U.S. patent application number 12/488020 was filed with the patent office on 2010-02-04 for inhibitors of cytosolic phospholipase a2.
Invention is credited to Lihren Chen, James D. Clark, Valerie Clerin, Katherine L. Lee, Suzana Marusic, John C. McKew, Kevin Pong, Richard Vargas, Cara Williams.
Application Number | 20100029645 12/488020 |
Document ID | / |
Family ID | 37188759 |
Filed Date | 2010-02-04 |
United States Patent
Application |
20100029645 |
Kind Code |
A1 |
McKew; John C. ; et
al. |
February 4, 2010 |
Inhibitors of Cytosolic Phospholipase A2
Abstract
This invention provides chemical inhibitors of the activity of
various phospholipase enzymes, particularly cytosolic phospholipase
A.sub.2 enzymes (cPLA.sub.2), more particularly including
inhibitors of cytosolic phospholipase A.sub.2 alpha enzymes
(cPLA.sub.2.alpha.). In some embodiments, the inhibitors have the
Formula I: ##STR00001## wherein the constituent variables are as
defined herein.
Inventors: |
McKew; John C.; (Arlington,
MA) ; Lee; Katherine L.; (Newton, MA) ; Chen;
Lihren; (Bedford, MA) ; Vargas; Richard;
(Brighton, MA) ; Clark; James D.; (Acton, MA)
; Williams; Cara; (Methuen, MA) ; Clerin;
Valerie; (Watertown, MA) ; Marusic; Suzana;
(Reading, MA) ; Pong; Kevin; (Robbinsville,
NJ) |
Correspondence
Address: |
WYETH LLC;PATENT LAW GROUP
5 GIRALDA FARMS
MADISON
NJ
07940
US
|
Family ID: |
37188759 |
Appl. No.: |
12/488020 |
Filed: |
June 19, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11442199 |
May 26, 2006 |
7557135 |
|
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12488020 |
|
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60685564 |
May 27, 2005 |
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Current U.S.
Class: |
514/235.2 ;
514/254.09; 514/415; 544/143; 544/373; 548/491 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 11/00 20180101; A61P 25/16 20180101; A61P 9/10 20180101; A61P
19/00 20180101; A61P 11/06 20180101; C07D 209/14 20130101; A61P
25/00 20180101; A61P 7/02 20180101; A61P 25/28 20180101; A61P 29/00
20180101; C07D 209/18 20130101; A61P 19/02 20180101; A61P 3/00
20180101 |
Class at
Publication: |
514/235.2 ;
548/491; 544/373; 544/143; 514/415; 514/254.09 |
International
Class: |
A61K 31/5377 20060101
A61K031/5377; C07D 209/04 20060101 C07D209/04; C07D 403/02 20060101
C07D403/02; C07D 413/02 20060101 C07D413/02; A61K 31/405 20060101
A61K031/405; A61K 31/497 20060101 A61K031/497; A61P 29/00 20060101
A61P029/00; A61P 19/02 20060101 A61P019/02; A61P 25/00 20060101
A61P025/00; A61P 7/02 20060101 A61P007/02 |
Claims
1. A compound having the Formula I: ##STR00018## wherein: n.sub.1
is 1 or 2; n.sub.2 is 1 or 2; n.sub.3 is 1 or 2; n.sub.5 is 0, 1 or
2; X.sup.2 is a bond, O, --CH.sub.2-- or SO.sub.2; each R.sub.5 is
independently H or C.sub.1-3 alkyl; R.sub.6 is H or C.sub.1-6
alkyl; R.sub.7 is selected from the group consisting of OH,
benzyloxy, CH.sub.3, CF.sub.3, OCF.sub.3, C.sub.1-3 alkoxy,
halogen, COH, CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl)
quinoline-5-yl, quinoline-8-yl, 3,5-dimethylisoxazol-4-yl,
thiophene-3-yl, pyridin-4-yl pyridine-3-yl, --CH.sub.2-Q, and
phenyl optionally substituted by from one to three independently
selected R.sub.30 groups; R.sub.8 is selected from the group
consisting of H, OH, NO.sub.2, CF.sub.3, C.sub.1-3 alkoxy, halogen,
CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl), quinoline-5-yl
quinoline-8-yl, 3,5-dimethylisoxazol-4-yl, thiophene-3-yl,
--CH.sub.2-Q, and phenyl substituted by from one to three
independently selected R.sub.30 groups; Q is OH, dialkylamino,
##STR00019## R.sub.20 is selected from the group consisting of H,
C.sub.1-3 alkyl and CO(C.sub.1-3 alkyl); and R.sub.30 is selected
from the group consisting of dialkylamino, CN and OCF.sub.3;
provided that: a) when each R.sub.5 is H, R.sub.6 is H, n.sub.5 is
0, and R.sub.8 is H, then R.sub.7 cannot be chlorine; b) when each
R.sub.5 is H, R.sub.6 is H, n.sub.5 is 0, X.sup.2 is O or
--CH.sub.2--, and R.sub.8 is H, then R.sub.7 cannot be CH.sub.3; c)
when each R.sub.5 is H, and R.sub.6 is H, then R.sub.7 and R.sub.8
cannot both be fluorine; d) when each R.sub.5 is H, R.sub.6 is H,
and X.sup.2 is O, then R.sub.7 and R.sub.8 cannot both be chlorine;
e) when each R.sub.5 is H, R.sub.6 is H, X.sup.2 is O, and R.sub.8
is NO.sub.2, then R.sub.7 cannot be fluorine; and f) when each
R.sub.5 is H, R.sub.6 is H, X.sup.2 is SO.sub.2, and R.sub.8 is H,
then R.sub.7 cannot be fluorine or chlorine; or a pharmaceutically
acceptable salt thereof.
2. A compound or salt of claim 1 wherein X.sup.2 is CH.sub.2.
3. A compound or salt of claim 2 wherein n.sub.3 is 1.
4. A compound or salt of claim 2 wherein n.sub.1 is 1.
5. A compound or salt of claim 2 wherein n.sub.2 is 1.
6. A compound or salt of claim 2 wherein n.sub.3 is 1; n.sub.1 is
1; and n.sub.2 is 1.
7. A compound or salt of claim 2 wherein R.sub.6 is H.
8. A compound or salt of claim 2 wherein: n.sub.3 is 1; n.sub.1 is
1; n.sub.2 is 1; R.sub.6 is H; R.sub.7 is selected from the group
consisting of CH.sub.3, CF.sub.3, OCF.sub.3, halogen, COOCH.sub.3,
COH, CH.sub.2OH, diethylaminomethyl, quinoline-5-yl,
quinoline-8-yl, 3,5-dimethylisoxazol-4-yl, thiophene-3-yl,
pyridin-4-yl, pyridine-3-yl, phenyl, 4-dimethylamino-phen-1-yl,
2-trifluoromethoxy-phen-1-yl, 2-cyano-phen-1-yl,
morpholine-1-yl-methyl, piperazine-1-yl methyl,
4-acetyl-piperazine-1-yl methyl, and 4-methyl-piperazine-1-yl
methyl; and R.sub.8 is selected from the group consisting of H,
halogen, CF.sub.3 and NO.sub.2.
9. A compound or salt of claim 8 wherein R.sub.5 is H.
10. A compound or salt of claim 8 wherein R.sub.5 is CH.sub.3.
11. A compound or salt of claim 1 wherein X.sup.2 is O.
12. A compound or salt of claim 11 wherein n.sub.3 is 1.
13. A compound or salt of claim 11 wherein n.sub.1 is 1.
14. A compound or salt of claim 11 wherein n.sub.2 is 1.
15. A compound or salt of claim 11 wherein R.sub.6 is H.
16. A compound or salt of claim 11 wherein n.sub.3 is 1; n.sub.1 is
1; and n.sub.2 is 1.
17. A compound or salt of claim 11 wherein: n.sub.3 is 1; n.sub.1
is 1; n.sub.2 is 1; R.sub.6 is H; R.sub.7 is selected from the
group consisting of benzyloxy, OH, halogen, CH.sub.3 and CF.sub.3;
and R.sub.8 is selected from the group consisting of H, halogen,
and NO.sub.2.
18. A compound or salt of claim 17 wherein R.sub.5 is H.
19. A compound or salt of claim 17 wherein R.sub.5 is CH.sub.3.
20. A compound or salt of claim 17 wherein R.sub.7 is CF.sub.3, and
R.sub.8 is H.
21. A compound or salt of claim 1 wherein X.sup.2 is SO.sub.2.
22. A compound or salt of claim 1 wherein X.sup.2 is SO.sub.2, and
n.sub.5 is 2.
23. A compound or salt of claim 22 wherein n.sub.3 is 1.
24. A compound or salt of claim 22 wherein n.sub.1 is 1.
25. A compound or salt of claim 22 wherein n.sub.2 is 1.
26. A compound or salt of claim 22 wherein n.sub.3 is 1; n.sub.1 is
1; and n.sub.2 is 1.
27. A compound or salt of claim 22 wherein: n.sub.3 is 1; n.sub.1
is 1; n.sub.2 is 1; R.sub.6 is H; R.sub.7 is CF.sub.3; and R.sub.8
is H.
28. A compound or salt of claim 1, wherein n.sub.1 is 1; n.sub.2 is
1 or 2; n.sub.3 is 1, n.sub.5 is 0; X.sup.2 is CH.sub.2, each
R.sub.5 and each R.sub.6 is H; and R.sub.7 and R.sub.8 are
independently selected from the group consisting of H, F, CF.sub.3,
OCF.sub.3, OH, quinoline-5-yl and quinoline-8-yl, provided that
R.sub.7 and R.sub.8 are not both H.
29. A compound or salt of claim 28, wherein n.sub.2 is 1.
30. A compound of Formula II: ##STR00020## wherein: X is a bond or
O; n.sub.1 is 1 or 2; n.sub.2 is 1 or 2; n.sub.6 is 1 or 2; R.sub.5
is H or CH.sub.3; R.sub.6 is H or C.sub.1-6 alkyl; and R.sub.8 is
selected from the group consisting of H, OH, NO.sub.2, CF.sub.3,
OCF.sub.3, OCH.sub.3, halogen, COCH.sub.3, COOCH.sub.3,
dimethylamino, diethylamino and CN; or a pharmaceutically
acceptable salt thereof.
31. A compound or salt of claim 30 wherein n.sub.1 is 1.
32. A compound or salt of claim 30 wherein n.sub.2 is 1.
33. A compound or salt of claim 30 wherein n.sub.6 is 2.
34. A compound or salt of claim 30 wherein R.sub.5 is H.
35. A compound or salt of claim 30 wherein n.sub.1 is 1; n.sub.2 is
1; and n.sub.6 is 2.
36. A compound or salt of claim 30 wherein R.sub.6 is H.
37. A compound or salt of claim 30 wherein R.sub.5 is H; R.sub.6 is
H, n.sub.1 is 1, n.sub.2 is 1; and n.sub.6 is 2.
38. A compound or salt of claim 37 wherein R.sub.8 is selected from
the group consisting of H, CF.sub.3, OCF.sub.3 and halogen.
39. A compound or salt of claim 37 wherein R.sub.8 is H.
40. A compound of claim 1 that is selected from the group
consisting of: a)
4-{2-[2-[2-({[2-(benzyloxy)benzyl]sulfonyl}amino)ethyl]-5-chloro-1-(di-
phenylmethyl)-1H-indol-3-yl]ethoxy}benzoic acid; b)
4-{2-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}-
ethyl)-1H-indol-3-yl]ethoxy}benzoic acid; c)
4-{2-[5-chloro-2-(2-{[(2,6-dibromobenzyl)sulfonyl]amino}ethyl)-1-(dipheny-
lmethyl)-1H-indol-3-yl]ethoxy}benzoic acid; d)
4-(2-{1-benzhydryl-5-chloro-2-[2-methyl-6-nitro-phenylmethanesulfonylamin-
o)-ethyl-1-H-indol-3-yl}-ethoxy)-benzoic acid; e)
4-(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]sulfo-
nyl}amino)ethyl]-1H-indol-3-yl}ethoxy)benzoic acid; f)
4-(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)ben-
zyl]sulfonyl}amino)ethyl]-1H-indol-3-yl}ethoxy)benzoic acid; g)
4-{3-[5-chloro-2-(2-{[(2,6-dibromobenzyl)sulfonyl]amino}ethyl)-1-(dipheny-
lmethyl)-1H-indol-3-yl]propyl}benzoic acid; h)
4-{3-[5-chloro-2-(2-{[(2,6-dichlorobenzyl)sulfonyl]amino}ethyl)-1-(diphen-
ylmethyl)-1H-indol-3-yl]propyl}benzoic acid; i)
4-(3-{1-benzhydryl-5-chloro-2-[2-(2-methyl-6-nitro-phenylmethanesulfonyla-
mino)-ethyl-1H-indol-3-yl}propyl)-benzoic acid; j)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]sulfo-
nyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; k)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-(methyl{[2-(trifluoromethyl)benzyl-
]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; l)
4-{3-[2-[2-({[2,6-bis(trifluoromethyl)benzyl]sulfonyl}amino)ethyl]-5-chlo-
ro-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid; m)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(methoxycarbonyl)benzyl]sulfo-
nyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; n)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)ben-
zyl]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; o)
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({2-[2-(trifluoromethyl)phenyl]et-
hyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic acid; p)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoic acid; q)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(morpholin-4-ylmethyl)benzyl]-
sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; r)
4-{3-[5-chloro-2-{2-[({2-[(diethylamino)methyl]benzyl}sulfonyl)amino]ethy-
l}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid; s)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(hydroxymethyl)benzyl]sulfony-
l}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; t)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(piperazin-1-ylmethyl)benzyl]-
sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; u)
4-{3-[2-{2-[({2-[(4-acetylpiperazin-1-yl)methyl]benzyl}sulfonyl)amino]eth-
yl}-5-chloro-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid;
v)
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({2-[(4-methylpiperazin-1-yl)meth-
yl]benzyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic acid;
w)
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({1-[2-(trifluoromethyl)phenyl]et-
hyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic acid; x)
4-{3-[2-(2-{[(2-bromobenzyl)sulfonyl]amino}ethyl)-5-chloro-1-(diphenylmet-
hyl)-1H-indol-3-yl]propyl}benzoic acid; y)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethoxy)benzyl]sulf-
onyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; z)
4-{3-[5-chloro-2-(2-{[(3-chloro-6-fluoro-2-methylbenzyl)sulfonyl]amino}et-
hyl)-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid; aa)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-nitro-6-(trifluoromethyl)benz-
yl]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; ab)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-fluorobenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoic acid; ac)
4-{3-[2-(2-{[(biphenyl-2-ylmethyl)sulfonyl]amino}ethyl)-5-chloro-1-(diphe-
nylmethyl)-1H-indol-3-yl]propyl}benzoic acid; ad)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-pyridin-4-ylbenzyl)sulfonyl]a-
mino}ethyl)-1H-indol-3-yl]propyl}benzoic acid; ae)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-pyridin-3-ylbenzyl)sulfonyl]a-
mino}ethyl)-1H-indol-3-yl]propyl}benzoic acid; af)
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(3-thienyl)benzyl]sulfonyl}am-
ino)ethyl]-1H-indol-3-yl}propyl)benzoic acid; ag)
4-{3-[5-chloro-2-[2-({[2-(3,5-dimethylisoxazol-4-yl)benzyl]sulfonyl}amino-
)ethyl]-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid; ah)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-quinolin-5-ylbenzyl)sulfonyl]-
amino}ethyl)-1H-indol-3-yl]propyl}benzoic acid; ai)
4-{3-[5-chloro-2-{2-[({[4'-(dimethylamino)biphenyl-2-yl]methyl}sulfonyl)a-
mino]ethyl}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid;
aj)
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({[2'-(trifluoromethoxy)biphenyl--
2-yl]methyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic
acid; ak)
4-{3-[5-chloro-2-[2-({[(2'-cyanobiphenyl-2-yl)methyl]sulfonyl}amino)ethyl-
]-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid; al)
3-{4-[(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]s-
ulfonyl}amino)ethyl]-1H-indol-3-yl}ethyl)sulfony]phenyl}propanoic
acid; am)
3-(4-{[2-(5-chloro-1-(diphenylmethyl)-2-{2-[({1-[2-(trifluoromethyl)p-
henyl]ethyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)ethyl]sulfonyl}phenyl)prop-
anoic acid; an)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}-
ethyl)-1H-indol-3-yl]propyl}benzoic acid; and ao)
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-quinolin-5-ylbenzyl)sulfonyl]-
amino}ethyl)-1H-indol-3-yl]propyl}benzoic acid; or a
pharmaceutically acceptable salt thereof.
41. A method of treating inflammation caused or potentiated by
prostaglandins, leukotrienes, or platelet activation factor, in a
mammal, the method comprising administering to a mammal in need
thereof a pharmaceutically acceptable amount of a compound of claim
1, or a pharmaceutically acceptable salt thereof.
42. A method of treating pain caused or potentiated by
prostaglandins, leukotrienes, or platelet activation factor, in a
mammal, the method comprising administering to a mammal in need
thereof a pharmaceutically acceptable amount of a compound of claim
1, or a pharmaceutically acceptable salt thereof.
43. A method of treatment of asthma in a mammal, the method
comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound of the claim 1, or
a pharmaceutically acceptable salt thereof.
44. A method of treatment of arthritic and rheumatic disorders in a
mammal, the method comprising administering to a mammal in need
thereof a pharmaceutically effective amount of a compound of claim
1, or a pharmaceutically acceptable salt thereof.
45. A method of claim 44 wherein the disorder is rheumatoid
arthritis.
46. A method of claim 44 wherein the disorder is
osteoarthritis.
47. A method of claim 44 wherein the disorder is juvenile
arthritis.
48. A method of treating or preventing a disease or disorder in a
mammal, or preventing progression of symptoms such a disease or
disorder, wherein the disease or disorder is selected from the
group consisting of stroke, atherosclerosis, multiple sclerosis,
Parkinson's disease, central nervous system damage resulting from
stroke, central nervous system damage resulting from ischemia, and
central nervous system damage resulting from trauma, the method
comprising administering to a mammal in need thereof a
pharmaceutically acceptable amount of a compound of claim 1, or a
pharmaceutically acceptable salt thereof.
49. A method of claim 48 wherein the disease or disorder is
stroke.
50. A method of claim 48 wherein the disease or disorder is
atherosclerosis.
51. A method of claim 48 wherein the disease or disorder is
multiple sclerosis.
52. A method of claim 48 wherein the disease or disorder is
Parkinson's disease.
53. A method of claim 48 wherein the disease or disorder is central
nervous system damage resulting from stroke, from ischemia, or from
trauma.
54. A method for treating or preventing venous or arterial
thrombosis in a mammal, or preventing progression of a symptom of
said thrombosis, the method comprising administering to a mammal in
need thereof a pharmaceutically acceptable amount of a compound of
claim 1, or a pharmaceutically acceptable salt thereof.
55. A method of claim 54, wherein the thrombosis is
atherothrombosis.
56. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority benefit of U.S. Provisional
Application Ser. No. 60/685,564, filed May 27, 2005, the entire
content of which is incorporated herein by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to chemical inhibitors of the
activity of various phospholipase enzymes, particularly cytosolic
phospholipase A.sub.2 enzymes (cPLA.sub.2), more particularly
including inhibitors of cytosolic phospholipase A.sub.2 alpha
enzymes (cPLA.sub.2.alpha.).
BACKGROUND OF THE INVENTION
[0003] Leukotrienes and prostaglandins are important mediators of
inflammation, each of which contributes to the development of an
inflammatory response in a different way. Leukotrienes recruit
inflammatory cells such as neutrophils to an inflamed site, promote
the extravasation of these cells and stimulate release of
superoxide and proteases which damage the tissue. Leukotrienes also
play a pathophysiological role in the hypersensitivity experienced
by asthmatics [See, e.g. B. Samuelson et al., Science, 237:1171-76
(1987)]. Prostaglandins enhance inflammation by increasing blood
flow and therefore infiltration of leukocytes to inflamed sites.
Prostaglandins also potentiate the pain response induced by
stimuli.
[0004] Prostaglandins and leukotrienes are unstable and are not
stored in cells, but are instead synthesized [W. L. Smith, Biochem.
J., 259:315-324 (1989)] from arachidonic acid in response to
stimuli. Prostaglandins are produced from arachidonic acid by the
action of COX-1 and COX-2 enzymes. Arachidonic acid is also the
substrate for the distinct enzyme pathway leading to the production
of leukotrienes.
[0005] Arachidonic acid, which is fed into these two distinct
inflammatory pathways, is released from the sn-2 position of
membrane phospholipids by phospholipase A.sub.2 enzymes
(hereinaffer PLA.sub.2). The reaction catalyzed by PLA.sub.2 is
believed to represent the rate-limiting step in the process of
lipid mediator biosynthesis, including but not limited to the
production of inflammatory prostaglandins and leukotrienes. When
the phospholipid substrate of PLA.sub.2 is of the phosphotidyl
choline class with an ether linkage in the sn-1 position, the
lysophospholipid produced is the immediate precursor of platelet
activating factor (hereafter called PAF), another potent mediator
of inflammation [S. I. Wasserman, Hospital Practice, 15:49-58
(1988)].
[0006] Most anti-inflammatory therapies have focused on preventing
production of either prostaglandins or leukotrienes from these
distinct pathways, but not on all of them. For example, ibuprofen,
aspirin, and indomethacin are all NSAIDs which inhibit the
production of prostaglandins by COX-1/COX-2 inhibition, but have no
direct effect on the inflammatory production of leukotrienes from
arachidonic acid in the other pathways. Conversely, zileuton
inhibits only the pathway of conversion of arachidonic acid to
leukotrienes, without directly affecting the production of
prostaglandins. None of these widely-used anti-inflammatory agents
affects the production of PAF.
[0007] Consequently the direct inhibition of the activity of
PLA.sub.2 has been suggested as a useful mechanism for a
therapeutic agent, i.e., to interfere with the inflammatory
response. [See, e.g., J. Chang et al, Biochem. Pharmacol.,
36:2429-2436 (1987)].
[0008] A family of PLA.sub.2 enzymes characterized by the presence
of a secretion signal sequenced and ultimately secreted from the
cell have been sequenced and structurally defined. These secreted
PLA.sub.2s have an approximately 14 kD molecular weight and contain
seven disulfide bonds which are necessary for activity. These
PLA.sub.2 are found in large quantities in mammalian pancreas, bee
venom, and various snake venom. [See, e.g., references 13-15 in
Chang et al, cited above; and E. A. Dennis, Drug Devel. Res.,
10:205-220 (1987).] However, the pancreatic enzyme is believed to
serve a digestive function and, as such, should not be important in
the production of the inflammatory mediators whose production must
be tightly regulated.
[0009] The primary structure of the first human non-pancreatic
PLA.sub.2 has been determined. This non-pancreatic PLA.sub.2 is
found in platelets, synovial fluid, and spleen and is also a
secreted enzyme. This enzyme is a member of the aforementioned
family. [See, J. J. Seilhamer et al, J. Biol. Chem., 264:5335-5338
(1989); R. M. Kramer et al, J. Biol. Chem., 264:5768-5775 (1989);
and A. Kando et al, Biochem. Biophys. Res. Comm., 163:42-48
(1989)]. However, it is doubtful that this enzyme is important in
the synthesis of prostaglandins, leukotrienes and PAF, since the
non-pancreatic PLA.sub.2 is an extracellular protein which would be
difficult to regulate, and the next enzymes in the biosynthetic
pathways for these compounds are intracellular proteins. Moreover,
there is evidence that PLA.sub.2 is regulated by protein kinase C
and G proteins [R. Burch and J. Axelrod, Proc. Natl. Acad. Sci.
U.S.A., 84:6374-6378 (1989)] which are cytosolic proteins which
must act on intracellular proteins. It would be impossible for the
non-pancreatic PLA.sub.2 to function in the cytosol, since the high
reduction potential would reduce the disulfide bonds and inactivate
the enzyme.
[0010] A murine PLA.sub.2 has been identified in the murine
macrophage cell line, designated RAW 264.7. A specific activity of
2 .mu.mols/min/mg, resistant to reducing conditions, was reported
to be associated with the approximately 60 kD molecule. However,
this protein was not purified to homogeneity. [See, C. C. Leslie et
al, Biochem. Biophys. Acta., 963:476-492 (1988)]. The references
cited above are incorporated by reference herein for information
pertaining to the function of the phospholipase enzymes,
particularly PLA.sub.2.
[0011] A cytosolic phospholipase A.sub.2 alpha (hereinafter
"cPLA.sub.2a") has also been identified and cloned. See, U.S. Pat.
Nos. 5,322,776 and 5,354,677, which are incorporated herein by
reference as if fully set forth. The enzyme of these patents is an
intracellular PLA.sub.2 enzyme, purified from its natural source or
otherwise produced in purified form, which functions
intracellularly to produce arachidonic acid in response to
inflammatory stimuli.
[0012] Bioactive metabolites of arachidonic acid, the eicosanoids,
are recognized as important modulators of platelet signaling.
Inhibitors of the eicosaniod pathway (e.g., aspirin) reduce the
formation of thromboxane A.sub.2 (TXA.sub.2), a labile and potent
platelet agonist, resulting in depression of platelet function,
thrombus formation, and proven clinical benefit in reducing
morbidity and mortality.
[0013] Platelets play a central role in several biological
processes, including thrombosis. [See S. P. Jackson and S. M.
Schoenwaelder, Nature Reviews, Drug Discovery Vol. 2, 1-15, October
2003; D. L. Bhatt and E. J. Topol, Nature Reviews, Drug Discovery
Vol. 2, 15-28, January 2003]. Accordingly, recent efforts have been
made to characterize platelet receptors and signaling pathways. In
addition, a number of rodent models have been developed to enable
the study of potential therapeutics in thrombosis. [See B.
Nieswandt et al., J. Thrombosis and Haemostasis, 3: 1725-1736
(2005).
[0014] Inhibitors of cytosolic phospholipase A.sub.2 are disclosed
in U.S. Pat. No. 6,797,708, which is incorporated herein by
reference in its entirety.
[0015] Now that several phospholipase enzymes have been identified,
it would be desirable to identify chemical inhibitors of the action
of specific phospholipase enzymes, which inhibitors could be used
to treat inflammatory conditions, particularly where inhibition of
production of prostaglandins, leukotrienes and PAF are all desired
results. There remains a need in the art for an identification of
such anti-inflammatory agents for therapeutic use in a variety of
disease states.
SUMMARY OF THE INVENTION
[0016] In some embodiments, the invention provides compounds having
the Formula I:
##STR00002##
wherein:
[0017] n.sub.1 is 1 or 2;
[0018] n.sub.2 is 1 or 2;
[0019] n.sub.3 is 1 or 2;
[0020] n.sub.5 is 0, 1 or 2;
[0021] X.sup.2 is a bond, O, --CH.sub.2-- or SO.sub.2;
[0022] each R.sub.5 is independently H or C.sub.1-3 alkyl;
[0023] R.sub.6 is H or C.sub.1-8 alkyl;
[0024] R.sub.7 is selected from the group consisting of OH,
benzyloxy, CH.sub.3, CF.sub.3, OCF.sub.3, C.sub.1-3 alkoxy,
halogen, COH, CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl),
quinoline-5-yl, quinoline-8-yl, 3,5-dimethylisoxazol-4-yl,
thiophene-3-yl, pyridin-4-yl, pyridine-3-yl, --CH.sub.2-Q, and
phenyl optionally substituted by from one to three independently
selected R.sub.30 groups;
[0025] R.sub.8 is selected from the group consisting of H, OH,
NO.sub.2, CF.sub.3, OCF.sub.3, C.sub.1-3 alkoxy, halogen,
CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl), quinoline-5-yl,
quinoline-8-yl, 3,5-dimethylisoxazol-4-yl, thiophene-3-yl,
--CH.sub.2-Q, and phenyl substituted by from one to three
independently selected R.sub.30 groups;
[0026] Q is OH, dialkylamino,
##STR00003##
[0027] R.sub.20 is selected from the group consisting of H,
C.sub.1-3 alkyl and CO(C.sub.1-3 alkyl); and
[0028] R.sub.30 is selected from the group consisting of
dialkylamino, CN and OCF.sub.3;
[0029] provided that:
[0030] a) when each R.sub.5 is H, R.sub.6 is H, n.sub.5 is 0, and
R.sub.8 is H, then R.sub.7 cannot be chlorine;
[0031] b) when each R.sub.5 is H, R.sub.6 is H, n.sub.5 is 0,
X.sup.2 is O or --CH.sub.2--, and R.sub.8 is H, then R.sub.7 cannot
be CH.sub.3;
[0032] c) when each R.sub.5 is H, and R.sub.6 is H, then R.sub.7
and R.sub.8 cannot both be fluorine;
[0033] d) when each R.sub.5 is H, R.sub.6 is H, and X.sup.2 is O,
then R.sub.7 and R.sub.8 cannot both be chlorine;
[0034] e) when each R.sub.5 is H, R.sub.6 is H, X.sup.2 is O, and
R.sub.8 is NO.sub.2, then R.sub.7 cannot be fluorine; and
[0035] f) when each R.sub.5 is H, R.sub.6 is H, X.sup.2 is
SO.sub.2, and R.sub.8 is H, then R.sub.7 cannot be fluorine or
chlorine.
[0036] In some preferred embodiments, compounds are provided having
the Formula II:
##STR00004##
wherein:
[0037] X is a bond or O;
[0038] n.sub.1 is 1 or 2;
[0039] n.sub.2 is 1 or 2;
[0040] n.sub.6 is 1 or 2;
[0041] R.sub.5 is H or CH.sub.3;
[0042] R.sub.6 is H or C.sub.1-6 alkyl; and
[0043] R.sub.8 is selected from the group consisting of H, OH,
NO.sub.2, CF.sub.3, OCF.sub.3, OCH.sub.3, halogen, COCH.sub.3,
COOCH.sub.3, dimethylamino, diethylamino and CN.
[0044] The present invention also provides methods of treating
inflammation caused or potentiated by prostaglandins, leukotrienes,
or platelet activation factor, in a mammal, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof.
[0045] The present invention further provides methods of treating
pain caused or potentiated by prostaglandins, leukotrienes, or
platelet activation factor, in a mammal, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof.
[0046] The present invention further provides methods of treating
or preventing a disease or disorder in a mammal, or preventing
progression of symptoms such a disease or disorder, wherein the
disease or disorder is selected from the group consisting of
asthma, stroke, atherosclerosis, multiple sclerosis, Parkinson's
disease, arthritic disorders, rheumatic disorders, central nervous
system damage resulting from stroke, central nervous system damage
resulting from ischemia, central nervous system damage resulting
from trauma, inflammation caused or potentiated by prostaglandins,
inflammation caused or potentiated by leukotrienes, pain, and
inflammation caused or potentiated by platelet activation factor,
in a mammal, the method comprising administering to a mammal in
need thereof a pharmaceutically acceptable amount of a compound of
the invention, or a pharmaceutically acceptable salt thereof.
[0047] The present invention also provides methods for treating or
preventing venous or arterial thrombosis in a mammal, or preventing
progression of symptoms of thrombosis, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof. In some embodiments, the
thrombosis is atherothrombosis.
[0048] The present invention also provides pharmaceutical
compositions comprising a compound of the invention, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0049] Also provided in accordance with the present invention are
pharmaceutically acceptable salts, and prodrugs, of the compounds
disclosed herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0050] FIG. 1 shows the in vitro inhibition of platelet aggregation
in human blood by the compounds of Examples 14 and 25, as
determined by the platelet function analyzer (PFA-100.RTM.).
[0051] FIG. 2 shows the improved blood flow and reduction of
thrombus formation by the compounds of Examples 14 and 15, in a rat
model of acute thrombosis
[0052] FIG. 3 shows the reduction of serum thromboxane B.sub.2
levels in rats subjected to ferric chloride induced thrombosis by
the compounds of Examples 14 and 25.
DETAILED DESCRIPTION OF THE INVENTION
[0053] In some embodiments, the invention provides compounds having
the Formula I:
##STR00005##
wherein:
[0054] n.sub.1 is 1 or 2;
[0055] n.sub.2 is 1 or 2;
[0056] n.sub.3 is 1 or 2;
[0057] n.sub.5 is 0, 1 or 2;
[0058] X.sup.2 is a bond, O, --CH.sub.2-- or SO.sub.2;
[0059] each R.sub.5 is independently H or C.sub.1-3 alkyl;
[0060] R.sub.6 is H or C.sub.1-6 alkyl;
[0061] R.sub.7 is selected from the group consisting of OH,
benzyloxy, CH.sub.3, CF.sub.3, OCF.sub.3, C.sub.1-3 alkoxy,
halogen, COH, CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl),
quinoline-5-yl, quinoline-8-yl, 3,5-dimethylisoxazol-4-yl,
thiophene-3-yl, pyridin-4-yl, pyridine-3-yl, --CH.sub.2-Q, and
phenyl optionally substituted by from one to three independently
selected R.sub.30 groups;
[0062] R.sub.6 is selected from the group consisting of H, OH,
NO.sub.2, CF.sub.3, OCF.sub.3, C.sub.1-3 alkoxy, halogen,
CO(C.sub.1-3 alkyl), CO(OC.sub.1-3 alkyl), quinoline-5-yl,
quinoline-8-yl, 3,5-dimethylisoxazol-4-yl, thiophene-3-yl,
--CH.sub.2-Q, and phenyl substituted by from one to three
independently selected R.sub.30 groups;
[0063] Q is OH, alkylamino,
##STR00006##
[0064] R.sub.20 is selected from the group consisting of H,
C.sub.1-3 alkyl and CO(C.sub.1-3 alkyl); and
[0065] R.sub.30 is selected from the group consisting of
dialkylamino, CN and OCF.sub.3; provided that:
[0066] a) when each R.sub.5 is H, R.sub.6 is H, n.sub.5 is 0, and
R.sub.8 is H, then R.sub.7 cannot be chlorine;
[0067] b) when each R.sub.5 is H, R.sub.6 is H, n.sub.5 is 0,
X.sup.2 is 0 or --CH.sub.2--, and R.sub.8 is H, then R.sub.7 cannot
be CH.sub.3;
[0068] c) when each R.sub.5 is H, and R.sub.6 is H, then R.sub.7
and R.sub.8 cannot both be fluorine;
[0069] d) when each R.sub.5 is H, R.sub.6 is H, and X.sup.2 is O,
then R.sub.7 and R.sub.8 cannot both be chlorine;
[0070] e) when each R.sub.5 is H, R.sub.6 is H, X.sup.2 is O, and
R.sub.8 is NO.sub.2, then R.sub.7 cannot be fluorine; and
[0071] f) when each R.sub.5 is H, R.sub.6 is H, X.sup.2 is
SO.sub.2, and R.sub.8 is H, then R.sub.7 cannot be fluorine or
chlorine.
[0072] In some embodiments, X.sup.2 is CH.sub.2. In some further
embodiments, n.sub.3 is 1. In some further embodiments, n.sub.1 is
1. In still further embodiments, n.sub.2 is 1.
[0073] In some embodiments, n.sub.3 is 1; n.sub.1 is 1; and n.sub.2
is 1. In some such embodiments, R.sub.6 is H. In some such
embodiments, n.sub.3 is 1; n.sub.1 is 1; n.sub.2 is 1; R.sub.6 is
H; R.sub.7 is selected from the group consisting of CH.sub.3,
CF.sub.3, OCF.sub.3, halogen, COOCH.sub.3, COH, CH.sub.2OH,
diethylaminomethyl, quinoline-5-yl, quinoline-8-yl,
3,5-dimethylisoxazol-4-yl, thiophene-3-yl, pyridin-4-yl,
pyridine-3-yl, phenyl, 4-dimethylamino-phen-1-yl,
2-trifluoromethoxy-phen-1-yl, 2-cyano-phen-1-yl,
morpholine-1-yl-methyl, piperazine-1-yl methyl,
4-acetyl-piperazine-1-yl methyl, and 4-methyl-piperazine-1-yl
methyl; and R.sub.8 is selected from the group consisting of H,
halogen, CF.sub.3 and NO.sub.2. In some such embodiments, R.sub.5
is H. In some further embodiments, R.sub.5 is CH.sub.3.
[0074] In some embodiments of the compounds of Formula I, X.sup.2
is O. In some such embodiments, n.sub.3 is 1. In some such
embodiments, n.sub.1 is 1. In some such embodiments, n.sub.2 is 1.
In some embodiments, R.sub.6 is H. In some such embodiments,
n.sub.3 is 1; n.sub.1 is 1; and n.sub.2 is 1. In some such
embodiments, n.sub.3 is 1; n.sub.1 is 1; n.sub.2 is 1; R.sub.6 is
H; R.sub.7 is selected from the group consisting of benzyloxy, OH,
halogen, CH.sub.3 and CF.sub.3; and R.sub.8 is selected from the
group consisting of H, halogen, and NO.sub.2. In some such
embodiments, R.sub.5 is H. In some further embodiments, R.sub.5 is
CH.sub.3. In some preferred embodiments, R.sub.7 is CF.sub.3, and
R.sub.8 is H.
[0075] In some embodiments, X.sup.2 is SO.sub.2. In some such
embodiments, n.sub.5 is 2. In some further such embodiments,
n.sub.3 is 1. In some further such embodiments, n.sub.1 is 1. In
some further such embodiments, n.sub.2 is 1. In some embodiments,
R.sub.6 is H. In some further such embodiments, n.sub.3 is 1;
n.sub.1 is 1; and n.sub.2 is 1. In some embodiments, X.sup.2 is
SO.sub.2; n.sub.3 is 1; n.sub.1 is 1; n.sub.2 is 1; R.sub.6 is H;
R.sub.7 is CF.sub.3; and R.sub.8 is H.
[0076] In some embodiments, n.sub.1 is 1; n.sub.2 is 1 or 2;
n.sub.3 is 1, n.sub.5 is 0; X.sup.2 is CH.sub.2, each R.sub.5 and
each R.sub.6 is H; and R.sub.7 and R.sub.8 are independently
selected from the group consisting of H, F, CF.sub.3, OCF.sub.3,
OH, quinoline-5-yl and quinoline-8-yl, provided that R.sub.7 and
R.sub.8 are not both H.
[0077] In some preferred embodiments, compounds are provided having
the Formula II:
##STR00007##
wherein:
[0078] X is a bond or O;
[0079] n.sub.1 is 1 or 2;
[0080] n.sub.2 is 1 or 2;
[0081] n.sub.6 is 1 or 2;
[0082] R.sub.5 is H or CH.sub.3;
[0083] R.sub.6 is H or C.sub.1-6 alkyl; and
[0084] R.sub.8 is selected from the group consisting of H, OH,
NO.sub.2, CF.sub.3, OCF.sub.3, OCH.sub.3, halogen, COCH.sub.3,
COOCH.sub.3, dimethylamino, diethylamino and CN.
[0085] In some embodiments, n.sub.1 is 1. In some further
embodiments, n.sub.2 is 1. In some further embodiments, n.sub.6 is
2. In some further embodiments, R.sub.5 is H. In some further
embodiments, R.sub.6 is H. In some further embodiments, n.sub.1 is
1; n.sub.2 is 1; and n.sub.6 is 2.
[0086] In some preferred embodiments, R.sub.5 is H; R.sub.6 is H;
n.sub.1 is 1, n.sub.2 is 1; and n.sub.6 is 2. In some such
embodiments, R.sub.8 is selected from the group consisting of H,
CF.sub.3, OCF.sub.3 and halogen, preferably H.
[0087] The present invention also provides methods of treating
inflammation caused or potentiated by prostaglandins, leukotrienes,
or platelet activation factor, in a mammal, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof.
[0088] The present invention further provides methods of treating
pain caused or potentiated by prostaglandins, leukotrienes, or
platelet activation factor, in a mammal, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof.
[0089] The present invention also provides methods for treating or
preventing venous or arterial thrombosis in a mammal, or preventing
progression of symptoms of thrombosis, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof. In some embodiments, the
thrombosis is atherothrombosis.
[0090] The present invention also provides pharmaceutical
compositions comprising a compound of the invention, or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0091] Also provided in accordance with the present invention are
pharmaceutically acceptable salts, and prodrugs, of the compounds
disclosed herein.
[0092] Compounds of the present invention may be used in a
pharmaceutical composition when combined with a pharmaceutically
acceptable carrier. Such a composition may also contain (in
addition to a compound or compounds of the present invention and a
carrier) diluents, fillers, salts, buffers, stabilizers,
solubilizers, and other materials well known in the art. The term
"pharmaceutically acceptable" means a non-toxic material that does
not interfere with the effectiveness of the biological activity of
the active ingredient(s). The characteristics of the carrier will
depend on the route of administration. The pharmaceutical
composition may further contain other anti-inflammatory agents.
Such additional factors and/or agents may be included in the
pharmaceutical composition to produce a synergistic effect with
compounds of the present invention, or to minimize side effects
caused by the compound of the present invention.
[0093] The pharmaceutical compositions of the invention may be in
the form of a liposome or micelles in which compounds of the
present invention are combined, in addition to other
pharmaceutically acceptable carriers, with amphipathic agents such
as lipids which exist in aggregated form as micelles, insoluble
monolayers, liquid crystals, or lamellar layers in aqueous
solution. Suitable lipids for liposomal formulation Include,
without limitation, monoglycerides, diglycerides, sulfatides,
lysolecithin, phospholipids, saponin, bile acids, and the like.
Preparation of such liposomal formulations is within the level of
skill in the art, as disclosed, for example, in U.S. Pat. Nos.
4,235,871; 4,501,728; 4,837,028; and 4,737,323, all of which are
incorporated herein by reference.
[0094] As used herein, the terms "pharmaceutically effective
amount" or "therapeutically effective amount" as used herein means
the total amount of each active component of the pharmaceutical
composition or method that is sufficient to show a meaningful
patient benefit, i.e., treatment, healing, prevention, inhibition
or amelioration of a physiological response or condition, such as
an inflammatory condition or pain, or an increase in rate of
treatment, healing, prevention, inhibition or amelioration of such
conditions. When applied to an individual active ingredient,
administered alone, the term refers to that ingredient alone. When
applied to a combination, the term refers to combined amounts of
the active ingredients that result in the therapeutic effect,
whether administered in combination, serially or
simultaneously.
[0095] Each of the methods of treatment or use of the present
invention, as described herein, comprises administering to a mammal
in need of such treatment or use a pharmaceutically or
therapeutically effective amount of a compound of the present
invention, or a pharmaceutically acceptable salt form thereof.
Compounds of the present invention may be administered in
accordance with the method of the invention either alone or in
combination with other therapies such as treatments employing other
anti-inflammatory agents, cytokines, lymphokines or other
hematopoietic factors. When co-administered with one or more other
anti-inflammatory agents, cytokines, lymphokines or other
hematopoietic factors, compounds of the present invention may be
administered either simultaneously with the other anti-inflammatory
agent(s), cytokine(s), lymphokine(s), other hematopoietic
factor(s), thrombolytic or anti-thrombotic factors, or
sequentially. If administered sequentially, the attending physician
will decide on the appropriate sequence of administering compounds
of the present invention in combination with other
anti-inflammatory agent(s), cytokine(s), lymphokine(s), other
hematopoietic factor(s), thrombolytic or anti-thrombotic
factors.
[0096] Administration of compounds of the present invention used in
the pharmaceutical composition or to practice the method of the
present invention can be carried out in a variety of conventional
ways, such as oral ingestion, inhalation, or cutaneous,
subcutaneous, or intravenous injection.
[0097] When a therapeutically effective amount of compounds of the
present invention is administered orally, compounds of the present
invention will be in the form of a tablet, capsule, powder,
solution or elixir. When administered in tablet form, the
pharmaceutical composition of the invention may additionally
contain a solid carrier such as a gelatin or an adjuvant. The
tablet, capsule, and powder contain from about 5 to 95% compound of
the present invention, and preferably from about 10% to 90%
compound of the present invention. When administered in liquid
form, a liquid carrier such as water, petroleum, oils of animal or
plant origin such as peanut oil, mineral oils, phospholipids,
tweens, triglycerides, including medium chain triglycerides,
soybean oil, or sesame oil, or synthetic oils may be added. The
liquid form of the pharmaceutical composition may further contain
physiological saline solution, dextrose or other saccharide
solution, or glycols such as ethylene glycol, propylene glycol or
polyethylene glycol. When administered in liquid form, the
pharmaceutical composition contains from about 0.5 to 90% by weight
of compound of the present invention, and preferably from about 1
to 50% compound of the present invention.
[0098] When a therapeutically effective amount of compounds of the
present invention is administered by intravenous, cutaneous or
subcutaneous injection, compounds of the present invention will be
in the form of a pyrogen-free, parenterally acceptable aqueous
solution. The preparation of such parenterally acceptable protein
solutions, having due regard to pH, isotonicity, stability, and the
like, is within the skill in the art. A preferred pharmaceutical
composition for intravenous, cutaneous, or subcutaneous injection
should contain, in addition to compounds of the present invention,
an isotonic vehicle such as Sodium Chloride Injection, Ringer's
Injection, Dextrose Injection, Dextrose and Sodium Chloride
Injection, Lactated Ringer's Injection, or other vehicle as known
in the art. The pharmaceutical composition of the present invention
may also contain stabilizers, preservatives, buffers, antioxidants,
or other additives known to those of skill in the art.
[0099] The amount of compound(s) of the present invention in the
pharmaceutical composition of the present invention will depend
upon the nature and severity of the condition being treated, and on
the nature of prior treatments the patient has undergone.
Ultimately, the attending physician will decide the amount of
compound of the present invention with which to treat each
individual patient. Initially, the attending physician will
administer low doses of compound of the present invention and
observe the patient's response. Larger doses of compounds of the
present invention may be administered until the optimal therapeutic
effect is obtained for the patient, and at that point the dosage is
not increased further. It is contemplated that the various
pharmaceutical compositions used to practice the method of the
present invention should contain about 0.1 .mu.g to about 100 mg
(preferably about 0.1 mg to about 50 mg, more preferably about 1 mg
to about 2 mg) of compound of the present invention per kg body
weight.
[0100] The duration of intravenous therapy using the pharmaceutical
composition of the present invention will vary, depending on the
severity of the disease being treated and the condition and
potential idiosyncratic response of each individual patient. It is
contemplated that the duration of each application of the compounds
of the present invention will be in the range of 12 to 24 hours of
continuous intravenous administration. Ultimately the attending
physician will decide on the appropriate duration of intravenous
therapy using the pharmaceutical composition of the present
invention.
[0101] A lipid based oral formulation of this invention has been
prepared by blending 50% PHOSAL.RTM. 53MCT (American Lecithin
Company), 5% Polysorbate 80, 15% LABRASOL.RTM. Caprylocaproyl
macrogol-8 glycerides (Gattefosse Corp.), 15% Propylene Carbonate
and 15% active cPLA2 inhibiting compound(s) of this invention, each
percentage listed being by weight.
[0102] Pharmaceutically acceptable salts of the compounds of
Formula (I) having an acidic moiety can be formed from organic and
inorganic bases. Suitable salts with bases are, for example, metal
salts, such as alkali metal or alkaline earth metal salts, for
example sodium, potassium, or magnesium salts; or salts with
ammonia or an organic amine, such as morpholine, thiomorpholine,
piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, for
example ethyl-tert-butyl-, diethyl-, diisopropyl-, triethyl-,
tributyl- or dimethylpropylamine, or a mono-, di-, or trihydroxy
lower alkylamine, for example mono-, di- or triethanolamine.
[0103] The present invention also includes prodrugs of the
compounds described herein. As used herein, "prodrug" refers to a
moiety that releases a compound of the invention when administered
to a mammalian subject. Prodrugs can be prepared by modifying
functional groups present in the compounds in such a way that the
modifications are cleaved, either in routine manipulation or in
vivo, to the parent compounds. Examples of prodrugs include
compounds of the invention as described herein that contain one or
more molecular moieties appended to a hydroxyl, amino, sulfhydryl,
or carboxyl group of the compound, and that when administered to a
mammalian subject, cleaves in vivo to form the free hydroxyl,
amino, sulfhydryl, or carboxyl group, respectively. Examples of
prodrugs include, but are not limited to, acetate, formate and
benzoate derivatives of alcohol and amine functional groups in the
compounds of the invention. Preparation and use of prodrugs is
discussed in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery
Systems," Vol. 14 of the A.C.S. Symposium Series, and in
Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987, both
of which are hereby incorporated by reference in their
entirety.
[0104] As used herein, the term "alkyl" is meant to refer to a
saturated hydrocarbon group which is straight-chained or branched.
Example alkyl groups include methyl (Me), ethyl (Et), propyl (e.g.,
n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl,
t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl) and the
like. Alkyl groups can contain from 1 to about 20, 1 to about 10, 1
to about 8, 1 to about 6, 1 to about 4, or 1 to about 3 carbon
atoms. In some embodiments, alkyl groups can be substituted with up
to four substituent groups, as described below. As used herein, the
term "lower alkyl" is intended to mean alkyl groups having up to
six carbon atoms.
[0105] As used herein, "hydroxy" or "hydroxyl" refers to OH.
[0106] As used herein, "halo" or "halogen" includes fluoro, chloro,
bromo, and iodo.
[0107] As used herein, "cyano" refers to CN.
[0108] As used herein, "alkoxy" refers to an --O-alkyl group.
Example alkoxy groups include methoxy, ethoxy, propoxy (e.g.,
n-propoxy and isopropoxy), t-butoxy, and the like. An alkoxy group
can contain from 1 to about 20, 1 to about 10, 1 to about 8, 1 to
about 6, 1 to about 4, or 1 to about 3 carbon atoms.
[0109] As used herein, "benzyloxy" refers to an --O-benzyl
group.
[0110] At various places in the present specification substituents
of compounds of the invention are disclosed in groups or in ranges.
It is specifically intended that the invention include each and
every individual subcombination of the members of such groups and
ranges. For example, the term "C.sub.1-6 alkyl" is specifically
intended to individually disclose methyl, ethyl, propyl, isopropyl,
n-butyl, sec-butyl, isobutyl, etc.
[0111] The compounds of the present invention can contain an
asymmetric atom (also referred as a chiral center), and some of the
compounds can contain one or more asymmetric atoms or centers,
which can thus give rise to optical isomers (enantiomers) and
diastereomers. The present invention includes such optical isomers
(enantiomers) and diastereomers (geometric isomers); as well as the
racemic and resolved, enantiomerically pure R and S stereoisomers,
as well as other mixtures of the R and S stereoisomers and
pharmaceutically acceptable salts thereof. Optical isomers can be
obtained in pure form by standard procedures known to those skilled
in the art, and include, but are not limited to, diastereomeric
salt formation, kinetic resolution, and asymmetric synthesis. It is
also understood that this invention encompasses all possible
regioisomers, and mixtures thereof, which can be obtained in pure
form by standard separation procedures known to those skilled in
the art, and include, but are not limited to, column
chromatography, thin-layer chromatography, and high-performance
liquid chromatography.
[0112] The novel compounds of the present invention can be prepared
in a variety of ways known to one skilled in the art of organic
synthesis. The compounds of the present invention can be
synthesized using the methods as hereinafter described below,
together with synthetic methods known in the art of synthetic
organic chemistry or variations thereon as appreciated by those
skilled in the art.
[0113] The compounds of present invention can be conveniently
prepared in accordance with the procedures outlined in the schemes
below, from commercially available starting materials, compounds
known in the literature, or readily prepared intermediates, by
employing standard synthetic methods and procedures known to those
skilled in the art. Standard synthetic methods and procedures for
the preparation of organic molecules and functional group
transformations and manipulations can be readily obtained from the
relevant scientific literature or from standard textbooks in the
field. It will be appreciated that where typical or preferred
process conditions (i.e., reaction temperatures, times, mole ratios
of reactants, solvents, pressures, etc.) are given, other process
conditions can also be used unless otherwise stated. Optimum
reaction conditions may vary with the particular reactants or
solvent used, but such conditions can be determined by one skilled
in the art by routine optimization procedures. Those skilled in the
art of organic synthesis will recognize that the nature and order
of the synthetic steps presented may be varied for the purpose of
optimizing the formation of the compounds of the invention.
[0114] The processes described herein can be monitored according to
any suitable method known in the art. For example, product
formation can be monitored by spectroscopic means, such as nuclear
magnetic resonance spectroscopy (e.g., .sup.1H or .sup.13C)
infrared spectroscopy, spectrophotometry (e.g., UV-visible), or
mass spectrometry, or by chromatography such as high performance
liquid chromatography (HPLC) or thin layer chromatography.
[0115] Preparation of compounds can involve the protection and
deprotection of various chemical groups. The need for protection
and deprotection, and the selection of appropriate protecting
groups can be readily determined by one skilled in the art. The
chemistry of protecting groups can be found, for example, in
Greene, et al., Protective Groups in Organic Synthesis, 2d. Ed.,
Wiley & Sons, 1991, which is incorporated herein by reference
in its entirety.
[0116] The reactions of the processes described herein can be
carried out in suitable solvents which can be readily selected by
one of skill in the art of organic synthesis. Suitable solvents can
be substantially nonreactive with the starting materials
(reactants), the intermediates, or products at the temperatures at
which the reactions are carried out, i.e., temperatures which can
range from the solvent's freezing temperature to the solvent's
boiling temperature. A given reaction can be carried out in one
solvent or a mixture of more than one solvent. Depending on the
particular reaction step, suitable solvents for a particular
reaction step can be selected.
[0117] Although not wishing to be limited to any source,
publications and literatures such as WO 200044723; Li, J. P.;
Newlander, K. A.; Yellin, T. O. Synthesis, 1988, 73-76; Gilchrist,
T. L.; Roberts, T. G. J. Chem. Soc. Perkin. Trans 11983, 1283-1292
are useful and recognized references of organic synthesis known to
those in the art. Each of the foregoing is incorporated herein by
reference in its entirety.
[0118] The invention compounds are prepared using conventional
techniques known to those skilled in the art of organic synthesis.
The starting materials used in preparing the compounds of the
invention are known, made by known methods or are commercially
available.
[0119] Those skilled in the art of organic synthesis will recognize
that the nature and order of the synthetic steps presented may be
varied for the purpose of optimizing the formation of the compounds
of the invention.
EXAMPLES
Preparation of Compounds of the Invention
[0120] The following describes the preparation of representative
compounds of this invention in greater detail. The following
examples are offered for illustrative purposes, and are not
intended to limit the invention in any manner. Those of skill in
the art will readily recognize a variety of noncritical parameters
which can be changed or modified to yield essentially the same
results.
[0121] Mass spectral data is reported as the mass-to-charge ratio,
m/z; and for high resolution mass spectral data, the calculated and
experimentally found masses, [M+H].sup.+, for the neutral formulae
M are reported. Nuclear magnetic resonance data is reported as
.delta. in parts per million (ppm) downfield from the standard,
tetramethylsilane; along with the solvent, nucleus, and field
strength parameters. The spin-spin homonuclear coupling constants
are reported as J values in hertz; and the multiplicities are
reported as a: s, singlet; d, doublet; t, triplet; q, quartet;
quintet; or br, broadened.
General Synthetic Scheme(s) for Preparation of Compounds
[0122] Compounds of the invention can be prepared by the procedures
of Methods A-E, shown below:
##STR00008##
[0123] As shown above in Method A, an initial indole may be
alkylated at the C3 position (the carbon atom at the 3-position of
the indole moiety) with aldehydes or the corresponding acetals in
the presence of a Lewis or Bronsted acid, such as boron triflouride
etherate or trifluoroacetic acid. The indole nitrogen may then be
alkylated by treatment with a strong base such as sodium
bis(trimethylsilyl) amide, n-BuLi, sodium hydride or potassium
hydride in a solvent such as DMF, DMSO or THF followed by exposure
to the appropriate alkyl halide. The resulting product can be
treated with carbon tetrabromide in carbon tetrachloride and a
catalytic amount of benzoyl peroxide to effect dibromination of the
C2 methyl group. The dibromide can then either be stirred with
silver carbonate in acetone water or poured into DMSO and stirred.
Both of these procedures generate the aldehyde which is then
subjected to the nitro aldol reaction with nitromethane and
ammonium acetate at reflux. The resulting vinyl nitro intermediate
is reduced to the amine upon treatment with zinc mercury amalgam in
a mixture of THF and conc. HCl at reflux. This amine can then be
treated with the requisite sulfonyl chloride under biphasic
conditions, aqueous sodium bicarbonate/dichloromethane, or in
organic solvent with the addition of a hindered organic amine base.
The final hydrolysis can be accomplished under basic conditions
with sodium hydroxide in water and methanol and THF at room
temperature or at elevated temperature. Alternatively it may be
cleaved by treatment with sodium thiomethoxide in a solvent such as
THF or DMF at elevated temperatures (50.degree. C.-100.degree.
C.).
##STR00009##
[0124] As shown above in Method B, a halide is placed in a vessel
with a boronic acid, a base (for example KF), a palladium source
(for example Pd(OAc).sub.2) a ligand (for example PPh.sub.3), and a
suitable degassed solvent, for example DMF, MeOH, water or a
combination thereof. The mixture is then heated either thermally or
in a microwave reactor. Standard workup yields the protected
(ester) product, which is then hydrolyzed in base to afford the
free acid product.
##STR00010##
[0125] As shown in Method C above, a formyl containing compound is
treated with an amine, an acid source if necessary, and a suitable
reducing agent, such as NaBH(OAc).sub.3. The reaction is allowed to
stir at room temperature, or can be heated if necessary. Standard
workup yields the protected (ester) product, which is then
hydrolyzed in base to afford the free acid product
##STR00011## ##STR00012## ##STR00013##
[0126] As shown in Method D above, an appropriately substituted
halo amine is reacted with trifluoroacetic anhydride to yield an
intermediate that could be treated with a Pd(II) catalyst in the
presence of a base such as triethylamine, CuI and a suitable
alkyne, under heat to yield the desired indole intermediate. The
primary alcohol is protected as a silyl ether using a silyl
chloride such as t-butyldiphenyl silyl chloride and a base such as
imidazole. The protected indole is then treated with oxalyl
chloride followed by methanol which produces the desired oxalate
ester, the indole nitrogen of which can be alkylated using a
suitable base such as cesium carbonate in refluxing acetonitrile
and a halide. The oxalate can then be reduced via the action of a
suitable reducing agent such as borane. The resulting primary
alcohol is converted to a halide using, for example, CBr.sub.4 and
a phosphine, which can then be displaced with a nucleophile such as
a thiophenol. The resulting thioether can be oxidized by a variety
of oxidizing agents including oxone and TPAP/NMO. The resulting
sulfone can be deprotected via the action of a fluoride source such
as TBAF, CsF or HF. The resulting alcohol can be converted to a
halide or mesylate, for example using methane sulfonyl chloride and
an organic base, which can then be displaced by sodium azide in
DMF. The resulting alkyl azide can be reduced under the action of
triphenyl phosphine and wet THF. The amine can be sulfonylated by
the action of a sulfonyl chloride under either biphasic
Schotten-Baumann conditions (aq. bicarbonate and dichloromethane)
or under anhydrous conditions consisting of dichloromethane and an
organic base such as Hunigs base. The resulting ester intermediate
is hydrolyzed using a base, such as NaOH, KOH or LiOH, and a
mixture of solvents including an alcoholic solvent, water and
tetrahydrofuran.
##STR00014## ##STR00015##
[0127] As shown in Method E above, a starting amino acid is
esterified and N-alkylated in one pot (using for example a diazo
reagent or trimethylsilyldiazo reagent). This N-alkyl ester is then
sulfonylated with a sulfonyl chloride using either Schotten-Baumann
conditions or organic solvents and organic bases. Finally, the
N-alkyl ester is hydrolyzed to the desired product using a base,
such as NaOH, and a suitable solvent system, such as THF and an
alcohol.
[0128] The following compounds were prepared in accordance with
Methods A-E above.
Example 1
4-{2-[2-[2-({[2-(benzyloxy)benzyl]sulfonyl}amino)ethyl]-5-chloro-1-(diphen-
ylmethyl)-1H-indol-3-yl]ethoxy}benzoic acid
[0129] Step 1: To 4-hydroxy-benzoic acid methyl ester (1.0 eq) in
DMF (0.83 M) was added K.sub.2CO.sub.3 (2.0 eq) followed by
2-bromo-1,1-diethoxy-ethane and the reaction mixture was stirred at
110.degree. C. for 2 days. TLC showed a new spot. The reaction
mixture was diluted with ethyl acetate, washed with 1N NaOH, water,
and brine, dried over sodium sulfate, and solvent was removed to
afford desired product in 84% yield. This material was used in the
next step without further purification.
[0130] Step 2: To the above product (1.0 eq) and 5-chloro-2-methyl
indole (1.0 eq) in CH.sub.2Cl.sub.2 (0.12 M) was added
triethylsilane (3.0 eq) followed by trifluoroacetic acid (3.0 eq).
After being stirred overnight at room temperature, water and
trifluoroacetic acid (1.0 eq) were added to the reaction mixture,
which was stirred at room temperature for two days, diluted with
CH.sub.2Cl.sub.2, washed with 1N NaOH, water, and brine, and dried
over sodium sulfate. Trituration of the material with
CH.sub.2Cl.sub.2 and hexanes afforded the C3 alkylated indole in
92% yield
[0131] Step 3: To the indole from above (1.0 eq) in DMF (0.36 M) at
25.degree. C. was added NaH (1.2 eq, 60% dispersion in oil). The
brown solution was stirred at 0 to -5.degree. C. for 1 h, and then
bromodiphenylmethane was added (1.1 eq). The reaction mixture was
stirred overnight, and then quenched with water, diluted with ethyl
acetate, washed with water and brine, dried over sodium sulfate and
purified by column chromatography to yield 72% of the desired
product.
[0132] Step 4: To the N-alkylated indole from above (1.0 eq) in
CCl.sub.4 (0.2 M) was added N-bromosuccinimide (2.0 eq) and a
catalytic amount of benzoyl peroxide. The solution was heated to
reflux for 3 h, cooled to 25.degree. C., filtered, and the solid
was washed with CCl.sub.4. The filtrate was concentrated to a foam
which was dried in vacuo. The foam was dissolved in acetone, and
Ag.sub.2CO.sub.3 (1.1 eq.) was added followed by water, and the
reaction mixture was stirred overnight at room temperature, and
then filtered and washed with acetone. The filtrate was
concentrated to a residue, to which was added water. This mixture
was extracted with ethyl acetate, washed with brine, and dried over
sodium sulfate. Chromatographic purification of the residue gave
the desired product in 85% yield.
[0133] Step 5: To the above aldehyde (1.0 equiv) in
CH.sub.3NO.sub.2 (0.2 M) was added ammonium acetate (4 equiv) and
the resulting mixture was heated to reflux for 4 h. The reaction
mixture was then diluted with EtOAc and washed with brine. The
aqueous phase was extracted with EtOAc. The combined organic
extracts were washed with brine, dried over sodium sulfate, and
concentrated until an orange crystalline solid precipitated. The
mixture was refrigerated overnight and the nitroolefin (76% yield)
was collected by filtration. Evaporation of the solution phase and
purification of the residue by column chromatography (gradient
elution 100% toluene.fwdarw.1% EtOAc-toluene) afforded an
additional amount of the nitroolefin (23% yield).
[0134] Step 6: Zinc dust (20 equiv) was suspended in 5% aqueous HCl
solution (8 M Zn/5% HCl). To this mixture was added HgCl.sub.2
(0.28 equiv). The mixture was shaken for 10 min, the aqueous phase
was decanted and replaced with fresh 5% HCl, and again the mixture
was shaken for 5 min and the aqueous phase was removed. The
zinc-mercury amalgam thus generated was then added to a mixture of
the nitroolefin (1.0 equiv) and conc. HCl (80 equiv) in THF (0.04 M
nitroolefin/THF). The mixture was maintained at a gentle reflux for
1 h. The formation of product was followed by TLC analysis. The
mixture was cooled to room temperature and the solids were removed
by filtration through Celite. Conc. NH.sub.4OH was added to the
solution phase and the mixture was concentrated on the rotary
evaporator. The residue was dissolved in CH.sub.2Cl.sub.2 and conc.
NH.sub.4OH. The aqueous phase was extracted with CH.sub.2Cl.sub.2,
and the organic phase was washed with brine, dried over sodium
sulfate, and concentrated. Purification by column chromatography
afforded the desired product (65% yield).
[0135] Step 7: Sodium sulfite (4.2 g) was added to a stirred
mixture of 1-benzyloxy-2-bromomethyl-benzene (8.9 g),
tetrabutylammonium iodide (59 mg) and water (150 ml). The mixture
was warmed to reflux for overnight. As the mixture cooled to
0.degree. C., it was acidified by 6N HCl. Extraction by ethyl
acetate (100 ml.times.6) was performed (some remained in the
aqueous layer). The combined organic phases were dried over
MgSO.sub.4. The filtrate was concentrated on vacuo. The product was
triturated by ethyl ether to give
(2-Benzyloxy-phenyl)-methanesulfonic acid (678 mg, 8%). .sup.1H NMR
(400 MHz, DMSO-D6): .delta. 3.82 (s, 2H) 5.09 (s, 2H) 6.86 (t,
J=7.45 Hz, 1H) 6.96 (d, J=8.08 Hz, 1H) 7.14 (t, J=7.83 Hz, 1H) 7.32
(d, J=7.33 Hz, 1H) 7.38 (t, J=7.33 Hz, 2H) 7.46 (d, J=9.09 Hz, 1H)
7.52 (d, J=7.07 Hz, 2H).
[0136] Step 8: Tetrahydrofuran (10 ml),
(2-Benzyloxy-phenyl)-methanesulfonic acid (138 mg), and
N,N-dimethylformamide (2 drops) was cooled to -78.degree. C. and
oxalyl chloride (315 mg) was added slowly. The reaction mixture was
stirred for 3 h from -78.degree. C. to 0.degree. C. The reaction
mixture was clarified by filtration. The filtrate was washed with
iced-water and heptane, and dried to give
(2-benzyloxy-phenyl)-methanesulfonyl chloride (114 mg, 77%).
.sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 5.06 (s, 2H) 5.15 (s,
2H) 7.04 (m, 2H) 7.42 (m, 7H).
[0137] Step 9: To methyl
4-{2-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]ethoxy}benzoat-
e (1.0 equiv, Step 6) and sat. NaHCO.sub.3 (0.14 M) in
CH.sub.2Cl.sub.2 (0.07 M) was added
(2-benzyloxy-phenyl)-methanesulfonyl chloride (1.0 equiv, step 8).
After 16 h the mixture was poured into saturated sodium bicarbonate
and extracted with CH.sub.2Cl.sub.2. The combined organic phase was
washed with brine, dried over sodium sulfate and purified by column
chromatography to afford 77% of the desired product.
[0138] Step 10. The resulting ester was hydrolyzed by stirring with
1N NaOH (5 equiv) in THF (0.07 M) and enough MeOH to produce a
clear solution. The reaction was monitored by TLC (10%
MeOH--CH.sub.2Cl.sub.2) for the disappearance of starting material.
When the reaction was complete, the mixture was concentrated,
diluted with H.sub.2O, and acidified to pH 2-4 using 1 M HCl. The
aqueous phase was extracted with EtOAc and the organic phase was
washed with brine, dried over sodium sulfate, and concentrated to
afford the title acid in 97% yield. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.86 (d, J=14.40 Hz, 2H) 2.92-3.04 (m, 2H) 3.13
(t, J=6.69 Hz, 2H) 4.12-4.23 (m, 2H) 4.28 (s, 2H) 4.34-4.45 (m, 1H)
4.90 (s, 2H) 6.47 (d, J=8.84 Hz, 1H) 6.73-6.93 (m, 6H) 6.95-7.08
(m, 4H) 7.16-7.36 (m, 13H) 7.53 (d, J=1.77 Hz, 1H) 7.92-8.04 (m,
2H); HRMS calc for [C.sub.46H.sub.41ClN.sub.2O.sub.6.S+H.sup.-]
783.2301 found 783.2292; purity H.sub.2O/MeOH 97%, H.sub.2O/MeCN
95%.
Example 2
4-{2-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]ethoxy}benzoic acid
[0139] Step 1: To
4-{2-[2-[2-({[2-(benzyloxy)benzyl]sulfonyl}amino)ethyl]-5-chloro-1-(diphe-
nylmethyl)-1H-indol-3-yl]ethoxy}benzoic acid (Step 9, Example 1,
109 mg, 0.14 mmol) was added THF and MeOH. 10% of Pd/C (11 mg) was
added. The mixture was stirred at room temperature under H.sub.2 (1
atm) overnight and filtered through celite, concentrated, and
column chromatographed (35% EtOAc/hex) to give
4-(2-{1-benzhydryl-5-chloro-2-[2-(2-hydroxy-phenylmethanesulfonylamino)-e-
thyl]-1H-indol-3-yl}-ethoxy)-benzoic acid methyl ester (74 mg,
76%), an off-white solid.
[0140] Step 2: The ester intermediate was hydrolyzed according to
Step 10 Example 1 to afford the title acid in 85% yield. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 2.87-3.01 (m, 2H) 3.00-3.11 (m,
2H) 3.18 (t, J=6.57 Hz, 2H) 4.17 (s, 2H) 4.19-4.30 (m, 2H) 4.52 (t,
J=5.81 Hz, 1H) 6.52 (d, J=8.84 Hz, 1H) 6.75-6.90 (m, 6H) 6.99 (dd,
J=7.45, 1.64 Hz, 1H) 7.01-7.13 (m, 4H) 7.13-7.22 (m, 1H) 7.27-7.37
(m, 6H) 7.53 (d, J=2.02 Hz, 1H) 7.91-8.04 (m, 2H); HRMS calc for
[C.sub.39H.sub.35ClN.sub.2O.sub.6.S+H.sup.-] 695.1977 found
695.1984.
Example 3
4-{2-[5-chloro-2-(2-{[(2,6-dibromobenzyl)sulfonyl]amino}ethyl)-1-(diphenyl-
methyl)-1H-indol-3-yl]ethoxy}benzoic acid
[0141] Step 1. To a solution of 2,6-dibromotoluene (5.38 g, 21.53
mmol) in benzene (1.54 M) was added N-bromosuccinimide (4.21 g,
23.68 mmol) and benzoyl peroxide (0.52 g, 2.15 mmol). The mixture
was then heated to reflux overnight. The mixture was cooled to rt,
diluted with H.sub.2O and extracted with EtOAc. The combined
organic phase was washed with brine, dried over MgSO.sub.4 and
concentrated to afford 7.65 g of the benzyl bromide, a brown solid.
1H NMR (400 MHz, CDCl.sub.3) .delta. 4.83 (s, 2H), 7.01 (t, J=8.0
Hz, 1H), 7.55 (d, J=8.1 Hz, 2H).
[0142] Step 2. To a solution of the 2,6-dibromobenzyl bromide (1.0
equiv, Step 1) in DMF (1.30 M) was added potassium thioacetate (1.2
equiv.) and the mixture was allowed to stir at rt for 3-4 h. The
reaction was monitored by LC/MS for disappearance of starting
material. The mixture was diluted with H.sub.2O and extracted with
EtOAc. The combined organic phase was washed with brine, dried over
MgSO.sub.4 and concentrated to afford 6.70 g (89%) of the benzyl
thioacetate as a brown oil.
[0143] Step 3. To a solution of the thioacetate (1.0 equiv, 6.70 g,
20.7 mmol) in AcOH (0.19M) and H.sub.2O (0.91M) was added sodium
acetate (6.7 equiv.). Chlorine was then bubbled through the
reaction mixture vigorously for a period of 3045 min. The mixture
was then concentrated, diluted with ether, washed with H.sub.2O and
brine, dried with MgSO.sub.4 and concentrated to afford 5.30 g
(74%) of the desired 2,6-dibromophenyl-methanesulfonyl chloride, a
brown solid. 1H NMR (400 MHz, CDCl.sub.3) .delta. 5.55 (s, 2H),
7.17 (t, J=8.0 Hz, 1H), 7.67 (d, J=8.1 Hz, 2H)
[0144] Step 4.
4-{2-[2-(2-Amino-ethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]-ethoxy}-benz-
oic acid methyl ester (Example 1, Step 6, 126 mg, 0.23 mmol) was
reacted with 2,6-dibromophenyl-methanesulfonyl chloride (Step 3)
according to the procedure in Example 1, Step 9 to afford 203 mg of
the desired sulfonamide as a white solid in quantitative yield.
[0145] Step 5. Using the procedure in Example 1, Step 10, the
sulfonamide ester (175 mg, 0.206 mmol) was hydrolyzed to afford the
146 mg (85%) of the title product, a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.87-3.03 (m, 2H), 3.06-3.14 (m, 2H), 3.22 (t,
J=6.9 Hz, 2H), 4.23 (t, J=6.4 Hz, 2H), 4.53 (t, J=5.9 Hz, 1H), 4.72
(s, 2H), 6.51 (d, J=8.8 Hz, 1H), 6.82 (dd, J=9.0, 2.1 Hz, 1H), 6.87
(d, J=8.8 Hz, 2H), 6.92 (s, 1H), 6.97 (t, J=8.0 Hz, 1H), 7.05-7.12
(m, J=6.2, 2.9 Hz, 4H), 7.29-7.34 (m, 6H), 7.49 (d, J=8.1 Hz, 2H),
7.54 (d, J=2.0 Hz, 1H), 8.00 (d, J=8.8 Hz, 2H).
Example 4
4-(2-{1-benzhydryl-5-chloro-2-[2-methyl-6-nitro-phenylmethanesulfonylamino-
)-ethyl-1-H-indol-3-yl}ethoxy)-benzoic acid
[0146] Step 1. To a solution of 2-methyl-6-nitrophenylbenzoic acid
(3.02 g, 16.67 mmol) in thionyl chloride (0.56 M) was added DMF
(cat.) and the mixture was heated to reflux for 5.5 h. The mixture
was then cooled to room temperature and concentrated. The residue
was then taken up in THF (30 mL) and added slowly over 20 min to a
slurry of NaBH.sub.4 in THF (30 mL) which pre-cooled to 0.degree.
C. The mixture was stirred at rt for 2 h and then quenched by
addition of H.sub.2O followed by 4M HCl. The mixture was extracted
with EtOAc. The combined organic phase was washed with sat.
NaHCO.sub.3 and brine, dried over MgSO.sub.4 and concentrated to
afford 2.52 g (90%) of the benzyl alcohol, an orange solid. 1H NMR
(400 MHz, CDCl.sub.3) .delta. 2.55 (s, 3H), 4.70 (s, 2H), 7.35 (t,
J=7.8 Hz, 1H), 7.48 (d, J=7.6 Hz, 1H), 7.70 (d, J=8.3 Hz, 1H).
[0147] Step 2. To a solution of the benzyl alcohol (2.52 g, 15.07
mmol) in CH.sub.2Cl.sub.2 (0.12M) cooled to -78.degree. C. and
under argon was slowly added BBr.sub.3, 1.0M in CH.sub.2Cl.sub.2,
(23 mL, 22.6 mmol). The mixture was stirred at room temperature
overnight and then diluted with H.sub.2O (150 mL). The layers were
separated and the organic phase was washed with brine, dried over
MgSO.sub.4 and concentrated to afford 2.97 g (86%) of
2-methyl-6-nitrobenzyl bromide, a brown solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.53 (s, 3H), 4.72 (s, 2H), 7.36 (t, J=7.8 Hz,
1H), 7.46 (d, J=7.6 Hz, 1H), 7.75 (d, J=8.1 Hz, 1H).
[0148] Step 3. 2-Methyl-6-nitrobenzyl bromide (Step 2, 1.5 g, 6.5
mmol) was reacted with potassium thioacetate according to the
procedure in Example 3, Step 2, to afford 1.44 g of the benzyl
thioacetate, a brown oil.
[0149] Step 4. Following the procedure in Example 3, Step 3, the
benzyl thioacetate (1.44 g, 6.39 mmol) was oxidized to afford 1.35
g (84%) of (2-methyl-6-nitrophenyl)methanesulfonyl chloride, a
orange solid. 1H NMR (400 MHz, CDCl.sub.3) .delta. 2.62-2.65 (m,
3H), 5.68 (s, 2H) Broad, 7.54 (t, J=7.8 Hz, 1H), 7.58-7.60 (m, 1H),
7.91 (d, J=7.8 Hz, 1H).
[0150] Step 5. Using the procedure in Example 1, Step
9,4-{2-[2-(2-amino-ethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]-ethoxy}-be-
nzoic acid methyl ester (Example 1, Step 6, 255 mg, 0.47 mmol) was
reacted with the sulfonyl chloride from Step 4 to afford 318 mg of
the sulfonamide, a yellow solid in 90% yield.
[0151] Step 6. The sulfonamide ester (101 mg, 0.13 mmol) was
hydrolyzed according to Example 1, Step 10 to afford the 87 mg
(91%) of the title product, a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.48 (s, 3H), 2.87-2.99 (m, 2H), 3.03-3.10 (m,
2H), 3.22 (t, J=6.6 Hz, 2H), 4.23 (t, J=6.6 Hz, 2H), 4.33 (t, J=5.9
Hz, 1H), 4.77 (s, 2H), 6.51 (d, J=8.8 Hz, 1H), 6.82 (dd, J=8.8, 2.0
Hz, 1H), 6.88 (d, J=8.8 Hz, 2H), 6.91 (s, 1H), 7.04-7.12 (m, 4H),
7.29-7.35 (m, 7H), 7.42 (d, J=7.3 Hz, 1H), 7.54 (d, J=2.0 Hz, 1H),
7.66 (d, J=7.6 Hz, 1H), 7.99 (d, J=8.8 Hz, 2H);
Example 5
4-(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]sulfon-
yl}amino)ethyl]-1H-indol-3-yl}ethoxy)benzoic acid
[0152] Step 1: A mixture of 2-(trifluoromethyl)benzyl bromide (25
g, 0.14 mol) sodium sulfite (19.1 g, 0.15 mol), tetrabutylammonium
iodide (0.224 g, 0.6 mmol) and H.sub.2O (930 mL) was heated to
reflux for 2 d. The mixture was cooled to room temperature and the
aqueous phase was decanted from the oily residue and concentrated
on the rotovap to dryness to afford the desired sodium salt (22.2
g, 60%), as a white solid, which was used without further
purification.
[0153] Step 2: (2-Trifluoromethylphenyl)methanesulfonic acid sodium
salt (22.2 g, 84 mmol) was suspended in MeOH (950 mL) and cooled to
-20.degree. C. At that temp with continued cooling HCl (g) was
bubbled through the mixture for 5 min. The resulting white
suspension was stirred at room temperature for 1.5 h, then cooled
in an ice-bath. The resulting suspension was filtered and the
collected solid allowed to air-dry overnight to afford
(2-trifluoromethylphenyl)methanesulfonic acid (20.3 g,
.about.100%), a white solid, which was used without further
purification
[0154] Step 3: To a suspension of
(2-trifluoromethylphenyl)methanesulfonic acid (20.3 g, 84 mmol) in
THF (1.9 L) and DMF (5.0 mL) at -20.degree. C. was added oxalyl
chloride (44.7 mL, 0.5 mol) slowly dropwise over 1 hr. The bath
temperature was maintained below 0.degree. C. for 4 h, at which
point the reaction was evaporated to a volume of .about.250 mL and
diluted with 500 mL of ethyl acetate. This solution was washed with
brine in a separatory funnel and dried over magnesium sulfate. The
solution was then evaporated to a brown oil. This oil was taken up
in 500 mL of pet ether (30-50.degree.) and heated with a heat gun
until the oil went into solution. The solution was then placed into
a dry-ice acetone bath to cool resulting in formation of a white
crystalline material. This material was collected via filtration
and dried to afford 19 g (85%) of
(2-trifluoromethylphenyl)methanesulfonyl chloride as a white
solid.
[0155] Step 4: As outlined in Step 9, Example 1, methyl
4-{2-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]ethoxy}benzoat-
e (Step 9, Example 1, 0.15 g, 0.28 mmol) was reacted with
2-(trifluoromethylphenyl)methanesulfonyl chloride (0.145 g, 0.50
mmol) to afford 0.220 g of the sulfonamide, a white foam, in 75%
yield. 1H NMR (400 MHz, CDCl.sub.3) 2.73-2.88 (m, 2H), 2.96-3.09
(m, 2H), 3.16 (t, J=6.6 Hz, 2H), 3.88 (s, 3H), 4.19 (t, J=6.6 Hz,
2H), 4.23 (t, J=6.4 Hz, 1H), 4.34 (s, 2H), 6.51 (d, J=8.8 Hz, 1H),
6.77-6.84 (m, 3H), 6.86 (s, 1H), 6.98-7.12 (m, 4H), 7.27-7.35 (m,
6H), 7.36-7.47 (m, 2H), 7.53 (d, J=1.5 Hz, 1H), 7.59-7.69 (m, 2H),
7.95 (d, J=8.8 Hz, 2H).
[0156] Step 5: Using the procedure in Step 10 Example 1, the
sulfonamide ester (137 mg, 0.18 mmol) was hydrolyzed to afford 86
mg (64%) of the title product, a white powder. .sup.1H NMR (400
MHz, DMSO-d.sub.6) 3.04 (s, 4H), 3.18 (t, J=6.6 Hz, 2H), 4.23 (t,
J=5.9 Hz, 2H), 4.42 (s, 2H), 6.48 (d, J=8.8 Hz, 1H), 6.81 (dd,
J=9.0, 2.1 Hz, 1H), 6.98 (d, J=9.1 Hz, 2H), 7.03-7.18 (m, 5H),
7.29-7.42 (m, 6H), 7.48-7.62 (m, 3H), 7.66 (d, J=2.0 Hz, 2H), 7.72
(d, J=7.8 Hz, 1H), 7.85 (d, J=8.8 Hz, 2H), 12.49 (s, 1H); HRMS:
calcd for C.sub.40H.sub.34ClF.sub.3N.sub.2O.sub.5S+H+, 747.19018;
found (ESI-FTMS, [M+H].sup.1+), 747.1886; HPLC purity
H.sub.2O/CH.sub.3CN: 96.2%, H.sub.2O/MeOH: 95.4%.
Example 6
4-(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)benz-
yl]sulfonyl}amino)ethyl]-1H-indol-3-yl}ethoxy)benzoic acid
[0157] Step 1: Using the procedure described in Example 5, Step
1,2-fluoro-6-(trifluoromethylphenyl)benzyl bromide (15 g, 61 mmol)
afforded 2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid
sodium salt (15 g, 89%), a white solid. .sup.1HNMR (400 MHz,
DMSO-d.sub.6) 4.02 (s, 2H), 7.26-7.66 (m, 3H).
[0158] Step 2: Using the procedure described in Example 5, Step
2,2-fluoro-6-(trifluoromethylphenyl)methanesulfonic sodium salt (15
g, 53 mmol) afforded
2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid (15 g), a
pale orange oil which was used without further purification.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) 4.12 (s, 2H), 7.39-7.73 (m,
3H).
[0159] Step 3: Using the procedure described in Example 5, Step
3,2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid (15 g, 53
mmol) afforded 11 g of crude product which was purified by
low-temperature crystallization from hexanes to afford
2-fluoro-6-(trifluoromethylphenyl)methanesulfonyl chloride (9.0 g,
62%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.31 (s, 2H),
7.38-7.51 (m, 1H), 7.58-7.68 (m, 2H).
[0160] Step 4: As outlined in Step 9, Example 1, methyl
4-{2-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]ethoxy}benzoat-
e (Step 6, Example 1, 0.12 g, 0.22 mmol) was reacted with
2-fluoro-6-(trifluoromethylphenyl)methanesulfonyl chloride (0.074
g, 0.27 mmol) to afford 0.164 g of the sulfonamide, a white foam,
in 95% yield. .sup.1H NMR (400 MHz, CDCl.sub.3). .delta. 2.83-3.03
(m, 2H), 3.07-3.17 (m, 2H), 3.21 (t, J=6.6 Hz, 2H), 3.88 (s, 3H),
4.22 (t, J=6.6 Hz, 2H), 4.31 (t, J=6.3 Hz, 1H), 4.43 (s, 2H), 6.52
(d, J=9.1 Hz, 1H), 6.76-6.89 (m, 3H), 6.92 (s, 1H), 7.07 (dd,
J=6.1, 4.3 Hz, 4H), 7.23 (t, J=8.6 Hz, 2H), 7.28-7.34 (m, 5H),
7.38-7.52 (m, 2H), 7.54 (d, J=1.8 Hz, 1H), 7.95 (d, J=9.1 Hz,
2H).
[0161] Step 5: Using the procedure in Step 10 Example 1, the
sulfonamide ester (136 mg, 0.17 mmol) was hydrolyzed to afford 130
mg (97%) of the title product, a white powder. .sup.1H NMR (400
MHz, DMSO-d.sub.6) 3.00-3.15 (m, 4H), 3.17 (t, J=6.4 Hz, 2H), 4.22
(t, J=6.6 Hz, 2H), 4.45 (s, 2H), 6.46 (d, J=8.8 Hz, 1H), 6.79 (dd,
J=8.8, 2.3 Hz, 1H), 6.97 (d, J=9.1 Hz, 2H), 7.03-7.13 (m, 5H),
7.16-7.41 (m, 6H), 7.48-7.70 (m, 4H), 7.74-7.90 (m, 3H), 12.56 (s,
1H); HRMS: calcd for C.sub.40H.sub.33ClF.sub.4N.sub.2O.sub.5S+H+,
765.18076; found (ESI-FTMS, [M+H].sup.1+), 765.1814; HPLC purity
H.sub.2O/CH.sub.3CN: 96.6%, H.sub.2O/MeOH: 97.9%.
Example 7
4-{3-[5-chloro-2-(2-{[(2,6-dibromobenzyl)sulfonyl]amino}ethyl)-1-(diphenyl-
methyl)-1H-indol-3-yl]propyl}benzoic acid
[0162] Step 1: A mixture of methyl-4-iodobenzoate (5.3 g, 20.2
mmol), allyl alcohol (1.78 g, 30.3 mmol), NaHCO.sub.3 (4.24 g, 50.5
mmol), Pd(OAc).sub.2 (0.14 g, 0.60 mmol), (n-Bu).sub.4NBr (6.55 g,
20.2 mmol) and 4-A molecular Sieves (4.1 g) in anhydrous DMF (69
mL) was stirred at room temperature for 4 days. The reaction
mixture was filtered through celite and the filtrate poured onto
water and extracted with EtOAc. The organic layer was washed with
brine, dried (Na.sub.2SO.sub.4), and concentrated under vacuum.
Flash chromatography (silica gel, 10-20% EtOAc-hexanes) gave 2.11 g
(85% based on the recovered starting material) of the desired
4-(3-oxo-propyl)-benzoic acid methyl ester as a clear oil.
[0163] Step 2: To a solution of 5-chloro-2-methylindole (0.86 g,
5.2 mmol) and 4-(3-oxo-propyl)-benzoic acid methyl ester (1.0 g,
5.2 mmol) in methylene chloride (50 mL), was added TFA (1.78 g,
15.6 mmol), followed by triethylsilane (1.81 g, 15.6 mmol). The
reaction mixture was stirred overnight, quenched with sat.
NaHCO.sub.3 solution (50 mL), and the organic layer was washed with
sat. NaHCO.sub.3 solution, water, brine, and dried
(Na.sub.2SO.sub.4). The solvent was removed under reduced pressure,
and the residue was purified by flash column chromatography with
10-20% EtOAc/hexanes to yield the desired product (1.67 g) in 94%
yield.
[0164] Step 3: To a solution of the product from step 2 (1.66 g,
4.86 mmol) in DMF (20 mL) was added NaH (60% in mineral oil, 0.24
g, 5.83 mmol) under N.sub.2 atmosphere. The mixture was stirred for
1 h at room temperature, followed by the dropwise addition of
benzhydryl bromide (1.8 g, 7.29 mmol) in DMF (5 mL). This reaction
mixture was stirred overnight at room temperature. Water (500 mL)
was added, and the mixture was extracted with EtOAc, washed with
brine, dried (Na.sub.2SO.sub.4), and concentrated under reduced
pressure to a brown syrup, which was purified by silica-gel
chromatography using 10% EtOAc/hexanes as eluent to isolate the
N-benzhydrylindole as a white solid (1.47 g) in 59% yield.
[0165] Step 4: The product from above (1.46 g, 2.87 mmol) was
dissolved in CCl.sub.4 (14.5 mL), followed by the addition of NBS
(1.02 g, 5.73 mmol) and benzoyl peroxide (2 mg). The reaction
mixture was heated to reflux for 1 h (until all the starting
material disappeared by TLC analysis). This mixture was cooled to
room temperature, filtered and the solid was washed with CCl.sub.4.
The filtrate was evaporated to a brown residue, which was dissolved
in acetone (40 mL) and water (4 mL). Ag.sub.2CO.sub.3 (1.75 g, 3.16
mmol) was then added to this solution and after being stirred
overnight at room temperature, it was filtered through celite, the
solvent was evaporated under reduced pressure, and water was added
to the residue. It was extracted with EtOAc, washed with brine,
dried (Na.sub.2SO.sub.4), and evaporated to a syrup, which was
purified by 10% EtOAc/hexanes to isolate the 2-formyl indole (1.13
g) in 75% yield.
[0166] Step 5: To a solution of the 2 formyl indole from above
(0.52 g, 1 mmol) in CH.sub.3NO.sub.2 (6.2 mL) was added NH.sub.4OAc
(0.077 g, 1 mmol), the mixture was heated to reflux for 1 h,
NH.sub.4OAc (0.077 g, 1 mmol) was then added, heating at reflux was
continued for an additional 1 h, NH.sub.4OAc (0.077 g, 1 mmol) was
added again and the heating continued for further 1 h. The reaction
mixture was allowed to cool to room temperature and EtOAc (50 mL)
was added, followed by water (100 mL). The aqueous layer was
extracted with EtOAc, and the combined organic layers were washed
with brine, dried (Na.sub.2SO.sub.4), and evaporated to a yellow
foam, which was subjected to chromatographic purification using 10%
EtOAc/hexanes as an eluent to yield the nitroolefin as a yellow
foam (0.38 g) in 68% yield.
[0167] Step 6: Zn(Hg) was prepared by adding HgCl.sub.2 (3.4 g, 7.2
mmol) to a mixture of Zn-dust (34.7 g, 530.4 mmol) and 5% HCl (38
mL) in a 100 mL beaker. The mixture was stirred vigorously for 10
min. The aqueous phase was decanted, 38 mL of 5% HCl was added to
the Zn(Hg) and the mixture was stirred for 10 min. The aqueous
phase was decanted. The Zn(Hg) solid was added to the vinyl nitro
compound from Step 5 (15 g, 26.57 mmol) in THF (660 mL) and conc.
HCl (64.5 mL). This mixture was stirred at room temperature for 1
h, then heated to reflux for 15 min. The reaction mixture was
cooled to room temperature and filtered through celite. Aq.
NH.sub.4OH solution (200 mL) was added to the filtrate, the
resulting mixture was stirred for 15 min and the mixture was
concentrated to remove THF. The aqueous layer was extracted with
CH.sub.2Cl.sub.2 and the combined organic layer was washed with
brine, dried (Na.sub.2SO.sub.4) and concentrated to a brown foam,
which was purified by column chromatography by eluting the column
with CHCl.sub.3 in the beginning to remove non-polar impurities
then with 2% MeOH/CHCl.sub.3 to isolate the desired amine in 46%
yield (6.1 g).
[0168] Step 7. As outlined in step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Step 6, 128 mg, 0.24 mmol) was reacted with
2,6-dibromo-phenyl-methanesulfonyl chloride (Step 3, Example 3) to
afford 203 mg of the sulfonamide, a tan solid in 100% yield.
[0169] Step 8. Using the procedure in step 10 Example 1, the
sulfonamide ester (175 mg, 0.206 mmol) was hydrolyzed to afford the
133 mg (77%) of the title product, a yellow solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.91-2.02 (m, 2H), 2.75 (t, J=8.1 Hz, 4H),
2.86-2.94 (m, 2H), 2.94-3.03 (m, 2H), 4.46-4.54 (m, 1H), 4.70 (s,
2H), 6.49 (d, J=9.1 Hz, 1H), 6.79 (dd, J=9.0, 1.9 Hz, 1H), 6.87 (s,
1H), 6.96 (t, J=8.1 Hz, 1H), 7.04-7.11 (m, J=6.2, 2.4 Hz, 4H),
7.25-7.34 (m, 8H), 7.40 (d, J=1.8 Hz, 1H), 7.48 (d, J=7.8 Hz, 2H),
8.00 (d, J=7.8 Hz, 2H).
Example 8
4-{3-[5-chloro-2-(2-{[(2,6-dichlorobenzyl)sulfonyl]amino}ethyl)-1-(dipheny-
lmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0170] Step 1: Using the conditions in Example 3, Step
1,2,6-dichlorobenzyl bromide (3.32 g, 13.84 mmol) was reacted with
potassium thioacetate to afford 2.92 g (90%) of the benzyl
thioacetate.
[0171] Step 2: Using the procedure in Example 3, Step 2, the benzyl
thioacetate (2.90 g, 12.33 mmol) was oxidized to afford 1.7 g (53%)
of the sulfonyl chloride, a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.43 (s, 2H), 7.32-7.39 (m, 1H), 7.43-7.50 (m,
2H).
[0172] Step 3: As outlined in Step 9, Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 149 mg, 0.28 mmol) was reacted with
2,6-dichloro-phenyl-methanesulfonyl chloride to afford 170 mg of
the sulfonamide, a yellow solid in 80% yield.
[0173] Step 4. Using the procedure in Step 10 Example 1, the
sulfonamide ester (145 mg, 0.19 mmol) was hydrolyzed to afford the
140 mg (99%) of the tide product, a tan solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.89-2.01 (m, 2H), 2.71-2.80 (m, 4H), 2.84-2.92
(m, 2H), 2.95-3.03 (m, 2H), 4.31 (t, J=6.2 Hz, 1H), 4.60 (s, 2H),
6.49 (d, J=9.1 Hz, 1H), 6.80 (dd, J=8.8, 2.0 Hz, 1H), 6.87 (s, 1H),
7.01-7.10 (m, 4H), 7.12-7.19 (m, 1H), 7.25-7.34 (m, 10H), 7.41 (d,
J=2.0 Hz, 1H), 8.00 (d, J=8.1 Hz, 2H);
Example 9
4-(3-{1-benzhydryl-5-chloro-2-[2-(2-methyl-6-nitro-phenylmethanesulfonylam-
ino)-ethyl-1H-indol-3-yl}-propyl)-benzoic acid
[0174] Step 1: As outlined in Step 9, Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 255 mg, 0.47 mmol) was reacted with
2-methyl-6-nitro-phenyl-methanesulfonyl chloride (Example 4, Step
4) to afford 180 mg of the sulfonamide, a yellow solid in 51%
yield.
[0175] Step 2: Using the procedure in Step 10 Example 1, the
sulfonamide ester (60 mg, 0.080 mmol) was hydrolyzed to afford the
48 mg (81%) of the title product, a white solid. 1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.89-2.01 (m, 2H), 2.49 (s, 3H), 2.75 (q, J=7.2
Hz, 4H), 2.82-2.89 (m, 2H), 2.90-2.98 (m, 2H), 4.10-4.18 (m, 2H),
4.76 (s, 2H) broad, 6.48 (d, J=8.8 Hz, 1H), 6.79 (dd, J=8.8, 2.3
Hz, 1H), 6.86 (s, 1H), 7.02-7.11 (m, J=6.6, 2.5 Hz, 4H), 7.27-7.35
(m, 8H), 7.38-7.47 (m, 2H), 7.67 (d, J=7.8 Hz, 1H), 8.00 (d, J=8.3
Hz, 2H)
Example 10
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]sulfon-
yl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0176] Step 1: To a suspension of
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoic
acid (prepared as described in U.S. Pat. No. 6,797,708 B2,
incorporated herein by reference in its entirety) (10.0 g, 19 mmol)
in CH.sub.3CN (100 mL) and MeOH (25 mL) was added
(trimethylsilyl)diazomethane (2.0 M soln. in hexanes, 9.6 mL, 19
mmol). After 16 h the mixture was filtered and concentrated to
afford the methyl ester (8.8 g, ca. 86%), an orange foam, which was
used without purification.
[0177] Step 2: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 1, Step 1, 9.1 g, 17 mmol) was reacted with
(2-trifluoromethylphenyl)methanesulfonyl chloride (Example 5, Step
3, 4.8 g, 17 mmol) to afford 6.1 g of the sulfonamide, a white foam
in 47% yield. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.88-2.00
(m, 2H), 2.64-2.77 (m, 6H), 2.83-2.95 (m, 2H), 3.90 (s, 3H), 4.05
(t, J=5.9 Hz, 1H), 4.33 (s, 2H), 6.49 (d, J=8.8 Hz, 1H), 6.70-6.88
(m, 2H), 7.04 (dd, J=6.4, 2.7 Hz, 4H), 7.24 (s, 1H), 7.28-7.35 (m,
7H), 7.36-7.49 (m, 3H), 7.55-7.71 (m, 2H), 7.95 (d, J=8.1 Hz, 2H).
In addition, the N-methyl sulfonamide byproduct (0.70 g, 5%) was
obtained as a pale yellow foam. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.82-2.02 (m, 2H), 2.56 (s, 3H), 2.63-2.78 (m, 4H),
2.79-2.89 (m, 2H), 2.89-3.01 (m, 2H), 3.90 (s, 3H), 4.29 (s, 2H),
6.42 (d, J=8.8 Hz, 1H), 6.77 (dd, J=8.8, 2.0 Hz, 1H), 6.84 (s, 1H),
6.98-7.11 (m, 4H), 7.21-7.28 (m, 2H), 7.28-7.35 (m, 6H), 7.37-7.51
(m, 3H), 7.63 (d, J=7.1 Hz, 1H), 7.70 (d, J=8.6 Hz, 1H), 7.95 (d,
J=8.3 Hz, 2H).
[0178] Step 3: Using the procedure in Step 10 Example 1, the methyl
ester (2.6 g, 3.4 mmol) was hydrolyzed to afford 2.25 g (88%) of
the title product, a yellow solid.
[0179] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 1.81-1.97 (m,
2H), 2.66-2.79 (m, 4H), 2.95 (s, 4H), 4.41 (s, 2H), 6.45 (d, J=8.8
Hz, 1H), 6.78 (dd, J=8.8, 2.0 Hz, 1H), 7.01-7.14 (m, 5H), 7.24-7.42
(m, 8H), 7.46 (d, J=2.0 Hz, 1H), 7.50-7.66 (m, 4H), 7.73 (d, J=7.8
Hz, 1H), 7.85 (d, J=8.3 Hz, 2H), 12.77 (s, 1H); HRMS: calcd for
C.sub.41H.sub.36ClF.sub.3N.sub.2O.sub.4S+H+, 745.21092; found
(ESI-FTMS, [M+H].sup.1+), 745.2132; Anal. Calcd for
C.sub.41H.sub.36ClF.sub.3N.sub.2O.sub.4S: C, 66.08; H, 4.87; N,
3.76. Found: C, 66.07; H, 4.57; N, 3.67.
Example 11
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-(methyl{[2-(trifluoromethyl)benzyl]-
sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0180] Step 1: Using the procedure in Step 10 Example 1, the N-Me
sulfonamide ester isolated from Example 10, Step 2 as a side
product (0.66 g, 0.85 mmol) was hydrolyzed to afford 0.30 g (46%)
of the title product, a pale yellow powder. .sup.1H NMR (400 MHz,
DMSO-d) .delta. 1.76-1.93 (m, 2H), 2.63-2.81 (m, 9H), 3.31 (s, 2H),
4.46 (s, 2H), 6.46 (d, J=8.8 Hz, 1H), 6.78 (dd, J=8.8, 2.3 Hz, 1H),
6.98-7.13 (m, 5H), 7.23-7.43 (m, 8H), 7.46 (d, J=2.0 Hz, 1H),
7.51-7.66 (m, 3H), 7.72 (d, J=7.8 Hz, 1H), 7.86 (d, J=8.3 Hz, 2H),
12.75 (br s, 1H); HRMS: calcd for
C.sub.42H.sub.38ClF.sub.3N.sub.2O.sub.4S+H+, 759.22657; found
(ESI-FTMS, [M+H].sup.1+), 759.2269; HPLC purity
H.sub.2O/CH.sub.3CN: 96.2%, H.sub.2O/MeOH: 95.7%.
Example 12
4-{3-[2-[2-({[2,6-bis(trifluoromethyl)benzyl]sulfonyl}amino)ethyl]-5-chlor-
o-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0181] Step 1: 2,6-Bis(trifluoromethyl)benzoyl fluoride. Using the
procedure described by W. Dmowski and K. Piasecka-Maciejewska,
Tetrahedron Lett. 1998, 54, 6781-6792, incorporated herein by
reference in its entirety, 7.0 g of 2,6-bis(trifluoromethyl)benzoic
acid was converted to the acid fluoride (7.0 g, 100%), an orange
solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.17 (t, J=8.0
Hz, 1H), 8.40 (d, J=8.0 Hz, 2H).
[0182] Step 2: 2,6-Bis(trifluoromethylphenyl)benzyl alcohol. Using
the procedure described by W. Dmowski and K. Piasecka-Maciejewska,
Tetrahedron Lett. 1998, 54, 6781-6792, incorporated herein by
reference in its entirety, 7.0 g of 2,6-bis(trifluoromethyl)benzoyl
fluoride was converted to the alcohol (6.6 g, 100%), a pale yellow
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.95 (s, 2H), 7.59 (t, J=8.0
Hz, 1H), 7.94 (d, J=7.8 Hz, 2H).
[0183] Step 3: 2,6-Bis(trifluoromethylphenyl)benzyl bromide. To a
solution of 2,6-bis(trifluoromethylphenyl)benzyl alcohol (6.6 g, 28
mmol) and 1,3-bis(diphenylphosphino)propane (6.9 g, 17 mmol) in
CH.sub.2Cl.sub.2 (50 mL) at 0.degree. C. was slowly added carbon
tetrabromide (11 g, 33 mmol). The mixture was stirred overnight at
room temperature then added via pipette to 200 mL Et.sub.2O. The
mixture was filtered through Celite and concentrated. The yellow
oil was suspended in 2% EtOAc-hex and filtered through a pad of
SiO.sub.2 to afford the bromide (7.2 g, 84%), a colorless oil.
.sup.1HNMR (400 MHz, CDCl.sub.3) .delta. 4.78 (s, 2H), 7.59 (t,
J=7.9 Hz, 1H), 7.92 (d, J=7.9 Hz, 2H).
[0184] Step 4: 2,6-Bis(trifluoromethylphenyl)methanesulfonic acid
sodium salt. A mixture of bis(trifluoromethylphenyl)benzyl bromide
(7.2 g, 23 mmol), sodium sulfite (3.1 g, 25 mmol),
tetrabutylammonium iodide (0.043 g, 0.1 mmol) and H.sub.2O (20 mL)
was heated to reflux for 2 d. The mixture was cooled to room
temperature and the aqueous phase was decanted from the oily
residue and concentrated on the rotovap to dryness to afford
2,6-bis(trifluoromethylphenyl)methanesulfonic acid sodium salt
hydrobromide (3.2 g, 32%), a white solid, which was used without
purification.
[0185] Step 5: 2,6-Bis(trifluoromethylphenyl)methanesulfonic acid.
2,6-Bis(trifluoromethylphenyl)methanesulfonic acid sodium salt
(0.19 g, 0.44 mmol) was suspended in MeOH (5 mL) and cooled at
-20.degree. C. while HCl was bubbled through the mixture for 5 min.
The resulting white suspension was stirred at room temperature for
1.5 h, filtered through Celite, and concentrated to afford
2,6-bis(trifluoromethylphenyl)methanesulfonic acid (0.14 g, 100%),
an orange solid which was used without further purification.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) 4.25 (s, 2H), 7.64 (t, J=8.5
Hz, 1H), 7.96 (d, J=7.8 Hz, 2H).
[0186] Step 6: 2,6-Bis(trifluoromethylphenyl)methanesulfonyl
chloride. To a suspension of
2,6-bis(trifluoromethylphenyl)methanesulfonic acid (0.14 g, 0.44
mmol) in THF (10 mL) and DMF (0.05 mL) at -20.degree. C. was added
oxalyl chloride (0.24 mL, 2.7 mmol) slowly dropwise. The bath
temperature was maintained below 0.degree. C. for 4 h, then the
reaction mixture was filtered through Celite and washed with THF
(10 mL) and concentrated to .about.5 mL total volume. The mixture
was cooled to -40.degree. C. and H.sub.2O (0.3 mL) was added
slowly. The mixture was extracted with EtOAc (2.times.10 mL),
washed with sat. NaHCO.sub.3 (20 mL), H.sub.2O (20 mL), and brine
(20 mL), dried (Na.sub.2SO.sub.4) and concentrated to afford 99 mg
of crude product which was purified by low-temperature
crystallization from hexanes to afford
2,6-bis(trifluoromethylphenyl)methanesulfonyl chloride (33 mg,
23%), a white powder. Concentration of the mother liquors afforded
additional product (57 mg, 40%). .sup.1H NMR (400 MHz, CDCl.sub.3)
5.56 (s, 2H), 7.70 (t, J=8.0 Hz, 1H), 7.97 (d, J=8.0 Hz, 2H).
[0187] Step 7: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 5, Step 6, 148 mg, 0.27 mmol) was reacted with
2,6-bis(trifluoromethylphenyl)methanesulfonyl chloride (90 mg, 0.27
mmol) to afford 137 mg of the sulfonamide, a pale yellow foam in
60% yield. .sup.1H NMR (400 MHz, CDCl.sub.3) 1.83-2.03 (m, 2H),
2.68-2.78 (m, 4H), 2.79-2.91 (m, 2H), 2.92-3.03 (m, 2H), 3.89 (s,
3H), 4.21 (t, J=6.4 Hz, 1H), 4.66 (s, 2H), 6.51 (d, J=9.1 Hz, 1H),
6.81 (dd, J=8.7, 2.1 Hz, 1H), 6.87 (s, 1H), 7.00-7.11 (m, 4H),
7.21-7.28 (m, 4H), 7.28-7.35 (m, 4H), 7.41 (d, J=1.5 Hz, 1H), 7.59
(t, J=7.7 Hz, 1H), 7.89 (d, J=7.8 Hz, 2H), 7.95 (d, J=8.1 Hz,
2H).
[0188] Step 8: Using the procedure in Step 10 Example 1, the
sulfonamide ester (119 mg, 0.14 mmol) was hydrolyzed to afford 97
mg (83%) of the title product, a yellow solid. .sup.1H NMR (400
MHz, DMSO-d.sub.6) 1.75-1.95 (m, 2H), 2.73 (q, J=7.5 Hz, 4H), 2.97
(s, 4H), 4.67 (s, 2H), 6.45 (d, J=8.8 Hz, 1H), 6.79 (dd, J=8.8, 2.0
Hz, 1H), 7.04 (s, 1H), 7.06-7.16 (m, 4H), 7.27-7.43 (m, 8H), 7.47
(d, J=2.0 Hz, 1H), 7.75 (t, J=5.2 Hz, 1H), 7.77-7.91 (m, 3H), 8.10
(d, J=8.1 Hz, 2H), 12.78 (s, 1H); HRMS: calcd for
C.sub.42H.sub.35ClF.sub.6N.sub.2O.sub.4S+H+, 813.19830; found
(ESI-FTMS, [M+H].sup.1+), 813.1965. HPLC purity
H.sub.2O/CH.sub.3CN: 95.5%, H.sub.2O/MeOH: 96.8%.
Example 13
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(methoxycarbonyl)benzyl]sulfon-
yl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0189] Step 1: To a mixture of methyl 2-methylbenzoate (5.0 g,
0.033 mmol) and N-bromosuccinimide (5.9 g, 0.033 mmol) in CCl.sub.4
(50 mL) was added benzoyl peroxide (0.04 g, 0.00016 mmol). The
mixture was heated to reflux for 1.5 h, cooled to room temperature,
filtered through Celite, and concentrated to afford methyl
2-(bromomethyl)benzoate (7.2 g, ca. 94% mass recovery), which was
contaminated with ca. 14% unreacted starting material and was used
without purification.
[0190] Step 2: A mixture of the crude bromide from Step 1 (7.2 g,
0.031 mmol) and thiourea (2.6 g, 35 mmol) in MeOH (40 mL) was
heated to reflux for 4 h, cooled to room temperature, and
concentrated to afford methyl
2-({[amino(imino)methyl]thio}methyl)benzoate hydrobromide (10 g,
ca. 100%), which was used without purification.
[0191] Step 3: The isothiouronium salt from Step 2 (10 g, 0.031
mmol) was suspended in H.sub.2O (100 mL) and cooled to 0.degree. C.
Chlorine gas was bubbled into the mixture for 30 min. The ice bath
was removed and the reaction mixture was poured into a separatory
funnel and diluted with EtOAc (250 mL). The organic phase was
separated and washed with sat. NaHCO.sub.3 (100 mL), H.sub.2O (100
mL), and brine (100 mL), dried (MgSO.sub.4) and concentrated to
afford an orange solid (6.48 g). The crude product was
recrystallized from 20% EtOAc-hexanes at -78.degree. C. to afford
3.63 g (47%) of methyl 2-[(chlorosulfonyl)methyl]benzoate, as pale
yellow crystals.
[0192] Step 4: To a suspension of
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoic
acid (0.40 g, 0.76 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added
bis(trimethylsilyl)trifluoroacetamide (0.30 mL, 0.29 g, 1.1 mmol).
The mixture was heated to reflux for 30 min, then cooled to
35.degree. C. Pyridine (0.16 mL, 0.15 g, 2.0 mmol) was added,
followed by a solution of methyl 2-[(chlorosulfonyl)methyl]benzoate
from Step 3 (0.29 g, 1.1 mmol) in CH.sub.2Cl.sub.2 (2 mL). After 5
h, the mixture was cooled to room temperature. A solution of conc.
HCl (0.17 mL) in H.sub.2O (5 mL) was added and the mixture was
stirred for 45 min. The aqueous phase was separated and extracted
with CH.sub.2Cl.sub.2 (50 mL). The combined organic extracts were
washed with H.sub.2O (25 mL) and brine (25 mL), dried (MgSO.sub.4)
and concentrated to afford a gold foam (0.40 g). Purification by
prep HPLC afforded the title compound (70 mg, 12%), a pale yellow
foam. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 1.87-2.10 (m, 2H), 2.85
(t, J=6.6 Hz, 4H), 3.03 (s, 2H), 3.48 (s, 2H), 3.86 (s, 3H), 4.94
(s, 2H), 6.57 (d, J=8.8 Hz, 1H), 6.92 (dd, J=8.8, 2.3 Hz, 1H), 7.14
(s, 1H), 7.17-7.29 (m, 4H), 7.43-7.57 (m, 10H), 7.57-7.69 (m, 3H),
7.90-7.97 (m, 1H), 8.00 (d, J=8.3 Hz, 2H), 12.93 (s, 1H), HRMS:
calcd for [C.sub.42H.sub.39ClN.sub.2O.sub.6S.sub.1-H].sup.1-,
733.2144; found (ESI-FTMS, [M-H].sup.1-), 733.2141; HPLC purity
H.sub.2O/CH.sub.3CN: 95.3%, H.sub.2O/MeOH: 100%.
Example 14
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)benz-
yl]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0193] Step 1: Using the procedure described in Example 5, Step
1,2-fluoro-6-(trifluoromethylphenyl)benzyl bromide (15 g, 61 mmol)
afforded 2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid
sodium salt (15 g, 89%), a white solid. .sup.1HNMR (400 MHz,
DMSO-d.sub.6) 4.02 (s, 2H), 7.26-7.66 (m, 3H).
[0194] Step 2: Using the procedure described in Example 5, Step
2,2-fluoro-6-(trifluoromethylphenyl)methanesulfonic sodium salt (15
g, 53 mmol) afforded
2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid (15 g), a
pale orange oil which was used without further purification.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 4.12 (s, 2H), 7.39-7.73
(m, 3H).
[0195] Step 3: Using the procedure described in Example 5, Step
3,2-fluoro-6-(trifluoromethylphenyl)methanesulfonic acid (15 g, 53
mmol) afforded 11 g of crude product which was purified by
low-temperature crystallization from hexanes to afford
2-fluoro-6-(trifluoromethylphenyl)methanesulfonyl chloride (9.0 g,
62%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.31 (s, 2H),
7.38-7.51 (m, 1H), 7.58-7.68 (m, 2H).
[0196] Step 4: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 0.12 g, 0.22 mmol) was reacted with
2-fluoro-6-(trifluoromethylphenyl)methanesulfonyl chloride, 0.074
g, 0.27 mmol) to afford 0.127 g of the sulfonamide, a pale yellow
foam, in 73% yield. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
1.79-2.02 (m, 2H), 2.74 (t, J=8.0 Hz, 4H), 2.82-2.92 (m, 2H),
2.92-3.02 (m, 2H), 3.89 (s, 3H), 4.15 (t, J=5.8 Hz, 1H), 4.42 (d,
2H), 6.50 (d, J=8.6 Hz, 1H), 6.80 (dd, J=8.8, 2.0 Hz, 1H), 6.87 (s,
1H), 7.07 (dd, J=6.4, 2.7 Hz, 4H), 7.19-7.28 (m, 5H), 7.29-7.35 (m,
5H), 7.39-7.56 (m, 2H), 7.95 (d, J=8.3 Hz, 2H).
[0197] Step 4: Using the procedure in Step 10 Example 1, the
sulfonamide ester (115 mg, 0.15 mmol) was hydrolyzed to afford 101
mg (89%) of the title product, a pale yellow powder. .sup.1H NMR
(400 MHz, DMSO-d.sub.6) 0.80-1.95 (m, 2H), 2.63-2.78 (m, 4H),
2.88-3.14 (m, 4H), 4.45 (s, 2H), 6.43 (d, J=8.8 Hz, 1H), 6.76 (dd,
J=8.8, 2.3 Hz, 1H), 6.96-7.15 (m, 5H), 7.20-7.41 (m, 8H), 7.45 (d,
J=2.3 Hz, 1H), 7.50-7.59 (m, 1H), 7.59-7.66 (m, 2H), 7.71 (t, J=5.6
Hz, 1H), 7.83 (d, J=8.3 Hz, 2H), 12.73 (s, 1H); HRMS: calcd for
C.sub.41H.sub.35ClF.sub.4N.sub.2O.sub.4S+H+, 763.20149; found
(ESI-FTMS, [M+H].sup.1+), 763.1998; HPLC purity
H.sub.2O/CH.sub.3CN: 95.4%, H.sub.2O/MeOH: 96.4%.
Example 15
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({2-[2-(trifluoromethyl)phenyl]eth-
yl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic acid
[0198] Step 1: 2-(Trifluoromethyl)phenethyl alcohol (5.0 g, 26
mmol) was treated with CBr.sub.4 as in Example 12, Step 3 to afford
the bromide (6.6 g, 100%) which was used without purification.
[0199] Step 2: The bromide from Step 1 (1.5 g, 5.9 mmol) was
treated with thiourea as in Example 13, Step 2 to afford the
isothiouronium salt (2.2 g) a wet white solid, which was used
without purification.
[0200] Step 3: The isothiouronium salt from Step 2 (2.2 g,
.about.5.9 mmol) was suspended in H.sub.2O and treated with
Cl.sub.2 gas as in Example 13, Step 3 and an orange oil (1.15 g)
was obtained. To the crude product was added hexanes (75 mL) and
the mixture was heated at 60.degree. C. for 4 h. The hexanes
soluble fraction was decanted and cooled to -78.degree. C. to
afford a white solid (0.13 g, ca. 7% yield, 2 steps) which was used
without purification.
[0201] Step 4: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 98 mg, 0.18 mmol) was reacted with
(2-trifluoromethylphenyl)ethanesulfonyl chloride from Step 3, (75
mg, 0.28 mmol) to afford 70 mg of the sulfonamide, a white foam in
50% yield. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.88-2.03 (m,
2H), 2.70-2.83 (m, 4H), 2.88-3.07 (m, 6H), 3.07-3.23 (m, 2H), 3.90
(s, 3H), 4.07 (t, J=6.2 Hz, 1H), 6.51 (d, J=8.8 Hz, 1H), 6.72-6.86
(m, 1H), 6.90 (s, 1H), 7.07 (d, J=6.8 Hz, 4H), 7.17-7.32 (m, 9H),
7.35 (t, J=7.6 Hz, 1H), 7.41 (d, J=2.0 Hz, 1H), 7.48 (t, J=6.9 Hz,
1H), 7.63 (d, J=7.8 Hz, 1H), 7.95 (d, J=8.3 Hz, 2H).
[0202] Step 5: Using the procedure in Step 10 Example 1, the
sulfonamide ester (70 mg, 0.09 mmol) was hydrolyzed to afford 54 mg
(78%) of the title product, a white powder. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 1.82-2.22 (m, 2H), 2.68-2.90 (m, 4H), 2.99-3.24 (m,
8H), 6.50 (d, J=8.8 Hz, 1H), 6.83 (dd, J=8.8, 2.3 Hz, 1H), 7.15
(appar d, J=6.8 Hz, 5H), 7.32-7.44 (m, 8H), 7.45-7.55 (m, 3H), 7.59
(t, J=5.7 Hz, 1H), 7.65 (t, J=7.3 Hz, 1H), 7.75 (d, J=7.8 Hz, 1H),
7.83-7.98 (m, J=8.3 Hz, 2H), 12.83 (s, 1H); HRMS: calcd for
C.sub.42H.sub.38ClF.sub.3N.sub.2O.sub.4S+H+, 759.22657; found
(ESI-FTMS, [M+H].sup.1+), 759.2277; HPLC purity
H.sub.2O/CH.sub.3CN: 96.0%, H.sub.2O/MeOH: 98.0%.
Example 16
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}et-
hyl)-1H-indol-3-yl]propyl}benzoic acid
[0203] Step 1: To .alpha.-bromo-o-tolunitrile (10 g, 51 mmol) in
CH.sub.2Cl.sub.2 at 0.degree. C. was added DIBAL-H (1M in hexane,
55 mL, 55 mmol) and the reaction mixture was stirred at the same
temperature for 3.5 h, then poured into a solution of cold 5% HBr
at 0.degree. C. The mixture was stirred for 15 min, then the layers
were separated and the aqueous layer was extracted with
CH.sub.2Cl.sub.2 and the combined organic layers were washed with
NaHCO.sub.3 and water, dried over MgSO.sub.4 and evaporated to
yield a dark liquid (9.4 g). The material was used directly in the
next step without further purification.
[0204] Step 2: (2-Formyl-phenyl)-methanesulfonic acid sodium salt:
Using the procedure described in Example 5, Step
1,2-bromomethyl-benzaldehyde (1.58 g, 7.94 mol) afforded
(2-formyl-phenyl)-methanesulfonic acid sodium salt (1.40 g, 80%),
an off white solid.
[0205] Step 3: (2-Formyl-phenyl)-methanesulfonic acid: Using the
procedure described in Example 5, Step 3,
(2-formyl-phenyl)-methanesulfonic acid sodium salt (1.40 g, 6.30
mmol) afforded (2-formyl-phenyl)-methanesulfonic acid (418 mg,
33%), a pale yellow solid.
[0206] Step 4: (2-Formyl-phenyl)-methanesulfonyl chloride: Using
the procedure described in Example 5, Step 4,
(2-formyl-phenyl)methanesulfonic acid (418 mg, 2.09 mmol) afforded
(2-formyl-phenyl)-methanesulfonyl chloride (367 mg, 80%).
.sup.1HNMR (400 MHz, CDCl.sub.3) .delta. 10.15. (s, 1H), 7.92 (dd,
1H), 7.74-7.61 (m, 3H), 5.67 (s, 2H).
[0207] Step 5: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 63 mg, 0.12 mmol) was reacted with
(2-formyl-phenyl)-methanesulfonyl chloride (36 mg, 0.16 mmol) to
afford 34 mg (40%) of the sulfonamide, a yellow solid.
[0208] Step 6: Using the procedure in Step 10 Example 1, the
sulfonamide ester (28 mg, 0.039 mmol) was hydrolyzed to afford the
17 mg (62%) of the title product, a white solid.
Example 17
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(morpholin-4-ylmethyl)benzyl]s-
ulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0209] Step 1: To methyl
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoate (Example 16, Step 5, 58 mg,
0.081 mmol) in DCE (2 mL) at 0.degree. C. were added morpholine
(0.0092 mL, 0.105 mmol) and NaBH(OAc).sub.3 (27 mg, 0.13 mmol) and
the reaction mixture was allowed to warm to rt overnight. The
reaction was quenched with sat. NaHCO.sub.3, extracted with EtOAc,
and dried over MgSO.sub.4. Purification by silica gel
chromatography (35% to 50% EtOAc/hexanes) gave the desired product
as a white solid (41 mg, 64%).
[0210] Step 2. Using the procedure in Step 10 Example 1, the
sulfonamide ester (18 mg, 0.039 mmol) was hydrolyzed to afford the
15 mg (83%) of the title product, a white solid.
Example 18
4-{3-[5-chloro-2-{2-[({2-[(diethylamino)methyl]benzyl}sulfonyl)amino]ethyl-
}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0211] Step 1: As outlined in Example 17, Step 1 methyl
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoate (Example 16, Step 5, 58 mg,
0.081 mmol) was reacted with HNEt.sub.2 (0.022 mL, 0.21 mmol) and
NaBH(OAc).sub.3 (56 mg, 0.26 mmol) in DCE (2 mL) to afford methyl
4-{3-[5-chloro-2-{2-[({2-[(diethylamino)methyl]benzyl}sulfonyl)amino]ethy-
l}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoate (26 mg, 41%)
and the side product methyl
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(hydroxymethyl)benzyl]sulfony-
lamino}ethyl]-1H-indol-3-yl}propyl)benzoate (8.6 mg, 15%), both as
white solids.
[0212] Step 2. Using the procedure in Step 10 Example 1, methyl
4-{3-[5-chloro-2-{2-[({2-[(diethylamino)methyl]benzyl}sulfonyl)amino]ethy-
l}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoate (20 mg, 0.026
mmol) was hydrolyzed to afford the 13 mg (66%) of the title
product, a white solid.
Example 19
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(hydroxymethyl)benzyl]sulfonyl-
}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0213] Step 1. Using the procedure in Step 10 Example 1, the side
product from Example 18 Step 1, methyl
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(hydroxymethyl)benzyl]sulfony-
l}amino)ethyl]-1H-indol-3-yl}propyl)benzoate (8.4 mg, 0.0012 mmol)
was hydrolyzed to afford the 5.0 mg (61%) of the title product, a
white solid.
Example 20
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(piperazin-1-ylmethyl)benzyl]s-
ulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid and Example
21 4
{3-[2-{2-[({2-[(4-acetylpiperazin-1-yl)methyl]benzyl}sulfonyl)amino]ethyl-
}-5-chloro-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0214] Step 1: As outlined in Example 17, Step 1, methyl
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoate (Example 16, Step 5, 45 mg,
0.063 mmol) was reacted with 1-acetylpiperazine (28 mg, 0.22 mmol)
and NaBH(OAc).sub.3 (26 mg, 0.12 mmol) in DCE (3 mL) to afford the
sulfonamide (39 mg, 75%) as a white foam.
[0215] Step 2. Using the procedure in Step 10 Example 1, the
sulfonamide (37 mg, 0.045 mmol) was hydrolyzed to afford, after
preparative HPLC separation, methyl
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(piperazin-1-ylmethyl)benzyl]-
sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoate (7.4 mg, 21%)
and methyl
4-{3-[2-{2-[({2-[(4-acetylpiperazin-1-yl)methyl]benzyl}sulfonyl)am-
ino]ethyl}-5-chloro-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoate
(7.7 mg, 21%), both as solids.
Example 22
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({2-[(4-methylpiperazin-1-yl)methy-
l]benzyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic
acid
[0216] Step 1: As outlined in Example 17, Step 1, methyl
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-formylbenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoate (Example 16, Step 5, 44 mg,
0.061 mmol) was reacted with 1-methylpiperazine (0.026 mL, 0.23
mmol) and NaBH(OAc).sub.3 (34 mg, 0.16 mmol) in DCE (3 mL) to
afford the sulfonamide (41 mg, 84%) as white solid.
[0217] Step 2: Using the procedure in Step 10 Example 1, the
sulfonamide (39 mg, 0.026 mmol) was hydrolyzed to afford the 27 mg
(69%) of the title product, a white solid.
Example 23
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({1-[2-(trifluoromethyl)phenyl]eth-
yl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic acid
[0218] Step 1: To .alpha.-methyl-2-trifluoromethyl benzyl bromide
(10.0 g, 39.5 mmol) in DMF (50 mL) added potassium thioacetate (8.1
g, 71.1 mmol) according to the procedure outlined in Example 3,
Step 2. This afforded the thioacetate as an orange oil (10.49 g,
91%).
[0219] Step 2: The thioacetate from Step 1 (10.49 g, 36.1 mmol) and
sodium acetate (21.5 g, 155.4 mmol) was dissolved in a mixture of
acetic acid (137 mL) and water (31 mL) and chlorine gas was bubbled
in according to the procedure in Example 3, Step 3. This yielded
upon concentration and low-temperature recrystallization from
hexanes an off white solid that later melted into a pale orange oil
(4.9 g, 47%). 1H NMR (400 MHz, CDCl.sub.3) .delta. 2.01 (d, J=6.8
Hz, 3H) 5.32 (q, J=7.1 Hz, 1H) 7.56 (t, J=6.7 Hz, 1H) 7.67 (t,
J=7.8 Hz, 1H) 7.77 (d, J=7.8 Hz, 1H) 7.91 (d. J=8.1 Hz, 1H)
[0220] Step 3: Using the procedure in Example 1, Step 9,
4-{3-[2-(2-amino-ethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]-propyl}-benz-
oic acid methyl ester (Example 7, Step 6, 0.123 g, 0.23 mmol) was
reacted with 1-(2-trifluoromethyl-phenyl)-ethanesulfonyl chloride
(0.79 g, 0.29 mmol) to afford 0.042 g of a racemic mixture of
sulfonamides in 24% yield.
[0221] Step 4: The sulfonamide ester (0.042 g, 0.054 mmol) was
hydrolyzed according to Example 1, Step 10, to afford 0.035 g (85%)
of the title product, a pale orange solid.
[0222] 1H NMR (400 MHz, CDCl.sub.3) .delta. 1.68 (d, J=7.1 Hz, 3H)
1.93 (t, J=5.4 Hz, 2H) 2.02-2.08 (m, 1H) 2.63-2.77 (m, 6H) 2.86 (t,
J=7.6 Hz, 2H) 6.47 (d, J=8.8 Hz, 1H) 7.01-7.07 (m, 4H) 7.24-7.40
(m, 12H) 7.44 (t, J=7.5 Hz, 1H) 7.60 (d, J=7.6 Hz, 1H) 7.85 (d,
J=8.1 Hz, 1H) 8.00 (d, J=8.1 Hz, 2H)
Example 24
4-{3-[2-(2-{[(2-bromobenzyl)sulfonyl]amino}ethyl)-5-chloro-1-(diphenylmeth-
yl)-1H-indol-3-yl]propyl}benzoic acid
[0223] Step 1: Using the procedure in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 1.51 g, 2.81 mmol) was reacted with
2-bromo-phenyl-methanesulfonyl chloride to afford 1.06 g of the
sulfonamide, a white solid, in 49% yield.
[0224] Step 2: As described in example 1, step 10, the sulfonamide
ester (90 mg, 0.117 mmol) was hydrolyzed to afford the 81 mg (91%)
of the title product, a white solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.89-2.02 (m, 2H), 2.68-2.78 (m, 4H), 2.78-2.87 (m, 2H),
2.89-2.97 (m, 2H), 4.21 (t, J=5.1 Hz, 1H), 4.37 (s, 2H), 6.48 (d,
J=8.8 Hz, 1H), 6.79 (dd, J=8.8, 2.0 Hz, 1H), 6.84 (s, 1H),
7.00-7.08 (m, 4H), 7.09-7.17 (m, 1H), 7.17-7.24 (m, 1H), 7.25-7.34
(m, 8H), 7.36-7.45 (m, 2H), 7.49 (dd, J=8.1, 1.3 Hz, 1H), 8.01 (d,
J=8.3 Hz, 2H). HRMS: calcd for
C.sub.40H.sub.36BrClN.sub.2O.sub.4S+H+, 755.13404; found (ESI-FTMS,
[M+H].sup.1+), 755.1341.
Example 25
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethoxy)benzyl]sulfo-
nyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0225] Step 1: Using the procedure in Example 1, Step 9, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 164 mg, 0.305 mmol) was reacted with
2-trifluoromethoxy-phenyl-methanesulfonyl chloride to afford 109 mg
of the sulfonamide, a white solid in 46% yield.
[0226] Step 2: As described in Example 1, Step 10, the sulfonamide
ester (83 mg, 0.107 mmol) was hydrolyzed to afford the 80 mg (98%)
of the title product, a white solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.86-2.03 (m, 2H), 2.74 (q, J=7.6 Hz, 4H), 2.76-2.86 (m,
2H), 2.90-3.00 (m, 2H), 4.12 (t, J=6.2 Hz, 1H), 4.19 (s, 1H), 6.49
(d, J=8.8 Hz, 1H), 6.80 (dd, J=8.8, 2.3 Hz, 1H), 6.85 (s, 1H),
7.00-7.11 (m, 4H), 7.16-7.23 (m, 2H), 7.24-7.28 (m, 2H), 7.28-7.37
(m, 8H), 7.39-7.44 (m, 2H), 8.00 (d, 2H). HRMS: calcd for
C.sub.41H.sub.36ClF.sub.3N.sub.2O.sub.5S+H+, 761.20583; found
(ESI-FTMS, [M+H].sup.1+), 761.2057.
Example 26
4-{3-[5-chloro-2-(2-{([(3-chloro-6-fluoro-2-methylbenzyl)sulfonyl]amino}et-
hyl)-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0227] Step 1:
2,6-Difluoro-N-(2-hydroxy-1,1-dimethyl-ethyl)-benzamide. To a
0.degree. C. solution of 2-amino-2-methylpropanol (10.1 g, 113.3
mmol) in CH.sub.2Cl.sub.2 (75 mL) under nitrogen was added dropwise
a solution of 2,6-difluorobenzoyl chloride (10.0 g, 56.6 mmol) in
CH.sub.2Cl.sub.2 (50 mL). The reaction mixture was then warmed to
room temperature and stirred overnight. The reaction mixture was
diluted with H.sub.2O, and the aqueous phase was extracted with
CH.sub.2Cl.sub.2, dried (MgSO.sub.4) and concentrated. Purification
via trituration with hexanes afforded 12.05 g (93%) of the amide, a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.41 (s,
6H), 3.66-3.75 (m, 2H), 3.77-3.87 (m, 1H), 5.94 (s, 1H), 6.90-6.99
(m, 2H), 7.31-7.41 (m, 1H).
[0228] Step 2:
2-(2,6-Difluorophenyl)-4,4-dimethyl-4,5-dihydro-oxazole. To a
0.degree. C. solution of the amide from Step 1 (11.9 g, 51.9 mmol)
in CH.sub.2Cl.sub.2 (50 mL) was added thionyl chloride (6.4 ml,
88.3 mmol). The reaction mixture was allowed to warm to room
temperature. After 4 h, the reaction mixture was concentrated and
triturated with Et.sub.2O. The residue was taken up in H.sub.2O,
basified with 6N NaOH and extracted with EtOAc. The combined
organic phase was washed with brine, dried (MgSO.sub.4) and
concentrated to afford 9.42 g (86%) of the dihydrooxazole, a white
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.42 (s, 6H), 4.13
(s, 2H), 6.90-7.02 (m, 2H), 7.32-7.44 (m, 1H).
[0229] Step 3:
2-(2-Fluoro-6-methyl-phenyl)-4,4-dimethyl-4,5-dihydro-oxazole. To a
0.degree. C. solution of the dihydrooxazole from Step 2 (9.18 g,
43.5 mmol) in THF (140 mL) under argon was added dropwise
methylmagnesium chloride (3.0 M solution in THF, 43.5 mL, 130
mmol). After 2 h the ice bath was removed and the mixture was
stirred overnight at room temperature. The reaction mixture was
quenched with a saturated aq. NH.sub.4Cl solution and extracted
with EtOAc. The combined organic phase was washed with brine, dried
(MgSO.sub.4) and concentrated to afford 8.64 g (96%) of the
dihydrooxazole, a clear colorless oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 1.42 (s, 6H), 2.40 (s, 3H), 4.11 (s, 2H), 6.92
(t, J=9.0 Hz, 1H), 6.99 (d, J=7.6 Hz, 1H), 7.21-7.30 (m, 1H).
[0230] Step 4: 2-Fluoro-6-methyl-benzoic acid. To a solution of the
dihydrooxazole from Step 3 (8.43 g, 40.7 mmol) in CH.sub.3CN (70
mL) was added methyl iodide (9.2 mL, 146 mmol) and the mixture was
heated to reflux for 6 h. The reaction mixture was then cooled to
room temperature and stirred overnight. The reaction mixture was
concentrated and the residue was triturated with Et.sub.2O. The
residue was taken up in equal parts 20% NaOH and methanol and
heated to reflux for 6 h. The reaction mixture was cooled to room
temperature and concentrated to remove organic solvents. The
aqueous phase was washed several times with EtOAc and acidified to
pH 1. The aqueous phase was extracted with EtOAc. The combined
organic phase was washed with brine, dried (MgSO.sub.4) and
concentrated to afford 3.67 g (58%) of the benzoic acid, a white
solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 2.52 (s, 3H) 6.99
(t, J=9.09 Hz, 1H) 7.05 (d, J=7.83 Hz, 1H) 7.31-7.41 (m, 1H).
[0231] Step 5: To a solution of the acid from Step 4 (3.60 g, 23.4
mmol) in thionyl chloride (40 mL) was added DMF (0.42 mL) and the
mixture was heated to reflux for 5.5 h. The mixture was cooled to
room temperature and concentrated. The residue was taken up in THF
(40 mL) and added over 20 min to a 0.degree. C. slurry of
NaBH.sub.4 (3.53 g, 93.4 mmol) in THF (40 mL). The mixture was
stirred at room temperature for 2 h and then quenched by the
addition of H.sub.2O and 4 M HCl and extracted with EtOAc. The
combined organic phase was washed with sat. NaHCO.sub.3 and brine,
dried (MgSO.sub.4) and concentrated to afford 2.67 g (.about.75%)
of the benzyl alcohol, a pale yellow solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 2.42 (s, 3H), 4.72 (s, 2H), 6.88 (t, J=9.0 Hz,
1H), 6.96 (d, J=7.6 Hz, 1H), 7.07-7.20 (m, 1H).
[0232] Step 6: To a solution of the benzyl alcohol from Step 5
(2.65 g, 18.9 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added
1,3-bis(diphenylphosphino)propane (4.7 g, 11 mmol). The mixture was
cooled to 0.degree. C. and CBr.sub.4 (7.4 g, 22 mmol) was added
slowly. The mixture was stirred overnight at room temperature. The
mixture was diluted with CH.sub.2Cl.sub.2 (50 mL) and poured into
Et.sub.2O (75 mL). The mixture was filtered and the solution phase
was concentrated. The resulting product was again dissolved in
CH.sub.2Cl.sub.2 (75 mL) and poured into Et.sub.2O (100 mL).
Filtration and concentration afforded 3.27 g (85%) of the bromide,
an orange oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 2.42 (s,
3H), 4.56 (d, J=1.5 Hz, 2H), 6.91 (t, J=9.1 Hz, 1H), 6.97 (d, J=7.6
Hz, 1H), 7.08-7.24 (m, 1H).
[0233] Step 7: Using the procedure from Example 3, Step 2, the
benzyl bromide from Step 6 (3.27 g, 16.1 mmol) was reacted with
potassium thioacetate to afford 3.17 g (98%) of the benzyl
thioacetate, a brown oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
2.35 (s, 6H), 4.22 (d, J=1.5 Hz, 2H), 6.88 (t, J=9.0 Hz, 1H), 6.95
(d, J=7.6 Hz, 1H), 7.06-7.21 (m, 1H).
[0234] Step 8: Using the procedure from Example 3, Step 3, the
benzyl thioacetate (3.17 g, 16.0 mmol) was oxidized to afford 3.30
g (80%) of (3-chloro-6-fluoro-2-methyl-phenyl)-methanesulfonyl
chloride, a tan solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
2.52 (s, 3H), 5.10 (d, J=1.3 Hz, 2H), 7.02 (t, J=9.0 Hz, 1H), 7.48
(dd, J=9.1, 5.3 Hz, 1H).
[0235] Step 9: Using the procedure in Step 9, Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 163 mg, 0.303 mmol) was reacted with
(3-chloro-6-fluoro-2-methyl-phenyl)-methanesulfonyl chloride from
Step 8 to afford 102 mg of the sulfonamide, a white solid in 44%
yield.
[0236] Step 10: As described in Example 1, Step 10, the sulfonamide
ester (74 mg, 0.097 mmol) was hydrolyzed to afford the 65 mg (90%)
of the title product, a white solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.89-2.05 (m, 2H), 2.41 (s, 3H), 2.75 (q, J=7.6 Hz, 4H),
2.83-2.93 (m, 2H), 2.92-3.02 (m, 2H), 4.21-4.31 (m, 3H), 6.49 (d,
J=8.8 Hz, 1H), 6.72-6.83 (m, 2H), 6.87 (s, 1H), 7.01-7.13 (m, 4H),
7.24-7.35 (m, 9H), 7.41 (d, J=2.0 Hz, 1H), 8.00 (d, J=8.1 Hz, 2H).
HRMS: calcd for C.sub.41H.sub.37Cl.sub.2FN.sub.2O.sub.4S+H+,
743.19079; found (ESI-FTMS, [M+H].sup.1+), 743.1907.
Example 27
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-nitro-6-(trifluoromethyl)benzy-
l]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0237] Step 1: 2-Bromomethyl-1-nitro-3-trifluoromethyl-benzene. To
a solution of 2-methyl-1-nitro-3-trifluoromethyl-benzene (5.0 g,
24.4 mmol) in CCl.sub.4 (300 mL) was added N-bromosuccinimide (4.35
g, 24.4 mmol) and benzoyl peroxide (0.11 g, 0.45 mmol). The mixture
was heated to reflux and exposed to light (300 W) for 20 h. The
mixture was cooled to room temperature, filtered and concentrated.
Purification by column chromatography (EtOAc-hexanes) afforded 3.03
g (44%) of the benzyl bromide, a yellow oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.93 (s, 2H), 7.63 (t, J=8.1 Hz, 1H), 7.94 (d,
J=7.8 Hz, 1H), 8.06 (d, J=8.1 Hz, 1H).
[0238] Step 2: Using the procedure from Example 3, Step 2, the
benzyl bromide Step 1 (3.02 g, 10.6 mmol) was reacted with
potassium thioacetate to afford 2.71 g (91%) of the benzyl
thioacetate as a brown oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 2.34 (s, 3H), 4.55 (s, 2H), 7.58 (t, J=7.7 Hz, 1H), 7.93
(d, J=7.8 Hz, 1H), 7.98 (d, J=8.1 Hz, 1H).
[0239] Step 3: (2-Nitro-6-trifluoromethylphenyl)methanesulfonyl
chloride. Using the procedure from Example 3, Step 3, the benzyl
thioacetate (2.71 g, 9.70 mmol) was oxidized to afford 2.42 g (82%)
of the title product as a brown oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 5.82 (s, 2H) broad, 7.84 (t, J=8.1 Hz, 1H),
8.11 (d, J=7.8 Hz, 1H), 8.27 (d, J=8.1 Hz, 1H).
[0240] Step 4: Using the procedure in Step 9, Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 164 mg, 0.305 mmol) was reacted with
(2-Nitro-6-trifluoromethylphenyl)methanesulfonyl chloride from Step
3 to afford 119 mg of the sulfonamide as a yellow solid in 49%
yield.
[0241] Step 5: As described in Example 1, Step 10, the sulfonamide
ester (94 mg, 0.117 mmol) was hydrolyzed to afford the 90 mg (92%)
of the title product, a white solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.90-2.04 (m, 2H), 2.76 (q, J=7.5 Hz, 4H), 2.80-2.90 (m,
2H), 2.93-3.01 (m, 2H), 4.24 (t, J=6.2 Hz, 1H), 4.87 (s, 2H) broad,
6.51 (d, J=8.8 Hz, 1H), 6.81 (dd, J=9.0, 2.1 Hz, 1H), 6.86 (s, 1H),
7.08 (dd, J=6.8, 2.5 Hz, 4H), 7.23-7.36 (m, 9H), 7.42 (d, J=2.0 Hz,
1H), 7.61 (t, J=8.1 Hz, 1H), 7.89-8.09 (m, 3H). HRMS: calcd for
C.sub.41H.sub.35ClF.sub.3N.sub.3O.sub.6S+H+, 790.19599; found
(ESI-FTMS, [M+H].sup.1+), 790.1944.
Example 28
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-fluorobenzyl)sulfonyl]amino}et-
hyl)-1H-indol-3-yl]propyl}benzoic acid
[0242] Step 1. Using the procedure in Step 9, Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6,163 mg, 0.303 mmol) was reacted with
(2-nitro-6-trifluoromethyl-phenyl)-methanesulfonyl chloride to
afford 15 mg of the sulfonamide, a white solid in 7% yield.
[0243] Step 2. As described in Example 1, Step 10, the sulfonamide
ester (14 mg, 0.020 mmol) was hydrolyzed to afford the 12 mg (88%)
of the title product, a white solid. 1H NMR (400 MHz, CDCl.sub.3)
.delta. 1.88-2.03 (m, 2H), 2.66-2.78 (m, 4H), 2.81-2.90 (m, 2H),
2.90-3.00 (m, 2H), 4.12-4.20 (m, 3H), 6.49 (d, J=8.8 Hz, 1H), 6.80
(dd, J=8.8, 2.3 Hz, 1H), 6.86 (s, 1H), 6.94-7.01 (m, 1H), 7.02-7.12
(m, 5H), 7.23-7.36 (m, 10H), 7.40 (d, J=2.0 Hz, 1H), 8.00 (d, J=8.3
Hz, 2H). HRMS: calcd for C.sub.40H.sub.36ClFN.sub.2O.sub.4S+H+,
695.21411; found (ESI-FTMS, [M+H].sup.1+), 695.2128.
Example 29
4-{3-[2-(2-{[(biphenyl-2-ylmethyl)sulfonyl]amino}ethyl)-5-chloro-1-(diphen-
ylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0244] Step 1. The bromide from Example 24, Step 1 (83 mg, 0.108
mmol) was placed in a microwave reaction vessel with phenylboronic
acid (19.8 mg, 0.162 mmol), KF (9.4 mg, 0.162 mmol), Pd(OAc).sub.2
(3.4 mg, 0.015 mmol) and PPh.sub.3 (11.8 mg, 0.045 mmol). DME (0.12
M), MeOH (0.42 M), H.sub.2O (0.42 M) was added to the vessel and
the mixture was degassed under a stream of argon, capped, and
heated in the Smith Creator microwave at 120.degree. C. for 1 h.
The reaction mixture was cooled to room temperature, filtered
through celite (washing with EtOAc), and diluted with H.sub.2O. The
aqueous layer was extracted with EtoAc. The combined organic phase
was washed with H.sub.2O and brine, dried over MgSO.sub.4 and
concentrated. Purification of the crude product by column
chromatography (EtOAc-Hex.) afforded 75 mg (91%) of the Suzuki
product, a yellow solid.
[0245] Step 2. As described in Example 1, Step 10, the ester (70
mg, 0.091 mmol) was hydrolyzed to afford 46 mg (67%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.85-1.98 (m, 2H), 2.45-2.55 (m, 2H), 2.64-2.78 (m, 4H), 2.82-2.90
(m, 2H), 3.99 (t, J=6.3 Hz, 1H), 4.18 (s, 2H), 6.47 (d, J=8.8 Hz,
1H), 6.71-6.84 (m, 2H), 6.97-7.08 (m, 4H), 7.15-7.24 (m, 3H),
7.25-7.28 (m, 4H), 7.28-7.36 (m, 9H), 7.40 (d, J=2.0 Hz, 1H), 7.47
(dd, J=7.7, 1.1 Hz, 1H), 8.01 (d, J=8.3 Hz, 2H). HRMS: calcd for
C.sub.46H.sub.41ClN.sub.2O.sub.4S+H+, 753.25483; found (ESI-FTMS,
[M+H].sup.1+), 753.253.
Example 30
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-pyridin-4-ylbenzyl)sulfonyl]am-
ino}ethyl)-1H-indol-3-yl]propyl}benzoic acid
[0246] Step 1. The bromide from Example 24, Step 1 (77 mg, 0.10
mmol) was reacted with pyridine-4-boronic acid according to the
procedure in Example 29, Step 1 to afford 33 mg (43%) of the Suzuki
product, a white solid.
[0247] Step 2. As described in Example 1, Step 10, the ester (33
mg, 0.043 mmol) was hydrolyzed to afford 30 mg (91%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.90-2.01 (m, 2H), 2.62-2.78 (m, 6H), 2.90 (t, J=7.5 Hz, 2H), 4.02
(s, 2H), 4.56 (s, 1H) broad, 6.48 (d, J=8.8 Hz, 1H), 6.79 (dd,
J=8.8, 2.0 Hz, 1H), 6.83 (s, 1H), 6.99-7.10 (m, 4H), 7.19 (dd,
J=7.6, 1.3 Hz, 1H), 7.22 (dd, J=4.5, 1.5 Hz, 2H), 7.24-7.28 (m,
2H), 7.28-7.34 (m, 7H), 7.36-7.46 (m, 3H), 7.98 (d, J=8.3 Hz, 2H),
8.55 (d, J=5.8 Hz, 2H). HRMS: calcd for
C.sub.45H.sub.40ClN.sub.3O.sub.4S+H+, 754.25008; found (ESI-FTMS,
[M+H].sup.1+), 754.2505.
Example 31
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-pyridin-3-ylbenzyl)sulfonyl]am-
ino}ethyl)-1H-indol-3-yl]propyl}benzoic acid
[0248] Step 1. The bromide from Example 24, Step 1 (77 mg, 0.10
mmol) was reacted with pyridine-3-boronic acid according to the
procedure in Example 29, Step 1 to afford 59 mg (77%) of the Suzuki
product, a yellow solid.
[0249] Step 2. As described in Example 1, Step 10, the ester (54
mg, 0.070 mmol) was hydrolyzed to afford 44 mg (83%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.80-1.93 (m, 2H), 2.53-2.62 (m, 2H), 2.67 (t, J=7.5 Hz, 2H), 2.82
(s, 2H) broad, 2.95-3.03 (m, 2H), 4.09 (s, 2H), 5.61 (dd, J=4.9,
3.4 Hz, 1H), 6.41 (d, J=8.8 Hz, 1H), 6.76 (dd, J=9.0, 2.1 Hz, 1H),
6.89 (s, 1H), 7.01-7.12 (m, 5H), 7.22-7.36 (m, 9H), 7.36-7.47 (m,
3H), 7.55-7.62 (m, 1H), 7.68-7.74 (m, 1H), 7.89 (d, J=8.3 Hz, 2H),
8.60 (dd, J=5.1, 1.5 Hz, 1H), 8.90 (d, J=2.3 Hz, 1H). HRMS: calcd
for C.sub.45H.sub.40ClN.sub.3O.sub.4S+H+, 754.25008; found
(ESI-FTMS, [M+H].sup.1+), 754.2505.
Example 32
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(3-thienyl)benzyl]sulfonyl}ami-
no)ethyl]-1H-indol-3-yl}propyl)benzoic acid
[0250] Step 1. The bromide from Example 24, Step 1 (77 mg, 0.10
mmol) was reacted with thiophene-3-boronic acid according to the
procedure in Example 29, Step 1 to afford 67 mg (87%) of the Suzuki
product, a yellow solid.
[0251] Step 2. As described in Example 1, Step 10, the ester (62
mg, 0.080 mmol) was hydrolyzed to afford 51 mg (83%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.88-2.00 (m, 2H), 2.54-2.64 (m, 2H), 2.68-2.81 (m, 4H), 2.88-2.99
(m, 2H), 4.11 (t, J=6.3 Hz, 1H), 4.20 (s, 2H), 6.49 (d, J=8.6 Hz,
1H), 6.80 (dd, J=8.8, 2.0 Hz, 1H), 6.82 (s, 1H), 7.00 (dd, J=4.9,
1.4 Hz, 1H), 7.05 (dd, J=6.7, 2.4 Hz, 4H), 7.13 (dd, J=3.0, 1.3 Hz,
1H), 7.20-7.28 (m, 4H), 7.28-7.34 (m, 8H), 7.38-7.44 (m, 2H),
7.97-8.04 (m, 2H). HRMS: calcd for
C.sub.44H.sub.39ClN.sub.2O.sub.4S.sub.2+H+, 759.21125; found
(ESI-FTMS, [M+H].sup.1+), 759.2099
Example 33
4-{3-[5-chloro-2-[2-({[2-(3,5-dimethylisoxazol-4-yl)benzyl]sulfonyl}amino)-
ethyl]-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0252] Step 1. The bromide from Example 24, Step 1, (77 mg, 0.10
mmol) was reacted with 3,5 dimethylisooxazole-4-boronic acid
according to the procedure in Example 29, Step 1 to afford 36 mg
(46%) of the Suzuki product, a yellow solid.
[0253] Step 2. As described in Example 1, Step 10, the ester (36
mg, 0.046 mmol) was hydrolyzed to afford 32 mg (90%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.88-2.04 (m, 5H), 2.14 (s, 3H), 2.68-2.78 (m, 4H), 2.78-2.85 (m,
2H), 2.96 (t, J=7.5 Hz, 2H), 3.82-3.97 (m, 2H), 4.18-4.27 (m, 1H),
6.49 (d, J=8.8 Hz, 1H), 6.80 (dd, J=8.8, 2.0 Hz, 1H), 6.83 (d,
J=11.4 Hz, 1H), 7.06 (dd, J=3.7, 1.6 Hz, 4H), 7.12 (dd, J=7.5, 1.1
Hz, 1H), 7.26-7.34 (m, 10H), 7.34-7.40 (m, 1H), 7.41 (d, J=2.0 Hz,
1H), 7.99 (d, J=8.3 Hz, 2H). HRMS: calcd for
C.sub.45H.sub.42ClN.sub.3O.sub.5S+H+, 772.26065; found (ESI-FTMS,
[M+H].sup.1+), 772.2595.
Example 34
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-quinolin-8-ylbenzyl)sulfonyl]a-
mino}ethyl)-1H-indol-3-yl]propyl}benzoic acid
[0254] Step 1. The bromide from Example 24, Step 1, (77 mg, 0.10
mmol) was reacted with 8-quinolineboronic acid according to the
procedure in Example 29, Step 1 to afford 67 mg (82%) of the Suzuki
product, a white solid.
[0255] Step 2. As described in Example 1, Step 10, the ester (60
mg, 0.073 mmol) was hydrolyzed to afford 42 mg (72%) of the title
compound, a yellow solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.68-1.85 (m, 1H), 1.99-2.13 (m, 1H), 2.23-2.37 (m, 1H), 2.43-2.53
(m, 3H), 2.56-2.85 (m, 4H), 3.91 (d, J=14.1 Hz, 1H), 4.28 (d,
J=14.1 Hz, 1H), 4.83 (t, J=4.7 Hz, 1H), 6.39 (d, J=8.8 Hz, 1H),
6.72-6.80 (m, 2H), 6.94-7.01 (m, 2H), 7.01-7.09 (m, 2H), 7.19-7.27
(m, 4H), 7.27-7.31 (m, 4H), 7.32-7.37 (m, 1H), 7.37-7.44 (m, 4H),
7.47-7.56 (m, 3H), 7.75-7.91 (m, 3H), 8.22 (dd, J=8.3, 1.8 Hz, 1H),
8.94 (dd, J=4.3, 1.8 Hz, 1H). HRMS: calcd for
C.sub.49H.sub.42ClN.sub.3O.sub.4S+H+, 804.26573; found (ESI-FTMS,
[M+H].sup.1+), 804.2641.
Example 35
4-{3-[5-chloro-2-{2-[({[4'-(dimethylamino)biphenyl-2-yl]methyl}sulfonyl)am-
ino]ethyl}-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0256] Step 1. The bromide from Example 24, Step 1 (77 mg, 0.10
mmol) was reacted with 4-(dimethylamino)-phenylboronic acid
according to the procedure in Example 29, Step 1 to afford 51 mg
(ca. 52%) of the Suzuki product, a white solid.
[0257] Step 2. As described in Example 1, Step 10, the ester (51
mg, 0.063 mmol) was hydrolyzed to afford 17 mg (ca. 41%) of the
title compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.87-1.98 (m, 2H), 2.44-2.53 (m, 2H), 2.64-2.70 (m, 2H), 2.70-2.77
(m, 2H), 2.79-2.89 (m, 8H), 4.01-4.07 (m, 1H), 4.28 (s, 2H), 6.46
(d, J=8.8 Hz, 1H), 6.59 (d, J=8.8 Hz, 2H), 6.76-6.81 (m, 2H),
6.99-7.07 (m, 6H), 7.16-7.23 (m, 2H), 7.25-7.33 (m, 9H), 7.39 (d,
J=2.3 Hz, 1H), 7.44-7.49 (m, 1H), 8.00 (d, J=8.3 Hz, 2H). HRMS:
calcd for (C.sub.48H.sub.46ClN.sub.3O.sub.4S+2H+)/2, 398.65215;
found (ESI-FTMS, [M+2H].sup.2+), 398.6504
Example 36
4-[3-(5-chloro-1-(diphenylmethyl)-2-{2-[({[2'-(trifluoromethoxy)biphenyl-2-
-yl]methyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)propyl]benzoic
acid
[0258] Step 1. The bromide from Example 24, Step 1 (77 mg, 0.10
mmol) was reacted with 2-(trifluoromethoxy)phenylboronic acid
according to the procedure in Example 29, Step 1 to afford 36 mg
(ca. 36%) of the Suzuki product, a white solid.
[0259] Step 2. As described in Example 1, Step 10, the ester (36
mg, 0.042 mmol) was hydrolyzed to afford 23 mg (ca. 75%) of the
title compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.80-1.91 (m, 2H), 2.54 (q, J=7.2 Hz, 2H), 2.59-2.70 (m, 4H),
2.78-2.87 (m, 2H), 3.90 (q, J=14.1 Hz, 2H), 4.05-4.11 (m, 1H), 6.40
(d, J=8.8 Hz, 1H), 6.67-6.76 (m, 2H), 6.92-7.02 (m, 4H), 7.07-7.16
(m, 3H), 7.16-7.30 (m, 12H), 7.31-7.36 (m, 2H), 7.93 (d, J=8.3 Hz,
2H). HRMS: calcd for C.sub.47H.sub.40ClF.sub.3N.sub.2O.sub.5S+H+,
837.23713; found (ESI-FTMS, [M+H].sup.1+), 837.2375.
Example 37
4-{3-[5-chloro-2-[2-({[(2'-cyanobiphenyl-2-yl)methyl]sulfonyl}amino)ethyl]-
-1-(diphenylmethyl)-1H-indol-3-yl]propyl}benzoic acid
[0260] Step 1. The bromide from Example 24, Step 1 (73 mg, 0.095
mmol) was reacted with 2-cyanophenylboronic acid to afford 23 mg
(30%) of the Suzuki product, a yellow solid.
[0261] Step 2. As described in Example 1, Step 10, the ester (19
mg, 0.024 mmol) was hydrolyzed to afford 10 mg (53%) of the title
compound, a white solid. 1H NMR (400 MHz, CDCl.sub.3) .delta.
1.87-2.01 (m, 2H), 2.62-2.79 (m, 6H), 2.92 (t, J=7.6 Hz, 2H),
3.91-4.14 (m, 3H), 6.47 (d, J=8.8 Hz, 1H), 6.75-6.85 (m, 2H),
7.01-7.08 (m, 4H), 7.22-7.28 (m, 3H), 7.28-7.36 (m, 8H), 7.36-7.44
(m, 4H), 7.49-7.59 (m, 1H), 7.63-7.69 (m, 1H). 8.00 (d, J=8.3 Hz,
2H). HRMS: calcd for C.sub.47H.sub.40ClN.sub.3O.sub.4S+H+,
778.25008; found (ESI-FTMS, [M+H].sup.1+), 778.2489.
Example 38
3-{4-[(2-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethyl)benzyl]su-
lfonyl}amino)ethyl]-1H-indol-3-yl}ethyl)sulfonyl]phenyl}propanoic
acid
[0262] Step 1: 2-Bromo-4-chloroaniline (1.0 eq) was dissolved in
CH.sub.2Cl.sub.2 (0.25 M), then triethylamine and trifluoroacetyl
anhydride (1.1 eq each) were added. The resulting mixture was
stirred at room temperature for 1 hour. The solvent was evaporated
and the residue was purified by flash chromatography with
CH.sub.2Cl.sub.2 as eluent to give the amide in 97% yield. m/z
(M-H).sup.- 300.0.
[0263] Step 2: N-(2-Bromo-4-chlorophenyl)-2,2,2-trifluoroacetamide
(Step 1, 1.0 eq) was mixed with 3-butyn-1-ol (2.0 eq),
dichlorobis(triphenylphosphine)palladium(II) (2.5% eq),
triethylamine (3.0 eq), CuI (5% eq) in DMF (0.2 M) in a sealed
vessel under N.sub.2 and heated to 120.degree. C. for 4 hours. The
reaction mixture was then diluted with ethyl acetate, washed with
brine and dried over Na.sub.2SO.sub.4. Purification by flash column
chromatography with 2% MeOH/CH.sub.2Cl.sub.2 afforded the alkyne in
67% yield. m/z (M-H).sup.- 194.09
[0264] Step 3: 2-(5-Chloro-1H-indol-2-yl)ethanol (step 2, 1.0 eq)
and imidazole (2.0 eq) were dissolved in DMF (0.3 M) at room
temperature with stirring before tert-butylchlorodiphenylsilane
(1.2 eq) was added. The resulting mixture was stirred overnight at
room temperature before it was quenched with a saturated aqueous
sodium bicarbonate solution and extracted with ethyl acetate. The
organic phase was washed with water and brine and dried over
Na.sub.2SO.sub.4. Purification by flash chromatography with
CH.sub.2Cl.sub.2 as eluent afforded the silyl ether as a brown gum
in over 90% yield. m/z (M-H).sup.- 433.0
[0265] Step 4:
2-({[tert-Butyl(diphenyl)silyl]oxy}ethyl)-5-chloro-1H-indole (Step
3, 1.0 eq) was dissolved in ether (0.4 M) and the solution was
cooled to 0.degree. C. Oxalyl chloride (1.2 eq) was added to the
above cold solution with vigorous stirring. The reaction mixture
was stirred at 0.degree. C. for 1 hour before EtOH was added,
followed by NEt.sub.3. The resulting mixture was then diluted with
more EtOH before it was poured into water and extracted with EtOAc.
The organic phase washed with brine, dried over Na.sub.2SO.sub.4,
and concentrated to give the ketoester as yellow solid in 70%
yield. m/z (M-H).sup.- 533.0
[0266] Step 5:
Ethyl[2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-chloro-1H-indol-3-yl](o-
xo)acetate
[0267] (Step 4, 1 eq), Ph.sub.2CHBr (1.5 eq) and Cs.sub.2CO.sub.3
(1.5 eq) were mixed in dry acetonitrile (0.1M). The mixture was
heated to reflux for 2 hours. The reaction mixture was cooled to
room temperature, diluted with water and extracted with EtOAc. The
organic phase was concentrated and the residue was chromatographed
with CH.sub.2Cl.sub.2 as eluent to give the N-benzhydryl indole as
an orange gum in 45% yield. m/z (M+H).sup.+ 701.3
[0268] Step 6: To a solution of
ethyl[1-benzhydryl-2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-chloro-1H--
indol-3-yl] (oxo)acetate (Step 5, 1 eq) in THF (0.1M) was added
BH.sub.3Me.sub.2S (2M in THF) (2 eq). The resulting mixture was
heated to reflux overnight under N.sub.2. The reaction mixture was
cooled to room temperature, then quenched slowly with 1N NaOH,
extracted with EtOAc, and washed with brine. Concentration afforded
the alcohol in 65% yield. m/z (M+H).sup.+ b 645.0
[0269] Step 7: To a solution of
2-[1-benzhydryl-2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-chloro-1H-ind-
ol-3-yl]ethanol (Step 6, 1 eq) in CH.sub.2Cl.sub.2 (0.08M) was
added 1,3-bis(diphenylphosphino)-propane (DPPP, 0.75 eq). The
solution was cooled to 0.degree. C. under N.sub.2, then CBr.sub.4
(1.25 eq) was added. The reaction temperature was allowed to return
to room temperature over 2 h. The solvent was evaporated, and the
residue was purified using a short silica gel column with
CH.sub.2Cl.sub.2 as eluent to give the bromide in quantitative
yield. m/z (M+H).sup.+ 708.0
[0270] Step 8:
1-Benzhydryl-3-(2-bromoethyl)-2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-
-chloro-1H-indole (Step 7, 1 eq) was mixed with
methyl-3-(4-mercaptolphenyl)propionate (1.5 eq) and K.sub.2CO.sub.3
(1.5 eq) in DMF (0.1 M). The resulting mixture was stirred at room
temperature under N.sub.2 for 2 h, then diluted with water and
extracted with EtOAc. The organic extract was washed with brine,
concentrated, and purified by flash chromatography
(CH.sub.2Cl.sub.2 as eluent) to give the thioether as a brownish
gum in 80% yield. m/z (M+H) 823.0
[0271] Step 9: Methyl
3-[4-({2-[1-benzhydryl-2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-chloro-
-1H-indol-3-yl]ethyl}sulfanyl)phenyl]propanoate (Step 8, 1 eq) was
dissolved in acetonitrile (0.1 M), then molecular sieves (powder, 4
A,) and 4-methylmorpholine N-oxide (NMO) (4 eq) were added under
N.sub.2. After 5 min, n-Pr.sub.4NRuO.sub.4 (TPAP) (5% eq) was
added. The resulting mixture was heated at 40.degree. C. for 1.5 h.
The mixture was concentrated and the residue was purified by flash
chromatography with CH.sub.2Cl.sub.2, then 1%
EtOAc/CH.sub.2Cl.sub.2 as eluent to give the sulfone as a white
foam in 44% yield. m/z (M+H).sup.+ 855.1
[0272] Step 10: Methyl
3-(4-{2-[1-benzhydryl-2-({[tert-butyl(diphenyl)silyl]oxy}ethyl)-5-chloro--
1H-indol-3-yl]ethoxy}phenyl)propanoate (Step 9, 1 eq) was dissolved
in THF (0.1 M) and cooled to 0.degree. C. then treated with
nBu.sub.4NF (1 M in THF) (1.2 eq). The resulting mixture was
stirred at 0.degree. C. for 5 min, then warmed to room temperature
and stirred for 30 min. The solvent was evaporated and the residue
was purified by flash chromatography with EtOAc/CH.sub.2Cl.sub.2
(1:9 to 1:4) as eluent to give the alcohol as a white foam in 90%
yield. m/z (M+H).sup.+ 616.20
[0273] Step 11: Methyl
3-[4-{2-[1-benzhydryl-5-chloro-2-(hydroxyethyl)-1H-indol-3-yl]ethyl}-sulf-
onyl)phenyl] propanoate (Step 10, 1 eq) in CH.sub.2Cl.sub.2 (0.02
M) was treated at 0.degree. C. with MeSO.sub.2Cl (2.0 eq) and
Et.sub.3N (2.5 eq) and stirred for 1 hour. The ice-bath was removed
and the reaction mixture was stirred for 1 hour at room temperature
before it was diluted with CH.sub.2Cl.sub.2, washed with
NaH.sub.2PO.sub.4, brine and dried over Na.sub.2SO.sub.4.
Evaporation of the solvent afforded the mesylate in quantitative
yield. m/z (M+H).sup.+ 695.0
[0274] Step 12: Methyl
3-(4-{[2-(1-benzhydryl-5-chloro-2-{2-[(methylsulfonyl)oxy]ethyl}-1H-indol-
-3-yl)ethyl]sulfonyl}phenyl)propanoate (Step 11, 1.0 eq) was
dissolved in DMF (0.03 M) and treated with NaN.sub.3 (3.0 eq). The
resulting mixture was heated to 60.degree. C. and stirred for 2
hours, then cooled to room temperature, diluted with water,
extracted with ethyl acetate, washed with brine and dried with
Na.sub.2SO.sub.4. Evaporation of solvent afforded the azide in
quantitative yield. m/z (M+H).sup.+ 641.1
[0275] Step 13: Methyl
3-[4-({2-[2-(2-azidoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]ethyl}sulf-
onyl)phenyl]propanoate (Step 12, 1 eq) was dissolved in THF (0.1
M), and treated with triphenylphosphine (1.1 eq). After 2 days
water was added, and the mixture was stirred overnight,
concentrated, and purified by flash chromatography using 4%
MeOH:CH.sub.2Cl.sub.2 as eluent to give the amine in 71% yield. m/z
(M+H).sup.+ 615.2
[0276] Step 14: As outlined in Step 9, Example 1, ethyl
3-[4-({2-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]ethyl}sulf-
onyl)phenyl]propanoate (Step 13, 200 mg, 0.32 mmol) was reacted
with (2-trifluoromethylphenyl)methanesulfonyl chloride (Example 5,
Step 3, 110 mg, 0.42 mmol) to afford 250 mg of the sulfonamide, a
pale yellow foam, in 93% yield. .sup.1HNMR (400 MHz, CDCl.sub.3)
1.23 (t, J=7.2 Hz, 3H), 2.62-2.71 (m, 2H), 2.76-2.93 (m, 4H),
2.98-3.17 (m, 4H), 3.27-3.38 (m, 2H), 4.11 (q, J=7.2 Hz, 2H), 4.35
(s, 2H), 4.57 (t, J=5.3 Hz, 1H), 6.43 (d, J=9.1 Hz, 1H), 6.77 (dd,
J=8.8, 2.0 Hz, 1H), 6.81 (s, 1H), 7.18 (d, J=2.0 Hz, 1 H),
7.24-7.35 (m, 10H), 7.41 (d, J=8.6 Hz, 3H), 7.49 (t, J=8.3 Hz, 1H),
7.60-7.77 (m, 2H), 7.88 (d, J=8.6 Hz, 2H).
[0277] Step 15: Using the procedure in Step 10 Example 1, the
sulfonamide ester (220 mg, 0.26 mmol) was hydrolyzed to afford 200
mg (92%) of the title product, a white foam. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 2.65 (t, J=7.6 Hz, 2H), 2.91-3.13 (m, 8H), 3.60 (dd,
J=9.7, 5.4 Hz, 2H), 4.46 (s, 2H), 6.48 (d, J=8.8 Hz, 1H), 6.83 (dd,
J=8.7, 2.1 Hz, 1H), 7.05-7.16 (m, 5H), 7.19 (d, J=2.3 Hz, 1H),
7.33-7.47 (m, 6H), 7.53-7.72 (m, 6H), 7.80 (d, J=7.6 Hz, 1H), 7.94
(d, J=8.3 Hz, 2H), 12.26 (s, 1H); HRMS: calcd for
C.sub.42H.sub.38ClF.sub.3N.sub.2O.sub.6S.sub.2+H+, 823.18847; found
(ESI-FTMS, [M+H].sup.1+), 823.1887; HPLC purity
H.sub.2O/CH.sub.3CN: 100%, H.sub.2O/MeOH: 100%.
Example 39
3-(4-{[2-(5-chloro-1-(diphenylmethyl)-2-{2-[({1-[2-(trifluoromethyl)phenyl-
]ethyl}sulfonyl)amino]ethyl}-1H-indol-3-yl)ethyl]sulfonyl}phenyl)propanoic
acid
[0278] Step 1: Using the procedure in Example 1, Step 9, ethyl
3-[4-({2-[2-(2-aminoethyl)-5-chloro-1-(diphenylmethyl)-1H-indol-3-yl]ethy-
l}sulfonyl)phenyl]propanoate (Example 38, Step 14) was reacted with
1-(2-trifluoromethyl-phenyl)-ethanesulfonyl chloride (0.13 g, 0.46
mmol) to afford
3-{4-[2-(1-benzhydryl-5-chloro-2-{2-[1-(2-trifluoromethyl-pheny-
l)-ethanesulfonylamino]-ethyl}-1H-indol-3-yl)-ethanesulfonyl]-phenyl}-prop-
ionic acid ethyl ester (0.110 g, 40%).
[0279] Step 2: The sulfonamide ester (0.11 g, 0.13 mmol) was
hydrolyzed according to Example 1, Step 10 to afford 0.068 g (64%)
of the title product, a white solid.
[0280] 1H NMR (400 MHz, CDCl.sub.3) .delta. 1.67 (d, J=6.8 Hz, 3H)
2.57-2.72 (m, 4H) 2.80 (t, J=6.8 Hz, 2H) 2.84-2.94 (m, 2H) 3.03 (t,
J=6.4 Hz, 2H) 3.20-3.31 (m, 2H) 5.82-5.88 (m, 1H) 6.37 (s, 1H)
6.73-6.81 (m, 2H) 6.98 (d, J=4.4 Hz, 2H) 7.05 (d, J=5.4 Hz, 2H)
7.24-7.49 (m, 11H) 7.63 (d, J=7.8 Hz, 1H) 7.80 (d, J=7.8 Hz, 1H)
7.88 (d, J=8.6 Hz, 2H).
Example 40
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}e-
thyl)-1H-indol-3-yl]propyl}benzoic acid
[0281] Step 1: Using the procedure described in Example 5, Step
1,2-benzyloxybenzylbromide (ref. J. Med. Chem. 2006, 49, 31-34, R.
V. Somu et al.) (32.2 g, 116 mmol) afforded
(2-benzyloxy-phenyl)-methanesulfonic acid sodium salt (30 g, 86%),
a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.82 (s,
2H) 5.09 (s, 2H) 6.81-6.91 (m, 1H) 6.96 (d, J=7.58 Hz, 1H)
7.08-7.18 (m, 1H) 7.26-7.34 (m, 1H) 7.34-7.41 (m, 2H) 7.45 (dd,
4=1.77 Hz, 1H) 7.52 (d, J=7.07 Hz, 2H).
[0282] Step 2: Using the procedure described in Example 5, Step 2,
(2-benzyloxy-phenyl)-methanesulfonic acid sodium salt (30 g, 99
mmol) afforded (2-benzyloxy-phenyl)-methanesulfonic acid (15 g), a
white solid which was used without further purification. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 3.81 (s, 2H) 5.08 (s, 2H)
6.80-6.92 (m, 1H) 6.95 (d, J=7.83 Hz, 1H) 7.07-7.17 (m, 1H) 7.31
(d, J=6.82 Hz, 1H) 7.34-7.42 (m, 2H) 7.45 (dd, 1H) 7.52 (d, J=7.33
Hz, 2H).
[0283] Step 3: Using the procedure described in Example 5, Step 3,
(2-benzyloxy-phenyl)-methanesulfonic acid (7 g, 25.15 mmol)
afforded (2-benzyloxy-phenyl)-methanesulfonyl chloride (2.6 g,
35%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.06 (s, 2H) 5.15
(s, 2H) 7.00-7.10 (m, 2H) 7.30-7.50 (m, 7H).
[0284] Step 4: As outlined in Step 9 Example 1, methyl
4-{3-[2-(2-aminoethyl)-1-benzhydryl-5-chloro-1H-indol-3-yl]propyl}benzoat-
e (Example 7, Step 6, 3.92 g, 7.3 mmol) was reacted with
(2-benzyloxy-phenyl)-methanesulfonyl chloride (2.6 g, 8.76 mmol) to
afford 4.1 g of methyl
4-{-[2-[2-({[2-(benzyloxy)benzyl]sulfonyl}amino)ethyl]-5-chloro-1-(diphen-
ylmethyl)-1H-indol-3-yl]propyl}benzoate, a white foam, in 59%
yield. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.80-1.99 (m, 2H)
2.49-2.78 (m, 6H) 2.85 (t, J=8.84 Hz, 2H) 3.89 (s, 3H) 3.96-4.05
(m, 1H) 4.26 (s, 2H) 4.90 (s, 2H) 6.45 (d, J=8.84 Hz, 1H) 6.73-6.82
(m, 2H) 6.83-6.93 (m, 2H) 6.94-7.08 (m, 4H) 7.16-7.34 (m, 15H) 7.39
(d, J=2.02 Hz, 1H) 7.85-7.98 (m, 2H).
[0285] Step 5: Methyl
4-{3-[2-[2-({[2-(benzyloxy)benzyl]sulfonyl}amino)ethyl]-5-chloro-1-(diphe-
nylmethyl)-1H-indol-3-yl]propyl}benzoate (5.1 g, 6.4 mmol) was
reacted with hydrogen in the presence of palladium on carbon (0.5
g) to afford a mixture of methyl
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}-
ethyl)-1H-indol-3-yl]propyl}benzoate and methyl
4-{3-[1-(diphenylmethyl)-2-(2-{[(2-hydroxybenzyl)sulfonyl]amino}ethyl)-1H-
-indol-3-yl]propyl}benzoate (3:1) as a white foam in 74% overall
yield.
[0286] Step 6: Using the procedure in Step 10, Example 1, the
sulfonamide ester mixture (3.35 g) was hydrolyzed and purified by
preparative HPLC to afford 1.18 g (36%) of the title product, a
white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 1.89-2.01
(m, 2H) 2.64-2.96 (m, 8H) 4.16 (s, 2H) 4.17-4.25 (m, 1H) 6.50 (d,
J=8.84 Hz, 1H) 6.74-6.89 (m, 4H) 6.95 (dd, J=1.64 Hz, 1H) 7.01-7.13
(m, 4H) 7.11-7.23 (m, 1H) 7.23-7.38 (m, 8H) 7.41 (d, J=2.02 Hz, 1H)
7.90-8.04 (m, 2H); HRMS: calcd for
C.sub.40H.sub.37ClN.sub.2O.sub.5S+H+, 693.21845; found (ESI-FTMS,
[M+H].sup.1+), 693.21709; HPLC purity (CH.sub.3CN--H.sub.2O): 7.24
min, 100.0%. HPLC purity (MeOH--H.sub.2O): 8.12 min, 100.0%.
Example 41
4-{3-[5-chloro-1-(diphenylmethyl)-2-(2-{[(2-quinolin-5-ylbenzyl)sulfonyl]a-
mino}ethyl)-1H-indol-3-yl]propyl}benzoic acid
[0287] Step 1. The bromide from Example 24, Step 1 was reacted with
5-quinolineboronic acid according to the procedure in Example 29,
Step 1 to afford the Suzuki product.
[0288] Step 2. As described in Example 1, Step 10, the ester was
hydrolyzed and the product purified by preparative HPLC to afford
the title compound, a white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
1.77-1.91 (m, 2H), 2.35-2.70 (m, 6H), 2.76 (t, J=7.2 Hz, 2H), 3.65
(d, J=13.9 Hz, 1H), 3.89 (d, J=13.9 Hz, 1H), 4.00 (t, J=5.3 Hz,
1H), 6.39 (d, J=9.1 Hz, 1H), 6.63-6.79 (m, 2H), 6.86-7.04 (m, 4H),
7.08-7.24 (m, 10H), 7.24-7.40 (m, 4H), 7.43-7.51 (m, 1H), 7.55 (dd,
J=8.6, 7.1 Hz, 1H), 7.62 (d, J=8.3 Hz, 1H), 7.84-7.94 (m, 2H), 8.05
(d, J=8.6 Hz, 1H), 8.83 (dd, J=4.3, 1.8 Hz, 1H). HRMS: calcd for
C.sub.49H.sub.42ClN.sub.3O.sub.4S+H+, 804.26573; found (ESI-FTMS,
[M+H].sup.1+), 804.2663.
[0289] An alternative method for preparing intermediate compounds
of the general formula:
##STR00016##
[0290] where X is halogen, preferably chlorine, is disclosed in
U.S. patent application Ser. No. 11/064,241, filed Feb. 23, 2005,
which is incorporated by reference herein in its entirety. Briefly,
the method involves the formation of sulfonic acid prior to
conversion to the sulfonyl halide, according to the general scheme
below:
##STR00017##
wherein L is a leaving group; Ar represents a 2,6-disubstituted
phenyl moiety; R represents a (CHR.sub.5).sub.n2 moiety, and M is a
group I or group II metal ion. In accordance with the scheme,
sulfonic acids of Formula IV can be converted to sulfonyl halides
by reaction with a halogen substitution reagent (i.e., a reagent
that can convert a non-halogen substituent such as, for example, H
or OH, to a halogen substituent; i.e., convert a sulfonic acid
moiety to a sulfonyl halide moiety), for example SOCl.sub.2,
POCl.sub.3, CCl.sub.4/triphenylphosphine, oxalyl chloride or oxalyl
bromide, preferably oxalyl chloride. The halogen substitution agent
is preferably used in excess quantity, particularly if there is
residual solvent in either the starting material, solvents or both.
When oxalyl chloride is used as the halogen substitution agent, it
can be used in a range from about 1 to about 6 equivalents; about 2
to about 4 equivalents or about 3 to about 3.5 equivalents with
respect to the amount of sulfonic acid reagent (compound of Formula
IV). One skilled in the art will recognize that the amount of
halogen substitution agent used will depend, inter alia, on the
amount of water in the starting material or solvent and the nature
and reactivity of the starting material and solvents.
[0291] Suitable solvents for the halogen substitution reaction
(step 3 of the scheme above) include any organic solvent that can
at least partially dissolve the compound of Formula IV. Preferred
solvents include non-polar or weakly polar solvents, including
acetonitrile, aromatic hydrocarbons such as benzene and toluene,
and halogenated solvents such as 1,2-dichloroethane and methylene
chloride. More preferred solvents are ethers. Suitable ethers
include tetrahydrofuran, dioxane, diethyl ether, dibutyl ether,
diisopropyl ether or mixtures thereof and the like. A more
preferred ether is tetrahydrofuran.
[0292] The halogen substitution reaction can be carried out at any
suitable temperature, for example at about -40.degree. C. to about
room temperature, preferably below about -10.degree. C.
[0293] The sulfonyl halide-forming step (step 3 of the scheme
above) can also be carried out in the presence of an acyl transfer
catalyst, such as a tertiary amide (e.g., dimethylformamide). The
acyl transfer catalyst can be provided in an amount sufficient to
accelerate the reaction rate. The acyl transfer catalyst is present
in less than about one equivalent relative to the amount of
sulfonic acid reagent, preferably in an amount of about 0.01 to
about 0.5 equivalents; even more preferred, about 0.1 to about 0.2
equivalents, relative to the amount of sulfonic acid reagent.
[0294] The compounds of Formula I can be isolated from the reaction
mixture by precipitation and filtration. Any of numerous well known
methods for inducing precipitation can be used. In some preferred
embodiments, an anti-solvent such as water or a solvent containing
water can be added to the reaction mixture to induce precipitation.
Use of water as an anti-solvent can reduce decomposition rate of
the sulfonyl halide product relative to the decomposition rate
observed when an organic solvent such as heptane is used, resulting
in improved yields. Precipitation can be facilitated by lowering
the temperature of the reaction mixture to, for example, to below
about -20.degree. C.
[0295] As shown in the scheme above, sulfonic acids of Formula IV
can be prepared by reacting sulfonic acid salts (sulfonate salts)
of Formula III with a protic acid. Suitable protic acids are of
sufficient strength so as to be capable of converting a sulfonate
salt to its corresponding acid according to the processes of the
invention. For example, the protic acid can be a strong inorganic
acid such as HCl, HBr, H.sub.3PO.sub.4, HNO.sub.3, HClO.sub.4,
H.sub.2SO.sub.4, and the like. Alternatively, the protic acid can
be an organic acid, such as formic, methanesulfonic acid, p-toluene
sulfonic acid, benzenesulfonic acid, trifluoroacetic acid and other
strong organic acids. The protic acid can be provided in gaseous
form. Preferably, the inorganic acid is HCl, more preferably
gaseous HCl that is added to the reaction solvent containing the
sulfonate salt. The protic acid is advantageously provided in
excess molar equivalents relative to the sulfonic acid salt of
Formula III.
[0296] Formation of the sulfonic acid compound of Formula IV can be
carried out in any suitable solvent. For example, organic solvents
in which the compound of Formula III is at least partially soluble
are suitable. The solvent can be chosen such that it poorly
dissolves metal halide salts, such as NaCl or KCl, thereby
thermodynamically driving the reaction by precipitation of metal
halide salt. The solvent can contain an alcohol, such as methanol,
ethanol, isopropanol, and the like, or a mixture thereof,
preferably methanol. The solvent can also contain water. Reaction
temperature can be readily determined by the skilled artisan. For
example, the reaction can be carried out at a temperature below
room temperature, such as about -20 to about 10.degree. C.,
preferably at about 0 or below about 10.degree. C.
[0297] The sulfonic acid compound of Formula IV can be isolated
according to routine methods, such as precipitating the product
from the reaction mixture.
[0298] The sulfonic acid salt (sulfonate salt) compound of Formula
III can be prepared by reacting a compound of Formula II: Ar--R-L
(wherein Ar, R and L are defined hereinabove) with a Group I or II
metal sulfite salt optionally in the presence of a phase transfer
catalyst as shown in step 1 of the scheme above. Any Group I or II
metal sulfite salt is suitable, for example, Li.sub.2SO.sub.3,
Na.sub.2SO.sub.3, K.sub.2SO.sub.3, MgSO.sub.3, CaSO.sub.3, and the
like. Group I or II metal sulfite salts can be provided in molar
excess of, for example, about 2 eq, to about 1 eq, relative to the
amount of compound of Formula II. Suitable metal salts include
Na.sub.2SO.sub.3, K.sub.2SO.sub.3 and Na.sub.2SO.sub.3.
[0299] The formation of the sulfonate salt compounds of Formula III
can be carried out in the presence of a phase transfer catalyst,
for example a quaternary ammonium halide, such as tetrabutyl
ammonium iodide. The phase transfer catalyst can be provided in an
amount suitable to accelerate the reaction rate, for example in
about 0.1 to 2% or more preferably 0.5 to 1% by weight.
[0300] Any suitable solvent can be employed, such as solvent that
can at least partially dissolve Group I or II metal sulfite salts,
such as water, in an amount of from about 50%, more preferably
about 75%, even more preferably more than about 90%, still more
preferably more than about 95%, and yet more preferably more than
about 99% water. The reaction can also be carried out at any
suitable temperature, preferably an elevated temperature, for
example about 100.degree. C.
[0301] Isolation of the compound of Formula III from the reaction
mixture can be carried out by any routine method, such as
precipitation from the reaction mixture by, for example, treatment
of the reaction mixture with a water-soluble inorganic salt such as
NaCl or KCl, more preferably NaCl. Isolation of the compound of
Formula III can be further facilitated by the addition to the
reaction mixture of an organic solvent that is not substantially
miscible with water, such as ethyl acetate, ethers (e.g. ethyl
ether and the like), alkanes (e.g., hexanes, petroleum ether,
etc.), aromatics (e.g., benzene, toluene, xylene, etc.), and the
like, with ethyl acetate being most preferred. The reaction mixture
can also be cooled (e.g., less than about 10.degree. C.) to help
induce precipitation.
Biological Test Procedures
GLU Micelle Assay
[0302] The assay was carried in a 96-well format using a
fluorescent plate reader with a 355 nM excitation filter and a 460
nM emission filter (Lab Systems Fluoroscan II, Helsinki, Finland).
The assay buffer contained 940 .mu.M Triton X-100, 50 mM Hepes pH
7.4, 0.3 mM EDTA, 1 mM CaCl.sub.2 and 300 mM KCl. DTPC
(1,2-O-tetradecyl-sn-glycero-3-phosphocholine, Avanti) at a final
concentration of 120 .mu.M was added the day of the experiment and
GLU (7-Hydroxycoumarinyl-.gamma.-linolenate, Biomol Research Lab,
Inc.) at a final concentration of 90 .mu.M was added immediately
prior to each assay.
[0303] Compounds (10 .mu.L) dissolved in DMSO were placed in
duplicate wells of a black 96-well plate. Wells corresponding to
the positive and negative controls contained DMSO without
inhibitors. Just prior to the experiment, 200 .mu.L assay buffer
containing 90 .mu.M GLU and 120 .mu.M DTPC was added to all wells
in the assay plate. Assay buffer (50 .mu.L) was added to the
negative, and 50 .mu.L cPLA.sub.2.alpha. solution (5 mg/mL in assay
buffer) was added to all other wells to initiate the reaction. The
final concentration of enzyme was 1 .mu.g/ml. The content of each
well was mixed gently during the addition of the enzyme, and the
plate was rapidly transferred to the fluorescent plate reader. The
increase in fluorescence was read every 4 min for 84 min. The slope
of the resulting line was determined and the inhibition was
calculated using the equation below:
zi Percent Inhibition=[1-(slope with inhibitor-slope negative
control)/(slope positive control-slope negative
control)].times.100
Rat Whole Blood Assay
[0304] Fresh blood was collected in heparinized tubes by cardiac
puncture of male Sprague-Dawley rats. Aliquots of blood (0.6 mL)
were incubated with either 6 .mu.L solvent (DMSO), or 6 .mu.L of
test compounds at various concentrations for 15 minutes at
37.degree. C. This was followed by incubation of the blood with 6
.mu.L of calcium ionophore, A23187 (Sigma C-7522) diluted in DMSO
for 10 min at 37.degree. C. The final concentration of A23187 was 5
.mu.M. DMSO (6 .mu.L) was added in the unstimulated controls. The
reactions were stopped by mixing 60 .mu.L cold EDTA to give a final
concentration of 20 mM. The blood was centrifuged at 6,500 rpm for
10 min on a microcentrifuge to obtain plasma. A 70 .mu.L aliquot of
plasma was mixed with 400 .mu.L cold methanol for protein
precipitation. After incubation at -80.degree. C. for 30 min, the
supernatant was obtained by centrifuging at 6,500 rpm for 10 min,
and was assayed for TXB.sub.2 according to the manufacturer's
procedure (Assay Designs, Inc.'s ELISA kit #900-002).
[0305] Results of the GLU Micelle Assay and the Rat Whole Blood
Assay for compounds of the invention is shown in Table 1,
below:
TABLE-US-00001 GLU Micelle Rat Whole Blood TXB.sub.2 Example #
IC.sub.50 (uM) IC.sub.50 (uM) 1 0.26 0.14 2 0.19 0.12 3 0.054 0.06
4 0.054 0.02 5 0.026 0.02 6 0.092 0.08 7 0.018 0.03 8 0.024 0.02 9
0.022 0.02 10 0.009 0.02 11 0.28 0.38 12 0.021 0.04 13 0.026 0.03
14 0.03 0.02 15 0.068 0.12 16 0.023 0.05 17 0.01 0.02 18 0.025 0.04
19 0.022 0.03 20 0.014 0.17 21 0.0059 0.03 22 0.021 0.08 23 0.105
0.04 24 0.008 0.03 25 0.013 0.03 26 0.022 0.03 27 0.038 0.03 28
0.03 0.03 29 0.018 0.05 30 0.021 0.06 31 0.016 0.04 32 0.013 0.05
33 0.022 0.02 34 0.016 0.03 35 0.027 0.05 36 0.031 0.07 37 0.025
0.03 38 0.007 0.01 39 0.068 0.02 40 0.072 0.06 41 0.022 --
Effect of cPLA.sub.2 Inhibitor in Models of Thrombosis
[0306] The effect of administration of cPLA.sub.2 inhibitors in
models for thrombosis was determined by the following
procedures.
Platelet Function Analyzer (PFA-100.RTM.) Study
[0307] Human platelet aggregation was studied using the platelet
function analyzer (PFA-100.RTM.). Human blood was collected from
volunteers who had denied taking any platelet inhibitory
medications over the previous two weeks. Blood was collected in
3.2% sodium citrate Vacutainer tubes (Becton Dickinson). Tubes were
inverted 5 times and the blood was transferred to 15 ml
polypropylene conical tubes. 5 .mu.l of respective inhibitor
dissolved in 100% DMSO
(4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)be-
nzyl]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid,
Example 14;
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethoxy)benzyl]sulf-
onyl}amino) ethyl]-1H-indol-3-yl}propyl)benzoic acid, Example 25;
4-{3-[1-benzhydryl-5-chloro-2-(2-{[(3,4-dichlorobenzyl)sulfonyl]amino}eth-
yl)-1H-indol-3-yl]propyl}benzoic acid, Compound C) was added to 1
ml aliquot of whole human blood, to give the respective inhibitor
concentration and a final DMSO concentration of 0.5%. Tubes were
inverted 10 times to mix, and allowed to sit at room temperature
for 10 minutes prior to run in PFA-100. The manufacturers protocol
was followed for the PFA-100 using Collagen/Epinephrine cartridges
(0.5% DMSO alone in whole blood gave closure times of 125+/-13.9
seconds). Maximum closure time is 300 seconds, as set by the
manufacturer.
[0308] The results are shown in FIG. 1. Compound C or the compound
of Example 14 or Example 25 was allowed to incubate with whole
human blood prior to challenge testing in the PFA-100. All
compounds were efficacious in the platelet function assay. At a
concentration of 1.25 .mu.g/ml, Compound C and the compound of
Example 14 led to an increased closure time, while the compound of
Example 25 was efficacious at concentrations as low as 0.3
.mu.g/ml. These data show that these three compounds inhibit
platelet aggregation in human blood, in vitro.
FeCl.sub.3-induced model of arterial thrombosis
[0309] Two hours prior to induction of vascular injury, Sprague
Dawley outbreed rats (80-100 gram of body weight) received
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-fluoro-6-(trifluoromethyl)ben-
zyl]sulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid
(Example 14) or
4-(3-{5-chloro-1-(diphenylmethyl)-2-[2-({[2-(trifluoromethoxy)benzyl]s-
ulfonyl}amino)ethyl]-1H-indol-3-yl}propyl)benzoic acid (Example 25)
at a dose of 25 mg/kg by oral gavage. The total volume of gavage
was 0.5 ml. The control group of animals had been treated with only
vehicle. Fifteen minutes before vascular injury rats were
anesthetized by an intramuscular injection of a ketamine/xylazyne
mixture. Following anesthesia the left carotid artery was dissected
and exposed for further measurements. For induction of
prothrombotic injury, a round piece of filter paper (2 mm in
diameter) soaked in 10% of FeCl.sub.3 solution was applied onto the
wall of the exposed vessel. After 5 minutes the filter paper was
removed and the 1PRB perivasular Doppler flow probe (Transonic
Systems Inc.) was secured around the carotid artery to measure
blood flow. Blood flow was recorded for a total period of 30
minutes using Transonic Flow Meter (model TS420, Transonic Systems
Inc.) and Windaq data acquisition software.
[0310] The results, shown in FIG. 2, show that both compounds are
efficacious in the rat ferric chloride thrombosis model when dosed
orally at 25 mg/kg.
Thromboxane B.sub.2 Levels in Rats with FeCl.sub.3 Induced
Thrombosis
[0311] Blood was collected from rats that were dosed with vehicle,
the compound of Example 14 or the compound of Example 25, and
subjected to the ferric chloride Injury protocol above. Blood was
collected from the vena cava and blood coagulation was allowed to
take place for 1 hour at 37 degree Celsius. Serum was then isolated
and serum thromboxane B2 levels were determined by ELISA.
[0312] The results are shown in FIG. 3. These data show that both
compounds provided a reduction in serum Thromboxane B.sub.2
levels.
Effect of cPLA.sub.2 Inhibitor in an Animal Model of Multiple
Sclerosis.
[0313] The effect of administration of a cPLA.sub.2 inhibitor in an
animal model of multiple sclerosis was determined by the following
procedure.
[0314] Six groups of B6 mice were immunized with MOG/CFA and
injected with pertussis toxin to induce experimental autoimmune
encephalomyelitis (EAE), an animal model of Multiple sclerosis.
Three groups of mice were treated with vehicle,
4-{3-[1-benzhydryl-5-chloro-2-(2-{[(2,6-dimethylbenzyl)sulfonyl]amino}eth-
yl)-1H-indol-3-yl]propyl}benzoic acid (Compound A), or
4-{2-[1-benzhydryl-5-chloro-2-(2-{[(3,4-dichlorobenzyl)sulfonyl]amino}-et-
hyl)-1H-indol-3-yl]ethoxy}benzoic acid (Compound B) from the day of
Immunization (orally, 100 mg/kg, twice/day). Another three groups
of mice were treated with vehicle, Compound A or Compound B
starting on the day of EAE onset (day 15) (orally, 100 mg/kg,
twice/day). On this day, over 20% of the animals showed first
clinical signs of EAE and the treatment started in all the animals
in these groups. The results are shown in the Table below, wherein
mean clinical score is a mean of clinical evaluation of each animal
for that particular day.
Animals are Scored as Follows:
[0315] 0--no clinical signs of EAE (no paralysis) [0316]
1--paralysis of tail [0317] 2--paralysis of tail and partial hind
leg paralysis [0318] 3--paralysis of tail and complete hind leg
paralysis [0319] 4--paralysis of tail, complete hind leg paralysis
and a partial front leg paralysis [0320] 5--moribund animal (all
four limbs paralyzed, lack of responsiveness, these mice were
immediately euthanized).
TABLE-US-00002 [0320] Com- Com- Com- Days Vehicle pound pound
Vehicle pound Compound After Control, A, B, Control, A, B, Immun.
Day 1 Day 1 Day 1 Onset Onset Onset 11 0 0 0 0 0 0 12 0 0 0 0 0 0
13 0 0 0 0 0 0 14 0 0 0 0 0 0 15 0.23 0 0 0.37 0.33 0.10 16 0.50 0
0 0.97 0.60 0.45 17 0.63 0 0.06 1.30 0.80 0.80 18 0.93 0 0.06 1.63
1.03 1.05 19 1.03 0 0.11 1.63 1.13 1.10 20 1.47 0 0.39 2.23 1.50
1.00 21 1.63 0.06 0.44 2.40 1.57 1.20 22 2.13 0.09 0.50 2.43 1.60
1.10 23 2.53 0.16 0.67 2.47 1.37 1.40 24 2.73 0.19 0.56 2.47 1.43
1.35 25 2.67 0.25 0.56 2.30 1.23 1.25 26 2.73 0.28 0.56 2.17 0.87
1.40 27 2.63 0.38 0.72 2.17 0.87 1.55
[0321] In addition, the compounds of Examples 14 and 25 were also
found to be efficacious in the mouse experimental autoimmune
encephalomyelitis (EAE) model of multiple sclerosis. As shown by
the data in the Table below, these compounds led to a delayed onset
of disease and reduced severity of disease when administered orally
in doses as low as 2.5 mg/kg.
TABLE-US-00003 Example 14 Example 25 Days After 2.5 mg/kg 2.5 mg/kg
Immunization Vehicle Control Day 1 Day 1 11 0 0 0 12 0.08 0 0 13
0.50 0 0 14 1.13 0 0.14 15 1.48 0.14 0.20 16 2.00 0.32 0.55 17 2.00
0.50 0.55 18 2.19 0.55 0.55 19 3.31 1.32 0.84 20 3.48 1.41 1.27 21
3.60 1.77 1.68 22 3.60 1.89 1.89 23 3.58 2.00 1.93 24 3.60 2.05
2.00 25 3.71 1.90 1.98 26 3.71 1.89 1.95 27 3.71 1.89 1.93
[0322] These results show that treatment of mice with cPLA.sub.2
inhibitors of Examples 14, 25, Compound A and Compound B can
prevent EAE when administered from the time of immunization and
reduce clinical severity of EAE in mice which have already
developed EAE or are close to developing clinical signs of the
disease.
Effect of cPLA.sub.2 Inhibitor in Atherosclerosis
[0323] The effect of administration of a cPLA.sub.2 inhibitor in
the apolipoprotein E (ApoE) knockout mouse model of atherosclerosis
was determined by the following procedure.
ApoE KO Mouse Model
[0324] The apolipoprotein E (ApoE) knockout mouse was created by
gene targeting in embryonic stem cells to disrupt the ApoE gene.
ApoE is a glycoprotein that is responsible for the uptake of
chylomicrons and VLDL particles by the liver, thereby preventing
the accumulation of cholesterol rich remnants in the blood stream.
As a result of the homozygous inactivation of the ApoE gene, ApoE
KO mice exhibit high levels of cholesterol, which in turn induces
the formation atherosclerotic plaques in areas of singularities
along the arterial tree, specifically at the aortic sinus where
high hemodynamic disturbances prevail and at branching sites along
the aorta.
cPLA.sub.2 in Atherosclerosis
[0325] Cytosolic phospholipase A2 (cPLA.sub.2) preferentially
mediates the release of arachidonic acid upon cell activation.
Metabolites of arachidonic acid, the eicosanoids, are recognized as
important modulators of inflammatory processes. Decreased
biosynthesis of pro-inflammatory eicosanoids has been shown to
inhibit atherosclerotic lesion progression in humans and mice,
thereby suggesting a potential role of cPLA.sub.2 in
atherosclerosis (see Ranke et al., Circulation 1993; 87(6)
1873-1879; Paul et al., Life Sciences 2000; 68(4):457-465; Cyrus et
al., Circulation 2002; 106(10) 1282-1287; Pratic et al., PNAS 2001;
98(6): 3358-3363; Burleigh et al., Circulation 2002; 105(15):
1816-1823; Cayatte et al., ATVB 2000; 20(7): 1724-1728; Aiello et
al., ATVB 2002; 22(3): 443-449; Subbanagounder et al., Circ. Res.
1999; 85(4): 311-318). In addition, cPLA.sub.2 expression has been
detected in human atherosclerotic arteries but not in normal
healthy human arteries (see Schafer Elinder et al., ATVB 1997;
17(10):2257-2263).
Effect of an Inhibitor of cPLA.sub.2 on Atherosclerosis in Mice
[0326] Six week old male ApoE KO mice were treated with
4-{3-[1-benzhydryl-5-chloro-2-(2-{[(3,4-dichlorobenzyl)sulfonyl]amino}eth-
yl)-1H-indol-3-yl]propyl}benzoic acid (Compound C). Mice were fed a
normal chow diet supplemented with Compound C at 1.3 mg/g and 3.3
mg/g (resulting in .about.250 ng/mL and .about.500 ng/mL maximum
drug exposure, respectively) or vehicle for 20 weeks. Serum
thromboxane B2 levels were significantly decreased after 9 and 20
weeks of treatment when compared to control animals, as shown in
the table below:
TABLE-US-00004 Serum Thromboxane B2 Levels Compound C Compound C %
Decrease vs Control (1.3 mg/g) (3.3 mg/g) after 9 weeks of
treatment 52.5 61.2 after 20 weeks of treatment 36.6 49.5
[0327] In addition, atherosclerotic plaque burden at the aortic
sinus was decreased by 32.7% (349582.+-.132685 vs 519220.+-.100694
.mu.m.sup.2, p<0.05) and 45.6% (282697.+-.146462 vs
519220.+-.100694 .mu.m.sup.2, p<0.001) in animals that were
administered the compound at 1.3 mg/g and 3.3 mg/g, respectively,
when compared to control animals. Further, as shown in the table
below, reduction in percent lesion area along the aorta was not
significant (ns), demonstrating the role of this cPLA.sub.2
inhibitor in affecting disease specifically in regions of highest
hemodynamic disturbances.
TABLE-US-00005 Atherosclerotic LesionsAlong the Aorta Compound C
Compound C (1.3 mg/g) (3.3 mg/g) % Decrease vs Control 32.4 ns 35.2
ns
[0328] As shown in the Table below, atherosclerotic lesion
complexity was reduced in animals treated with Compound C when
compared to control animals, as attested by increased frequency of
early-stage lesions and decreased frequency of advanced stage
lesions at the aortic sinus. The Table shows percent of total
animals with Stage 1 (fibrofatty lesion), Stage 2 (early fibrous
plaque), Stage 3 (advanced fibrous plaque), Stage 4 (stable
complicated lesion) and Stage 5 (unstable complicated lesion) for
animals dosed with vehicle, Compound C at 1.3 mg/g and Compound C
at 3.3 mg/g.
TABLE-US-00006 Athersclerotic Lesion Complexity Stage 1 Stage 2
Stage 3 Stage 4 Stage 5 Vehicle 11% 56% 33% Cpd. C 33% 56% 11% 1.3
mg/g Cpd. C 9% 18% 37% 27% 9% 3.3 mg/g
Thromboxane B.sub.2 Levels in the ApoE KO Mouse Model of
Atherosclerosis
[0329] ApoE KO mice were fed a normal chow diet supplemented with
Compound C or the compound of Example 10 at 3.3 mg/g chow or
vehicle for two days. Blood was collected through the retro-orbital
siunus and left to coagulate at 37.degree. C. for one hour. Serum
was then isolated and assayed for thromboxane B2 by ELISA.
Thromboxane concentrations (ng/mL) were found to be: Vehicle:
76.1.+-.17.3; Compound C (3.3 mg/g): 33.5.+-.11.6; Compound of
Example 10 (3.3 mg/g): 1.4.+-.0.7.
[0330] In a separate experiment, ApoE KO mice were dosed with
vehicle or the compound of Example 25 at 10 mg/kg by oral gavage.
Blood was collected through the retro-orbital siunus and left to
coagulate at 37.degree. C. for one hour. Serum was then isolated
and assayed for thromboxane B2 by ELISA. Thromboxane concentrations
(ng/mL) were found to be: Vehicle: 267.9.+-.34.3; Compound of
Example 25: 9.4.+-.5.0.
Effect of cPLA.sub.2 Inhibitor in Models of Stroke
[0331] The effect of administration of a cPLA.sub.2 inhibitor in
models for stroke was determined by the following procedures.
Cerebellar Granule Neuron Cultures
[0332] Primary cerebellar granule neurons were isolated from P5-8
rat pups. Briefly, cerebelli were collected and pooled in ice-cold
phosphate buffer saline (PBS) without Ca.sup.2+ and Mg.sup.2+. The
tissue was finely chopped and transferred to an enzymatic
dissociation media containing 20 IU/ml papain in Earle's balanced
salt solution (Worthington Biochemical, Freehold, N.J.) and
incubated for 30 minutes at 37.degree. C. After enzymatic
dissociation, the papain solution was aspirated and the tissue
mechanically triturated with a fire-polished Pasteur pipette in
complete media [Neurobasal Medium with B-27 supplement (Gibco,
Grand Island, N.Y.), penicillin/streptomycin, aphidicolin,
glutamate, potassium chloride] containing 2,000 IU/ml DNase and 10
mg/ml ovomucoid protease inhibitor. Single-cell suspensions in
complete media were plated on pre-coated poly-L-ornithine/laminin
24-well plates (Becton-Dickinson, Bedford, Mass.) at a density
of
5.0.times.10.sup.5 cells/well. Cells were maintained for two weeks
prior to experimentation.
Oxygen-Glucose Deprivation (OGD) in Cultured Neurons
[0333] Cultures were treated with Compound A at various
concentrations, 60 minutes before OGD. Media was removed and
replaced with deoxygenated buffer in an anaerobic chamber (80%
nitrogen, 10% hydrogen, 10% carbon dioxide gas mixture). Fresh
Compound A, Example 34 or Example 41 was added to the cultures and
maintained in the anaerobic chamber for 2 hours. At the end of the
incubation, fresh media was exchanged and fresh Compound A was
added. Cultures were maintained for an additional 24 hours in a
normoxic incubator. Cell death was determined by measuring lactate
dehydrogenase release into the media 24 hours later (Roche
Biochemicals). In the table below, values are shown for the
control, OGD, various concentrations of Compound A, and MK801, a
NMDA receptor antagonist, which is a positive control.
TABLE-US-00007 Neuroprotection by cPLA2 inhibitors Against OGD
Control OGD 0.1 .mu.M 0.3 .mu.M 1 .mu.M 3 .mu.M Control Avg. 14 51
St. Dev, 3 5 Cpd. A Avg. 38 31 27 20 St. Dev. 2 5 4 2 Ex. 34 Avg.
35 28 21 18 St. Dev. 4 3 3 2 Ex. 41 Avg. 40 33 30 24 St. Dev. 6 4 5
4
[0334] It can be seen from these data that administration of
Compound A, the compound of Example 34, or the compound of Example
41 was effective in protecting cultured neurons from OGD-induced
cell death. At concentrations as low as 0.1 .mu.M, statistically
significant reduction in percent cell death was observed for these
compounds.
Effect of cPLA.sub.2 Inhibitor in Models of Parkinson's Disease
[0335] The effect of administration of a cPLA.sub.2 inhibitor in a
model for Parkinson's Disease was determined by the following
procedures.
Dopaminergic Neuron Cultures
[0336] Primary dopaminergic neurons were isolated from E15 rat
embryos as described in Pong K., et al., (1997) J. Neurochem. 69
986-994. Briefly, the ventral mesencephalon was isolated and tissue
was pooled in ice-cold phosphate buffer saline (PBS) without
Ca.sup.2+ and Mg.sup.2+. The tissue was transferred to an enzymatic
dissociation media containing 20 IU/ml papain in Earle's balanced
salt solution (Worthington Biochemical, Freehold, N.J.) and
incubated for 30 minutes at 37.degree. C. After enzymatic
dissociation, the papain solution was aspirated and the tissue
mechanically triturated with a fire-polished Pasteur pipette in
complete media [Neurobasal Medium with B-27 supplement (Gibco,
Grand Island, N.Y.), penicillin/streptomycin, aphidicolin,
glutamate] containing 2,000 IU/ml DNase and 10 mg/ml ovomucoid
protease inhibitor. Single-cell suspensions in complete media were
plated on pre-coated poly-L-ornithine/laminin 24-well plates
(Becton-Dickinson, Bedford, Mass.) at a density of
5.0.times.10.sup.5 cells/well. Cells were maintained for one week
prior to experimentation.
MPP.sup.+ Exposure in Dopaminergic Neurons
[0337] Cultures were treated with various concentrations of
Compound A, Compound B, Compound C and GDNF (glial-cell line
derived neurotrophic factor, a positive control) hours before
exposure to the neurotoxin MPP.sup.+, the toxic metabolite of MPTP.
Cultures were exposed to 10 .mu.M MPP.sup.+ for 60 minutes. After
the exposure, fresh media was exchanged and fresh compound was
added. Dopaminergic neuron viability was determined 24 hours later
by measuring .sup.3H-dopamine uptake as described in Pong et al.,
1997, supra. The results are shown in the Table below:
TABLE-US-00008 Neuroprotectin by cPLA2 inhibitors against MPP.sup.+
Control 10 .mu.M MPI 0.3 .mu.M 1 .mu.M 3 .mu.M 10 .mu.M Avg. 100
56.6 St. Dev, 8.2 2.2 Cpd. A Avg. 76.9 83.4 78.8 81.1 St. Dev. 3.4
5.7 3.3 6.4 Cpd. B Avg. 74 71.6 78.6 83.2 St. Dev. 3 2 5.5 5.1 Cpd.
C Avg. 68.6 70.6 75.9 79.6 St. Dev. 3.8 3.6 2.6 1.3
[0338] It can be seen from these data that administration of these
compounds were effective to protect dopaminergic neuron viability
against MPP.sup.+.
Effects of cPLA.sub.2 Inhibitor in Models of Osteoarthritis,
Rheumatoid Arthritis and Pain
[0339] In vivo pharmacology studies using the compound of Example
10 were conducted to demonstrate the effectiveness of oral
administration in models of inflammation and peripheral pain
including the carrageenan paw edema model (See Winter, C. A., et
al., Proc Soc Exp Biol Med 1962; 111:544-547), the collagen induced
arthritis model (See Trentham, D. E., et al., J. Exp. Med. 146;
828-833), and the Complete Freund's Adjuvant (CFA)-induced model of
hyperalgesia (See Stein C, et al., Pharmacology Biochemistry &
Behavior, 1988; 31:445-451). The in vivo inhibition of
prostaglandins and leukotrienes was also measured in the
CFA-challenged paws in the hyperalgesia model.
Carrageenan Paw Edema Assay
[0340] The carrageenan paw edema assay is an acute model of
inflammation that is particularly useful for in vivo assessment of
compounds that effect the production of prostaglandins. In
particular, NSAIDs inhibit edema in a characteristic dose response
fashion in this model, and the activity of NSAIDs in this model
correlates well with the activity observed in man (See Mukherjee A,
et al., Inflamm Res. 1996; 45:531-540). Therefore, the compound of
Example 10 was tested in the carrageenan paw edema model. In this
model, the compound was administered orally 2 hours before
sub-plantar injection of carrageenan and the inhibition of paw
swelling was determined over the next three hours. Paw edema was
statistically significantly decreased at doses as low as 3 mg/kg
and an approximate ED50 (based on a maximal inhibition of 50%) was
determined to be 7.5 mg/kg.
[0341] These data demonstrate that compound of Example 10 works in
a classic model of in vivo inflammation that has been used to
predict the effectiveness of both NSAIDs and COX-2 inhibitors.
Effect of Compound of Example 10 in the Collagen-Induced Arthritis
Model
[0342] The compound of Example 10 was tested in the mouse CIA
model, which has many immunologic and pathologic similarities to
human rheumatoid arthritis (See Trentham, D. E., et al., supra).
Arthritis was induced in DBA/1LacJ mice by intradermal injection of
an emulsion of bovine type II collagen and CFA followed by a boost
with an intradermal injection of bovine type II collagen emulsified
in incomplete Freund's Adjuvant 21 days after the initial
immunization. Compound efficacy was assessed in a semi-therapeutic
dosing regimen that was initiated when 10% of the animals showed
disease symptoms. At that point animals were randomly assigned to
treatment groups and administered the compound of Example 10 (100
mg/kg) PO BID for 28 days. Control groups received celecoxib,
vehicle alone or were left untreated. All animals were scored daily
in a blinded fashion for visual signs of disease symptoms.
[0343] The mean scores of the group treated with the compound of
Example 10 were compared to the vehicle-control group values using
the Student's t-test. During treatment starting on day 10, the
group treated with the compound of Example 10 (100 mg/kg BID)
showed a statistically significant decrease in the disease severity
scores in all experiments; and the number of animals without
disease symptoms was greatest in the groups that were treated with
the compound of Example 10.
[0344] After completion of the experiments, paws were processed for
histology. Two board certified veterinary pathologist evaluated the
slides in a blinded fashion. Each paw was assigned a numerical
score for both arthritis severity and the general number of joints
affected. Mice treated with the compound of Example 10 (100 mg/kg
BID) had the lowest group mean severity scores and the highest
percentage of unaffected (grade 0) paws; vehicle-treated and
untreated mice had the highest group mean severity scores and the
highest combined percentage of grade 3 and grade 4 affected paws.
Mice treated with the compound of Example 10 (100 mg/kg) had an
average severity grade of 0.9/2.1 (pathologist 1/pathologist 2)
whereas vehicle treated animals had an average severity score of
2.1/3.0. Similar results were seen in an additional experiment at
100 mg/kg.
Effect of Compound of Example 10 in Rat Hyperalgesia Models
[0345] The sensitivity of peripheral sensory neurons can be
enhanced such that they respond to both noxious and non-noxious
stimuli resulting in chronic pain (See Julius, D. et al., Nature.
2001; 413:203; Woolf, C. J., et al., Science. 200; 288:1765).
Prostaglandins and leukotrienes at the site of inflammation and
tissue damage are partially responsible for this potentiation of
the pain response. Prostaglandins promote the phosphorylation of
ion channels, increasing the excitability and lowering the pain
threshold of sensory neurons. Analogously, leukotriene B.sub.4 and
related arachidonate metabolites of 12-LO bind to and activate the
capsaicin receptor (or VR.sub.1) ion channel on neurons that
respond to heat and low pH (See Piomelli D., TRENDS in
Pharmacological Sciences, January 2001; 22(1):17-29). The effect of
the compound of Example 10 was measured in the CFA-induced
hyperalgesia model, and lipid mediator production was measured at
peripheral and central sites.
[0346] The animals were dosed with vehicle, the compound of Example
10, naproxen or celecoxib, and then CFA was immediately injected
into the hind footpad. To assess hyperalgesia, pressure was applied
to the left hind paw at a slow and constant rate using a digital
force gauge. Measurements were taken at 0 and 6 hours. The
application of the force was stopped when the animal vocalized, or
struggled. Readings were taken prior to dosing and CFA injection,
and repeated six hours after the CFA injection. Two independent
experiments were run and the data were analyzed separately. The
compound of Example 10 appeared to provide a statistically
significant decrease In pain compared to the vehicle control group
at 25 mg/kg.
[0347] The paws were collected at the end of each experiment (6
hours) and the levels of PGE.sub.2, LTB.sub.4 and TXB.sub.2 were
measured in the exudates. PGE.sub.2 levels in the paw were
significantly inhibited by the compound of Example 10 (at 25 mg/kg)
and by the celecoxib and naproxen controls. TXB.sub.2 levels were
also significantly inhibited by celecoxib, however, the inhibition
with the compound of Example 10 and naproxen was greater than the
inhibition with celecoxib, suggesting a COX-1 dependent component
to the synthesis. As expected, LTB.sub.4 levels were significantly
inhibited by the compound of Example 10, but there was evidence of
substrate shunting to the 5-lipoxygenase pathway as levels actually
increased with naproxen and celecoxib.
[0348] In summary, the compound of Example 10 was active in models
of osteoarthritis, rheumatoid arthritis and pain. The compound
significantly inhibited edema at a dose of 3 mg/kg and was at 50%
of the maximum effect at .about.7.5 mg/kg in the carrageenan paw
edema model. Daily treatment with the compound of Example 10 (100
mg/kg BID) for 28 days produced a significant reduction of disease
in the semi-therapeutic collagen-induced arthritis model based on
both clinical and histological assessment. The compound was also
effective at 25 mg/kg in the CFA model of hyperalgesia.
[0349] The compound of Example 10 was also effective at inhibiting
the production of both prostaglandins and leukotrienes using in
vivo models. The production of COX-2 dependent PGE2, COX-1
dependent Thromboxane and 5-LO dependent leukotriene B4 was
inhibited in paws challenged with CFA.
Effect of Compound of Example 10 in Rodent and Sheep Models of
Asthma
[0350] Asthma has been defined as a chronic inflammatory disorder
of the airways in which many cells and cellular elements play a
role. In susceptible individuals this inflammation causes recurrent
or persistent episodes of wheezing, breathlessness, chest tightness
and coughing, particularly at night or in the early morning. These
episodes are usually associated with widespread but variable
airflow obstruction that often resolves spontaneously or with
treatment. The inflammation also causes an associated increase in
the existing bronchial hyperresponsiveness to a variety of stimuli.
Preclinical models of asthma have provided insight into the
underlying mechanisms of disease pathology and have been
instrumental in the development of asthma therapeutics. In
particular, rodent models of allergen-induced pulmonary
inflammation are useful for in vivo assessment of compounds that
inhibit the inflammation associated with allergic asthma and have
been used extensively to evaluate the efficacy of glucocorticoids,
leukotriene receptor antagonists, 5-L0 inhibitors, and
phosphodiesterase 4 inhibitors (See Kumar, R. K. et al, J Pharmacol
Exp Ther. 2003; 307:349-355; Wu, A. Y. et al. Clin Exp Allergy.
2003; 33:359-366; Bell, R. L. et al., J. Pharmacol Exp Ther 1997;
280:1366-1373; and Henderson, W. R., Jr., et al., J Exp Med. 1996;
184:1483-1494).
[0351] The compound of Example 10 was tested in both rat and mouse
models of allergen-induced pulmonary inflammation.
[0352] In addition to the allergen induced pulmonary inflammation
models in rodents, allergen induced changes in lung function are
often evaluated in allergic sheep. Ascaris sensitized sheep that
are challenged via the airways with Ascaris suum antigen exhibit
features of reversible airway narrowing and AHR. Studies performed
in this animal model present strong evidence that the release of
arachidonic acid metabolites plays an important role in the
development of late bronchial responses to antigen challenge (See
Abraham, W. M., et al, Respiration. 1989; 56:48-56). Thus, the
compound of Example 10 was evaluated for effects on allergen
induced changes in lung function In a sheep model of asthma.
Rat Antigen Induced Pulmonary Inflammation Model
[0353] The efficacy of the compound of Example 10 was evaluated in
a Brown Norway rat model in which ovalbumin (OVA)-sensitized
animals were challenged via the airways with an aerosol of
ovalbumin (OVA) on days 1 and 2. OVA sensitized rats were
challenged via aerosol on day 1 and day 2. The compound of Example
10 was administered at 30 mg/kg PO BID 1 hour prior to challenge
and 10 hours after challenge over the 2 day challenge period.
Dexamethasone was administered at 3 mg/kg IP 1 hour prior to
challenge on day 1 and day 2. Animals were sacrificed on day 3 and
bronchoalveolar cavities ravaged for analysis of cellular influx.
Oral BID administration at 30 mg/kg over the 2 day challenge period
statistically significantly inhibited Bronchoalveolar lavage fluid
(BALF) eosinophil influx in 8 out of 8 independent studies. The
compound of Example 10 also statistically significantly attenuated
the total numbers of inflammatory cells within the BALF at the dose
tested but had no significant effect on the influx of lymphocytes
or neutrophils in this model.
Effect of Compound of Example 10 in a Sheep Model of
Antigen-Induced Early and Late-Phase Bronchoconstriction and
AHR
[0354] Allergen-induced reversible airway narrowing and AHR are two
hallmark features of allergic asthma that can be examined in vivo
in a sheep model of asthma. Studies performed in this animal model
present strong evidence that the release of arachidonic acid
metabolites plays an important role in the development of late
bronchial responses to antigen challenge (See Abraham, W. M., et
al., Respiration. 1989; 56:48-56). The release of leukotrienes
through the LO pathway during the acute bronchial constriction
after inhalation of Ascaris suum antigen represents the key factor
for the initiation of the subsequent events, namely the late-phase
response and the bronchial hyperreactivity. The 5-LO Inhibitor
zileuton blocks antigen-induced late airway responses,
inflammation, and AHR in this model, whereas a continuous IV
infusion of the selective LTD.sub.4 receptor antagonist,
montelukast, attenuates both the early and late-phase asthmatic
responses (See Abraham, W. M., et al., Eur J Pharmacol 1992;
217:119-126; Jones, T. R. et al., Can J Physiol Pharmacol. 1995;
73:191-201). In addition, oral administration of a dual
LTD.sub.4/TXB.sub.2 inhibitor can inhibit both the early and
late-phase response, as well as AHR to carbachol and histamine (See
Abraham, W. M., et al., J Pharmacol Exp Ther. 1988; 247:1004-1011).
PAF has also been implicated in the late-phase response in this
model providing further support for the concept that a more
complete blockade of lipid mediators by a cPLA.sub.2.alpha.
antagonist may provide better clinical efficacy compared with
current anti-leukotrienes (See Abraham, W. M., et al., J Appl
Physiol. 1989; 66:2351-2357).
[0355] The compound of Example 10 was administered at 3 mg/kg BID
(PO) 24 h prior to challenge, 2 h prior to challenge and 8 hr post
challenge. The mean % increase in airway resistance for 3
individual sheep over the ensuing 8 h period was determined.
Complete blockade of the late asthmatic response was observed
[0356] The following day, airway hyperresponsiveness (AHR) was
assessed in these same treated sheep by determining the cumulative
carbachol concentration that increased specific lung resistance by
400%. Treatment with the compound of Example 10 resulted in
complete blockade of airway hyperresponsiveness. In an extended
dosing regimen, compound of Example 10 was administered at 3 mg/kg
PO BID for 4 days before challenge, 2 hours before challenge on the
fifth day and 8 hr post challenge.
[0357] The mean % increase in airway resistance for 5 individual
sheep over the ensuing 8 h period was determined, and in this more
extended dose regimen, there was a modest but statistically
significant inhibition of the early asthmatic response in addition
to a complete blockade of the late-phase response and a complete
blockade of AHR to aerosolized carbachol.
[0358] The foregoing data show that the compound of Example 10 is a
potent inhibitor of allergen induced pulmonary inflammation,
bronchoconstriction and AHR in animal models of asthma.
[0359] The compounds of the invention inhibit cPLA2 activity that
is required for supplying arachidonic acid substrate to
cyclooxygenase-1 or 2 and 5-lipoxygenase, which in turn initiate
the production of prostaglandins and leukotrienes respectively. In
addition, cPLA.sub.2 activity is essential for producing the
lyso-phospholipid that is the precursor to PAF. Thus these
compounds are useful in the treatment and prevention of disease
states in which leukotrienes, prostaglandins or PAF are involved.
Moreover, in diseases where more than one of these agents plays a
role, a cPLA.sub.2 inhibitor would be expected to be more
efficacious than leukotriene, prostaglandin or PAF receptor
antagonists and also more effective than cyclooxygenase or
5-lipoxygenase inhibitors.
[0360] Therefore, the compounds, pharmaceutical compositions and
regimens of the present invention are useful in treating and
preventing the disorders treated by cyclooxygenase-2,
cycloxygenase-1, and 5-lipoxygenase inhibitors and also antagonists
of the receptors for PAF, leukotrienes or prostaglandins. Diseases
treatable by compounds of this invention include but are not
limited to: pulmonary disorders including diseases such as asthma,
chronic bronchitis, and related obstructive airway diseases;
allergies and allergic reactions such as allergic rhinitis, contact
dermatitis, allergic conjunctivitis, and the like; inflammation
such as arthritis or inflammatory bowel diseases, skin disorders
such as psoriasis, atopic eczema, acne, UV damage, burns and
dermatitis; cardiovascular disorders such as atherosclerosis,
angina, myocardial ischaemia, hypertension, platelet aggregation,
and the like; and renal insufficiency induced by immunological or
chemical. The drugs may also be cytoprotective, preventing damage
to the gastrointestinal mucosa by noxious agents. The compounds
will also be useful in the treatment of adult respiratory distress
syndrome, endotoxin shock and ischeamia induced injury including
myocardial or brain injury.
[0361] The methods of treatment, inhibition, alleviation or relief
of asthma of this invention include those for Extrinsic Asthma
(also known as Allergic Asthma or Atopic Asthma), Intrinsic Asthma
(also known as Nonallergic Asthma or Nonatopic Asthma) or
combinations of both, which has been referred to as Mixed Asthma.
The methods for those experiencing or subject to Extrinsic or
Allergic Asthma include incidents caused by or associated with many
allergens, such as pollens, spores, grasses or weeds, pet danders,
dust, mites, etc. As allergens and other irritants present
themselves at varying points over the year, these types of
incidents are also referred to as Seasonal Asthma. Also included in
the group of Extrinsic Asthmas is bronchial asthmas and allergic
bronchopulminary aspergillosis.
[0362] Intrinsic Asthmas that may be treated or alleviated by the
present methods include those caused by infectious agents, such as
cold and flu viruses in adults and respiratory syncytial virus
(RSV), rhinovirus and influenza viruses common in children. Also
included are the asthma conditions which may be brought about in
some asthmatics by exercise and/or cold air. The methods are useful
for Intrinsic Asthmas associated with industrial and occupational
exposures, such as smoke, ozone, noxious gases, sulfur dioxide,
nitrous oxide, fumes, including isocyanates, from paint, plastics,
polyurethanes, varnishes, etc., wood, plant or other organic dusts,
etc. The methods are also useful for asthmatic Incidents associated
with food additives, preservatives or pharmacological agents.
Common materials of these types are food coloring such as
Tartrazine, preservatives like bisulfites and metabisulfites, and
pharmacological agents such as aspirin and non-steroidal
anti-inflammatory agents (NSAIDs). Also included are methods for
treating, inhibiting or alleviating the types of asthma referred to
as Silent Asthma or Cough Variant Asthma.
[0363] A further method of treatment of asthma of this invention
comprises administering to a mammal in need of such treatment a
pharmaceutically effective amount of a compound of this invention,
as described above, and a pharmaceutically effective amount of one
or more additional anti-asthma agents.
[0364] Anti-asthma agents useful with these combinations include
long-term-control medications, such as corticosteroids
(glucocorticoids), cromolyn sodium (disodium cromoglycate--DSCG),
nedocromil, methylxanthines (such as theophylline) and leukotriene
modifiers. Useful leukotriene modifiers include leukotriene
receptor antagonists, such as zafirlukast (ACCOLATE.RTM.) and
monetlukast (SINGULAIR.RTM.), and 5-lipoxygenase inhibitors, such
as zileuton (ZYFLO.RTM.). Useful corticosteroids include inhaled
products, such as Beclomethasone dipropionate, Budesonide,
Flunisolide, Fluticasone, and Triamcinolone, as well as the
pharmaceutically acceptable salt forms thereof. Also useful are
systemic corticosteroids such as prednisone, prednisolone and
methylprednisolone.
[0365] Also useful are quick-relief anti-asthma medications, such
as long-acting beta.sub.2-agonists, short-acting
beta.sub.2-agonists, anticholinergics and systemic corticosteroids.
.beta.-Adrenergic agents which may be used include epinephrine,
isoproterenol, metaproterenol, terbutaline, isoetharine, albuterol,
bitolterol and perbuterol. Useful anticholinergic agents include
atropine (and its derivative ipatropium bromide) and
glycopyrrolate. The compounds of this invention may also be used to
treat asthma in conjunction with allergy immunotherapies, which
also referred to in the art as hyposensitization therapies. These
compounds may be administered according to the dosages and regimens
known in the art.
[0366] Additional anti-asthma agents which may be used in the
combinations of this invention include pranlukast, anakinra,
seratrodast, olopatadine hydrochloride, cromoglicate lisetil,
ramatroban, interleukin4 receptor (Immunex), urodilatin, colforsin
daropate, salbutamol, LCB-2183, andolast, ciclesonide, budesonide,
formoterol, omalizumab, tranilast, saredutant, CDP-835 (anti-IL-5
Mab), fexofenadine HCl,
N-(1-(Chlorophenyl)-1-methylethyl)-3-(imidazol-1-yl)
propylaminedihydrochloride (BTS-71-321), cilomilast, bimosiamose,
Corticotropin-releasing factor, clenoliximab, tiotropium bromide,
2H-1,2-Benzoselenazine, 3,4-dihydro-4,4-dimethyl (BXT-51072),
atreleuton, (R)-salbutamol,
8-Methoxyquinoline-5-(N-(2,5-dichloropyridin-3-yl)) carboxamide
(D-4418), triamcinolone acetonide, KW-4490 (KF-19514), LAX-300
(LX-109), IDEC-152 (ST-152; anti-CD23 antibody), cytokine Traps,
anandamide, SRL-172, salmeterol+Fluticasone, KCA-757,
2-Pyridinecarboxylic acid,
6-(2-(3,4-diethoxyphenyl)-4-thiazolyl)-(OPC-6535), PM-56D9,
salbutamol, CT-2820 (PDEIV inhibitors), beclometasone, nepadutant,
ketotifen fumarate, DHEAS (PB-005), Pharmaprojects No. 5163, No.
5278 and No. 5297, salbutamol sulfate, EPI-2010 (EpiGenRx),
mepolizumab, Benzamide,
N-(5-(3-((4-chlorophenyl)sulfonyl)propyl)-2-(1H-tetrazol-5-ylmethoxy)phen-
yl)-3-((4-(1,1-dimethylethyl)-2-thiazolyl)methoxy)-, monosodium
salt (YM-158),
2-(4-ethoxycarbonylaminobenzyl)-6-(3,4-dimethoxyphenyl)-2,3,4,5-
-tetrahydro-pyridazin-3-one Pharmaprojects (No. 5450), Sch-205528,
L-826141 (Pharmaprojects No. 5477), Budesonide, duramycin,
4,4-Bis(4-quinolin-2-ylmethoxy)phenyl)pentanoic acid sodium salt
(VML-530), IL-9 inhibitor, beclometasone dipropionate, formoterol,
cyclo(MePhe-Leu-Asp-Val-D-Arg-D-Arg) (ZD-7349), salbutamol,
Ethanaminium,2-(((2-acetyl-4-((1-oxohexadecyl)amino) phenoxy)
hydroxyphosphinyl)oxy)-N, N,N-trimethyl-, inner salt (CPR-2015),
PD-168787 (CI-1018), cathepsin S inhibitors, SB-240683 (anti-IL-4
Mab), BIIL-284, APC-2059, budesonide+formoterol, Bay-16-9996 (IL-4
antagonist), beclometasone, GW-328267, VLA-4 antagonists,
4-hydroxy-1-methyl-3-octyloxy-7-sinapinoylamino-2(1H)-quinolinone
(TA-270), CpG-7909 (ProMune), DNK-333A (Pharmaprojects No. 6070),
AWD-12-281, LM-1507 (LM-1484), formoterol, MOL-6131, cathepsin S
inhibitors, CS-615, ibudilast,
2-{N-(4-(4-Chlorophenylsulfonylamino)butyl)-N-{3-(2-(4-cyclobutylthiazol--
2-yl)ethyl)benzyl}sulfamoyl}benzoic acid (S-36527), and
2-{N-(4-(4-Chlorophenylsulfonylamino)butyl)-N-{3-((4-isopropylthiazol-2-y-
l)methyloxy)benzyl}sulfamoyl}benzoic acid (S-36496).
[0367] The methods herein are also useful for treatment and
alleviation of Intrinsic Asthma associated with gastroesophageal
reflux (GERD), which can stimulate bronchoconstriction. GERD, along
with retained bodily secretions, suppressed cough, and exposure to
allergens and irritants in the bedroom can contribute to asthmatic
conditions and have been collectively referred to as Nighttime
Asthma or Nocturnal Asthma. In methods of treatment, inhibition or
alleviation of asthma associated with GERD, a pharmaceutically
effective amount of the compounds of this invention may be used as
described herein in combination with a pharmaceutically effective
amount of an agent for treating GERD. These agents include, but are
not limited to, proton pump inhibiting agents like PROTONIX.RTM.
brand of delayed-release pantoprazole sodium tablets, PRILOSEC.RTM.
brand omeprazole delayed release capsules, ACIPHEX.RTM. brand
rebeprazole sodium delayed release tablets or PREVACID.RTM. brand
delayed release lansoprazole capsules.
[0368] The compounds of this invention can be used as an
antipyretic agent. The compounds of this Invention may be utilized
in methods of treating pain, particularly the pain associated with
inflammation. Specific methods include, but are not limited to,
those for treating centrally mediated pain, peripherally mediated
pain, musculo-skeletal pain, lumbosacral pain, structural or soft
tissue injury related pain, progressive disease related pain, such
as oncology and degenerative disorders, neuropathic pain, which can
include both acute pain, such as acute injury or trauma, pre and
post-surgical, migraine pain, dental pain, etc., chronic pains,
such as neuropathic pain conditions of diabetic peripheral
neuropathy, post-herpetic neuralgia and fibromyalgia, and
inflammatory conditions such as osteoarthritis or rheumatoid
arthritis, sequela to acute injury or trauma and cancer-related
pain.
[0369] The compounds of this invention can be used to alleviate,
inhibit, relieve and/or treat arthritic disorders in a mammal
including, but not limited to, rheumatoid arthritis,
spondyloarthropathies, gouty arthritis, infectious arthritis,
osteoarthritis (which includes erosive osteoarthritis and is also
known as osteoarthrosis or degenerative joint disease or DJD),
systemic lupus erythematosus and juvenile arthritis. Each of these
methods comprises administering to a mammal in need of such action
a pharmaceutically effective amount of a substituted indole of this
invention, as described herein, or a pharmaceutically acceptable
salt or ester form thereof.
[0370] In addition, the compounds of this invention can be used to
alleviate, inhibit, relieve and/or treat arthritic conditions
associated with spondylitis, including ankylosing spondylitis,
reactive arthritis (Reiter's syndrome), psoriatic arthritis,
arthritis associated with chronic inflammatory bowel disease and
AIDS-related seronegative spondyloarthropathy.
[0371] This invention also provides methods for treating,
alleviating or inhibiting rheumatic disease and disorders. These
methods are useful for treatment of systemic lupus erythematosus,
systemic sclerosis and forms of scleroderma, polymyositis,
dermatomyositis, necrotizing vasculitis and other vasculopathies,
hypersensitivity vasculitis (including Henoch-Schonlein purpura),
Wegener's granulomatosis, Giant cell arteritis, mucocutaneous lymph
node syndrome (Kawasaki disease), Behcet's syndrome,
Cryoglobulinemia, juvenile dermatomyositis, Sjogren's syndrome,
overlap syndromes (includes mixed connective tissue disease),
polymyalgia rheumaticqa, erythema nodosum, relapsing
polychondritis, tendonitis (tenosynovitis), Bicipital tendenitis,
bursitis, Olecranon bursitis, adhesive capsulitis of the shoulder
(frozen shoulder) trigger finger, and Whipple's disease.
[0372] The methods of this invention are also useful for treatment,
alleviation or inhibition of metabolic and endocrine diseases with
rheumatic states, including gout, pseudogout, chondrocalcinosis,
amyloidosis, scurvy, specific enzyme deficiency states (including
Fabry's disease, alkaptonuria, ochonosisi, Lesch-Nyhan syndrome,
and Gaucher's disease), hyperlipoproteinemias (types II, IIa, IV),
Ehlers-Danlos syndrome, Marfan's syndrome, pseudoxanthoma
elasticum, Wilson's disease. Also treatable with the present
methods are the rheumatic states associated with endocrine
diseases, such as diabetes mellitus, acromegaly,
hyperparathyroidism, myositis ossificans progressiva, hypermobility
syndromes, arthrogryposis multiplex congenita, and thyroid diseases
such as thyroiditis, hypothyroidism and hyperthyroidism. These
methods may also be used for rheumatic conditions associated with
neoplasms such as primary neoplasms (synovioma), metastatic
neoplasms, multiple myeloma, leukemia and lymphomas, pigmented
villonodular synovitis, osteochondromatosis and others. Also
included among the methods of this invention are relief from the
rheumatic conditions associated with neuropathic disorders
including, Charcot's joints, hand-arm vibration syndrome (also
known as vibration-induced white finger or Raynaud's phenomenon),
repetitive stress syndromes, reflex sympathetic dystrophy and
compression neuropathies, such as peripheral entrapment (including
carpal tunnel syndrome, pronator syndrome, thoracic outlet
syndromes and tarsal tunnel syndrome), radiculopathy and spinal
stenosis.
[0373] This invention further provides a method of alleviation,
inhibition, relief or treatment of arthritic disorders in a mammal,
the method comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a chemical inhibitor of
phospholipase enzymes, particularly phospholipase A.sub.2 enzymes,
as defined herein and a pharmaceutically effective amount of an
anti-rheumatic agent.
[0374] Combinations for the treatment of arthritic disorders may
include commercially available anti-rheumatic agents such as, but
not limited to, naproxen, which is commercially available in the
form of EC-NAPROSYN.RTM. delayed release tablets, NAPROSYN.RTM.,
ANAPROX.RTM. and ANAPROX.RTM. DS tablets and
NAPROSYN.RTM.suspension from Roche Labs, CELEBREX.RTM. brand of
celecoxib tablets, VIOXX.RTM. brand of rofecoxib, CELESTONE.RTM.
brand of betamethasone, CUPRAMINE.RTM. brand penicillamine
capsules, DEPEN.RTM. brand titratable penicillamine tablets,
DEPO-MEDROL brand of methylprednisolone acetate injectable
suspension, ARAVA.RTM. leflunomide tablets, AZULFIDIINE
EN-tabs.RTM. brand of sulfasalazine delayed release tablets,
FELDENE.RTM. brand piroxicam capsules, CATAFLAM.RTM. diclofenac
potassium tablets, VOLTAREN.RTM. diclofenac sodium delayed release
tablets, VOLTAREN.RTM.-XR diclofenac sodium extended release
tablets, ENBREL.RTM. etanerecept products, (should we add other
biologics use in RA) and other commercially available antirheumatic
agents.
[0375] Also useful are GENGRAF.RTM. brand cyclosprine capsules,
NEORAL.RTM. brand cyclosprine capsules or oral solution,
IMURAN.RTM. brand azathioprine tablets or IV injection,
INDOCIN.RTM. brand indomethacin capsules, oral suspension and
suppositories, PEDIAPED.RTM. prednisolone sodium phosphate oral
solution, PLAQUENIL.RTM. brand hydroxychloroquine sulfate,
PRELONE.RTM. brand prednisolone syrup, REMICADE.RTM. infliximab
recombinant for IV injection, and SOLU-MEDROL.RTM.
methylprednisolone sodium succinate for injection.
[0376] Also useful in the combinations of this invention are gold
compounds and products useful in the treatment of arthritis and
rheumatic conditions, such as auranofin or MYOCHRISYINE.RTM. gold
sodium thiomalate injection.
[0377] Each of these products may be administered according to the
pharmaceutically effective dosages and regimens known in the art,
such as those described for the products in the Physicians' Desk
Reference, 55 Edition, 2001, published by Medical Economics Co.,
Inc., Montvale, N.J.
[0378] The compounds of this invention may also be administered in
the methods of this invention with analgesic and anti-inflammatory
agents such as NSAIDs and aspirin and other salicylates. Examples
of useful agents include ibuprofen (MOTRIN.RTM., ADVIL.RTM.),
naproxen (NAPROSYN.RTM.), sulindac (CLINORIL.RTM., diclofenac
(VOLTAREN.RTM.), piroxicam (FELDENE.RTM.) ketoprofen (ORUDIS.RTM.),
diflunisal (DOLOBID.RTM.), nabumetone (RELAFEN.RTM.), etodolac
(LODINE.RTM.), oxaprozin (DAYPRO.RTM.), indomethacin
(INDOCIN.RTM.), melicoxam (MOBICOX.RTM.), valdecoxib and
eterocoxib. Aspirin is anti-inflammatory when given in high doses,
otherwise it is just a pain killer like acetaminophen
(TYLENOL.RTM.).
[0379] Suitable cyclooxygenase 2 (COX-2) inhibitors-for use with
the methods of this invention include, but are not limited to,
2-(4-ethoxy-phenyl)-3-(4-methanesulfonyl-phenyl)-pyrazolo[1,5-b]pyridazin-
e, CDC-501, celecoxib, COX-189,
4-(2-oxo-3-phenyl-2,3-dihydrooxazol-4-yl)benzenesulfonamide,
CS-179, CS-502, D-1367, darbufelone, DFP, DRF-4367, flosulide,
JTE-522
(4-(4-cyclohexyl-2-methyl-5-oxazolyl)-2-fluorobenzenesulfonamide),
L-745337, L-768277, L-776967, L-783003, L-791456, L-804600,
meloxicam, MK663 (etoricoxib), nimesulide, NS-398, parecoxib,
1-Methylsulfonyl-4-(1,1-dimethyl-4-(4-fluorophenyl)cyclopenta-2,4-dien-3--
yl)benzene,
4-(1,5-Dihydro-6-fluoro-7-methoxy-3-(trifluoromethyl)-(2)-benzothiopyran
o(4,3-c)pyrazol-1-yl)benzenesulfonamide,
4,4-dimethyl-2-phenyl-3-(4-methylsulfonyl)phenyl)cyclobutenone,
4-Amino-N-(4-(2-fluoro-5-trifluoromethyl)-thiazol-2-yl)-benzene
sulfonamide,
1-(7-tert-butyl-2,3-dihydro-3,3-dimethyl-5-benzo-furanyl)-4-cyclopropyl
butan-1-one, Pharmaprojects No. 6089 (Kotobuki Pharmaceutical),
RS-1 13472, RWJ-63556, S-2474, S-33516, SC-299, SC-5755,
valdecoxib, UR-8877, UR-8813, UR-8880. Further suitable COX-2
inhibitors for use according to the invention include parecoxib,
MK663,
4-(4-cyclohexyl-2-methyl-5-oxazolyl)-2-fluorobenzenesulfonamide
(JTE-522), nimesulide, flosulide, DFP and
2-(4-ethoxy-phenyl)-3-(4-methanesulfonyl-phenyl)-pyrazolo[1,5-b]pyridazin-
e, and their physiologically acceptable salts, esters or
solvates.
[0380] Such compositions are also useful in the treatment of
menstrual cramps, preterm labor, tendonitis, bursitis, allergic
neuritis, cytomegalovirus infection, apoptosis, including
HIV-induced apoptosis, lumbago, liver disease including
hepatitis.
[0381] The methods are also useful in treating gastrointestinal
conditions such as inflammatory bowel disease, Crohn's disease,
gastritis, irritable bowel syndrome and ulcerative colitis and for
the prevention of treatment of cancer such as colorectal cancer.
The compounds and compositions of the present invention are also
useful for the prevention or treatment of benign and malignant
tumors/neoplasia including cancers such as colorectal cancer, brain
cancer, bone cancer, epithelial cell-derived neoplasia (epithelial
carcinoma) such as basal cell carcinoma, adenocarcinoma,
gastrointestinal cancer, including lip cancer, mouth cancer,
esophogeal cancer, small bowel cancer and stomach cancer, colon
cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian
cancer, cervical cancer, lung cancer, breast cancer, and skin
cancers, such as squamous cell and basal cell cancers, prostate
cancer, renal cell carcinoma, and other known cancers that effect
epithelial cells throughout the body. Neoplasias for which
compositions of the invention are contemplated to be particularly
useful are gastrointestinal cancer, Barrett's esophagus, liver
cancer, bladder cancer, pancreas cancer, ovarian cancer, prostatic
cancer, cervical cancer, lung cancer, breast cancer, and skin
cancer, such as squamous cell and basal cell cancers. The compounds
and methods of this invention can also be used to treat the
fibrosis occurring with radiation therapy. Such compositions can be
used to treat subjects having adenomatous polyps, including those
with familial adenomatous polyposis (FAP). Additionally, such
compositions can be used to prevent polyps from forming in patients
at risk of FAP. Compounds of this invention are useful in the
treatment of cancers because of their anti-angiogenic effects.
[0382] Further uses include treating inflammation in such diseases
as vascular diseases, migraine headaches, periarteritis nodosa,
thyroiditis, aplastic anemia, Hodgkin's disease, sclerodoma,
rheumatic fever, type I diabetes, neuromuscular junction disease
Including myasthenia gravis, white matter disease including
multiple sclerosis, sarcoidosis, nephrotic syndrome, Behcet's
syndrome, polymyositis, gingivitis, nephritis, hypersensitivity,
swelling occurring after injury including brain edema, myocardial
ischemia, and the like. Also included are treatments of ophthalmic
diseases, such as retinitis, conjunctivitis, retinopathies,
uveitis, ocular photophobia, and of acute injury to the eye tissue.
The compounds of this invention will be useful in the treatment of
post-operative inflammation including that following ophthalmic
surgery such as cataract surgery or refractive surgery. Also
included are treatments of pulmonary and upper respiratory tract
inflammation, such as that associated with viral infections and
cystic fibrosis, and in bone resorption such as that accompanying
osteoporosis. These compounds and compositions are useful for the
treatment of certain central nervous system disorders, such as
cortical dementias including Alzheimer's disease,
neurodegeneration, and central nervous system damage resulting from
stroke, ischemia and trauma. The compounds of this invention may
also be useful in the treatment of Parkinson's disease.
[0383] Methods of treating pain comprise administering to a mammal
subject to such pain a pharmaceutically effective amount of a
compound of this invention alone or in combination with one or more
additional pharmaceutically effective agents for the treatment of
pain or inflammation or the related underlying medical condition.
Examples of drug agents which may be combined with the present
compounds are analgesics, anti-angiogenic agents, anti-neoplastic
agents, These compounds may also be combined with anti-epileptic
compounds that have pain alleviating properties, such as gabapentin
and pregabalin.
[0384] One such combination method of this invention comprises
administering to a mammal in need thereof a pharmaceutically
effective amount of a compound of this invention and a
pharmaceutically effective amount of a nontoxic
N-methyl-D-aspartate (NMDA) receptor antagonist and/or an agent
that blocks at least one major intracellular consequence of NMDA
receptor activation. Examples of NMDA receptor antagonists useful
in these methods include dextromethorphan, dextrorphan, amantadine
and memantine, or the pharmaceutically acceptable salts
thereof.
[0385] Another method herein of treating inflammation and
inflammatory disorders comprises the co-administration to a mammal
in need thereof of an inhibitor of induced nitric oxide synthase
with a compound of this invention. Administration of this
combination is useful for prophylactic or therapeutic
administration in a mammal experiencing or subject to an abnormally
low level of nitric oxide synbthase (NOS) activity, particularly
those subject to hypertension or an elevated risk of pulmonary
hypertension, ischemic stroke, myocardial infarction, heart
failure, progressive renal disease, thrombosis, reperfusion injury,
or a nervous system degenerative disorder, such as Alzheimer's
disease, or those chronically exposed to hypoxic conditions.
[0386] The methods of this invention also include those for
treating or preventing a neoplasia disorder in a mammal, including
a human, in need of such treatment or prevention. The method
comprises treating the mammal with a therapeutically effective
amount of a compound of this invention in combination with an MMP
inhibitor. These two components may further be optionally combined
with one or more agents selected from an antiangiogenesis agent, an
antineoplastic agent, an adjunctive agent, an immunotherapeutic
agent, an analgesic agent; and/or a radiotherapeutic agent. One
such multiple component therapy comprises administering to the
mammal in need thereof a compound of this invention, a matrix
metalloproteinase inhibitor and an antineoplastic agent.
[0387] The methods and combinations of this invention may be used
for the treatment or prevention of neoplasia disorders including
acral lentiginous melanoma, actinic keratoses, adenocarcinoma,
adenoid cycstic carcinoma, adenomas, adenosarcoma, adenosquamous
carcinoma, astrocytic tumors, bartholin gland carcinoma, basal cell
carcinoma, bronchial gland carcinomas, capillary, carcinoids,
carcinoma, carcinosarcoma, cavernous, cholangiocarcinoma,
chondosarcoma, choriod plexus papilloma/carcinoma, clear cell
carcinoma, cystadenoma, endodermal sinus tumor, endometrial
hyperplasia, endometrial stromal sarcoma, endometrioid
adenocarcinoma, ependymal, epitheloid, Ewing's sarcoma,
fibrolamellar, focal nodular hyperplasia, gastrinoma, germ cell
tumors, glioblastoma, glucagonoma, hemangiblastomas,
hemangioendothelioma, hemangiomas, hepatic adenoma, hepatic
adenomatosis, hepatocellular carcinoma, insulinoma, intaepithelial
neoplasia, interepithelial, squamous cell neoplasia, invasive
squamous cell carcinoma, large cell carcinoma, leiomyosarcoma,
lentigo maligna melanomas, malignant melanoma, malignant
mesothelial tumors, medulloblastoma, medulloepithelioma, melanoma,
meningeal, mesothelial, metastatic carcinoma, mucoepidermoid
carcinoma, neuroblastoma, neuroepithelial adenocarcinoma nodular
melanoma, oat cell carcinoma, oligodendroglial, osteosarcoma,
pancreatic polypeptide, papillary serous adenocarcinoma, pineal
cell, pituitary tumors, plasmacytoma, pseudosarcoma, pulmonary
blastoma, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma,
sarcoma, serous carcinoma, small cell carcinoma, soft tissue
carcinomas, somatostatin-secreting tumor, squamous carcinoma,
squamous cell carcinoma, submesothelial, superficial spreading
melanoma, undifferentiated carcinoma, uveal melanoma, verrucous
carcinoma, vipoma, well differentiated carcinoma, and Wilm's
tumor.
[0388] Antineoplastic agents useful in the combination therapies
herein include anastrozole, calcium carbonate, capecitabine,
carboplatin, cisplatin, Cell Pathways CP-461, docetaxel,
doxorubicin, etoposide, fluorouracil, fluoxymestrine, gemcitabine,
goserelin, irinotecan, ketoconazole, letrozol, leucovorin,
levamisole, megestrol, mitoxantrone, paclitaxel, raloxifene,
retinoic acid, tamoxifen, thiotepa, topotecan, toremifene,
vinorelbine, vinblastine, vincristine, selenium (selenomethionine),
ursodeoxycholic acid, sulindac sulfone, exemestane and eflornithine
(DFMO),
1-[4-(2-Azepan-1-yl-ethoxy)-benzyl]-2-(4-hydroxy-phenyl)-3-methyl-1H-indo-
l-5-ol (also known as TSE-424) and
2-(4-Hydroxy-phenyl)-3-methyl-1-[4-(2-piperidin-1-yl-ethoxy)-benzyl]-1H-i-
n dol-5-ol (also known as ERA-923).
[0389] This invention also includes methods of utilizing the
compounds herein in combination with a proteinaceous interleukin-1
inhibitor, such as an IL-1 receptor antagonist (IL-Ira), for
preventing or treating inflammatory diseases in a mammal. Acute and
chronic interleukin-1 (IL-1)-mediated inflammatory diseases of
interest in these methods include, but is not limited to acute
pancreatitis; ALS; Alzheimer's disease; cachexia/anorexia; asthma;
atherosclerosis; chronic fatigue syndrome, fever; diabetes (e.g.,
insulin diabetes); glomerulonephritis; graft versus host rejection;
hemohorragic shock; hyperalgesia, inflammatory bowel disease;
inflammatory conditions of a joint, including osteoarthritis,
psoriatic arthritis and rheumatoid arthritis; ischemic injury,
including cerebral ischemia (e.g., brain injury as a result of
trauma, epilepsy, hemorrhage or stroke, each of which may lead to
neurodegeneration); lung diseases (e.g., ARDS); multiple myeloma;
multiple sclerosis; myelogenous (e.g., AML and CML) and other
leukemias; myopathies (e.g., muscle protein metabolism, esp. in
sepsis); osteoporosis; Parkinson's disease; pain; pre-term labor;
psoriasis; reperfusion injury; septic shock; side effects from
radiation therapy, temporal mandibular joint disease, tumor
metastasis; or an inflammatory condition resulting from strain,
sprain, cartilage damage, trauma, orthopedic surgery, infection or
other disease processes.
[0390] This invention also provides a method of administering one
or more of the compounds of this invention to a female in need
thereof to substantially prevent or reducing changes in the
female's reproductive system associated with onset or continuation
of labor. Also provided is a method of substantially preventing or
reducing uterine contractility either occurring during pregnancy or
associated with menorrhagia. These methods may optionally include
coadministration of a compound of this invention with a
progestogen, a progestin or a progestational agent.
[0391] Cytosolic phospholipase A.sub.2.alpha. (cPLA.sub.2.alpha.)
is a ubiquitously expressed enzyme that preferentially mediates the
release of arachidonic acid upon cell activation. Bioactive
metabolites of arachidonic acid, the eicosanoids, are recognized as
important modulators of platelet signaling. Inhibitors of the
eicosanoid pathway (e.g. aspirin) reduce the formation of
thromboxane A.sub.2 (TXA.sub.2), a labile and potent platelet
agonist, resulting in depression of platelet function, thrombus
formation, and proven clinical benefit in reducing morbidity and
mortality.
[0392] The compounds of the invention inhibit cPLA.sub.2 activity
that is required for supplying arachidonic acid substrate to
cyclooxygenase -1 or 2 and 5-lipoxygenase, which in turn initiate
the production of prostaglandins and leukotrienes respectively. In
addition, cPLA.sub.2 activity is essential for producing the
lyso-phospholipid that is the precursor to PAF. Thus these
compounds are useful in the treatment and prevention of disease
states in which leukotrienes, prostaglandins or PAF are involved.
Moreover, in diseases where more than one of these agents plays a
role, a cPLA.sub.2 inhibitor would be expected to be more
efficacious than leukotriene, prostaglandin or PAF receptor
antagonists and also more effective than cyclooxygenase or
5-lipoxygenase inhibitors.
[0393] Therefore, the compounds, pharmaceutical compositions and
regimens of the present invention are useful in treating and
preventing the disorders treated by cyclooxygenase-2,
cycloxygenase-1, and 5-lipoxygenase inhibitors and also antagonists
of the receptors for PAF, leukotrienes or prostaglandins.
[0394] This invention also provides methods for treating or
preventing venous or arterial thrombosis in a mammal, or preventing
progression of symptoms of thrombosis, the method comprising
administering to a mammal in need thereof a pharmaceutically
acceptable amount of a compound of the invention, or a
pharmaceutically acceptable salt thereof. In some embodiments, the
thrombosis is atherothrombosis.
[0395] Each of the methods of this invention comprises
administering to a mammal in need of such treatment a
pharmaceutically or therapeutically effective amount of a compound
of this invention. In the instances of combination therapies
described herein, it will be understood the administration further
includes a pharmaceutically or therapeutically effective amount of
the second pharmaceutical agent in question. The second or
additional pharmacological agents described herein may be
administered in the doses and regimens known in the art.
[0396] The compounds of this invention may also be used in
comparable veterinary methods of treatment, particularly for the
veterinary treatment, inhibition or alleviation of inflammation and
pain. These methods will be understood to be of particular interest
for companion mammals, such as dogs and cats, and for use in farm
mammals, such as cattle, horses, mules, donkeys, goats, hogs,
sheep, etc. These methods may be used to treat the types of
inflammation and pain experienced in veterinary medicine including,
but not limited to, pain and inflammation associated with
arthritis, joint imperfections, developmental joint defects, such
as hip dysplasia, tendonitis, suspensary ligament inflammation,
laminitis, curb and bursitis, or pain or inflammation associated
with surgery, accident, trauma or disease, such as Lyme Disease.
These compounds may also be used in the treatment of inflammation
of the air passages, such as in conditions of asthma, laryngitis,
tracheitis, bronchitis, rhinitis and pharyngitis
[0397] Each of these veterinary methods comprises administering to
the mammal in need thereof a pharmaceutically effective amount of a
compound of this invention, or a pharmaceutically acceptable salt
form thereof. The compounds of this invention may be used for human
or veterinary methods in conjunction with other medicaments or
dietary supplements known in the art for the treatment, inhibition
or alleviation of inflammation or pain. These may include aspirin
(including buffered aspirin, aspirin with Maalox and enteric coated
aspirin), COX-2 inhibitors, such as celecoxib, non-acetylated
carboxylic acids, such as magnesium salicylate, salicylamide or
sodium salicylate, acetic acids, such as doclofenac or etodolac,
propionic acids, such as ibuprofen, naproxen (available in
NAPROSYNO.RTM. and EQUIPROXEN.RTM. brands), ketoprofen,
RIMADYL.RTM. (carprofen), flunixin meglumine, fenamic acids, such
as tolfenamic acid, mefanamic acid, meclofenamic acid (ARQUEL.RTM.)
or niflumic acid, enolic acids, such as oxyphenbutazone,
phenylbutazone, piroxicam or dipyrone, or non-acidic compounds like
nabumetone. Also used in veterinary applications are
dimethylsulfoxide (DMSO), orgotein (such as PALOSEIN.RTM. brand of
orgotein), polysulfated glycosaminoglycans or PS-GAGs (such as
ADEQUAN.RTM. brand polysulfated glycosaminoglycan), hyaluronic acid
and its natural and synthetic analogues, Ketorolac trimethamine
(such as the TORADOL.RTM. brand), FELDENE.RTM. (piroxicam), or
METACAM.RTM. (meloxicam).
[0398] Dietary supplements used in human or veterinary applications
include glucosamines, chondroitin sulfate, methylsulfonylmethane
(MSM), and omega 3 fatty acids and other cold water fish oils. The
compounds and methods of this invention may also be used in
conjunction with human or veterinary physical therapy, massage,
chiropractic and acupuncture treatments and regimens. Each of these
medicaments and dietary supplements may be administered to the
mammal in question using regimens and effective dosages known in
the art.
[0399] It is intended that each of the patents, applications, and
printed publications including books mentioned in this patent
document be hereby incorporated by reference in their entirety.
[0400] As those skilled in the art will appreciate, numerous
changes and modifications may be made to the preferred embodiments
of the invention without departing from the spirit of the
invention. It is intended that all such variations fall within the
scope of the invention.
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