8.5 Hz, 2H), 7.33 (d, J=8.0
Hz, 2H), 7.26 (d, J=8.5 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.09 (t,
J=8.5 Hz, 1H), 5.20 (s, 2H), 2.97-2.94 (m, 2H), 2.91-2.88 (m,
2H).
Example 4
##STR00038##
[0196] Step A.
1-{6-[2-({4-[(1S,2S)-2-Phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid and
1-{6-[2-({4-[(1R,2R)-2-phenylcyclopropyl]benzyl}oxy)phenyl]pyridin-2-yl}--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0197] A vial was charged with the title compound from Example 2
Step A (600 mg, 1.10 mmol), racemic
trans-2-phenylcyclopropylboronic acid (356 mg, 2.20 mmol), and
tribasic potassium phosphate (769 mg, 3.62 mmol). The flask was
flushed with nitrogen, then toluene (5.00 mL) and water (0.198 mL,
10.98 mmol) were added.
[0198] Tetrakis(triphenylphosphine)palladium(0) (127 mg, 0.110
mmol) was added, and the reaction was capped, placed in a
pre-heated oil bath (100.degree. C.), and stirred vigorously. After
18 h, the reaction mixture was allowed to cool to ambient
temperature, then was purified by flash chromatography on silica
gel (0 to 20% EtOAc in hexanes, then 20 to 100% EtOAc in hexanes)
to provide Suzuki product: LCMS m/z 556.2 [M+H].sup.+. The
enantiomers were separated via preparative chiral HPLC (IA column,
30% IPA in heptane, 9 mL/min flow rate: first eluting enantiomer
t.sub.r=15.03 min; second eluting enantiomer t.sub.r=22.83 min. The
enantiopure ethyl esters were saponified separately with LiOH (1.5
mL, 2.0 M aqueous, 3.0 mmol) in dioxane (4 mL) at 50.degree. C.
After 1 h, the reaction mixtures were rendered acidic by addition
of 2 N HCl, then were diluted with 1,4-dioxane and DMF, and
purified by reversed phase HPLC (50 to 100% acetonitrile in water,
both 0.1% v/v with TFA) to provide the title compounds: LCMS m/z
544.4 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 8.29
(s, 1H), 8.15-8.09 (m, 2H), 7.73 (dd, J=7.5, 1.5 Hz, 1H), 7.70 (d,
J=7.0 Hz, 1H), 7.44-7.40 (m, 1H), 7.33 (d, J=8.0 Hz, 2H), 7.28-7.24
(m, 3H), 7.16-7.15 (m, 5H), 5.20 (s, 2H), 2.17 (t, J=7.0 Hz, 2H),
1.46-1.43 (m, 2H).
Example 5
##STR00039##
[0199] Step A. Ethyl
1-(6-{2-[(4-iodobenzyl)oxy]phenyl}pyridin-2-yl)-5-(trifluoromethyl)-1H-py-
razole-4-carboxylate
[0200] To a solution of the title compound from Example 1 Step B
(304 mg, 0.81 mmol) in DMF (2.7 mL) were added 4-iodobenzyl bromide
(359 mg, 1.21 mmol) and cesium carbonate (788 mg, 2.42 mmol). After
12 h, the reaction mixture was poured into sat. aq. NH.sub.4Cl and
extracted with EtOAc. The organic phase was separated, dried over
sodium sulfate, filtered, and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 25% EtOAc in hexanes,
then 25 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 594.4 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.13 (s, 1H), 8.08 (d, J=8.0 Hz, 1H), 7.94 (dd, J=7.5, 1.5 Hz, 1H),
7.88 (t, J=8.0 Hz, 1H), 7.68 (d, J=8.0 Hz, 2H), 7.54 (d, J=8.0 Hz,
1H), 7.39-7.35 (m, 1H), 7.14-7.12 (m, 1H), 7.11 (d, J=8.0 Hz, 2H),
7.02 (d, J=8.0 Hz, 1H), 5.09 (s, 2H), 4.39 (q, J=7.0 Hz, 2H), 1.39
(t, J=7.0 Hz, 3H).
Step B.
1-[6-(2-{[4-(4-Chlorophenoxy)benzyl]oxy}phenyl)pyridin-2-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0201] A vial was charged with copper (I) chloride (1.7 mg, 0.017
mmol), cesium carbonate (55.0 mg, 0.169 mmol), 4-chlorophenol (21.7
mg, 0.169 mmol), and the title compound from Example 5 Step A (50.0
mg, 0.084 mmol). 2,2,6,6-Tetramethyl-3,5-dione (0.007 mL, 0.034
mmol) was added and the mixture was flushed with nitrogen. Degassed
N-methylpyrrolidinone (0.170 mL) was added, and the vial was capped
and placed in a pre-heated oil bath (120.degree. C.). After 15 h,
the mixture was allowed to cool to ambient temperature, then was
filtered through a short plug of silica gel with DCM and
concentrated in vacuo: LCMS m/z 594.3 [M+H].sup.+. To a solution of
the unpurified coupling product in 1,4-dioxane (1 mL) was added
lithium hydroxide (0.5 mL, 2.0 M in water, 1.00 mmol), and the
resulting mixture was stirred at 50.degree. C. After 30 min, the
reaction mixture was rendered acidic by addition of aqueous
hydrochloric acid, then was diluted with dioxane and passed through
a 0.45 micron syringe filter. Purification by reverse phase HPLC
(50 to 100% acetonitrile in water, each with 0.1% v/v TFA) provided
the title compound: LCMS m/z 564.4 [M-H].sup.-; .sup.1H NMR (500
MHz, d.sub.6-DMSO) .delta. 8.27 (s, 1H), 8.15-8.09 (m, 2H), 7.74
(dd, J=8.0, 2.0 Hz, 1H), 7.70 (dd, J=7.5, 1.0 Hz, 1H), 7.47-7.42
(m, 4H), 7.29 (d, J=8.0 Hz, 2H), 7.12-7.09 (m, 2H), 7.04-7.00 (m,
3H), 5.22 (s, 2H).
Example 6
##STR00040##
[0202] Step A. Methyl 4-[4-(trifluoromethyl)phenoxy]benzoate
[0203] A flask was charged with methyl 4-hydroxybenzoate (500 mg,
3.29 mmol), copper (II) acetate (895 mg, 4.93 mmol),
4-trifluoromethylphenylboronic acid (2.50 g, 13.15 mmol), and 4
angstrom molecular sieves (500 mg). Dichloromethane (33 mL) and
triethylamine (1.83 mL, 13.15 mmol) were added, and the reaction
mixture was stirred rapidly, open to air. After 48 h, the reaction
mixture was filtered and concentrated in vacuo. Purification by
flash chromatography on silica gel (0 to 10% EtOAc in hexanes, then
10 to 100% EtOAc in hexanes) provided the title compound: LCMS m/z
297.5 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.07-8.05 (m, 2H), 7.63 (d, J=8.5 Hz, 2H), 7.12 (d, J=8.5 Hz, 2H),
7.06-7.04 (m, 2H), 3.92 (s, 3H).
Step B. 4-[4-(Trifluoromethyl)phenoxy]phenyl}methanol
[0204] To a cooled (-78.degree. C.) solution of the title compound
from Example 6 Step A (325 mg, 1.10 mmol) in THF (6 mL) was added
DIBAL-H (2.2 mL, 1.50 M in heptane, 3.29 mmol). After 30 min, the
reaction mixture was transferred to a 0.degree. C. bath and was
held at this temperature for 45 min, whereupon it was quenched by
addition of MeOH (0.5 mL). The resulting mixture was diluted with
ether and saturated aqueous sodium/potassium tartrate, and the
mixture was stirred vigorously until a clear phase separation was
achieved. The organic phase was then separated, dried over
anhydrous sodium sulfate, and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 60% EtOAc in hexanes,
then 60 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 251.6 [M-OH].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.57 (d, J=9.0 Hz, 2H), 7.40 (d, J=8.0 Hz, 1H), 7.06-7.03 (m, 4H),
4.71 (s, 2H).
Step C.
5-(Trifluoromethyl)-1-{6-[2-({4-[4-(trifluoromethyl)phenoxy]benzyl-
}oxy)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid
[0205] To a solution of the title compound from Example 1 Step B
(107 mg, 0.40 mmol), the title compound from Example 6 Step B (75.0
mg, 0.20 mmol), and triphenylphosphine (104 mg, 0.40 mmol) in DCM
(1 mL) was added diisopropyl azodicarboxylate (0.077 mL, 0.40 mmol)
and the resulting mixture was stirred at ambient temperature. After
18 h, the reaction mixture was concentrated in vacuo. Filtration
through a silica gel plug with DCM provided the title compound:
LCMS m/z 628.1 [M+H].sup.+. To a solution of the Mitsunobu product
in 1,4-dioxane (2 mL) was added lithium hydroxide (1.0 mL, 2.0 M
aqueous, 2.00 mmol), and the resulting mixture was stirred at
50.degree. C. After 30 min, the reaction mixture was rendered
acidic by addition of aqueous hydrochloric acid, then was diluted
with dioxane and passed through a 0.45 micron syringe filter.
Purification by reverse phase HPLC (50 to 100% acetonitrile in
water, each with 0.1% v/v TFA) provided the title compound: LCMS
m/z 598.2 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta.
8.28 (s, 1H), 8.16-8.10 (m, 3H), 7.75-7.70 (m, 4H), 7.51 (d, J=8.5
Hz, 2H), 7.47-7.44 (m, 1H), 7.30 (d, J=8.5 Hz, 1H), 7.15-7.10 (m,
5H), 5.25 (s, 2H).
Example 7
##STR00041##
[0206] Step A. Ethyl
1-[6-(2-{[4-(hydroxymethyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluorome-
thyl)-1H-pyrazole-4-carboxylate
[0207] To a solution of the title compound of Example 1 Step B (150
mg, 0.40 mmol) and 1,4-benzenedimethanol (165 mg, 1.19 mmol) in THF
(2 mL) were added triphenylphosphine (313 mg, 1.19 mmol), followed
by diisopropyl azodicarboxylate (0.232 mL, 1.19 mmol). The reaction
vial was capped and stirred at 60.degree. C. After 1.5 h, the
reaction mixture was allowed to cool to ambient temperature, then
was concentrated in vacuo. Purification by flash chromatography on
silica gel (0 to 40% EtOAc in hexanes, then 40 to 100% EtOAc in
hexanes) provided the title compound: LCMS m/z 498.1 [M+H].sup.+;
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.13 (d, J=8.0 Hz, 1H),
8.12 (s, 1H), 7.95 (dd, J=8.0, 1.5 Hz, 1H), 7.85 (t, J=7.5 Hz, 1H),
7.51 (d, J=8.5 Hz, 1H), 7.39-7.28 (m, 5H), 7.11 (t, J=7.5 Hz, 1H),
7.06 (d, J=7.5 Hz, 1H), 5.15 (s, 2H), 4.98-4.94 (m, 2H), 4.38 (q,
J=7.0 Hz, 2H), 1.39 (t, J=7.0 Hz, 3H).
Step B. Ethyl
5-(trifluoromethyl)-1-(6-{2-[(4-{[4-(trifluoromethyl)phenoxy]methyl}benzy-
l)oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylate
[0208] To a solution of the title compound from Example 7 Step A
(30.0 mg, 0.060 mmol) and 4-hydroxybenzotrifluoride (29.3 mg, 0.181
mmol) in THF (0.400 mL) were added triphenylphosphine (47.5 mg,
0.181 mmol) and diisopropyl azodicarboxylate (0.035 mL, 0.181
mmol). The resulting mixture was stirred at 60.degree. C. After 3.5
h, the mixture was allowed to cool to ambient temperature, then was
concentrated in vacuo. Purification by flash chromatography on
silica gel (0 to 50% EtOAc in hexanes, then 50 to 100% EtOAc in
hexanes) provided the title compound: LCMS m/z 642.3
[M+H].sup.+.
Step C.
5-(Trifluoromethyl)-1-(6-{2-[(4-{[4-(trifluoromethyl)phenoxy]methy-
l}benzyl)oxy]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid
[0209] To a solution of the title compound from Example 7 Step B
(15.0 mg, 0.023 mmol) 1,4-dioxane (0.500 mL) was added lithium
hydroxide (0.5 mL, 2.0 M in water, 1.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 30 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (30 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 614.2 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.29 (s, 1H), 8.14 (d, J=8.0 Hz, 1H), 8.10
(t, J=8.0 Hz, 1H), 7.73 (dd, J=7.5, 1.5 Hz, 1H), 7.70 (d, J=8.0 Hz,
1H), 7.65 (d, J=8.5 Hz, 2H), 7.46-7.41 (m, 5H), 7.27 (d, J=8.5 Hz,
1H), 7.18 (d, J=8.5 Hz, 2H), 7.09 (t, J=7.5 Hz, 1H), 5.25 (s, 2H),
5.18 (s, 2H).
Example 8
##STR00042##
[0210] Step A. Ethyl
1-[6-(2-methoxy-5-methylphenyl)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyraz-
ole-4-carboxylate
[0211] To a flask containing the title compound from Example 1 Step
A (1.50 g, 4.69 mmol) were added 2-methoxy-5-methylphenyl boronic
acid (0.779 g, 4.69 mmol) and trans-dichlorobis(triphenylphosphine)
palladium (II) (329 mg, 0.469 mmol). Acetonitrile (12 mL) and
sodium carbonate (11.7 mL, 1.0 M aqueous, 11.7 mmol) were added,
and the resulting mixture was degassed via nitrogen sparge. The
reaction mixture was stirred at 70.degree. C. for 18 h, then was
allowed to cool to room temperature and was poured into water. The
mixture was extracted with EtOAc, and the organic phase was
concentrated in vacuo. Purification by chromatography on silica gel
(0 to 20% EtOAc in hexanes, then 20 to 100% EtOAc in hexanes)
provided compound the title compound: LCMS m/z 406.4 [M+H].sup.+;
.sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.12 (s, 1H), 8.11 (d,
J=8.0 Hz, 1H), 7.91 (t, J=8.0 Hz, 1H), 7.77 (d, J=2.0 Hz, 1H), 7.52
(d, J=8.0 Hz, 1H), 7.19 (dd, J=8.0, 2.0 Hz, 1H), 6.91 (d, J=8.0 Hz,
1H), 4.39 (q, J=7.0 Hz, 2H), 3.87 (s, 3H), 2.35 (s, 3H), 1.40 (t,
J=7.0 Hz, 3H).
Step B. Ethyl
1-[6-(2-hydroxy-5-methylphenyl)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyraz-
ole-4-carboxylate
[0212] To a cooled (0.degree. C.) solution of the title compound
from Example 8 Step A in DCM (20 mL) was added boron tribromide
(11.7 mL, 1.0 M in DCM, 11.7 mmol). After 15 min, the reaction
mixture was allowed to warm to ambient temperature. After an
additional 2 h, the reaction mixture was cooled to 0.degree. C.,
then was quenched by dropwise addition of sat. aq. NaHCO.sub.3 (gas
evolution) and was extracted with DCM. The organic phase was
separated and concentrated in vacuo. Purification by flash
chromatography on silica gel (0 to 30% EtOAc in hexanes, then 30 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 392.6
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 11.78 (s,
1H), 8.17 (s, 1H), 8.07-8.03 (m, 2H), 7.60 (d, J=1.5 Hz, 1H), 7.48
(dd, J=7.0, 1.5 Hz, 1H), 7.17 (dd, J=8.0, 2.0 Hz, 1H), 6.94 (d,
J=8.0 Hz, 1H), 4.39 (q, J=7.0 Hz, 2H), 2.36 (s, 3H), 1.40 (t, J=7.0
Hz, 3H).
Step C. Ethyl 4-[4-(trifluoromethyl)cyclohex-1-en-1-yl]benzoate
[0213] To a cooled (-78.degree. C.) solution of 4-trifluoromethyl
cyclohexanone (3.00 grams, 18.1 mmol) in anhydrous THF (100 mL) was
added lithium bis(trimethylsilyl)amide (19.9 mL, 1.0 M in THF, 19.9
mmol) dropwise. After 10 min, a solution of
2-[N,N-bis(trifluoromethylsulfonyl)amino]5-chloropyridine (7.09 g,
18.1 mmol) in THF (20 mL) was added, and the resulting mixture was
allowed to warm slowly to ambient temperature overnight, at which
point it was quenched by pouring into sat. aq. NaHCO.sub.3. The
mixture was extracted with EtOAc. The organic phase was separated,
dried over anhydrous sodium sulfate, and concentrated in vacuo. The
resulting enol triflate was used without further purification. To a
flask containing the unpurified enol triflate were added
4-ethoxycarbonylphenylboronic acid (3.68 g, 18.69 mmol) and
trans-dichlorobis(triphenylphosphine) palladium (II) (633 mg, 0.903
mmol). Acetonitrile (90 mL) and sodium carbonate (45 mL, 1.0 M
aqueous, 45.0 mmol) were added, and the resulting mixture was
degassed via nitrogen sparge. The reaction mixture was stirred at
70.degree. C. for 1.5 h, then was allowed to cool to room
temperature and was poured into water. The mixture was extracted
with EtOAc, and the organic phase was concentrated in vacuo.
Purification by chromatography on silica gel (0 to 10% EtOAc in
hexanes, then 10 to 100% EtOAc) provided the title compound: LCMS
m/z 299.5 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.99 (d, J=8.5 Hz, 2H), 7.42 (d, J=8.5 Hz, 2H), 6.20-6.18 (m, 1H),
4.37 (q, J=7.0 Hz, 2H), 2.61-2.18 (m, 6H), 1.74-1.68 (m, 1H), 1.39
(t, J=7.0 Hz, 3H).
Step D. Ethyl 4-[cis-4-(trifluoromethyl)cyclohexyl]benzoate and
ethyl 4-[trans-4-(trifluoromethyl)cyclohexyl]benzoate
[0214] To a degassed solution of the title compound from Example 8
Step C (50.0 mg, 0.168 mmol) in i-PrOH (3 mL) was added 5% rhodium
on alumina (25.0 mg). The reaction mixture was fitted with a
hydrogen balloon attached to a 3-way adapter. The reaction flask
was then evacuated and back-filled with hydrogen. After this
process was repeated three times, the reaction mixture was placed
under a hydrogen atmosphere, and was stirred vigorously. After 1 h,
the reaction mixture was filtered through Celite, rinsing with
EtOAc. The mixture was concentrated in vacuo, yielding a 2:1
(cis:trans) mixture of isomers. Purification by flash
chromatography on silica gel (0 to 2% ether in hexanes) provided
the product as mixture of isomers (1:1 cis:trans). A small portion
was further purified for characterization. Analytical data for the
first eluting isomer on silica gel (trans cyclohexyl): LCMS m/z
301.5 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 7.89
(d, J=8.5 Hz, 2H), 7.38 (d, J=8.5 Hz, 2H), 4.29 (q, J=7.0 Hz, 2H),
2.67-2.61 (m, 1H), 2.39-2.33 (m, 1H), 1.97 (app d, J=13.0 Hz, 2H),
1.88 (app d, J=13.0 Hz, 2H), 1.59-1.51 (m, 2H), 1.46-1.39 (m, 2H),
1.31 (t, J=7.0 Hz, 3H). Analytical data for the second eluting
isomer on silica gel (cis cyclohexyl): LCMS m/z 301.5 [M+H].sup.+;
.sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 7.90 (d, J=8.0 Hz, 2H),
7.41 (d, J=8.0 Hz, 2H), 4.30 (q, J=7.0 Hz, 2H), 2.87-2.82 (m, 1H),
2.54-2.49 (m, 1H), 1.87-1.82 (m, 2H), 1.77-1.73 (m, 6H), 1.31 (t,
J=7.0 Hz, 3H).
Step E. 4-[cis-4-(Trifluoromethyl)cyclohexyl]phenyl methanol and
4-[trans-4-(trifluoromethyl)cyclohexyl]phenyl methanol
[0215] To a cooled (0.degree. C.) solution of the title compound
from Example 8 Step D (2.49 g, 8.28 mmol) in THF (55 mL) was added
DIBAL-H (33.1 mL, 1.0 M in toluene, 33.1 mmol). After 2 h, the
reaction mixture was quenched by addition of MeOH (5.0 mL). The
resulting mixture was diluted with ether and saturated aqueous
sodium/potassium tartrate, and the mixture was stirred vigorously
until a clear phase separation was achieved. The organic phase was
then separated, dried over anhydrous sodium sulfate, and
concentrated in vacuo to provide the title compound as a mixture of
diastereomers (1:1 cis:trans), which was used without further
purification: LCMS m/z 241.4 [M-OH].sup.+.
Step F.
1-{6-[5-Methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}o-
xy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid and
1-{6-[5-methyl-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)ph-
enyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0216] To a solution of the title compound from Example 8 Step B
(150 mg, 0.383 mmol), the title compound from Example 8 Step E (148
mg, 0.575 mmol), and triphenylphosphine (151 mg, 0.575 mmol) in DCM
(5 mL) was added diisopropyl azodicarboxylate (0.112 mL, 0.575
mmol), and the resulting mixture was stirred at ambient
temperature. After 18 h, the reaction mixture was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 8%
EtOAc in hexanes, then 8 to 100% EtOAc in hexanes) provided the
title compounds. The first eluting diastereomer is the trans
isomer: LCMS m/z 632.3 [M+H].sup.+. The second eluting diastereomer
is the cis isomer: LCMS m/z 632.3 [M+H].sup.+. To separate
solutions of the Mitsunobu products in 1,4-dioxane (2 mL) was added
lithium hydroxide (1.0 mL, 2.0 M in water, 2.00 mmol), and the
resulting mixtures were stirred at 50.degree. C. After 30 min, the
reaction mixtures were rendered acidic by addition of aqueous
hydrochloric acid, then were diluted with 1,4-dioxane and passed
through a 0.45 micron syringe filter. Purification by reverse phase
HPLC (60 to 100% acetonitrile in water, each with 0.1% v/v TFA)
provided the title compounds. Analytical data for the trans isomer:
LCMS m/z 604.6 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO)
.delta. 8.29 (s, 1H), 8.15-8.08 (m, 2H), 7.69 (d, J=7.0 Hz, 1H),
7.56 (d, J=2.0 Hz, 1H), 7.32 (d, J=8.0 Hz, 2H), 7.24-7.19 (m, 2H),
7.15 (d, J=8.5 Hz, 2H), 5.16 (s, 2H), 2.54-2.49 (m, 1H), 2.36-2.31
(m, 1H), 2.27 (s, 3H), 1.96-1.93 (m, 2H), 1.86-1.84 (m, 2H),
1.54-1.46 (m, 2H), 1.43-1.35 (m, 2H). Analytical data for the cis
isomer: LCMS m/z 604.6 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.27 (s, 1H), 8.14 (d, J=7.5 Hz, 1H), 8.10
(t, J=7.5 Hz, 1H), 7.69 (d, J=7.5 Hz, 1H), 7.57 (br s, 1H), 7.33
(d, J=8.0 Hz, 2 H0, 7.23 (d, J=8.0 Hz, 2H), 7.23-7.22 (m, 1H), 7.16
(d, J=8.5 Hz, 1H), 5.16 (s, 2H), 2.76-2.71 (m, 1H), 2.50-2.48 (m,
1H, obscured by DMSO signal), 1.82-1.78 (m, 2H), 1.74-1.71 (m,
6H).
Example 9
##STR00043##
[0217] Step A. Ethyl
1-[6-(5-chloro-2-hydroxyphenyl)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyraz-
ole-4-carboxylate
[0218] To a solution of the title compound from Example 1 Step B
(1.00 g, 2.65 mmol) in DCM (13 mL) was added
benzyltrimethylammonium tetrachloroiodate (1.13 g, 2.70 mmol) and
the resulting mixture was allowed to stir at ambient temperature.
After 24 h, the mixture was concentrated in vacuo. Purification by
chromatography on silica gel (0 to 18% EtOAc in hexanes, then 18 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 412.0
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 11.98 (s,
1H), 8.18 (s, 1H), 8.10 (t, J=8.0 Hz, 1H), 8.00 (d, J=8.0 Hz, 1H),
7.78 (br s, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.30 (dd, J=9.0, 2.0 Hz,
1H), 6.98 (d, J=9.0 Hz, 1H), 4.39 (q, J=7.0 Hz, 2H), 1.40 (t, J=7.0
Hz, 3H).
Step B. Ethyl
1-{6-[5-chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate and
ethyl
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0219] To a solution of the title compound from Example 9 Step A
(90.0 mg, 0.219 mmol), the title compound from Example 8 Step E
(85.0 mg, 0.328 mmol), and triphenylphosphine (86.0 mg, 0.328 mmol)
in DCM (1 mL) was added diisopropyl azodicarboxylate (0.064 mL,
0.328 mmol), and the resulting mixture was stirred at ambient
temperature. After 18 h, the reaction mixture was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 12%
EtOAc in hexanes then 12 to 100% EtOAc in hexanes) provided the
title compounds. The first eluting compound is the trans isomer:
LCMS m/z 652.1 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 8.15 (d, J=8.0 Hz, 1H), 8.13 (s, 1H), 7.97 (d, J=2.5 Hz,
1H), 7.88 (t, J=8.0 Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.31 (dd,
J=9.0, 2.5 Hz, 1H), 7.28 (d, J=8.0 Hz, 2H), 7.19 (d, J=8.0 Hz, 2H),
6.99 (d, J=8.5 Hz, 1H), 5.12 (s, 2H), 4.39 (q, J=7.0 Hz, 2H),
2.55-2.50 (m, 1H), 2.10-2.01 (m, 5H), 1.50-1.46 (m, 4H), 1.40 (t,
J=7.0 Hz, 3H). The second eluting compound is the cis isomer: LCMS
m/z 652.1 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.16 (d, J=8.0 Hz, 1H), 8.13 (s, 1H), 7.88 (t, J=8.0 Hz, 1H), 7.56
(d, J=8.0 Hz, 1H), 7.31 (dd, J=8.5, 2.5 Hz, 1H), 7.29 (d, J=8.5 Hz,
2H), 7.24 (d, J=8.5 Hz, 2H), 7.00 (d, J=8.5 Hz, 1H), 5.12 (s, 2H),
4.39 (q, J=7.0 Hz, 2H), 2.73-2.70 (m, 1H), 2.34-2.28 (m, 1H),
1.98-1.89 (m, 4H), 1.80-1.71 (m, 4H), 1.40 (t, J=7.0 Hz, 3H).
Step C.
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}o-
xy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid and
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)ph-
enyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0220] To separate solutions of the title compounds from Example 9
Step B in 1,4-dioxane (1 mL) was added lithium hydroxide (0.5 mL,
2.0 M aqueous, 1.00 mmol), and the resulting mixtures were stirred
at 50.degree. C. After 30 min, the reaction mixtures were rendered
acidic by addition of aqueous hydrochloric acid, then were diluted
with 1,4-dioxane and passed through a 0.45 micron syringe filter.
Purification by reverse phase HPLC (65 to 100% acetonitrile in
water, each with 0.1% v/v TFA) provided the title compounds.
Analytical data for the trans isomer: LCMS m/z 624.4 [M+H].sup.+;
.sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 8.29 (s, 1H), 8.19 (d,
J=7.5 Hz, 1H), 8.14 (t, J=8.0 Hz, 1H), 7.77-7.75 (m, 2H), 7.49 (dd,
J=9.0, 2.0 Hz, 1H), 7.34 (d, J=8.0 Hz, 2H), 7.31 (d, J=9.0 Hz, 1H),
7.21 (d, J=8.0 Hz, 2H), 5.22 (s, 2H), 2.52-2.48 (m, 1H, obscured by
residual DMSO peak), 2.35-2.32 (m, 1H), 1.96-1.94 (m, 2H),
1.87-1.84 (m, 2H), 1.54-1.46 (m, 2H), 1.44-1.35 (m, 2H). Analytical
data for the cis isomer: LCMS m/z 624.2 [M+H].sup.+; .sup.1H NMR
(500 MHz, d.sub.6-DMSO) .delta. 8.29 (s, 1H), 8.20 (d, J=8.0 Hz,
1H), 8.14 (t, J=8.0 Hz, 1H), 7.78-7.75 (m, 2H), 7.48 (dd, J=9.0,
3.0 Hz, 1H), 7.35 (d, J=8.0 Hz, 2H), 7.31 (d, J=9.0 Hz, 1H), 7.25
(d, J=8.0 Hz, 2H), 5.23 (s, 2H), 2.77-2.73 (m, 1H), 2.52-2.48 (m,
1H, obscured by residual DMSO peak), 1.82-1.78 (m, 2H), 1.74-1.71
(m, 6H).
Example 10
##STR00044##
[0221] Step A. Ethyl
4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzoate
[0222] To a cooled (-78.degree. C.) solution of
1,4-cyclohexanedione mono-ethylene ketal (1.00 g, 6.40 mmol) in
anhydrous THF (30 mL) was added lithium bis(trimethylsilyl)amide
(7.7 mL, 1.0 M in THF, 7.70 mmol) dropwise. After 10 min, a
solution of
2-[N,N-bis(trifluoromethylsulfonyl)amino]5-chloropyridine (2.51 g,
6.40 mmol) in THF (10 mL) was added, and the resulting mixture was
allowed to warm slowly to ambient temperature overnight, at which
point it was quenched by pouring into sat. aq. NaHCO.sub.3. The
mixture was extracted with EtOAc. The organic phase was separated,
dried over anhydrous sodium sulfate, and concentrated in vacuo. To
a flask containing the unpurified enol triflate (1.36 g, 4.72 mmol)
were added 4-ethoxycarbonylphenylboronic acid (1.10 g, 5.66 mmol)
and trans-dichlorobis(triphenylphosphine) palladium (II) (331 mg,
0.472 mmol). Acetonitrile (24 mL) and sodium carbonate (11.8 mL,
1.0 M aqueous, 11.8 mmol) were added and the resulting mixture was
degassed via nitrogen sparge. The reaction mixture was stirred at
70.degree. C. for 3 h, then was allowed to cool to ambient
temperature and was poured into water. The mixture was extracted
with EtOAc, and the organic phase was concentrated in vacuo.
Purification by chromatography on silica gel (0 to 30% EtOAc in
hexanes, then 30 to 100% EtOAc in hexanes) provided the title
compound: LCMS m/z 289.1 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.97 (d, J=8.5 Hz, 2H), 7.44 (d, J=8.5 Hz, 2H),
6.12-6.10 (m, 1H), 4.36 (q, J=7.0 Hz, 2H), 4.03 (s, 4H), 2.70-2.67
(m, 2H), 2.50-2.48 (m, 2H), 1.93 (t, J=6.5 Hz, 2H), 1.39 (t, J=7.0
Hz, 3H).
Step B. Ethyl 4-(1,4-dioxaspiro[4.5]dec-8-yl)benzoate
[0223] A solution of the title compound from Example 10 Step A (907
mg, 3.15 mmol) in EtOAc was degassed via nitrogen sparge.
Platinum(IV) oxide (225 mgs, 0.991 mmol) was then added. The
reaction flask was fitted with a 3-way adapter equipped with a
hydrogen balloon. After 3 vacuum/hydrogen cycles, the reaction
mixture was placed under a hydrogen atmosphere. After 1 h, the
reaction mixture was filtered through a pad of Celite, rinsing with
EtOAc, and concentrated in vacuo. The unpurified product was used
in the subsequent step: LCMS m/z 291.0 [M+H].sup.+.
Step C. [4-(1,4-Dioxaspiro[4.5]dec-8-yl)phenyl]methanol
[0224] To a cooled (-78.degree. C.) solution of the title compound
from Example 10 Step B (450 mg, 1.55 mmol) in THF (8 mL) was added
DIBAL-H (3.10 mL, 1.50 M in heptane, 4.65 mmol). After 30 min, the
reaction mixture was transferred to a 0.degree. C. bath and was
held at this temperature for 45 min, whereupon it was quenched by
addition of MeOH (0.63 mL, 15.5 mmol). The resulting mixture was
diluted with ether and saturated aqueous sodium/potassium tartrate,
and the mixture was stirred rapidly until a clear phase separation
was achieved. The organic phase was separated, dried over anhydrous
sodium sulfate, and concentrated in vacuo. Purification by flash
chromatography on silica gel (0 to 30% EtOAc in hexanes, then 30 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 231.1
[M-OH].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.29 (d,
J=7.5 Hz, 2H), 7.24 (d, J=7.5 Hz, 2H), 4.65 (s, 2H), 3.98 (s, 4H),
2.59-2.54 (m, 1H), 1.87-1.66 (m, 8H).
Step D. Ethyl
1-[6-(2-{[4-(1,4-dioxaspiro[4.5]dec-8-yl)benzyl]oxy}phenyl)pyridin-2-yl]--
5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0225] To a solution of the title compound from Example 1 Step B
(251 mg, 0.67 mmol), the title compound from Example 10 Step C (248
mg, 1.00 mmol), and triphenylphosphine (349 mg, 1.33 mmol) in DCM
(3 mL) was added diisopropyl azodicarboxylate (0.259 mL, 1.33
mmol). The resulting mixture was stirred at ambient temperature.
After 5 h, the reaction mixture was concentrated in vacuo.
Purification by flash chromatography on silica gel (0 to 25% EtOAc
in hexanes then 25 to 100% EtOAc in hexanes) provided the title
compound: LCMS m/z 608.06 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.15 (d, J=8.0 Hz, 1H), 8.12 (s, 1H), 7.97 (d,
J=8.0 Hz, 1H), 7.85 (t, J=8.0 Hz, 1H), 7.52 (d, J=7.5 Hz, 1H),
7.38-7.35 (m, 1H), 7.29 (d, J=7.5 Hz, 2H), 7.23 (d, J=7.5 Hz, 2H),
7.12-7.06 (m, 2H), 5.12 (s, 2H), 4.38 (q, J=7.0 Hz, 2H), 3.98 (s,
4H), 2.60-2.54 (m, 1H), 1.87-1.65 (m, 8H), 1.39 (t, J=7.0 Hz,
3H).
Step E. Ethyl
1-[6-(2-{[4-(4-oxocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylate
[0226] A solution of the title compound from Example 10 Step D (300
mg, 0.49 mmol) in acetic acid (1.8 mL) and water (0.6 mL) was
stirred at 80.degree. C. After 2 h, the mixture was allowed to cool
to ambient temperature, then was concentrated in vacuo. The
resulting oil was diluted with ether, then was washed successively
with water, saturated aqueous sodium bicarbonate, water, and brine.
The organic phase was dried over anhydrous sodium sulfate,
filtered, and concentrated in vacuo. Purification by flash
chromatography on silica gel (0 to 20% EtOAc in hexanes, then 20 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 564.2
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.14 (d,
J=8.0 Hz, 1H), 8.12 (s, 1H), 7.96 (dd, J=8.0, 1.5 Hz, 1H), 7.87 (t,
J=8.0 Hz, 1H), 7.54 (d, J=7.5 Hz, 1H), 7.40-7.36 (m, 1H), 7.32 (d,
J=8.0 Hz, 2H), 7.23 (d, J=8.0 Hz, 2H), 7.12 (t, J=7.5 Hz, 1H), 7.07
(d, J=7.5 Hz, 1H), 5.14 (s, 2H), 4.38 (q, J=7.0 Hz, 2H), 3.04
(dddd, J=12.0, 12.0, 3.5, 3.5 Hz, 1H), 2.54-2.50 (m, 4H), 2.25-2.21
(m, 2H), 1.99-1.91 (m, 2H), 1.39 (t, J=7.0 Hz, 3H).
Step F.
1-[6-(2-{[4-(4-Oxocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0227] To a solution of the title compound from Example 10 Step E
(20.0 mg, 0.035 mmol) in 1,4-dioxane (1.0 mL) was added lithium
hydroxide (0.500 mL, 2.0 M in water, 1.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 30 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (40 to
95% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 536.0 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.30 (s, 1H), 8.14 (d, J=7.5 Hz, 1H), 8.11
(t, J=7.5 Hz, 1H), 7.73-7.70 (m, 2H), 7.43 (t, J=8.0 Hz, 1H), 7.35
(d, J=8.0 Hz, 2H), 7.29-7.26 (m, 3H), 7.09 (t, J=7.5 Hz, 1H), 5.21
(s, 2H), 3.07-3.02 (m, 1H), 2.57 (td, J=14.0, 5.5 Hz, 2H),
2.26-2.24 (m, 2H), 2.05-2.03 (m, 2H), 1.90-1.82 (m, 2H).
Example 11
##STR00045##
[0228] Step A. Ethyl
1-[6-(2-{[4-(4,4-difluorocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylate
[0229] A Teflon vial was charged with a solution of the title
compound from Example 10 Step E (87 mg, 0.154 mmol) in DCM (0.75
mL). DAST (0.035 mL, 0.262 mmol) was added, followed by ethanol
(0.002 mL, 0.03 mmol), and the resulting mixture was stirred at
ambient temperature. After 4 h, the reaction mixture was quenched
by addition of saturated aqueous sodium bicarbonate and was
extracted with DCM. The organic phase was dried over anhydrous
sodium sulfate, filtered, and concentrated in vacuo. Purification
by chromatography on silica gel (0 to 25% EtOAc in hexanes, then 25
to 100% EtOAc in hexanes) provided the title compound: LCMS m/z
586.2 [M+H].sup.+.
Step B.
1-[6-(2-{[4-(4,4-Difluorocyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl-
]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0230] To a solution of the title compound from Example 11 Step A
(20.0 mg, 0.034 mmol) in 1,4-dioxane (1 mL) was added lithium
hydroxide (0.500 mL, 2.0 M in water, 1.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 30 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (50 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 558.2 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.29 (s, 1H), 8.15-8.09 (m, 2H), 7.73 (dd,
J=8.0, 1.5 Hz, 1H), 7.70 (d, J=7.0 Hz, 1H), 7.42 (t, J=7.5 Hz, 1H),
7.35 (d, J=7.0 Hz, 2H), 7.27 (d, J=8.5 Hz, 1H), 7.23 (d, J=7.0 Hz,
2H), 7.09 (t, J=8.5 Hz, 1H), 5.20 (s, 2H), 2.71-2.66 (m, 1H),
2.11-2.07 (m, 2H), 2.00-1.83 (m, 4H), 1.67-1.60 (m, 2H).
Example 12
##STR00046##
[0231] Step A.
1-[6-(2-{[4-(trans-4-Methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5--
(trifluoromethyl)-1H-pyrazole-4-carboxylic acid and
1-[6-(2-{[4-(cis-4-methoxycyclohexyl)benzyl]oxy}phenyl)pyridin-2-yl]-5-(t-
rifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0232] To a solution of the title compound from Example 10 Step E
(220 mg, 0.390 mmol) in THF (2 mL) was added sodium borohydride
(29.5 mg, 0.781 mmol), and the mixture was stirred at ambient
temperature. After 30 min, the reaction mixture was quenched by
addition of saturated aqueous ammonium chloride, then was extracted
with EtOAc. The organic phase was dried over anhydrous sodium
sulfate, filtered, and concentrated in vacuo. The resulting alcohol
was carried forward without further purification: LCMS m/z 566.1
[M+H].sup.+. To a solution of the unpurified reduction product (64
mg, 0.113 mmol) in DCM (0.400 mL) were added
2,6-di-tert-butylpyridine (0.038 mL, 0.170 mmol) and silver
trifluoromethanesulfonate (32.0 mg, 0.124 mmol). The resulting
solution was cooled to 0.degree. C., and iodomethane (0.009 mL,
0.136 mmol) was added. After 30 min, the reaction mixture was
allowed to warm to ambient temperature and was held at this
temperature for 1.5 h, whereupon it was filtered through Celite,
rinsing with DCM. The reaction mixture was concentrated in vacuo
and taken into the subsequent step without purification: LCMS m/z
580.3 [M+H].sup.+. To a solution of the crude methyl ether (ca. 66
mg, 0.113 mmol) in 1,4-dioxane (2 mL) was added lithium hydroxide
(1.00 mL, 2.0 M in water, 2.00 mmol), and the resulting mixture was
stirred at 50.degree. C. After 30 min, the reaction mixture was
rendered acidic by addition of aqueous hydrochloric acid, then was
diluted with 1,4-dioxane and passed through a 0.45 micron syringe
filter. The diastereomers (major isomer is the trans cyclohexyl)
could be separated upon purification by reverse phase HPLC (40 to
90% acetonitrile in water, each with 0.1% v/v TFA) to provide the
title compounds. Analytical data for the trans cyclohexyl isomer:
LCMS m/z 552.2 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO)
.delta. 8.30 (s, 1H), 8.15-8.09 (m, 2H), 7.73-7.70 (m, 2H), 7.42
(t, J=7.5 Hz, 1H), 7.32 (d, J=8.0 Hz, 2H), 7.26 (d, J=8.0 Hz, 1H),
7.20 (d, J=8.0 Hz, 2H), 7.08 (t, J=7.5 Hz, 1H), 5.19 (s, 2H), 3.24
(s, 3H), 3.16-3.13 (m, 1H), 2.50-2.47 (m, 1H), 2.07 (d, J=10 Hz,
2H), 1.79 (d, J=12.5 Hz, 2H), 1.46-1.43 (m, 2H), 1.25-1.17 (m, 2H).
Analytical data for the cis cyclohexyl isomer: LCMS m/z 552.2
[M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 8.29 (s,
1H), 8.16-8.10 (m, 2H), 7.73 (dd, J=7.5, 1.5 Hz, 1H), 7.70 (d,
J=7.5 Hz, 1H), 7.44-7.41 (m, 1H), 7.33 (d, J=8.0 Hz, 2H), 7.27 (d,
J=8.0 Hz, 1H), 7.18 (d, J=8.0 Hz, 2H), 7.09 (t, J=7.5 Hz, 1H), 5.19
(s, 2H), 3.35 (m, 1H, obscured by residual water peak), 3.23 (s,
3H), 2.56-2.52 (m, 1H), 1.95-1.92 (m, 2H), 1.69-1.61 (m, 2H),
1.52-1.46 (m, 2H), 1.23-1.17 (m, 2H).
Example 13
##STR00047##
[0233] Step A. Methyl
2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate
[0234] A round bottomed flask was charged with methyl
4-bromo-2-methylbenzoate (3.98 g, 17.37 mmol),
bis(pinacolato)diboron (4.85 g, 19.11 mmol), potassium acetate
(5.12 g, 52.1 mmol), and
dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (II)
dichloromethane adduct (0.426 g, 0.521 mmol). The flask was purged
with nitrogen. Anhydrous DMSO (100 mL) was added, and the resulting
suspension was degassed via nitrogen sparge. The mixture was then
placed in a pre-heated oil bath (80.degree. C.), and was held at
this temperature for 2 h, whereupon it was allowed to cool to
ambient temperature, then was poured into water. The aqueous phase
was extracted with ether, and the organic phase was washed with
brine. The organic phase was then separated, dried over anhydrous
sodium sulfate, filtered, and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 10% EtOAc in hexanes,
then 10 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 277.6 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.87 (d, J=7.5 Hz, 1H), 7.68 (s, 1H), 7.66 (d, J=7.5 Hz, 1H), 3.89
(s, 3H), 2.59 (s, 3H), 1.35 (s, 12H).
Step B. Methyl
4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-2-methylbenzoate
[0235] To a flask containing the enol triflate synthesized
according to Example 10 Step A (1.10 g, 3.82 mmol) were added the
title compound from Example 13 Step A (1.16 g, 4.20 mmol) and
trans-dichlorobis(triphenylphosphine) palladium (II) (134 mg, 0.191
mmol). Acetonitrile (15 mL) and sodium carbonate (9.54 mL, 1.0 M
aqueous, 9.54 mmol) were added, and the resulting mixture was
degassed via nitrogen sparge. The reaction mixture was stirred at
70.degree. C. for 15 h, then was allowed to cool to ambient
temperature and was poured into water. The mixture was extracted
with EtOAc, and the organic phase was concentrated in vacuo.
[0236] Purification by chromatography on silica gel (0 to 30% EtOAc
in hexanes, then 30 to 100% EtOAc in hexanes) provided the title
compound: LCMS m/z 289.4 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 7.87 (d, J=8.5 Hz, 1H), 7.27-7.25 (m, 2H),
6.09-6.07 (m, 1H), 4.03 (s, 4H), 3.88 (s, 3H), 2.68-2.65 (m, 2H),
2.60 (3, H), 2.49-2.47 (m, 2H), 1.94-1.91 (m, 2H).
Step C. Methyl 4-(1,4-dioxaspiro[4.5]dec-8-yl)-2-methylbenzoate
[0237] To a degassed solution of the title compound from Example 13
Step B (606 mg, 2.10 mmol) in EtOAc (15 mL) was added platinum(IV)
oxide (150 mg). The reaction mixture was fitted with a 3-way
adapter with a hydrogen balloon attached. The reaction flask was
then evacuated and back-filled with hydrogen. After this process
was repeated three times, the reaction mixture was placed under a
hydrogen atmosphere, then was stirred vigorously. After 45 min, the
reaction mixture was filtered through Celite, rinsing with EtOAc.
The mixture was then concentrated in vacuo to provide the title
compound, which was used without further purification: LCMS m/z
259.4 [M-CH.sub.3O].sup.+.
Step D. Methyl 2-methyl-4-(4-oxocyclohexyl)benzoate
[0238] A solution of the title compound from Example 13 Step C (610
mg, 2.10 mmol) in acetic acid (7.8 mL) and water (2.6 mL) was
stirred at 80.degree. C. After 2 h, the mixture was allowed to cool
to ambient temperature, then was concentrated in vacuo. The
resulting oil was diluted with ether, then was washed successively
with water, saturated aqueous sodium bicarbonate, water, and brine.
The organic phase was dried over anhydrous sodium sulfate,
filtered, and concentrated in vacuo. Purification by flash
chromatography on silica gel (0 to 50% EtOAc in hexanes, then 50 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 215.4
[M-CH.sub.3O].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 7.88
(d, J=8.5 Hz, 1H), 7.14-7.10 (m, 2H), 3.88 (s, 3H), 3.05-3.00 (m,
1H), 2.59 (s, 3H), 2.51 (app dd, J=8.5, 4.0 Hz, 4H), 2.24-2.20 (m,
2H), 1.97-1.92 (m, 2H).
Step E. Methyl 4-(trans-4-hydroxycyclohexyl)-2-methylbenzoate
[0239] To a solution of the title compound from Example 13 Step D
(232 mg, 0.942 mmol) in THF (5.0 mL) was added sodium borohydride
(71.3 mg, 1.88 mmol). After 45 min, the reaction mixture was
quenched by addition of sat. aq. NH.sub.4Cl. The mixture was
extracted with EtOAc, and the organic phase was dried over sodium
sulfate, filtered, and concentrated in vacuo to provide the title
compound, which was used without further purification: LCMS m/z
249.4 [M+H].sup.+.
Step F. Methyl 4-(trans-4-methoxycyclohexyl)-2-methylbenzoate
[0240] To a solution of the title compound from Example 13 Step E
(234 mg, 0.942 mmol) in DCM (4.7 mL) were added
2,6-di-tert-butylpyridine (0.318 mL, 1.41 mmol) and silver
trifluoromethanesulfonate (266 mg, 1.04 mmol). The resulting
solution was cooled to 0.degree. C., and iodomethane (0.071 mL,
1.13 mmol) was added. After 30 min, the reaction mixture was
allowed to warm to ambient temperature and was held at this
temperature for 3 h, whereupon it was filtered through Celite,
rinsing with DCM. The reaction mixture was concentrated in vacuo.
Purification by flash chromatography on silica gel (0 to 30% EtOAc
in hexanes, then 30 to 100% EtOAc in hexanes) provided the title
compound. Analytical data for the major isomer (trans cyclohexyl):
LCMS m/z 263.17 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 7.85 (d, J=8.5 Hz, 1H), 7.08-7.06 (m, 2H), 3.86 (s, 3H),
3.38 (s, 3H), 3.24-3.17 (m, 1H), 2.58 (s, 3H), 2.49 (dt, J=12.0,
3.5 Hz, 1H), 2.21-2.18 (m, 2H), 1.94-1.92 (m, 2H), 1.54-1.46 (m,
2H), 1.39-1.30 (m, 2H).
Step G. [4-(trans-4-Methoxycyclohexyl)-2-methylphenyl]methanol
[0241] To a cooled (-78.degree. C.) solution of the title compound
from Example 13 Step F (90.0 mg, 0.343 mmol) in THF (1.7 mL) was
added DIBAL-H (1.03 mL, 1.0 M in toluene, 1.03 mmol). After 30 min,
the reaction mixture was allowed to warm to 0.degree. C. After 2 h,
the reaction mixture was quenched by addition of MeOH (0.140 mL).
The resulting mixture was diluted with diethyl ether and saturated
aqueous sodium/potassium tartrate, and the mixture was stirred
rapidly until a clear phase separation was achieved. The organic
phase was then separated, dried over anhydrous sodium sulfate, and
concentrated in vacuo to provide the title compound, which was used
without further purification.
Step H. Ethyl
1-[6-(2-{[4-(trans)-4-methoxycyclohexyl)-2-methylbenzyl]oxy}phenyl)pyridi-
n-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0242] The title compound was prepared by Mitsunobu coupling of the
title compound from Example 13 Step G with the title compound from
Example 1 Step B, using chemistry described in Example 8 Step F:
LCMS m/z 594.7 [M+H].sup.+
Step I.
1-[6-(2-{[4-(trans)-4-Methoxycyclohexyl)-2-methylbenzyl]oxy}phenyl-
)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0243] The title compound was prepared by hydrolysis of the title
compound from Example 13 Step H, using chemistry described in
Example 1 Step D: LCMS m/z 566.7 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.29 (s, 1H), 8.08 (t, J=7.5 Hz, 1H), 8.03
(d, J=7.5 Hz, 1H), 7.71-7.68 (m, 2H), 7.46-7.42 (m, 1H), 7.32 (d,
J=8.0 Hz, 1H), 7.25 (d, J=7.5 Hz, 1H), 7.09 (t, J=7.5 Hz, 1H), 7.03
(s, 1H), 6.98 (d, J=8.0 Hz, 1H), 5.16 (s, 1H), 3.24 (s, 3H),
3.18-3.13 (m, 1H), 2.43-2.40 (m, 1H), 2.08-2.06 (m, 2H), 1.79-1.76
(m, 2H), 1.48-1.42 (m, 2H), 1.24-1.17 (m, 2H).
Example 14
##STR00048##
[0244] Step A. Ethyl
1-[6-(2-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]oxy}pheny-
l)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0245] A vial was charged with the product from Example 2 Step A
(300 mg, 0.549 mmol), bis(pinacolato)diboron (153 mg, 0.604 mmol),
bis(tricyclohexylphosphine)palladium(0) (25.0 mg, 0.037 mmol), and
potassium acetate (135 mg, 1.37 mmol). The mixture was flushed with
nitrogen, then degassed 1,4-dioxane (2.7 mL) was added. The vial
was capped and stirred for 15 h, whereupon it was diluted with
water and extracted with EtOAc. The organic phase was separated,
dried over sodium sulfate, filtered, and concentrated. Purification
by flash chromatography on silica gel (0 to 20% EtOAc in hexanes,
then 20 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 594.8 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
8.19 (d, J=8.0 Hz, 1H), 8.16 (s, 1H), 8.02 (d, J=7.0 Hz, 1H), 7.90
(t, J=8.0 Hz, 1H), 7.85 (d, J=7.0 Hz, 2H), 7.57 (d, J=8.0 Hz, 1H),
7.42 (d, J=7.0 Hz, 2H), 7.39-7.37 (m, 1H), 7.14 (t, J=7.5 Hz, 1H),
7.07 (d, J=8.0 Hz, 1H), 5.21 (s, 2H), 4.42 (q, J=7.0 Hz, 2H), 1.43
(t, J=7.0 Hz, 3H), 1.39 (s, 12H).
Step B. Ethyl
5-(trifluoromethyl)-1-{6-[2-({4-[6-(trifluoromethyl)pyridin-3-yl]benzyl}o-
xy)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylate
[0246] To a vial containing the title compound from Example 14 Step
A (14.0 mg, 0.024 mmol) were added
3-bromo-6-trifluoromethylpyridine (6.4 mg, 0.028 mmol) and
trans-dichlorobis(triphenylphosphine) palladium (II) (1.7 mg,
0.0024 mmol). Acetonitrile (0.500 mL) and sodium carbonate (0.059
mL, 1.0 M in water, 0.059 mmol) were added, and the resulting
mixture was degassed by a nitrogen sparge. The reaction vial was
capped and placed in a pre-heated oil bath (70.degree. C.). After 6
h, the reaction mixture was allowed to cool to room temperature,
then was purified by flash chromatography on silica gel: LCMS m/z
613.2 [M+H].sup.+.
Step C.
5-(Trifluoromethyl)-1-{6-[2-({4-[6-(trifluoromethyl)pyridin-3-yl]b-
enzyl}oxy)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid
[0247] To a solution of the title compound from Example 14 Step B
in 1,4-dioxane (0.500 mL) was added lithium hydroxide (0.5 mL, 2.0
M in water, 1.00 mmol), and the resulting mixture was stirred at
50.degree. C. After 30 min, the reaction mixture was rendered
acidic by addition of aqueous hydrochloric acid, then was diluted
with 1,4-dioxane and passed through a 0.45 micron syringe filter.
Purification by reverse phase HPLC (50 to 100% acetonitrile in
water, each with 0.1% v/v TFA) provided the title compound: LCMS
m/z 585.2 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta.
9.09 (d, J=2.0 Hz, 1H), 8.36 (dd, J=8.0, 2.0 Hz, 1H), 8.30 (s, 1H),
8.18 (d, J=7.0 Hz, 1H), 8.14 (t, J=8.0 Hz, 1H), 7.97 (d, J=8.0 Hz,
1H), 7.82 (d, J=8.5 Hz, 1H), 7.75 (dd, J=8.0, 2.0 Hz, 1H), 7.72 (d,
J=8.0 Hz, 1H), 7.60 (d, J=8.5 Hz, 1H), 7.47-7.43 (m, 1H), 7.30 (d,
J=8.0 Hz, 1H), 7.11 (t, J=7.5 Hz, 1H), 5.34 (s, 2H).
Example 15
##STR00049##
[0248] Step A. Ethyl
1-[6-(2-hydroxy-3-iodophenyl)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyrazol-
e-4-carboxylate
[0249] To a suspension of the title compound from Example 1 Step B
(2.00 g, 5.30 mmol) and silver sulfate (1.653 g, 5.30 mmol) in EtOH
(53 mL) was added iodine (1.35 g, 5.30 mmol). The resulting
suspension was stirred vigorously at ambient temperature. After 4
h, the reaction mixture was diluted with EtOAc, and the organic
phase was washed successively with water, sat. aq. sodium
bisulfite, and sat. aq. NaHCO.sub.3. The organic phase was then
concentrated in vacuo. The title compound was separated from the
para-iodo isomer upon purification by flash chromatography on
silica gel (0 to 15% EtOAc in hexanes, then 15% EtOAc in hexanes,
then 15 to 100% EtOAc in hexanes; the title compound is the later
eluting isomer): LCMS m/z 504.5 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 13.08 (s, 1H), 8.17 (s, 1H), 8.10 (t, J=8.0 Hz,
1H), 8.06 (d, J=8.0 Hz, 1H), 7.86 (dd, J=7.5, 1.5 Hz, 1H), 7.82
(dd, J=8.0, 1.5 Hz, 1H), 7.52 (d, J=7.5 Hz, 1H), 6.75 (t, J=8.0 Hz,
1H), 4.33 (q, J=7.0 Hz, 2H), 1.43 (t, J=7.0 Hz, 3H).
Step B. Ethyl
1-(6-{2-[(2,4-dimethylbenzyl)oxy]-3-iodophenyl}pyridin-2-yl)-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylate
[0250] To a solution of the title compound from Example 15 Step A
(200 mg, 0.397 mmol), 2,4-dimethylbenzyl alcohol (81.0 mg, 0.596
mmol), and triphenylphosphine (156 mg, 0.596 mmol) in DCM (2 mL)
was added diisopropyl azodicarboxylate (0.114 mL, 0.596 mmol), and
the resulting mixture was stirred at ambient temperature. After 18
h, the reaction mixture was concentrated in vacuo. Purification by
flash chromatography on silica gel (0 to 20% EtOAc in hexanes, then
20 to 100% EtOAc in hexanes) provided the title compound: LCMS m/z
594.8 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.17
(s, 1H), 8.02 (d, J=8.0 Hz, 1H), 7.92 (d, J=7.5 Hz, 1H), 7.85-7.81
(m, 2H), 7.60 (d, J=7.5 Hz, 1H), 7.13 (d, J=7.5 Hz, 1H), 7.05 (t,
J=7.5 Hz, 1H), 6.95-6.92 (m, 2H), 4.69 (s, 2H), 4.41 (q, J=7.0 Hz,
2H), 2.30 (s, 3H), 2.17 (s, 3H), 1.43 (t, J=7.0 Hz, 3H).
Step C. Ethyl
1-(6-{2-[(2,4-dimethylbenzyl)oxy]-3-methylphenyl}pyridin-2-yl)-5-(trifluo-
romethyl)-1H-pyrazole-4-carboxylate
[0251] A vial was charged with the product from Example 15 Step B
(40 mg, 0.064 mmol), trimethyl boroxine (49 mg, 50 wt. %, 0.193
mmol), and
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (5.3 mg, 0.006 mmol). Sodium carbonate
(0.161 mL, 1.0 M aqueous, 0.161 mmol) and THF (0.25 mL) were added,
and the resulting suspension was degassed by a nitrogen sparge. The
vial was then capped and placed in a pre-heated (65.degree. C.) oil
bath. After 18 h, the reaction mixture was allowed to cool to
ambient temperature, then was poured into water. The aqueous phase
was extracted with EtOAc, and the organic phase was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 20%
EtOAc in hexanes, then 20 to 100% EtOAc in hexanes) provided the
title compound: LCMS m/z 510.8 [M+H].sup.+.
Step D.
1-(6-{2-[(2,4-Dimethylbenzyl)oxy]-3-methylphenyl}pyridin-2-yl)-5-(-
trifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0252] To a solution of the title compound from Example 15 Step C
(12.0 mg, 0.023 mmol) in 1,4-dioxane (1.5 mL) was added lithium
hydroxide (0.5 mL, 2.0 M in water, 1.00 mmol), and the resulting
mixture was stirred at 50.degree. C. After 30 min, the reaction
mixture was rendered acidic by addition of aqueous hydrochloric
acid, then was diluted with 1,4-dioxane and passed through a 0.45
micron syringe filter. Purification by reverse phase HPLC (40 to
100% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 482.8 [M+H].sup.+; .sup.1H NMR (500 MHz,
d.sub.6-DMSO) .delta. 8.30 (s, 1H), 8.08 (t, J=8.0 Hz, 1H), 7.95
(d, J=8.0 Hz, 1H), 7.74 (d, J=8.0 Hz, 1H), 7.45 (dd, J=8.0, 1.5 Hz,
1H), 7.35-7.32 (m, 1H), 7.18 (t, J=7.5 Hz, 1H), 6.91-6.86 (m, 3H),
4.52 (s, 2H), 2.27 (s, 3H), 2.21 (s, 3H), 2.06 (s, 3H).
Example 16
##STR00050##
[0253] Step A. 2-(6-Chloropyrazin-2-yl)phenol
[0254] To a mixture of 2,6-dichloropyrazine (1.00 g, 6.71 mmol),
2-hydroxyphenylboronic acid (972 mg, 7.05 mmol) and trans
dichlorobis(triphenylphosphine) palladium (II) (471 mg, 0.671 mmol)
were added acetonitrile (20 mL) and sodium carbonate (13.4 mL, 1.0
M in water, 13.4 mmol), and the resulting mixture was degassed by a
nitrogen sparge. The reaction flask was equipped with a reflux
condenser, then was placed in a pre-heated oil bath (75.degree. C.)
and was stirred rapidly. After 5 h, the reaction mixture was
allowed to cool to room temperature, then was poured into water and
extracted with EtOAc. The organic phase was separated and
concentrated in vacuo. Purification by chromatography on silica gel
(0 to 25% EtOAc in hexanes, then 25 to 100% EtOAc in hexanes)
provided the title compound: LCMS m/z 207.6 [M+H].sup.+; .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 11.64 (s, 1H), 9.11 (s, 1H), 8.53
(s, 1H), 7.85 (dd, J=8.0, 1.5 Hz, 1H), 7.40 (t, J=8.0 Hz, 1H), 7.07
(d, J=8.5 Hz, 1H), 6.99 (t, J=8.0 Hz, 1H).
Step B. Ethyl
1-[6-(2-hydroxyphenyl)pyrazin-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-car-
boxylate
[0255] To a solution of the title compound from Example 16 Step A
(0.500 grams, 2.42 mmol) in ethanol (20 mL) was added hydrazine
hydrate (3.25 mL, 35% v/v, 36.3 mmol). The reaction flask was
equipped with a reflux condenser, and the reaction mixture was
stirred at 80.degree. C. After 12 h, the mixture was allowed to
cool to room temperature, whereupon a yellow solid precipitated.
The solid was triturated with hexanes, filtered, washed with water,
and dried in vacuo: LCMS m/z 203.2 [M+H].sup.+. To a solution of
the crude pyrazinyl hydrazine in acetonitrile (10 mL) were added
triethylamine (0.675 mL, 4.84 mmol) and ethyl
2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutyrate (0.872 g, 3.63
mmol). After 40 min at ambient temperature, the reaction mixture
was placed in a 60.degree. C. bath and was stirred for 30 min, at
which point the reaction mixture was allowed to cool to ambient
temperature, then was concentrated in vacuo. Purification by flash
chromatography on silica gel (0 to 60% EtOAc in hexanes, then 60 to
100% EtOAc in hexanes) provided the title compound: LCMS m/z 379.1
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 10.98 (s,
1H), 9.35 (s, 1H), 8.85 (s, 1H), 8.23 (s, 1H), 7.93 (dd, J=8.0, 1.5
Hz, 1H), 7.43 (t, J=8.0 Hz, 1H), 7.07 (d, J=8.5 Hz, 1H), 7.04 (t,
J=8.0 Hz, 1H).
Step C.
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}o-
xy)phenyl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid and
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)ph-
enyl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0256] To a solution of the title compound from Example 16 Step B
(100.0 mg, 0.264 mmol), the title compound from Example 8 Step E
(102 mg, 0.397 mmol), and triphenylphosphine (104 mg, 0.397 mmol)
in DCM (7 mL) was added diisopropyl azodicarboxylate (0.077 mL,
0.397 mmol), and the resulting mixture was stirred at ambient
temperature. After 18 h, the reaction mixture was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 20%
EtOAc in hexanes then 20 to 100% EtOAc in hexanes) provided the
title compounds. The first eluting compound is the trans isomer:
LCMS m/z 619.2 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 9.44 (s, 1H), 8.85 (s, 1H), 8.17 (s, 1H), 8.00 (dd, J=7.5,
1.5 Hz, 1H), 7.46-7.43 (m, 1H), 7.33 (d, J=8.5 Hz, 2H), 7.21 (d,
J=8.5 Hz, 2H), 7.16-7.12 (m, 2H), 5.18 (s, 2H), 4.40 (q, J=7.0 Hz,
2H), 2.54-2.51 (m, 1H), 2.11-2.00 (m, 5H), 1.50-1.45 (m, 4H), 1.42
(t, J=7.0 Hz, 3H). The second eluting compound is the cis isomer:
LCMS m/z 619.2 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3)
.delta. 9.46 (s, 1H), 8.86 (s, 1H), 8.18 (s, 1H), 8.01 (dd, J=7.5,
1.5 Hz, 1H), 7.47-7.44 (m, 1H), 7.35 (d, J=8.5 Hz, 2H), 7.27 (d,
J=8.5 Hz, 2H), 7.17-7.13 (m, 1H), 5.20 (s, 2H), 4.41 (q, J=7.0 Hz,
2H), 2.74-2.71 (m, 1H), 2.36-2.29 (m, 1H), 2.01-1.90 (m, 4H),
1.81-1.71 (m, 4H), 1.41 (t, J=7.0 Hz, 3H).
Step D. Ethyl
1-{6-[5-chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phen-
yl]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate and
ethyl
1-{6-[5-chloro-2-({4-[cis-4-(trifluoromethyl)cyclohexyl]benzyl}oxy)phenyl-
]pyrazin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0257] To separate solutions of the title compounds from Example 16
Step C in 1,4-dioxane (1 mL) was added lithium hydroxide (0.5 mL,
2.0 M aqueous, 1.00 mmol), and the resulting mixtures were stirred
at 50.degree. C. After 30 min, the reaction mixtures were rendered
acidic by addition of aqueous hydrochloric acid, then were diluted
with 1,4-dioxane and passed through 0.45 micron syringe filters.
Purification by reverse phase HPLC (40 to 100% acetonitrile in
water, each with 0.1% v/v TFA) provided the title compounds.
Analytical data for the trans isomer: LCMS m/z 591.5 [M+H].sup.+;
.sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta. 9.37 (s, 1H), 9.00 (s,
1H), 8.39 (s, 1H), 7.78 (dd, J=7.5, 1.5 Hz, 1H), 7.54-7.50 (m, 1H),
7.37-7.34 (m, 3H), 7.22 (d, J=8.0 Hz, 2H), 7.15 (t, J=7.5 Hz, 1H),
5.23 (s, 2H), 2.55-2.49 (m, 1H), 2.35-2.32 (m, 1H), 1.96-1.85 (m,
4H), 1.55-1.36 (m, 4H). Analytical data for the cis isomer: LCMS
m/z 591.5 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta.
9.38 (s, 1H), 9.00 (s, 1H), 8.38 (s, 1H), 7.78 (dd, J=7.5, 1.5 Hz,
1H), -54-7.50 (m, 1H), 7.39-7.34 (m, 3H), 7.25 (d, J=8.0 Hz, 2H),
7.15 (t, J=7.5 Hz, 1H), 5.24 (s, 2H), 2.76-2.73 (m, 1H), 2.52-2.48
(m, 1H), 1.82-1.77 (m, 2H), 1.75-1.70 (m, 6H).
Example 17
##STR00051##
[0258] Step A. Ethyl
1-[4-(2-methoxyphenyl)-1,3-thiazol-2-yl]-5-(trifluoromethyl)-1H-pyrazole--
4-carboxylate
[0259] A solution of ethyl
1-(aminocarbonothioyl)-5-hydroxy-5-(trifluoromethyl)-4,5-dihydro-1H-pyraz-
ole-4-carboxylate (470 mg, 1.65 mmol, prepared according to J.
Comb. Chem. 2002, 4, 23-32) and 2-bromo-2'-methoxyacetophenone (377
mg, 1.65 mmol) in ethanol (8 mL) was heated at 80.degree. C. After
1 h, the mixture was allowed to cool to ambient temperature, then
was concentrated in vacuo. Purification by flash chromatography on
silica gel (0 to 55% EtOAc in hexanes) provided the title compound
as an off-white solid: LCMS m/z 398.5 [M+H].sup.+; .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 8.26 (dd, J=7.6, 1.6 Hz, 1H), 8.13 (s,
1H), 8.03 (s, 1H), 7.37 (ddd, J=8.9, 6.5, 1.8 Hz, 1H), 7.12-7.01
(m, 2H), 4.42 (q, J=7.0 Hz, 2H), 4.02 (s, 3H), 1.42 (t, J=7.0 Hz,
3H).
Step B. Ethyl
1-[4-(2-hydroxyphenyl)-1,3-thiazol-2-yl]-5-(trifluoromethyl)-1H-pyrazole--
4-carboxylate
[0260] To a cooled (0.degree. C.) solution of the title compound
from Example 17 Step A (353 mg, 0.888 mmol) in DCM (6.3 ml) was
added BBr.sub.3 (2.67 ml, 1.0 M in DCM, 2.67 mmol) dropwise. After
the addition was complete, the mixture was allowed to warm up to
ambient temperature. After 1 h, the reaction mixture was quenched
by addition of sat. aq. NaHCO.sub.3 and extracted with DCM. The
organic phase was separated and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 60% EtOAc in hexanes)
provided the title compound as a yellow solid: LCMS m/z 384.5
[M+H].sup.+; .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 10.17 (s,
1H), 8.18 (s, 1H), 7.67 (dd, J=8.0, 1.9 Hz, 1H), 7.34 (t, J=8.5 Hz,
1H), 7.09 (dd, J=8.2, 0.9 Hz, 1H), 6.98 (t, J=7.4 Hz, 1H), 4.45 (m,
2H), 1.44 (t, J=7.1 Hz, 3H).
Step C.
5-(Trifluoromethyl)-1-[4-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]me-
thoxy}phenyl)-1,3-thiazol-2-yl]-1H-pyrazole-4-carboxylic acid
[0261] The title compound was prepared from the title compound from
Example 17 Step B by direct analogy to the procedures outlined in
Example 2 Steps A-C: LCMS m/z 590.5 [M+H].sup.+; .sup.1H NMR (500
MHz, d.sub.6-DMSO) .delta. 8.40 (s, 1H), 8.22 (s, 1H), 8.08 (dd,
J=7.5, 1.5 Hz, 1H), 7.93 (d, J=8.5 Hz, 2H), 7.83 (d, J=8.5 Hz, 2H),
7.81 (d, J=8.5 Hz, 2H), 7.68 (d, J=8.5 Hz, 2H), 7.40-7.37 (m, 1H),
7.31 (d, J=8.5 Hz, 1H), 7.11 (t, J=7.5 Hz, 1H), 5.43 (s, 2H).
Example 18
##STR00052##
[0262] Step A. Ethyl
1-(2-chloropyrimidin-4-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0263] To a solution 2-chloro-6-hydrazinopyrimidine (1.00 g, 6.92
mmol) and triethylamine (0.964 mL, 6.92 mmol) in acetonitrile (35
mL) was added ethyl
2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutyrate (1.35 mL, 6.97
mmol). After 45 min, the reaction mixture was placed in a
90.degree. C. oil bath. After 1 h, the reaction mixture was allowed
to cool to ambient temperature, then was concentrated in vacuo.
Purification by flash chromatography on silica gel (0 to 60% EtOAc
in hexanes, then 60 to 100% EtOAc in hexanes) provided the title
compound: LCMS m/z 321.0 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.80 (d, J=5.5 Hz, 1H), 8.12 (s, 1H), 7.76 (d,
J=5.5 Hz, 1H), 4.39 (q, J=7.0 Hz, 2H), 1.39 (t, J=7.0 Hz, 3H).
Step B.
1-[2-(2-{[4-(2-Phenylethyl)benzyl]oxy}phenyl)pyrimidin-4-yl]-5-(tr-
ifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0264] Suzuki coupling of the title compound from Example 18 Step A
with 2-methoxyphenylboronic acid, followed by treatment with
BBr.sub.3 (according to Example 8 Steps A and B) gave a pyrimidinyl
phenol which was processed to the title compound according to
Example 1 Steps C and D: LCMS m/z 545.3 [M+H].sup.+; .sup.1H NMR
(500 MHz, d.sub.6-DMSO) .delta. 9.20 (d, J=5.5 Hz, 1H), 8.42 (s,
1H), 7.88 (d, J=5.5 Hz, 1H), 7.69 (dd, J=8.0, 2.0 Hz, 1H),
7.50-7.47 (m, 1H), 7.27-2.08 (m, 11H), 5.13 (s, 2H), 2.82 (s,
4H).
Example 19
##STR00053##
[0265] Step A.
2-Chloro-6-(2-methoxy-5-methylphenyl)-4-nitropyridine
[0266] The title compound was prepared according to the procedure
described in Example 1 Step B, by reaction of
2-methoxy-5-methylphenylboronic acid with
2,6-dichloro-4-nitropyridine: LCMS m/z 279.5 [M+H].sup.+; .sup.1H
NMR (500 MHz, CDCl.sub.3) .delta. 8.19 (d, J=8.5 Hz, 1H), 7.52 (d,
J=2.1 Hz, 1H), 7.44 (d, J=8.5 Hz, 1H), 7.28 (dd, J=8.5, 2.1 Hz,
1H), 6.83 (d, J=8.5 Hz, 1H), 3.72 (s, 3H), 2.42 (s, 3H).
Step B. Di-tert-butyl
1-[6-(2-methoxy-5-methylphenyl)-4-nitropyridin-2-yl]hydrazine-1,2-dicarbo-
xylate
[0267] A mixture of the title compound from Example 19 Step A (1.5
g, 5.4 mmol), di-tert-butyl hydrazine-1,2-dicarboxylate (1.375 g,
5.92 mmol), DPPF (360 mg, 0.65 mmol), Pd.sub.2dba.sub.3 (0.4 g,
0.43 mmol), Cs.sub.2CO.sub.3 (1.90 g, 5.83 mmol) and 12 mL toluene
was stirred at 100.degree. C. After 20 h, the mixture was allowed
to cool to ambient temperature, then purification by flash
chromatography on silica gel using hexane-EtOAc (20:1 to 4:1 v/v)
as mobile phase provided the title compound: LCMS m/z 375.6
(observed [M+H].sup.+ for the ion corresponding to loss of one Boc
group); .sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 7.86 (d,
J=2.1 Hz, 1H), 7.26 (dd, J=8.4, 2.1 Hz, 1H), 7.07 (d, J=8.4, 2.1
Hz, 1H), 3.93 (s, 3H), 2.31 (s, 3H), 1.54 (s, 9H), 1.47 (s,
9H).
Step C.
2-Hydrazino-6-(2-methoxy-5-methylphenyl)-4-nitropyridine
[0268] A mixture of the title compound from Example 19 Step B (2.0
g, 2.2 mmol), 23 mL 1,4-dioxane and concentrated HCl (2 mL) was
stirred for 14 h. Concentrated HCl (8 mL) was added dropwise. After
1 h, concentrated HCl (4 mL) was added dropwise. After 4 h, the
reaction mixture was diluted with water and concentrated to a solid
form. The solid was further washed by ether twice to give the crude
title compound which was used without further purification: LCMS
m/z 275.5 [M+H].sup.+.
Step D. Ethyl
1-[6-(2-methoxy-5-methylphenyl)-4-nitropyridin-2-yl]-5-(trifluoromethyl)--
1H-pyrazole 4-carboxylate
[0269] Reaction of the title compound from Example 19 Step C with
ethyl 2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutyrate, according
to chemistry described in Example 1 Step A, provided the title
compound: LCMS m/z 451.6 [M+H].sup.+.
Step E. Ethyl
1-[4-amino-6-(2-methoxy-5-methylphenyl)pyridin-2-yl]-5-(trifluoromethyl)--
1H-pyrazole-4-carboxylate
[0270] A mixture of the title compound from Example 19 Step D (1.2
g, 2.7 mmol), DMF (15 mL), water (1.5 mL) and tin (II) chloride
dihydrate (1.8 g, 7.9 mmol) was heated at 70.degree. C. for 1.5 h,
leading to reduction of the nitro group to the corresponding
hydroxylamine. A second addition of tin (II) chloride dehydrate
(2.4 g, 10.6 mmol) followed by heating at 100.degree. C. overnight
led to little progress. Water was added, and the reaction mixture
was extracted with hexanes-EtOAc. The organic phase was separated
and passed through a pad of silica gel. Hydrogenation using
Pd-black (450 mg, 4.2 mmol) in EtOAc:EtOH (100 mL, 1:1 v/v) under
50 psi H.sub.2, followed by filtration and concentration, provided
the title compound: LCMS m/z 421.6 [M+H].sup.+; .sup.1H NMR (400
MHz, acetone-d.sub.6) .delta. 8.09 (s, 1H), 7.71 (d, J=2.1 Hz, 1H),
7.45 (d, J=1.9 Hz, 1H), 7.13 (dd, J=8.4 Hz, 2.1 Hz, 1H), 6.96 (d,
J=8.4 Hz, 1H), 6.84 (d, J=1.9 Hz, 1H), 4.31 (q, J=7.1 Hz, 2H), 3.82
(s, 3H), 2.26 (s, 3H), 1.34 (t, J=7.1 Hz, 3H).
Step F. Ethyl
1-[4-iodo-6-(2-methoxy-5-methylphenyl)pyridin-2-yl]-5-(trifluoromethyl)-1-
H-pyrazole-4-carboxylate
[0271] Tert-butyl nitrite (0.68 mL, 0.59 mmol) was added to a
solution of the title compound from Example 19 Step E (1.2 g, 2.9
mmol) and iodine (0.87 g, 3.4 mmol) in chloroform (20 mL). The
resulting mixture was heated at 60.degree. C. for 30 min, cooled
and quenched with aqueous sodium sulfite. Extraction with
hexane-EtOAc, followed by silica gel flash chromatography using
Hexane-EtOAc (20/1 to 7/1) provided the title compound: LCMS m/z
532.6 [M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.54
(d, J=1.1 Hz, 1H), 8.15 (s, 1H), 7.98 (d, J=1.1 Hz, 1H), 7.79 (d,
J=2.1 Hz, 1H), 7.25 (dd, J=8.5 Hz, 2.1 Hz, 1H), 6.94 (d, J=8.5 Hz,
1H), 4.43 (q, J=7.1 Hz, 2H), 3.93 (s, 3H), 2.39 (s, 3H), 1.42 (t,
J=7.1 Hz, 3H).
Step G. Ethyl
1-[6-(2-hydroxy-5-methylphenyl)-4-iodopyridin-2-yl]-5-(trifluoromethyl)-1-
H-pyrazole-4-carboxylate
[0272] A solution of the title compound from Example 19 Step F (1
g, 1.88 mmol) and iodotrimethylsilane (3.6 mL) in 15 mL chloroform
was heated at 80.degree. C. for 7 h. A second portion of
iodotrimethylsilane (2 mL) was added and heating was continued at
90.degree. C. overnight. The volatiles were removed in vacuo.
Toluene was added, and the volatiles were removed in vacuo.
Finally, dry MeOH was added, and the volatiles were removed in
vacuo. Silica gel flash chromatography using hexanes:EtOAc (20:1 to
pure EtOAc) gave the title compound: LCMS m/z 518.6 [M+H].sup.+;
.sup.1H NMR (400 MHz, acetone-d.sub.6) .delta. 8.87 (d, J=1.1 Hz,
1H), 8.24 (s, 1H), 8.12 (d, J=1.1 Hz, 1H), 7.89 (d, J=2 Hz, 1H),
7.18 (dd, J=8.5, 2.0 Hz, 1H), 6.84 (d, J=8.3 Hz, 1H), 4.37 (q,
J=7.1 Hz, 2H), 2.30 (s, 3H), 1.35 (t, J=7.1 Hz, 3H).
Step H.
1-(Chloromethyl)-4-[trans]-4-(trifluoromethyl)cyclohexyl]benzene
and
1-(chloromethyl)-4-[cis]-4-(trifluoromethyl)cyclohexyl]benzene
[0273] To a solution of the title compound from Example 8 Step E
(140 mg, 0.542 mmol) in chloroform (1.4 mL) was added thionyl
chloride (0.100 mL, 1.37 mmol). The mixture was stirred at ambient
temperature for 30 min, then was concentrated in vacuo.
Purification by flash chromatography on silica gel (0 to 15% EtOAc
in hexanes then 15 to 25% EtOAc in hexanes) gave the title
compounds, as a mixture of cis:trans isomers: LCMS m/z 241.6
[M-Cl].sup.+.
Step I. Ethyl
1-{4-iodo-6-[5-methyl-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]benzyl}o-
xy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0274] A mixture of the title compound from Example 19 Step G (150
mg, 0.289 mmol), the title compound from Example 19 Step H (100 mg,
0.361 mmol), Cs.sub.2CO.sub.3 (141 mg, 0.434 mmol) and DMF (1 mL)
was stirred at RT overnight. 2 N HCl was added and the reaction
mixture was extracted with mixture of hexanes and EtOAc. The
combined organic layer was concentrated and purified by prep TLC
(20% EtOAc in hexanes) to give the title compound: LCMS m/z 758.7
[M+H].sup.+
Step J.
1-{4-Methyl-6-[5-methyl-2-({-4-[trans-4-(trifluoromethyl)cyclohexy-
l]benzyl}oxy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carbox-
ylic acid
[0275] A mixture of the title compound from Example 19 Step I (20
mg, 0.026 mmol) Pd(PPh.sub.3).sub.4 (10.7 mg, 0.009 mmol),
K.sub.2CO.sub.3 (11 mg, 0.08 mmol), trimethyl boroxine (11.6 mg,
0.09 mmol), and dioxane (0.5 mL) was heated in a microwave reactor
at 140.degree. C. for 1 h, cooled, diluted with hexanes and passed
through a pad of silica gel eluted by dichloromethane. The solvent
was removed in vacuo. Treatment with a mixture of 1,4-dioxane (0.1
mL), MeOH (0.1 mL) and 3 N NaOH (0.1 mL) at 50.degree. C. for 15
min, followed by reverse phase HPLC using a YMC C-18 column (45 to
95% acetonitrile in water, each with 0.1% v/v TFA) provided the
title compound: LCMS m/z 618.8 [M+H].sup.+; .sup.1H NMR (400 MHz,
acetone-d.sub.6), .delta. 8.17 (s, 1H), 8.08 (s, 1H), 7.75 (d,
J=2.1 Hz, 1H), 7.48 (s, 1H), 7.41 (d, J=8.1 Hz, 2H), 7.27 (d, J=8.1
Hz, 2H), 7.20 (dd, J=8.4 Hz, 2.1 Hz, 1H), 7.12 (d, J=8.4 Hz, 1H),
2.43 (s, 3H), 2.29 (s, 3H), 2.59 (m, 1H), 1.96 (m, 2H), 1.67-1.39
(m, 4H).
[0276] The compounds in TABLE 1 were prepared using the chemistry
described in Examples 1-19.
TABLE-US-00002 TABLE 1 ##STR00054## Entry R1 R2 R3 R4 MS 20
--CF.sub.3 H H ##STR00055## 592.1 [M + H].sup.+ 21 H H H
##STR00056## 546.2 [M + H].sup.+ 22 H H H ##STR00057## 572.1 [M +
Na].sup.+ 23 H H H ##STR00058## 584.1 [M + H].sup.+ 24 Me H H
##STR00059## 598.7 [M + H].sup.+ 25 Me H H ##STR00060## 564.6 [M +
H].sup.+ 26 Me H H ##STR00061## 528.4 [M - H].sup.- 27 Br H H
##STR00062## 624.5 [M + H].sup.+ 29 cPr H H ##STR00063## 584.5 [M +
H].sup.+ 30 CF.sub.3 H H ##STR00064## 618.2 [M + H].sup.+ 31
CF.sub.3 H H ##STR00065## 652.3 [M + H].sup.+ 32 CF.sub.3 H H
##STR00066## 602.2 [M + H].sup.+ 33 H H H ##STR00067## 584.4 [M +
H].sup.+ 34 H H H ##STR00068## 568.1 [M + H].sup.+ 35 H H H
##STR00069## 618.0 [M + H].sup.+ 36 H H H ##STR00070## 562.5 [M +
H].sup.+ 37 H H H ##STR00071## 610.5 [M + H].sup.+ 38 Me H H
##STR00072## 510.6 [M + H].sup.+ 39 Me H H ##STR00073## 522.5 [M +
H].sup.+ 40 H H H ##STR00074## 552.2 [M + Na].sup.+ 41 H H H
##STR00075## 600.2 [M + H].sup.+ 42 Me H H ##STR00076## 599.6 [M +
H].sup.+ 43 H H H ##STR00077## 508.6 [M + H].sup.+ 44 H H H
##STR00078## 522.6 [M + H].sup.+ 45 H H H ##STR00079## 590.5 [M +
H].sup.+ 46 H H H ##STR00080## 590.5 [M + H].sup.+ 47 Me H I
##STR00081## 730.6 [M + H].sup.+ 48 Cl H I ##STR00082## 750.6 [M +
H].sup.+ 49 H H I ##STR00083## 692.9 [M + H].sup.+ 50 cPr H H
##STR00084## 624.2 [M + H].sup.+ 51 H H H ##STR00085## 598.2 [M +
H].sup.+ 52 H H H ##STR00086## 586.2 [M + Na].sup.+ 53 H H H
##STR00087## 598.2 [M + H].sup.+ 54 H H H ##STR00088## 564.2 [M +
H].sup.+ 55 H H H ##STR00089## 496.6 [M + H].sup.+ 56 H H H
##STR00090## 602.4 [M + H].sup.+ 57 H H H ##STR00091## 568.5 [M +
H].sup.+ 58 H H H ##STR00092## 618.4 [M + H].sup.+ 59 H H H
##STR00093## 552.4 [M + H].sup.+ 60 H H H ##STR00094## 620.4 [M +
H].sup.+ 61 H H H ##STR00095## 510.5 [M + H].sup.+ 62 F H H
##STR00096## 602.3 [M + H].sup.+ 63 F H H ##STR00097## 566.3 [M -
H].sup.- 64 F H H ##STR00098## 618.3 [M + H].sup.+ 65 F H H
##STR00099## 552.4 [M + H].sup.+ 66 F H H ##STR00100## 620.4 [M +
H].sup.+ 67 F H H ##STR00101## 602.4 [M + H].sup.+ 68 H H H
##STR00102## 562.6 [M + H].sup.+ 69 H H H ##STR00103## 584.0 [M +
H].sup.+ 70 H H H ##STR00104## 618.0 [M + H].sup.+ 71 Me H H
##STR00105## 598.2 [M + H].sup.+ 72 H H H ##STR00106## 548.2 [M +
H].sup.+ 73 H H H ##STR00107## 548.5 [M - H].sup.- 74 H H H
##STR00108## 564.4 [M - H].sup.- 75 H H H ##STR00109## 548.5 [M -
H].sup.- 76 H H H ##STR00110## 598.4 [M - H].sup.- 77 H H H
##STR00111## 568.5 [M - H].sup.- 78 H H H ##STR00112## 596.2 [M +
Na].sup.+ 79 H H H ##STR00113## 552.2 [M + H].sup.+ 80 H H H
##STR00114## 584.0 [M + H].sup.+ 81 H H H ##STR00115## 602.1 [M +
Na].sup.+ 82 H H H ##STR00116## 617.8 [M + H].sup.+ 83 H H H
##STR00117## 617.8 [M + H].sup.+ 84 Me H H ##STR00118## 612.2 [M +
H].sup.+ 85 Me H H ##STR00119## 580.1 [M + Na].sup.+ 86 Me H H
##STR00120## 600.0 [M + Na].sup.+ 87 Me H H ##STR00121## 590.0 [M +
H].sup.+ 88 Me H H ##STR00122## 482.1 [M + H].sup.+ 89 H H H
##STR00123## 619.1 [M + H].sup.+ 90 H H H ##STR00124## 566.1 [M +
H].sup.+ 91 H H H ##STR00125## 534.0 [M + H].sup.+ 92 H H H
##STR00126## 558.1 [M + Na].sup.+ 93 H H H ##STR00127## 544.1 [M +
Na].sup.+ 94 H H H ##STR00128## 468.0 [M + H].sup.+ 95 Me H H
##STR00129## 488.0 [M + H].sup.+ 96 Me H H ##STR00130## 556.1 [M +
Na].sup.+ 97 Me H H ##STR00131## 460.0 [M + H].sup.+ 98 Me H H
##STR00132## 511.1 [M + H].sup.+ 99 H H H ##STR00133## 625.4 [M +
H].sup.+ 100 H H H ##STR00134## 604.4 [M + H].sup.+ 101 H H H
##STR00135## 604.5 [M + H].sup.+ 102 Me H H ##STR00136## 518.0 [M +
Na].sup.+ 103 CF3 H H ##STR00137## 536.6 [M + H].sup.+ 104 CF3 H H
##STR00138## 564.6 [M + H].sup.+ 105 Me H H ##STR00139## 578.6 [M +
H].sup.+ 106 Me H H ##STR00140## 508.6 [M + H].sup.+ 107 CF3 H H
##STR00141## 584.5 [M + H].sup.+ 108 H H H ##STR00142## 548.0 [M +
H].sup.+ 109 H H H ##STR00143## 614.1 [M + H].sup.+ 110 H H H
##STR00144## 616.2 [M + Na].sup.+ 111 H H H ##STR00145## 566.2 [M +
Na].sup.+ 112 H H H ##STR00146## 570.0 [M + Na].sup.+ 113 H H H
##STR00147## 582.0 [M + Na].sup.+ 114 H H H ##STR00148## 538.8 [M +
H].sup.+ 115 H H H ##STR00149## 540.9 [M + H].sup.+ 116 H H H
##STR00150## 618.1 [M + H].sup.+ 117 H H H ##STR00151## 540.5 [M +
H].sup.+ 118 H H H ##STR00152## 572.5 [M + H].sup.+ 119 H H H
##STR00153## 640.2 [M + H].sup.+ 120 Me H H ##STR00154## 524.6 [M +
H].sup.+ 121 H H H ##STR00155## 564.2 [M + H].sup.+ 122 H H H
##STR00156## 624.2 [M + H].sup.+ 123 H H H ##STR00157## 510.6 [M +
H].sup.+ 124 Me H H ##STR00158## 530.7 [M + H].sup.+ 125 H H H
##STR00159## 546.6 [M + Na].sup.+ 126 Cl H H ##STR00160## 580.6 [M
+ Na].sup.+ 127 H H H ##STR00161## 536.7 [M + H].sup.+ 128 H H H
##STR00162## 536.9 [M + H].sup.+ 129 Cl H H ##STR00163## 568.7 [M -
H].sup.- 130 Cl H H ##STR00164## 568.8 [M - H].sup.- 131 H H H
##STR00165## 548.8 [M - H].sup.- 132 H H H ##STR00166## 548.8 [M -
H].sup.- 133 Cl H H ##STR00167## 616.6 [M + Na].sup.+ 134 Cl H H
##STR00168## 630.8 [M - H].sup.- 135 Cl H H ##STR00169## 626.7 [M -
H].sup.- 136 Me H H ##STR00170## 538.8 [M + H].sup.+ 137 Me H H
##STR00171## 550.9 [M + H].sup.+ 138 Me H H ##STR00172## 550.9 [M +
H].sup.+ 139 H H H ##STR00173## 608.8 [M + H].sup.+ 140 Me H H
##STR00174## 566.6 [M + H].sup.+ 141 Me H H ##STR00175## 586.6 [M +
H].sup.+ 142 H H H ##STR00176## 550.9 [M - H].sup.-
143 cPr H H ##STR00177## 508.8 [M + H].sup.+ 144 Cl H H
##STR00178## 500.7 [M - H].sup.- 145 H H H ##STR00179## 562.4 [M -
H].sup.- 146 H Cl H ##STR00180## 578.5 [M + H].sup.+ 147 H F H
##STR00181## 586.5 [M + Na].sup.+ 148 H F H ##STR00182## 620.5 [M +
Na].sup.+ 149 H F H ##STR00183## 602.6 [M + H].sup.+ 150 H H cPr
##STR00184## 508.8 [M + H].sup.+ 151 H H Cl ##STR00185## 500.7 [M -
H].sup.- 152 Me H Cl ##STR00186## 516.7 [M + H].sup.+ 153 I H H
##STR00187## 592.7 [M - H].sup.- 154 H H I ##STR00188## 592.7 [M +
H].sup.+ 155 Me H Cl ##STR00189## 530.7 [M + H].sup.+ 156 Me H Cl
##STR00190## 556.7 [M + H].sup.+ 157 Me H Cl ##STR00191## 522.6 [M
+ H].sup.+ 158 H H H ##STR00192## 580.0 [M + H].sup.+ 159 H H Me
##STR00193## 604.9 [M + H].sup.+ 160 H H Me ##STR00194## 604.9 [M +
H].sup.+ 161 --CF.sub.3 H H ##STR00195## 612.6 [M + H].sup.+ 162 Cl
H H ##STR00196## 558.5 [M + H].sup.+ 163 H H H ##STR00197## 508.6
[M + H].sup.+ 164 H H H ##STR00198## 524.5 [M + H].sup.+ 165 H H H
##STR00199## 516.6 [M + H].sup.+ 166 Cl H H ##STR00200## 578.4 [M -
H].sup.- 167 Cl H H ##STR00201## 550.2 [M + H].sup.+ 168 Cl H H
##STR00202## 618.2 [M + H].sup.+ 169 Cl H H ##STR00203## 584.2 [M +
H].sup.+ 170 Cl H H ##STR00204## 618.2 [M + H].sup.+ 171 H H H
##STR00205## 508.5 [M + H].sup.+ 172 H H H ##STR00206## 542.4 [M +
H].sup.+ 173 H H H ##STR00207## 517.5 [M + H].sup.+ 174 H H H
##STR00208## 551.4 [M + H].sup.+ 175 H H H ##STR00209## 585.4 [M +
H].sup.+ 176 Me H H ##STR00210## 558.5 [M + H].sup.+ 177 F H H
##STR00211## 562.5 [M + H].sup.+
[0277] The compounds in TABLE 2 were prepared using the chemistry
described in Examples 1-9.
TABLE-US-00003 TABLE 2 ##STR00212## Entry R1 R2 R3 R4 MS 178 H H Me
##STR00213## 490.2 [M + H].sup.+ 179 Cl H Me ##STR00214## 524.1 [M
+ H].sup.+ 180 H H Me ##STR00215## 462.2 [M + H].sup.+ 181 Cl H Me
##STR00216## 496.1 [M + H].sup.+ 182 H H Me ##STR00217## 496.1 [M +
H].sup.+ 183 H H Me ##STR00218## 529.9 [M + H].sup.+ 184 H H CF2H
##STR00219## 532.0 [M + H].sup.+ 185 H H CF2H ##STR00220## 566.2 [M
+ H].sup.+ 186 H H CF2H ##STR00221## 526.2 [M + H].sup.+ 187 Cl H
CF2H ##STR00222## 566.0 [M + H].sup.+ 188 Cl H CF2H ##STR00223##
600.0 [M + H].sup.+ 189 H H CF2H ##STR00224## 582.0 [M + H].sup.+
190 H H CF2H ##STR00225## 566.1 [M + H].sup.+ 191 Me H CF2H
##STR00226## 580.5 [M + H].sup.+ 192 Me H CF2H ##STR00227## 546.5
[M + H].sup.+ 193 H H CF2H ##STR00228## 580.5 [M + H].sup.+ 194 H H
CF2H ##STR00229## 546.5 [M + H].sup.+ 195 H H CF2H ##STR00230##
584.4 [M + H].sup.+ 196 H H CF2H ##STR00231## 550.4 [M + H].sup.+
197 H H Et ##STR00232## 544.5 [M + H].sup.+ 198 H H iPr
##STR00233## 558.5 [M + H].sup.+ 199 H H cPr ##STR00234## 556.5 [M
+ H].sup.+ 200 H H --CH.sub.2OMe ##STR00235## 560.5 [M + H].sup.+
201 H H Et ##STR00236## 558.5 [M + H].sup.+ 202 H H iPr
##STR00237## 572.5 [M + H].sup.+ 203 H H cPr ##STR00238## 570.5 [M
+ H].sup.+ 204 H H --CH.sub.2OMe ##STR00239## 574.5 [M + H].sup.+
205 H H CF2H ##STR00240## 572.6 [M + H].sup.+ 206 H H CF2H
##STR00241## 572.6 [M + H].sup.+ 207 H H Pr ##STR00242## 572.6 [M +
H].sup.+ 208 H H Ph ##STR00243## 606.6[M + H].sup.+ 209 Cl Cl CF2H
##STR00244## 648.4 [M + H].sup.+ 210 Cl H CF2H ##STR00245## 614.5
[M + H].sup.+ 211 Cl H CF2H ##STR00246## 606.5 [M + H].sup.+ 212 Cl
H CF2H ##STR00247## 606.5 [M + H].sup.+ 213 Cl Cl CF2H ##STR00248##
640.5 [M + H].sup.+ 214 H H CF2H ##STR00249## 587.0 [M + H].sup.+
215 H H CF2H ##STR00250## 587.0 [M + H].sup.+
[0278] The compounds in TABLE 3 were synthesized using chemistry
described in Examples 1-19.
TABLE-US-00004 TABLE 3 Entry R MS 216 ##STR00251## 589.5 [M +
H].sup.+ 217 ##STR00252## 589.5 [M + H].sup.+ 218 ##STR00253##
583.5 [M + H].sup.+ 219 ##STR00254## 549.5 [M + H].sup.+ 220
##STR00255## 669.9 [M + H].sup.+ 221 ##STR00256## 669.9 [M +
H].sup.+ 222 ##STR00257## 599.5 [M + H].sup.+ 223 ##STR00258##
604.5 [M + H].sup.+ 224 ##STR00259## 570.4 [M + H].sup.+ 225
##STR00260## 556.5 [M + H].sup.+ 226 ##STR00261## 608.5 [M +
H].sup.+ 227 ##STR00262## 574.4 [M + H].sup.+ 228 ##STR00263##
730.8 [M + H].sup.+ 229 ##STR00264## 618.8 [M + H].sup.+ 230
##STR00265## 730.8 [M + H].sup.+ 231 ##STR00266## 638.8 [M +
H].sup.+ 232 ##STR00267## 551.1 [M + H].sup.+ 233 ##STR00268##
517.2 [M + H].sup.+ 234 ##STR00269## 750.9 [M + H].sup.+ 235
##STR00270## 750.9 [M + H].sup.+
Example 236
##STR00271##
[0279] Step A. Ethyl
1-(6-{2-[(4-methoxyphenyl)ethynyl]phenyl}pyridin-2-yl)-5-(trifluoromethyl-
)-1H-pyrazole-4-carboxylate
[0280] To a cooled (0.degree. C.) solution of the title compound
from Example 1 Step B (374 mg, 0.991 mmol) and pyridine (0.241 mL,
2.97 mmol) in DCM (5 mL) was added trifluoromethanesulfonic
anhydride (0.251 mL, 1.487 mmol). The cooling bath was immediately
removed, and the reaction mixture was allowed to stir at ambient
temperature. After 45 min, the reaction mixture was poured into
water and extracted with DCM. The layers were separated and the
organic phase was concentrated in vacuo, yielding material that was
sufficiently pure for use in the next step: LCMS m/z 510.0
[M+H].sup.+. To a flask containing the unpurified triflate were
added copper (I) iodide (56.1 mg, 0.294 mmol),
trans-dichlorobis(triphenylphosphine) palladium (II) (68.9 mg,
0.098 mmol), tetrabutylammonium iodide (1.088 g, 2.94 mmol), and
4-methoxyphenylacetylene (195 mg, 1.47 mmol). The flask with
flushed with nitrogen, and acetonitrile (5 mL) was added. The
mixture was degassed with nitrogen, triethylamine (1.00 mL, 7.17
mmol) was added, and the resulting mixture was stirred at ambient
temperature. After 20 h, the reaction mixture was concentrated in
vacuo. Purification by flash chromatography on silica gel (0 to 30%
EtOAc in hexanes, then 30 to 100% EtOAc in hexanes) provided the
title compound: LCMS m/z 492.1 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.32 (d, J=8.0 Hz, 1H), 8.14 (s, 1H), 7.99 (t,
J=8.0 Hz, 1H), 7.89 (d, J=7.5 Hz, 1H), 7.65-7.63 (m, 2H), 7.47-7.39
(m, 2H), 7.33 (d, J=8.5 Hz, 2H), 6.86 (d, J=8.5 Hz, 2H), 4.38 (q,
J=7.0 Hz, 2H), 3.82 (s, 3H), 1.39 (t, J=7.0 Hz, 3H).
Step B. Ethyl
1-(6-{2-[2-(4-methoxyphenyl)ethyl]phenyl}pyridin-2-yl)-5-(trifluoromethyl-
)-1H-pyrazole-4-carboxylate
[0281] To a degassed solution of the title compound from Example
236 Step A (255 mg, 0.519 mmol) in EtOAc (3 mL) and EtOH (3 mL) was
added platinum(IV) oxide (175 mg). The reaction mixture was fitted
with a hydrogen balloon attached to a 3-way adapter. The reaction
flask was then evacuated and back-filled with hydrogen. After this
process was repeated three times, the reaction mixture was placed
under a hydrogen atmosphere, and was stirred vigorously. After 2 h,
the reaction mixture was filtered through Celite, rinsing with
EtOAc. The mixture was then concentrated in vacuo, and used without
further purification: LCMS m/z 496.2 [M+H].sup.+.
Step C. Ethyl
5-(trifluoromethyl)-1-(6-{2-[2-(4-{[(trifluoromethyl)sulfonyl]oxy}phenyl)-
ethyl]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylate
[0282] To a cooled (0.degree. C.) solution of the title compound
from Example 236 Step B (177 mg, 0.357 mmol) in DCM (3 mL) was
added boron tribromide (1.07 mL, 1.0 M in DCM, 1.07 mmol). After 45
min, the reaction mixture was quenched by addition of sat. aq.
NaHCO.sub.3, then was allowed to warm to ambient temperature. The
aqueous phase was extracted with DCM. The organic phase was dried
over sodium sulfate, filtered, and concentrated in vacuo. The
resulting phenol was used without further purification: LCMS m/z
482.2 [M+H].sup.+. To a cooled (0.degree. C.) DCM (5 mL) solution
of the product obtained above were added pyridine (0.087 mL, 1.07
mmol) and trifluoromethanesulfonic anhydride (0.091 mL, 0.536
mmol), and the resulting mixture was allowed to warm to ambient
temperature. After 1 h, the reaction mixture was quenched by
addition of sat. aq. NaHCO.sub.3 and extracted with DCM. The
organic phase was separated and concentrated in vacuo. Purification
by flash chromatography on silica gel (0 to 40% EtOAc in hexanes,
then 40 to 100% EtOAc in hexanes) provided the title compound: LCMS
m/z 614.2 [M+H].sup.+.
Step D.
5-(Trifluoromethyl)-1-[6-(2-{2-[4'-(trifluoromethyl)biphenyl-4-yl]-
ethyl}phenyl)pyridin-2-yl]-1H-pyrazole-4-carboxylic acid
[0283] A vial was charged with the title compound from Example 236
Step C (60.0 mg, 0.098 mmol), 4-trifluoromethylphenyl boronic acid
(24.2 mg, 0.127 mmol), and trans-dichlorobis(triphenylphosphine)
palladium (II) (6.9 mg, 0.010 mmol). Acetonitrile (0.400 mL) and
sodium carbonate (0.244 mL, 1.0 M aqueous, 0.244 mmol) were added,
and the mixture was degassed with nitrogen. The vial was then
capped and placed in a pre-heated oil bath (70.degree. C.). After
15 h, the mixture was diluted with water and DCM and the organic
phase was filtered through a short pad of silica gel and Celite
with DCM, then was concentrated in vacuo: LCMS m/z 610.4
[M+H].sup.+. To a solution of the unpurified Suzuki product in
1,4-dioxane (2.0 mL) was added lithium hydroxide (1.0 mL, 2.0 M in
water, 2.00 mmol), and the resulting mixture was stirred at
50.degree. C. After 1 h, the reaction mixture was rendered acidic
by addition of aqueous hydrochloric acid, then was diluted with
1,4-dioxane and passed through a 0.45 micron syringe filter.
Purification by reverse phase HPLC (60 to 100% acetonitrile in
water, each with 0.1% v/v TFA) provided the title compound: LCMS
m/z 582.4 [M+H].sup.+; .sup.1H NMR (500 MHz, d.sub.6-DMSO) .delta.
8.34 (s, 1H), 8.20 (t, J=8.0 Hz, 1H), 7.82-7.73 (m, 6H), 7.50 (d,
J=8.0 Hz, 2H), 7.44-7.34 (m, 4H), 6.99 (d, J=8.0 Hz, 2H), 2.97-2.93
(m, 2H), 2.78-2.74 (m, 2H).
Example 237
##STR00272##
[0284] Step A. Ethyl
1-(2'-fluoro-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carbo-
xylate
[0285] A mixture of the title compound from Example 1 Step A (100
mg, 0.3 mmol), (2-fluoropyridin-3-yl)boronic acid (66 mg, 0.47
mmol), trans dichlorobis(triphenylphosphine) palladium (II) (31.0
mg, 0.05 mmol), Na.sub.2CO.sub.3 (0.47 mL, 2.0 M aqueous, 0.94
mmol) and acetonitrile (1 mL) in a nitrogen-filled capped vial was
stirred at 100.degree. C. After 50 min, the mixture was allowed to
cool to ambient temperature, then was concentrated in vacuo.
Purification by flash chromatography on silica gel using
hexane:EtOAc (6:1 to 4:1 v/v) as mobile phase provided the title
compound: LCMS m/z 381.1 [M+H].sup.+; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.63 (m, 1H), 8.27 (m, 1H), 8.13 (s, 1H), 8.10
(d, J=7.9 Hz, 1H), 8.03 (t, J=7.9 Hz, 1H), 7.70 (d, J=7.8 Hz, 1H),
7.35 (m, 1H), 4.39 (q, J=7.1 Hz, 2H), 1.39 (t, J=7.1 Hz, 3H).
Step B.
5-(Trifluoromethyl)-1-(2'-{[4'-(trifluoromethyl)biphenyl-4-yl]meth-
oxy}-2,3'-bipyridin-6-yl)-1H-pyrazole-4-carboxylic acid
[0286] To a vial were added successively KOtBu (15.0 mg, 0.13
mmol), [4'-(trifluoromethyl)biphenyl-4-yl]methanol (US Patent
2004209936) (36.0 mg, 0.14 mmol) and DMF (0.3 mL). After 5 min, the
title compound from Example 237 Step A (20.0 mg, 0.05 mmol) was
added. After 30 min, the reaction mixture was treated with NaOH
(0.1 mL, 3 N aqueous, 0.3 mmol), MeOH (0.1 mL) and 1,4-dioxane (0.1
mL) at 50.degree. C. for 20 min. Reverse phase HPLC using a YMC
C-18 column (65 to 100% acetonitrile in water, each with 0.1% v/v
TFA) gave the title compound: LCMS m/z 584.9 [M+H].sup.+; .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.44 (dd, J=7.6, 1.9 Hz, 1H),
8.31 (d, J=7.8 Hz, 1H), 8.27 (dd, J=4.9, 2.0 Hz, 1H), 8.20 (s, 1H),
7.94 (t, J=7.9 Hz, 1H), 7.69 (s, 4H), 7.62-7.53 (m, 5H), 7.10 (dd,
J=7.5, 4.9 Hz, 1H), 5.63 (s, 2H).
Example 238
##STR00273##
[0287] Step A. Ethyl
1-(5'-bromo-2'-fluoro-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazol-
e-4-carboxylate
[0288] A mixture of the title compound from Example 1 Step A (2.0
g, 6.3 mmol), chloroform (20 mL) and 57% HI (20 mL) was heated at
100.degree. C. with vigorous stirring. After 22 h, the organic
phase was washed with brine, water, and aq. NaHCO.sub.3 and the
combined aqueous phases were extracted with DCM. The combined
organic phases were concentrated in vacuo. Purification by flash
chromatography on silica gel (hexanes-EtOAc, 9:1 to 4:1 v/v)
provided a mixture (.about.2:1) of the title compound (LCMS m/z
412.0 [M+H].sup.+) and the title compound from Example 1 Step A.
The crude mixture obtained above (1.95 g),
(5-bromo-2-fluoropyridin-3-yl)boronic acid (1.31 g, 5.96 mmol),
tetrakis(triphenylphosphine)palladium(0) (274 mg, 0.237 mmol),
Na.sub.2CO.sub.3 (9.5 mL, 2 M aqueous, 19 mmol) and acetonitrile
(25 mL) were stirred at 100.degree. C. for 30 min. Aqueous workup
and purification by silica gel flash chromatography using
hexane-triethylamine (19:1 to 7:1 v/v) as mobile phase provided the
title compound: LCMS m/z 461.0 [M+H].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 8.78 (dd, J=8.5, 2.5 Hz, 1H), 8.36 (m,
1H), 8.17 (s, 1H), 8.13 (m, 2H), 7.80 (m, 1H), 4.41 (q, J=7.1 Hz,
2H), 1.42 (t, J=7.1 Hz, 3H).
Step B. Ethyl
1-(2'-fluoro-5'-methyl-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazo-
le-4-carboxylate
[0289] A mixture of the title compound from Example 238 Step A (120
mg, 0.26 mmol), tetrakis(triphenylphosphine)palladium(0) (30.0 mg,
0.026 mmol), K.sub.2CO.sub.3 (72.0 mg, 0.52 mmol), trimethyl
boroxine (33.0 mg, 0.26 mmol), and dioxane (1.5 mL) was heated in a
microwave reactor at 140.degree. C. for 35 min, cooled, filtered
and purified by silica gel flash chromatography (hexanes-EtOAc,
93:7 to 85:15 v/v) to yield the title compound: LCMS m/z 395.1
[M+H].sup.+.
Step C.
1-(5'-Methyl-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]metho-
xy}-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0290] The title compound was prepared according to the procedure
described in Example 237 Step B, by reaction of the title compound
from Example 238 Step B with
[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methanol (PCT
Publication WO2005118542): LCMS m/z 612.9 [M+H].sup.+; .sup.1H NMR
(500 MHz, acetone-d.sub.6) .delta. 8.18 (m, 2H), 7.91 (d, J=8.2 Hz,
2H), 7.80 (m, 3H), 7.63 (m, 2H), 5.66 (s, 2H), 2.51 (s, 3H), 2.37
(s, 3H).
Example 239
##STR00274##
[0291] Step A. Ethyl
1-(5'-chloro-2'-fluoro-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazo-
le-4-carboxylate
[0292] A mixture of the title compound from Example 238 Step A (115
mg, 0.25 mmol), CuCl (74 mg, 0.75 mmol) and DMF (1 mL) was heated
in a microwave reactor at 170.degree. C. for 10 min, diluted with
DCM, filtered, concentrated and purified by silica gel flash
chromatography (hexanes:EtOAc, 95:5 to 85:15 v/v) to provide the
title compound: LCMS m/z 415.1 [M+H].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 8.64 (dd, J=8.4, 2.6 Hz, 1H), 8.26 (m,
1H), 8.17 (s, 1H), 8.13 (m, 2H), 7.80 (m, 1H), 4.41 (q, J=7.2 Hz,
2H), 1.42 (t, J=7.2 Hz, 3H).
Step B.
1-(5'-Chloro-2'-{[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]metho-
xy}-2,3'-bipyridin-6-yl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0293] The title compound was prepared according to the procedure
described in Example 237 Step B, by reaction of the title compound
from Example 239 Step A with
[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methanol (PCT
Publication WO2005118542): LCMS m/z 632.7 [M+H].sup.+; .sup.1H NMR
(500 MHz, acetone-d.sub.6) .delta. 8.22 (m, 2H), 7.92 (d, J=8.2 Hz,
2H), 7.83 (m, 3H), 7.63 (m, 2H), 5.70 (s, 2H), 2.52 (s, 3H).
Example 240
##STR00275##
[0294] Step A. Ethyl
1-[2'-fluoro-5'-(trifluoromethyl)-2,3'-bipyridin-6-yl]-5-(trifluoromethyl-
)-1H-pyrazole-4-carboxylate
[0295] A mixture of 3-chloro-2-fluoro-5-(trifluoromethyl)pyridine
(1.30 g, 6.52 mmol), bis(pinacolato)diboron (2.00 g, 7.87 mmol),
KOAc (1.52 g, 15.5 mmol), 260 mg
bis(tricyclohexylphosphine)palladium(0) (260 mg, 0.40 mmol) and
1,4-dioxane (10 mL) was heated at 100.degree. C. for 50 min. Water
was added and the reaction mixture was extracted with EtOAc. The
organic phase was dried over sodium sulfate, passed through a
silica pad and concentrated. Hexane was added and the reaction
mixture was filtered, and concentrated to give crude
2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluorometh-
yl)pyridine. A mixture of the title compound from Example 1 Step A
(1.00 g, 3.1 mmol), the crude compound obtained above
(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromet-
hyl)pyridine, 2.09 g), trans-dichlorobis(triphenylphosphine)
palladium (II) (154 mg, 0.22 mmol), CsF (1.43 g, 9.40 mmol),
Na.sub.2CO.sub.3 (3.9 mL, 2.0 M aqueous, 7.8 mmol) and acetonitrile
(15 mL) was stirred at 100.degree. C. for 45 min. Aqueous work up
with water, hexane and EtOAc, followed by silica gel flash
chromatography (hexanes:EtOAc, 9:1 to 8.5:1.5 v/v) gave the title
compound: LCMS m/z 449.1 [M+H].sup.+; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.96 (dd, J=8.7, 2.5 Hz, 1H), 8.56 (s, 1H),
8.16-8.12 (m, 2H), 8.08 (t, J=7.9 Hz, 1H), 7.79 (dd, J=7.8, 0.8 Hz,
1H), 4.39 (q, J=7.2 Hz, 2H), 1.40 (t, J=7.2 Hz, 3H).
Step B.
1-[2'-{[3-Methyl-4'-(trifluoromethyl)biphenyl-4-yl]methoxy}-5'-(tr-
ifluoromethyl)-2,3'-bipyridin-6-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carb-
oxylic acid
[0296] The title compound was prepared according to the procedure
described in Example 237 Step B, by reaction of the title compound
from Example 240 Step A with
[3-methyl-4'-(trifluoromethyl)biphenyl-4-yl]methanol (PCT
Publication WO2005118542): LCMS m/z 666.8 [M+H].sup.+; .sup.1H NMR
(500 MHz, acetone-d.sub.6) 6 (ppm) 8.78 (d, J=2.3 Hz, 1H), 8.71 (s,
1H), 8.47 (d, J=8.0 Hz, 1H), 8.26 (m, 2H), 8.22 (m, 1H), 7.92 (d,
J=8.2 Hz, 2H), 7.88 (m, 1H), 7.81 (d, J=8.5 Hz, 2H), 7.66 (s, 1H),
7.59 (dd, J=7.9, 1.7 Hz, 1H), 5.80 (s, 2H), 2.54 (s, 3H).
Example 241
##STR00276##
[0297] Step A. Ethyl
1-{6-[2-(hydroxymethyl)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazo-
le-4-carboxylate
[0298] A mixture of the title compound from Example 1 Step A (300
mg, 0.94 mmol), 2,1-benzoxaborol-1(3H)-ol (189 mg, 1.40 mmol),
trans-dichlorobis(triphenylphosphine) palladium (II) (94 mg, 0.14
mmol), Na.sub.2CO.sub.3 (1.4 mL, 2.0 M aqueous, 2.8 mmol) and
acetonitrile (1.5 mL) in a nitrogen-filled capped vial was stirred
at 100.degree. C. for 1.5 h, cooled, concentrated and purified by
silica gel flash chromatography (hexanes-EtOAc, 3:1 to 2:1 v/v) to
provide the title compound: LCMS 374.1 [M-OH].sup.+; .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.16 (s, 1H), 8.06 (t, J=7.9 Hz, 1H),
7.77 (d, J=7.6 Hz, 1H), 7.59-7.41 (m, 5H), 4.51 (s, 2H), 4.36 (q,
J=7.1 Hz, 2H), 1.40 (t, J=7.1 Hz, 3H).
Step B.
5-(Trifluoromethyl)-1-{6-[2-({[4'-(trifluoromethyl)biphenyl-4-yl]o-
xy}methyl)phenyl]pyridin-2-yl}-1H-pyrazole-4-carboxylic acid
[0299] DIAD (0.03 mL, 0.15 mmol) was added dropwise to a solution
of the title compound from Example 241 Step A (33.0 mg, 0.08 mmol),
4'-(trifluoromethyl)biphenyl-4-ol (33 mg, 0.14 mmol) and PPh.sub.3
(34 mg, 0.13 mmol) in THF (0.5 mL). The reaction mixture was aged
for 15 min, concentrated and treated with a mixture of 1,4-dioxane
(0.15 mL), MeOH (0.15 mL) and 3 N NaOH (0.15 mL) at 60.degree. C.
for 30 min. Reverse phase HPLC using a YMC C-18 column (45 to 100%
acetonitrile in water, each with 0.1% v/v TFA) provided the title
compound: LCMS m/z 584.2 [M+H].sup.+; .sup.1H NMR (500 MHz,
CDCl.sub.3) .delta. 8.23 (s, 1H), 8.05 (t, J=7.9 Hz, 1H), 7.78 (d,
J=7.8 Hz, 1H), 7.74-7.48 (m, 11H), 6.98 (d, J=8.7 Hz, 2H), 5.28 (s,
2H).
Example 242
##STR00277##
[0300] Step A.
5-(Trifluoromethyl)-1-(6-{2-[({5-[4-(trifluoromethyl)phenyl]pyridin-2-yl}-
oxy)methyl]phenyl}pyridin-2-yl)-1H-pyrazole-4-carboxylic acid
[0301] To a solution of 5-bromo-2-fluoropyridine (72 mg, 0.41 mmol)
and the title compound from Example 241 Step A (80 mg, 0.2 mmol) in
DMF (1 mL) was added KOtBu (15 mg, 0.22 mmol). After 25 minutes the
reaction mixture was quenched by addition of sat. aq. NH.sub.4Cl.
The reaction mixture was concentrated and purified by silica gel
flash chromatography (7% to 20% EtOAc in hexanes) to give a mixture
containing about 50% of the desired product, ethyl
1-[6-(2-{[(5-bromopyridin-2-yl)oxy]methyl}phenyl)pyridin-2-yl]-5-(trifluo-
romethyl)-1H-pyrazole 4-carboxylate, according to LCMS analysis:
LCMS m/z 548.9 [M+H].sup.+. A mixture of the material obtained
above (35 mg), 4-trifluoromethylphenylboronic acid (24 mg, 0.13
mmol), tetrakis(triphenylphosphine)palladium(0) (15.0 mg, 0.013
mmol), Na.sub.2CO.sub.3 (0.128 mL, 2 M aqueous, 0.256 mmol) and DME
(0.7 mL) in a nitrogen-filled capped vial was stirred at
112.degree. C. for 15 min, cooled, concentrated and treated with a
mixture of 3 N NaOH (0.1 mL), MeOH (0.1 mL) and 1,4-dioxane (0.1
mL) at 60.degree. C. for 30 min. Reverse phase HPLC using a YMC
C-18 column (45 to 100% acetonitrile in water, each with 0.1% v/v
TFA) gave the title compound: LCMS m/z 584.9 [M+H].sup.+; .sup.1H
NMR (500 MHz, d.sub.6-DMSO) .delta. 8.40 (d, J=2.4 Hz, 1H), 8.22
(m, 2H), 8.04 (dd, J=8.7, 2.4 Hz, 1H), 7.89 (d, J=7.8 Hz, 1H), 7.86
(d, J=8.0 Hz, 2H), 7.80 (d, J=8.0 Hz, 2H), 7.76 (d, J=8.0 Hz, 1H),
7.51 (m, 2H), 6.83 (d, J=8.5 Hz, 1H).
Example 243
##STR00278##
[0302] Step A. Ethyl
1-[6-(5-methyl-2-{[(trifluoromethyl)sulfonyl]oxy}phenyl)pyridin-2-yl]-5-(-
trifluoromethyl)-1H-pyrazole-4-carboxylate
[0303] To a cooled (-78.degree. C.) solution of the title compound
from Example 8 Step B (2.05 g, 5.24 mmol) and pyridine (1.06 mL,
13.1 mmol) in DCM (50 mL) was added triflic anhydride (1.06 mL,
6.29 mmol), and the reaction mixture was allowed to warm to ambient
temperature. After the reaction was complete, the mixture was
quenched with 2 N aqueous HCl and the aqueous phase was extracted
with hexanes:ethyl acetate (3:1 v/v). The organic phase was
separated, dried over sodium sulfate, passed through a pad of
silica gel, eluting with DCM, and concentrated in vacuo. The title
compound was used without further purification: LCMS m/z 524.6
[M+H].sup.+; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta. 8.14 (s,
1H), 8.02 (t, J=7.9 Hz, 1H), 7.81 (d, J=7.8 Hz, 1H), 7.71 (s, 1H),
7.68 (d, J=8.0 Hz, 1H), 7.30 (s, 2H), 4.39 (q, J=7.2 Hz, 2H), 2.44
(s, 3H), 1.39 (t, J=7.2 Hz, 3H).
Step B. Ethyl
1-(6-{2-[(4-methoxybenzyl)thio]-5-methylphenyl}pyridin-2-yl)-5-(trifluoro-
methyl)-1H-pyrazole-4-carboxylate
[0304] To a solution of title compound from Example 243 Step A
(2.74 g, 5.23 mmol), 4-methoxy .alpha.-toluenethiol (0.88 mL, 6.28
mmol) in 1,4-dioxane (75 mL) were added DIEA (1.83 mL, 10.5 mmol),
Xantphos (0.61 g, 1.05 mmol) and
tris(dibenzylideneacetone)dipalladium (0) (0.48 g, 0.52 mmol), and
the reaction mixture was heated at 90.degree. C. After 15 h, the
reaction mixture was allowed to cool to ambient temperature, then
was diluted with hexane. The resulting yellow solid was removed by
filtration, and the collected organic filtrate was concentrated in
vacuo. Purification by flash chromatography on silica gel (5 to 20%
ethyl acetate in hexanes) provided the title compound: LCMS m/z
528.6 [M+H].sup.+; .sup.1H NMR (500 MHz, CD.sub.2Cl.sub.2) .delta.
8.11 (s, 1H), 7.94 (t, J=7.9 Hz, 1H), 7.72 (d, J=7.8 Hz, 1H), 7.59
(d, J=8.0 Hz, 1H), 7.36 (m, 2H), 7.17 (d, J=8.0 Hz, 1H), 7.04 (d,
J=8.5 Hz, 2H) 6.74 (d, J=8.7 Hz, 2H) 4.35 (q, J=7.1 Hz, 2H), 3.87
(s, 2H), 3.73 (s, 3H), 2.37 (s, 3H), 1.37 (t, J=7.1 Hz, 3H).
Step C. Ethyl
1-[6-(2-mercapto-5-methylphenyl)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyra-
zole-4-carboxylate and diethyl
1,1'-{dithiobis[(5-methyl-2,1-phenylene)pyridine-6,2-diyl]}bis[5-(trifluo-
romethyl)-1H-pyrazole-4-carboxylate]
[0305] To a solution of the title compound from Example 243 Step B
(2.76 g, 5.23 mmol) in TFA (15 mL) was added anisole (1.71 mL, 15.7
mmol), and the resulting mixture was heated at 60.degree. C. After
15 h, the mixture was allowed to cool to ambient temperature then
was concentrated in vacuo. Purification by flash chromatography on
silica gel (5 to 20% ethyl acetate in hexanes) provided the title
compound as a mixture of monomer and disulfide dimer. LCMS m/z
408.6 [M+H].sup.+ (monomer), LCMS m/z 406.6 [M+H].sup.+ (dimer);
.sup.1H NMR (500 MHz, CD.sub.2Cl.sub.2) (dimer) .delta. 8.14 (s,
1H), 7.97 (t, J=7.9 Hz, 1H), 7.67 (d, J=7.8 Hz, 1H), 7.58 (d, J=8.1
Hz, 1H), 7.53 (d, J=8.1 Hz, 1H), 7.35 (d, J=1.1 Hz, 1H), 7.13 (dd,
J=8.1, 1.5 Hz, 1H), 4.36 (q, J=7.2 Hz, 2H), 2.35 (s, 3H), 1.37 (t,
J=7.1 Hz, 3H).
Step D. Ethyl
1-(6-{2-[(4-bromobenzyl)thio]-5-methylphenyl}pyridin-2-yl)-5-(trifluorome-
thyl)-1H-pyrazole-4-carboxylate
[0306] To a solution of the title compounds from Example 243 Step C
(mixture of monomer and dimer, 209 mg, ca. 0.54 mmol) and
4-bromobenzyl bromide (192 mg, 0.77 mmol) in DMF (3 mL) was added
cesium carbonate (501 mg, 1.54 mmol), and the resulting mixture was
stirred for 30 minutes, until the monomer was consumed. Next,
sodium borohydride (58 mg, 1.54 mmol) was added, and the reaction
mixture was stirred again for 45 minutes, resulting in cleavage of
the disulfide bond of the dimer and formation of the desired
product. Once the reaction reached completion, the mixture was
cooled to 0.degree. C. and quenched by addition of 2 N aqueous HCl.
The aqueous phase was extracted with a 3:1 mixture of ethyl acetate
in hexanes, and the organic phase was dried over sodium sulfate and
concentrated in vacuo. Purification by flash chromatography on
silica gel (0 to 15% ethyl acetate in hexanes) provided the title
compound: LCMS m/z 576.6 [M+H].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 8.12 (s, 1H), 7.96 (t, J=7.8 Hz, 1H),
7.70 (d, J=7.8 Hz, 1H), 7.60 (d, J=8.1 Hz, 1H), 7.36 (d, J=1.4 Hz,
1H), 7.32 (d, J=8.2 Hz, 3H), 7.16 (dd, J=7.9, 1.5 Hz, 1H), 6.97 (d,
J=8.5 Hz, 2H), 4.35 (q, J=7.1 Hz, 2H), 3.81 (s, 2H), 2.37 (s, 3H),
1.37 (t, J=7.2 Hz, 3H).
Step E.
1-{6-[5-Methyl-2-({[4'-(trifluoromethyl)biphenyl-4-yl]methyl}thio)-
phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0307] A solution of the title compound from Example 243 Step D (24
mg, 0.04 mmol), 4-(trifluoromethyl)phenylboronic acid (10.4 mg,
0.06 mmol), trans dichlorobis(triphenylphosphine) palladium (II)
(8.9 mg, 0.01 mmol), and cesium fluoride (19 mg, 0.13 mmol), in
acetonitrile (0.5 mL) was stirred for 5 min, then sodium carbonate
(0.13 mL, 1.0 M aqueous, 0.13 mmol) was added. The resulting
mixture was stirred at 90.degree. C. After 30 minutes, the reaction
mixture was allowed to cool to ambient temperature, then was
quenched with water and extracted with 30% ethyl acetate in hexane.
The organic phase was dried over sodium sulfate, passed through a
silica pad, eluting with DCM, and concentrated in vacuo. To a
solution of the crude reaction product in 1,4-dioxane (0.200 mL)
and methanol (0.030 mL) was added sodium hydroxide (0.040 mL, 1.0 M
aqueous, 0.040 mmol), and the reaction mixture was then heated at
50.degree. C. After 15 min, the reaction mixture was allowed to
cool to ambient temperature. The reaction mixture was concentrated
in vacuo then acidified with TFA (2 N in DMSO). Purification by
reverse phase HPLC (65 to 100% acetonitrile in water, each with
0.1% v/v TFA) provided the title compound: LCMS m/z 614.9
[M+H].sup.+; .sup.1H NMR (500 MHz, CD.sub.2Cl.sub.2) .delta. 8.13
(s, 1H), 7.96 (t, J=7.9 Hz, 1H), 7.74 (d, J=7.8 Hz, 1H), 7.69-7.65
(m, 4H), 7.60 (d, J=8.0 Hz, 1H), 7.46 (d, J=8.0 Hz, 2H), 7.41 (d,
J=8.0 Hz, 1H), 7.37 (s, 1H), 7.18 (d, J=8.2 Hz, 3H), 3.94 (s, 2H),
2.36 (s, 3H).
[0308] The compounds listed in TABLE 4 were prepared using
chemistry described in Examples 236-243.
TABLE-US-00005 TABLE 4 ##STR00279## Entry X R1 R2 MS ([M +
H].sup.+) 244 CH H ##STR00280## 549.9 245 CH H ##STR00281## 595.8
246 CH Cl ##STR00282## 617.8 247 CH H ##STR00283## 617.8 248 CH H
##STR00284## 597.9 249 CH Me ##STR00285## 615.8 250 CH Me
##STR00286## 611.8 251 CH H ##STR00287## 548.4 252 N H ##STR00288##
544.9 253 N H ##STR00289## 599.0 254 N Cl ##STR00290## 620.8 255 N
Cl ##STR00291## 602.8 256 N Me ##STR00292## 598.8 257 N Me
##STR00293## 582.9 258 N Cl ##STR00294## 582.9 259 N Cl
##STR00295## 602.8 260 N Cl ##STR00296## 636.7 261 N Cl
##STR00297## 598.8 262 N Cl ##STR00298## 652.8 263 N Cl
##STR00299## 578.9 264 N Cl ##STR00300## 598.8 265 N Cl
##STR00301## 619.9 266 N Me ##STR00302## 600.0 267 N CF.sub.3
##STR00303## 613.0 268 CH H ##STR00304## 614.9 269 CH H
##STR00305## 634.9 270 CH H ##STR00306## 618.9 271 CH H
##STR00307## 560.9 272 CH H ##STR00308## 580.9 273 CH H
##STR00309## 564.9 274 CH H ##STR00310## 584.8 275 CH Me
##STR00311## 562.7 276 CH Me ##STR00312## 628.9 277 CH Me
##STR00313## 595.0 278 CH Me ##STR00314## 580.9 279 CH Me
##STR00315## 632.9 280 CH Me ##STR00316## 618.9 281 CH Me
##STR00317## 596.8 282 CH Me ##STR00318## 596.8 283 (isomer A) CH
Me ##STR00319## 635.0 284 (isomer B) CH Me ##STR00320## 635.0 285
CH Me ##STR00321## 566.9 286 CH Me ##STR00322## 566.9 287 CH Me
##STR00323## 580.9 288 CH Me ##STR00324## 580.9 289 CH Me
##STR00325## 498.7 290 CH Me ##STR00326## 512.7 291 CH Cl
##STR00327## 648.7 292 CH Cl ##STR00328## 594.7 293 (isomer A) CH
Cl ##STR00329## 654.8 294 (isomer B) CH Cl ##STR00330## 654.7 295
CH Cl ##STR00331## 518.7 296 CH Cl ##STR00332## 532.8 297 CH Cl
##STR00333## 574.9 298 CH Cl ##STR00334## 532.8 299 CH Cl
##STR00335## 520.7
Example 300
##STR00336##
[0309] Step A. Ethyl
1-(6-{2-[(4-bromophenyl)(difluoro)methoxy]phenyl}pyridin-2-yl)-5-(trifluo-
romethyl)-1H-pyrazole-4-carboxylate
[0310] To a cooled (0.degree. C.) solution of the title compound
from Example 1 Step B (515 mg, 1.37 mmol) in DMF (5 mL) was added
NaH (35.0 mg, 1.46 mmol). The reaction vessel was then allowed to
warm to ambient temperature for 25 min, followed by addition of
1-bromo-4-[bromo(difluoro)methyl]benzene (445 mg, 1.56 mmol,
synthesized according to U.S. Pat. No. 6,939,990) and heated at
60.degree. C. After 24 h, the reaction mixture was cooled, quenched
by addition of 2 N HCl and extracted with hexane-EtOAc.
Purification by silica gel flash chromatography (5% EtOAc in
hexanes to 10% EtOAc in hexanes) yielded the title compound: LCMS
m/z 564.1 [M-F].sup.+; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.12 (s, 1H), 7.88-7.82 (m, 2H), 7.78 (d, J=7.1 Hz, 1H), 7.57 (dd,
J=7.8, 0.7 Hz, 1H), 7.50 (d, J=8.5 Hz, 2H), 7.46-7.41 (m, 2H),
7.39-7.32 (m, 3H), 4.37 (q, J=7.1 Hz, 2H), 1.38 (t, J=7.1 Hz,
3H).
Step B.
1-[6-(2-{Difluoro[4'-(trifluoromethyl)biphenyl-4-yl]methoxy}phenyl-
)pyridin-2-yl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0311] A mixture of the title compound from Example 300 Step A
(50.0 mg, 0.086 mmol), 4-trifluoromethylphenylboronic acid (35 mg,
0.18 mmol), trans-dichlorobis(triphenylphosphine) palladium (II)
(9.0 mg, 0.013 mmol), Na.sub.2CO.sub.3 (0.1 mL, 2.0 M aqueous, 0.2
mmol), and MeCN (1 mL) was stirred at 90.degree. C. for 35 min. The
crude mixture was dried concentrated in vacuo. Treatment with a
mixture of 0.1 mL each of 3 N NaOH, dioxane and MeOH at 50.degree.
C. for 10 min, followed by reverse phase HPLC using a YMC C-18
column (65 to 100% acetonitrile in water, each with 0.1% v/v TFA)
provided the title compound: LCMS m/z 599.9 [M-F].sup.+; .sup.1H
NMR (500 MHz, acetone-d.sub.6) .delta. 8.34 (m, 2H), 8.03 (d, J=7.6
Hz, 1H), 7.99 (d, J=8.2 Hz, 1H), 7.95-7.78 (m, 7H), 7.69 (d, J=8.5
Hz, 2H), 7.62 (m, 2H).
Example 301
##STR00337##
[0312] Step A.
1-{6-[2-(Difluoro{4-[trans-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)p-
henyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid and
1-{6-[2-(difluoro{4-[cis-4-(trifluoromethyl)cyclohexyl]phenyl}methoxy)phe-
nyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0313] A mixture of the title compound from Example 300 Step A (40
mg, 0.068 mmol),
4,4,5,5-tetramethyl-2-[4-(trifluoromethyl)cyclohex-1-en-1-yl]-1,3,2-dioxa-
borolane (38 mg, 0.14 mmol, prepared according to J. Med. Chem.,
2006, 49, 3719), trans-dichlorobis(triphenylphosphine) palladium
(II) (12 mg, 0.017 mmol), Na.sub.2CO.sub.3 (0.1 mL, 2.0 M aqueous,
0.2 mmol), CsF (32 mg, 0.2 mmol) and MeCN (0.5 mL) was stirred at
90.degree. C. for 40 min. The crude mixture was dried and polar
material was removed by preparative TLC (5:1 hexane:EtOAc). The
remaining material was dissolved in EtOH (0.5 mL) and hydrogenated
in the presence of about 12 mg Pd black for 6 hours. The resulting
two isomers were separated by preparative TLC (5:1 hexane:EtOAc).
For each respective isomer, treatment with a mixture of 3 N NaOH
(0.1 mL), dioxane (0.1 mL) and MeOH (0.1 mL) at 50.degree. C. for
15 min, followed by acidification with TFA (2 M in DMSO) and
reverse phase HPLC using a YMC C-18 column (65 to 100% acetonitrile
in water, each with 0.1% v/v TFA) gave the trans (from hydrolysis
of the faster-moving isomer on normal-phase TLC) and cis (from
hydrolysis of the slower-moving isomer on normal-phase TLC) title
compounds. Analytical data for the trans isomer: LCMS (ESI) m/z
605.8 [M-F].sup.+; .sup.1H NMR (500 MHz, acetone-d.sub.6) .delta.
8.27 (s, 1H), 8.19 (t, J=7.9 Hz, 1H), 8.01 (d, J=7.8 Hz, 1H), 7.82
(d, J=8.0 Hz, 1H), 7.34 (d, J=8.0 Hz, 2H), 2.65 (m, 1 H), 2.32 (m,
1H), 2.08 (m, 2H), 1.98 (m, 2H), 1.67-1.45 (m, 4H). Analytical data
for the cis isomer: LCMS m/z 605.8 [M-F].sup.+; .sup.1H NMR (400
MHz, acetone-d.sub.6) .delta. 8.20 (s, 1H), 8.14 (t, J=7.9 Hz, 1H),
7.96 (d, J=7.9 Hz, 1H), 7.83 (d, J=7.8 Hz, 1H), 7.77 (d, J=7.9 Hz,
1H), 7.46 (d, J=8.2 Hz, 2H), 7.33 (d, J=8.2 Hz, 2H), 2.80 (m, 1H),
2.43 (m, 1H), 1.82 (m, 6H).
Example 302
##STR00338##
[0314] Step A. 5-Bromo-2-(difluoromethyl)benzonitrile
[0315] A mixture of 4-bromo-1-(difluoromethyl)-2-fluorobenzene (5.0
g, 22 mmol) and KCN (4.34 g, 67 mmol) was heated at 150.degree. C.
in NMP (50 mL) for 16 h. After trituration with a mixture of water,
hexane, EtOAc and DCM, the organic layer was concentrated and
purified by silica gel flash chromatography using hexanes:DCM (4:1
to 4:1.5 v/v) as mobile phase to yield the title compound: .sup.1H
NMR (500 MHz, CD.sub.2Cl.sub.2) .delta. 7.97 (s, 1H), 7.93 (d,
J=8.5 Hz, 1H), 7.67 (d, J=8.5 Hz, 1H), 6.95 (t, J=54.5 Hz, 1H).
Step B. 5-Bromo-2-[bromo(difluoro)methyl]benzonitrile
[0316] A mixture of the title compound from Example 302 Step A
(2.44 g, 10.5 mmol), CCl.sub.3Br (8 mL) and Na.sub.2CO.sub.3 (480
mg, 4.5 mmol) in a sealed vessel was illuminated by a sunlamp for
30 h. Purification by silica gel flash chromatography using 8:1 to
6:1 to 4:1 hexanes:DCM as mobile phase yielded the title compound:
.sup.1H NMR (500 MHz, CD.sub.2Cl.sub.2) .delta. 8.03 (s, 1H), 7.92
(d, J=8.5 Hz, 1H), 7.65 (d, J=8.5 Hz, 1H).
Step C. 5-Bromo-2-[bromo(difluoro)methyl]benzaldehyde
[0317] To a cooled (-78.degree. C.) solution of the title compound
from Example 302 Step B (1.23 g, 3.95 mmol) in toluene (10 mL) was
added dropwise DIBAL-H (5.14 mL, 1.0 M in toluene, 5.14 mmol). The
reaction mixture was then immediately quenched by addition of HOAc
(1.5 mL), followed by MeOH (0.5 mL) and 2 N HCl. Extraction with
hexanes:EtOAc, followed by flash chromatography on silica gel
(hexanes:DCM, 8:1 to 3:1) to provide the title compound: .sup.1H
NMR (500 MHz, CD.sub.2Cl.sub.2) .delta. (ppm) 10.63 (t, J=2.2 Hz,
1H), 8.22 (d, J=1.6 Hz, 1H), 7.87 (d, J=8.2, 1.6 Hz, 1H), 7.61 (d,
J=8.2 Hz, 1H).
Step D. Ethyl
1-(6-{2-[(4-bromo-2-formylphenyl)(difluoro)methoxy]phenyl}pyridin-2-yl)-5-
-(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0318] By analogy to Example 300 Step A, reaction of the title
compound from Example 302 Step C with the title compound from
Example 1 Step B provided the title compound: LCMS m/z 592.5
[M-F].sup.+.
Step E. Ethyl
1-(6-{2-[(4-bromo-2-vinylphenyl)(difluoro)methoxy]phenyl}pyridin-2-yl)-5--
(trifluoromethyl)-1H-pyrazole-4-carboxylate
[0319] To a cooled (0.degree. C.) suspension of methyltriphenyl
phosphonium bromide (5.33 g, 14.9 mmol) in THF (100 mL) was added
dropwise n-BuLi (5.5 mL, 2.5 M in hexanes, 13.8 mmol). After aging
for 40 min, the reaction mixture was allowed to warm up to ambient
temperature for 25 min, at which point 7.4 mL of the supernatant
was taken and added to the title compound from Example 302 Step D
(270 mg, 0.442 mmol). After 30 min, the mixture was quenched by
addition of water and the aqueous phase was extracted with
hexane-EtOAc. The organic phase was separated and concentrated in
vacuo. Purification by silica gel flash chromatography (5% to 15%
EtOAc in hexanes) provided the title compound: LCMS m/z 590.5
[M-F].sup.+; .sup.1H NMR (500 MHz, CD.sub.2Cl.sub.2) .delta. 8.13
(s, 1H), 7.86 (m, 2H), 7.77 (m, 2H), 7.60 (d, J=8.0 Hz, 1H),
7.55-7.48 (m, 2H), 7.45-7.40 (m, 3H), 7.08 (m, 1H), 5.68 (d, J=17.2
Hz, 1H), 5.32 (d, J=11.0 Hz, 1H), 4.38 (q, J=7.2 Hz, 2H), 1.40 (t,
J=7.2 Hz, 3H).
Step F. 1-(6-{2-[{2-Ethyl-4-[4
(trifluoromethyl)cyclohexyl]phenyl}(difluoro)methoxy]phenyl}pyridin-2-yl)-
-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic acid
[0320] The title compound was prepared according to the procedure
described in Example 301, starting from the title compound from
Example 302 Step E: LCMS m/z 634.6 [M-F].sup.+; .sup.1H NMR (500
MHz, acetone-d.sub.6) .delta. 8.24 (s, 1H), 8.10 (m, 1H), 7.93 (d,
J=7.8 Hz, 1H), 7.87 (d, J=7.6 Hz, 1H), 7.78 (d, J=8.0 Hz, 1H), 7.58
(m, 2H), 7.50 (d, J=8.2 Hz, 1H), 7.48 (m, 1H), 7.27 (s, 1H), 7.15
(d, 1H), 2.76 (q, J=7.6 Hz, 2H), 2.63 (m, 1H), 2.31 (m, 1H), 2.07
(m, 2H), 1.99 (m, 2H), 1.67-1.46 (m, 4H), 1.16 (t, J=7.6 Hz,
3H).
Example 303
##STR00339##
[0321] Step A. 1-Bromo-2-[bromo(difluoro)methyl]benzene
[0322] A solution of 1-bromo-2-difluoromethylbenzene (9.6 g, 46.8
mmol) and N-bromosuccinimide (24.8 g, 139 mmol) in carbon
tetrachloride (100 mL) was irradiated with a sunlamp. After 3 days,
the reaction mixture was diluted with hexane, the precipitate was
filtered off, and the collected organic filtrate was concentrated
in vacuo. Purification by flash chromatography on silica gel (100%
hexanes) provided the title compound: .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 7.74 (d, J=8.0 Hz, 1H), 7.66 (dd, J=8.0,
1.5 Hz, 1H), 7.44 (t, J=7.7 Hz, 1H), 7.37 (td, J=7.7, 1.3 Hz,
1H).
Step B. (2-Bromophenyl)(difluoro)methyl
4'-(trifluoromethyl)biphenyl-4-yl
ether-4-[(2-bromophenyl)(difluoro)methoxy]-4'-(trifluoromethyl)biphenyl
[0323] To a cooled (0.degree. C.) solution of
4'-(trifluoromethyl)[1,1'-biphenyl]-4-ol (178 mg, 0.75 mmol) in DMF
was added sodium hydride (27.0 mg, 1.12 mmol). Once the hydrogen
evolution subsided, the reaction mixture was allowed to warm to
ambient temperature. The title compound from Example 303 Step A
(373 mg, 1.30 mmol) was then added to the reaction flask, and the
reaction mixture was stirred at 60.degree. C. After 15 h, the
reaction mixture was quenched by addition of 2 N aqueous HCl. The
aqueous phase was extracted with hexanes/ethyl acetate (3:1 v/v).
The organic phase was separated, dried over sodium sulfate and
concentrated in vacuo. Purification was by flash chromatography on
silica gel (0-20% dichloromethane in hexanes) provided the title
compound: LCMS m/z 423.5 [M-F].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 7.85 (dd, J=7.8, 1.6 Hz, 1H), 7.76 (d,
J=8.0 Hz, 1H), 7.72 (s, 4H), 7.65 (dd, J=6.8, 1.9 Hz, 2H), 7.45 (d,
J=8.7 Hz, 3H), 7.40 (td, J=7.7, 1.5 Hz, 1H).
Step C. Ethyl
1-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]-5-(triflu-
oromethyl)-1H-pyrazole-4-carboxylate
[0324] The title compound from Example 1 Step A (538 mg, 1.68
mmol), bis(pinacolato)diboron (513 mg, 2.02 mmol),
1-1'-bis(diphenylphosphino)ferrocene (93 mg, 0.17 mmol),
1-1'-bis(diphenylphosphino) ferrocene palladium (II) chloride
complex with dichloromethane (137 mg, 0.17 mmol) and potassium
acetate (495 mg, 5.05 mmol) were dissolved in DMSO (10 mL), and the
resulting mixture was heated at 100.degree. C. After 2 h, the
reaction mixture was allowed to cool to ambient temperature, then
was quenched with brine and extracted with ether. The organic layer
was separated, dried over sodium sulfate and concentrated in vacuo.
Purification by flash chromatography on silica gel (20% ethyl
acetate in hexanes) provided the title compound: LCMS (mass of the
boronic acid observed) m/z 330.5 [M+H].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 8.10 (s, 1H), 7.92 (m, 2H), 7.57 (dd,
J=6.6, 2.5 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 1.37 (t, J=7.1 Hz, 3H),
1.37 (s, 12H).
Step D.
1-{6-[2-(Difluoro{[4'-(trifluoromethyl)biphenyl-4-yl]oxy}methyl)ph-
enyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid
[0325] To a solution of the title compound from Example 303 Step B
(95.5 mg, 0.22 mmol) and the title compound from Example 303 Step C
(106 mg, 0.33 mmol) in acetonitrile (2.0 mL) were added
trans-dichlorobis(triphenylphosphine) palladium (II) (30 mg, 0.04
mmol), cesium fluoride (98 mg, 0.65 mmol), and sodium carbonate
(0.90 mL, 1.0 M aqueous, 0.90 mmol). The resulting mixture was
heated at 90.degree. C. After 1.5 h, the reaction mixture was
allowed to cool to ambient temperature, then was quenched with
brine and extracted with 30% ethyl acetate in hexanes. The organic
phase was separated and dried over sodium sulfate. The mixture was
then was passed through a pad of silica gel, eluting with DCM, and
concentrated in vacuo. To a solution of the crude product obtained
above in dioxane (0.50 mL) and methanol (0.050 mL) was added sodium
hydroxide (0.100 mL, 1.0 M aqueous, 0.100 mmol) and the resulting
mixture was stirred at 50.degree. C. After 15 min, the reaction
mixture was allowed to cool to ambient temperature and was
concentrated in vacuo. The mixture was then acidified with TFA (2 M
in DMSO). Purification by reverse phase HPLC (65 to 100%
acetonitrile in water, each with 0.1% v/v TFA) provided the title
compound: LCMS m/z 600.8 [M-F].sup.+; .sup.1H NMR (500 MHz,
CD.sub.2Cl.sub.2) .delta. 8.10 (s, 1H), 8.02 (t, J=7.9 Hz, 1H),
7.74-7.60 (m, 9H), 7.53 (dd, J=6.8, 1.9 Hz, 3H), 7.06 (d, J=8.7 Hz,
2H).
[0326] The compounds in TABLE 5 were prepared using chemistry
described in Examples 300-303.
TABLE-US-00006 TABLE 5 ##STR00340## Entry R1 R2 R3 MS [M -
F].sup.+) 304 Me H ##STR00341## 613.9 305 H H ##STR00342## 565.9
306 H H ##STR00343## 538.0 307 H H ##STR00344## 524.0 308 H H
##STR00345## 605.8 309 H H ##STR00346## 605.8 310 H H ##STR00347##
566.0 311 Me H ##STR00348## 504.0 312 Me H ##STR00349## 530.0 313
Me H ##STR00350## 510.1 314 Me H ##STR00351## 552.0 315 H H
##STR00352## 590.8 316 H H ##STR00353## 624.8 317 Cl H ##STR00354##
599.8 318 H H ##STR00355## 575.9 319 Cl H ##STR00356## 639.9 320 Cl
H ##STR00357## 639.9 321 Cl H ##STR00358## 595.9 322 H H
##STR00359## 561.9 323 Me H ##STR00360## 576.0 324 Cl H
##STR00361## 629.9 325 H H ##STR00362## 595.9 326 Me H ##STR00363##
582.6 327 Me Br ##STR00364## 698.5 328 H H ##STR00365## 562.9 329 H
H ##STR00366## 618.9
Example 330
##STR00367##
[0327] Step A. 2-Azido-6-chloropyridine
[0328] To a 250 ml round-bottom flask equipped with a mechanical
stirrer, a Claisen head and an addition funnel were added
2-chloro-6-hydrazinopyridine (4.00 g, 27.9 mmol), Et.sub.2O (20 mL)
and concentrated hydrochloric acid (12 mL, 146 mmol). A solution of
sodium nitrite (2.211 g, 32.0 mmol) in water (28 mL) was added
dropwise to the cooled (0.degree. C.) reaction mixture. The
insoluble starting material gradually dissolved. After 2 h, the
aqueous phase was extracted with ether (2.times.50 mL). The
combined organic phase was dried over MgSO.sub.4, and concentrated
in vacuo to provide the title compound as yellow crystals: LCMS m/z
155.14 [M+H].sup.+.
Step B. Ethyl
1-(6-chloropyridin-2-yl)-5-(trifluoromethyl)-1H-1,2,3-triazole-4-carboxyl-
ate
[0329] To a solution of the title compound from Example 330 Step A
(300 mg, 1.94 mmol) in acetonitrile (4 mL) was added ethyl
2-(ethoxymethylene)-4,4,4-trifluoro-3-oxobutyrate (0.284 mL, 1.94
mmol) followed by TEA (0.271 mL, 1.94 mmol), and the mixture was
heated at 70.degree. C. After 16 h, the reaction mixture was
concentrated in vacuo. Purification by flash chromatography on
silica gel (0 to 65% EtOAc in hexanes) provided the title compound
as a yellow oil: LCMS m/z 320.85 [M+H].sup.+; .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 8.03 (t, J=7.9 Hz, 1H), 7.77 (d, J=7.7 Hz, 1H),
7.64 (d, J=8.0 Hz, 1H), 4.55 (q, J=7.2 Hz, 2H), 1.49 (t, J=7.2 Hz,
3H).
Step C.
5-(Trifluoromethyl)-1-[6-(2-{[4'-(trifluoromethyl)biphenyl-4-yl]me-
thoxy}phenyl)pyridin-2-yl]-1H-1,2,3-triazole-4-carboxylic acid
[0330] The title compound was prepared from the title compound from
Example 330 Step B by direct analogy to the procedures outlined in
Example 2 steps A-C: LCMS m/z 585.2 [M+H].sup.+; .sup.1H NMR (500
MHz, d.sub.6-DMSO) .delta. 8.29 (d, J=8.0 Hz, 1H), 8.24 (t, J=8.0
Hz, 1H), 7.91-7.88 (m, 3H), 7.81 (d, J=8.5 Hz, 2H), 7.75-7.72 (m,
3H), 7.56 (d, J=8.5 Hz, 1H), 7.49-7.45 (m, 1H), 7.32 (d, J=8.0 Hz,
1H), 7.12 (t, J=7.5 Hz, 1H), 5.33 (s, 2H).
[0331] The compounds in TABLE 6 were prepared using chemistry
described in Examples 1 and 330.
TABLE-US-00007 TABLE 6 ##STR00368## Entry R1 R2 MS 331 H
##STR00369## 551.0 [M + H].sup.+ 332 Me ##STR00370## 599.5 [M +
H].sup.+ 333 Me ##STR00371## 613.5 [M + H].sup.+ 334 Me
##STR00372## 617.4 [M + H].sup.+
Measurement of Soluble Guanylyl Cyclase (cGC) Activation
(Cell-Based)
[0332] The activity of compounds for sGC was determined by
measuring their ability to activate heterologously expressed sGC in
CHO cells through the generation of intracellular cyclic guanine
monophosphate (cGMP).
[0333] Human sGC subunits .alpha.1 and .beta.1 were cloned from
cDNA and inserted into a mammalian expression vector using the
CMVie promoter using standard molecular biological methods. A
Stably transfected CHO cell line overexpressing both sGC subunits
was generated using standard cell biological methods.
[0334] Test compounds (5 ul) were dissolved in DMSO and diluted in
DMSO to 50 times the desired final concentrations for 3-fold serial
dilution dose response curves. The compounds were incubated with
3500-4000 cells in 5 ul phosphate-buffered saline (PBS) containing
1 nM IBMX (3-isobutyl-1-methylzanthine) at 37.degree. C. for 1 hr
in a 384-well plate (Greiner #784076) in the presence and absence
of [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). ODQ is used to
differentiate between Heme-dependent (HDA) and Heme Independent
(HIA) compounds. At the end of the incubation period, the reaction
is terminated and the cells are lysed. The level of intracellular
cGMP is determined by an HTRF-based assay kit (CisBio, 62GM2PEC),
which detects the displacement of a labeled cGMP from its specific
antibody. Inflection point, maximum % of activation, and EC.sub.50
were derived based on the plot of compound concentration vs. %
activation. Compounds were determined to have an inflection point
greater than 10 .mu.M and at least 20% activation.
Measurement of Soluble Guanylyl Cyclase (cGC) Activation
(Enzyme-Based)
[0335] Activities of test compounds against purified sGC were
evaluated in PCASA assay which is a cell-free enzymatic assay.
[0336] Human recombinant sGC enzyme with greater than 95% purity
was obtained from Axxora, LLC (San Diego, Calif.). Compounds were
incubated with 0.1 ng of sGC enzyme in presence of its substrate
GTP for 1 hr at 37.degree. C. At the end of the incubation period,
the reaction was stopped and the amount of cGMP generated was
measured by an HTRF-based assay (CisBio, 62GN2PEC), which detects
the displacement of a fluorophore-labeled cGMP from its specific
antibody.
[0337] Inflection point, maximum % of activation, and EC50 were
derived based on the plot of compound concentration vs %
activation. In this assay,
1-{6-[5-Chloro-2-({4-[trans-4-(trifluoromethyl)cyclohexyl]-benzyl}-
oxy)phenyl]pyridin-2-yl}-5-(trifluoromethyl)-1H-pyrazole-4-carboxylic
acid (Example 9) gave an inflection point of 11 nM and EC50 of 1.7
nM.
* * * * *