U.S. patent application number 12/672413 was filed with the patent office on 2012-07-26 for heteroaryl amide analogues.
This patent application is currently assigned to H. LUNDBECK A/S. Invention is credited to Rajagopal Bakthavatchalam, Scott M. Capitosti, David C. Ihle, David J. Wustrow, Jun Yuan.
Application Number | 20120190680 12/672413 |
Document ID | / |
Family ID | 39327396 |
Filed Date | 2012-07-26 |
United States Patent
Application |
20120190680 |
Kind Code |
A1 |
Bakthavatchalam; Rajagopal ;
et al. |
July 26, 2012 |
Heteroaryl Amide Analogues
Abstract
Compounds, pharmaceutical compositions, and methods of use are
disclosed for heteroaryl amide analogues of formula Ia and/or Ib:
##STR00001## In certain embodiments, the heteroaryl amide analogues
are agonists and/or ligands of dopamine receptors and may be
useful, inter alia, for the treatment of a condition responsive to
P2X.sub.7 receptor modulation, for example, pain, inflammation, a
neurological or neurodegenerative disorder, a cardiovascular
disorder, an ocular disorder or an immune system disorder.
Inventors: |
Bakthavatchalam; Rajagopal;
(Madison, CT) ; Ihle; David C.; (Worcester,
MA) ; Capitosti; Scott M.; (Perry, OH) ;
Wustrow; David J.; (Los Gatos, CA) ; Yuan; Jun;
(Boston, MA) |
Assignee: |
H. LUNDBECK A/S
Valby-Copenhagen
DK
|
Family ID: |
39327396 |
Appl. No.: |
12/672413 |
Filed: |
August 11, 2008 |
PCT Filed: |
August 11, 2008 |
PCT NO: |
PCT/US08/72760 |
371 Date: |
March 1, 2012 |
Current U.S.
Class: |
514/234.5 ;
514/255.05; 514/275; 514/333; 514/339; 514/397; 514/419; 544/120;
544/122; 544/331; 544/405; 546/256; 546/272.7; 548/312.1;
548/492 |
Current CPC
Class: |
A61P 11/00 20180101;
A61P 11/06 20180101; A61P 29/00 20180101; A61P 25/22 20180101; B64D
33/04 20130101; A61P 9/00 20180101; A61P 25/00 20180101; A61P 25/24
20180101; A61P 27/02 20180101; B64D 29/02 20130101; A61P 25/08
20180101; A61P 9/10 20180101; A61P 1/00 20180101; A61P 19/02
20180101; A61P 25/18 20180101; A61P 25/28 20180101 |
Class at
Publication: |
514/234.5 ;
544/331; 546/272.7; 544/122; 548/492; 544/405; 546/256; 548/312.1;
544/120; 514/275; 514/339; 514/419; 514/255.05; 514/333;
514/397 |
International
Class: |
A61K 31/506 20060101
A61K031/506; C07D 401/14 20060101 C07D401/14; C07D 413/14 20060101
C07D413/14; C07D 209/42 20060101 C07D209/42; C07D 403/06 20060101
C07D403/06; A61K 31/4439 20060101 A61K031/4439; A61K 31/5377
20060101 A61K031/5377; A61K 31/404 20060101 A61K031/404; A61K
31/497 20060101 A61K031/497; A61K 31/444 20060101 A61K031/444; A61K
31/4178 20060101 A61K031/4178; A61P 29/00 20060101 A61P029/00; A61P
25/00 20060101 A61P025/00; A61P 9/00 20060101 A61P009/00; A61P
27/02 20060101 A61P027/02; A61P 19/02 20060101 A61P019/02; A61P
9/10 20060101 A61P009/10; A61P 1/00 20060101 A61P001/00; A61P 25/28
20060101 A61P025/28; A61P 11/06 20060101 A61P011/06; A61P 11/00
20060101 A61P011/00; A61P 25/24 20060101 A61P025/24; A61P 25/22
20060101 A61P025/22; A61P 25/18 20060101 A61P025/18; A61P 25/08
20060101 A61P025/08; C07D 403/04 20060101 C07D403/04 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 20, 2007 |
FR |
0757712 |
Claims
1. A compound or salt or hydrate thereof according to formula Ia or
Ib: ##STR00171## wherein: U is CR.sub.1A; W is
--C(.dbd.O)NR.sub.4--, --NR.sub.4C(.dbd.O)-- or
--NR.sub.4--NR.sub.4--C(.dbd.O)--; each R.sub.4 is independently
hydrogen, C.sub.1-C.sub.6alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.2alkyl or taken together
with a substituent of X to form a 4- to 7-membered
heterocycloalkyl; X is absent or C.sub.1-C.sub.6alkylene that is
optionally substituted with 1 to 4 substituents selected from
R.sub.B, R.sub.C, R.sub.D, and R.sub.E; R.sub.B, R.sub.C, R.sub.D,
and R.sub.E are each independently hydroxy, --COOH,
C.sub.1-C.sub.8alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.2-C.sub.8alkyl ether, mono- or
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.0-C.sub.4alkyl, (4- to
7-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl and
phenylC.sub.0-C.sub.2alkyl; or any two of R.sub.B, R.sub.C,
R.sub.D, and R.sub.E taken together with the carbon atom or atoms
through which they are connected form a 3- to 7-membered cycloalkyl
or a 4- to 7-membered heterocycloalkyl; or any one of R.sub.B,
R.sub.C, R.sub.D, and R.sub.E taken together with R.sub.4 and the
atom or atoms through which they are connected form a 4- to
7-membered heterocycloalkyl; Y is C.sub.1-C.sub.8alkyl,
C.sub.3-C.sub.16cycloalkyl, 4- to 16-membered heterocycloalkyl, 6-
to 16-membered aryl or 5- to 16-membered heteroaryl, each of which
is optionally substituted with 1 to 6 substituents independently
chosen from hydroxy, halogen, cyano, amino, nitro, oxo,
aminocarbonyl, aminosulfonyl, COOH, C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoyl, C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl and
(C.sub.1-C.sub.6alkyl)sulfonylamino; Z.sub.1, Z.sub.2, Z.sub.3, and
Z.sub.4 are independently CR.sub.1 or N; R.sub.1A is hydrogen,
hydroxy, halogen, cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; each R.sub.1 is independently
hydrogen, hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl, COOH, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; and R.sub.A is a group of the
formula -L-A-M, wherein: L is absent or C.sub.1-C.sub.6alkylene
that is optionally modified by (i) the replacement of a
carbon-carbon single bond with a double or triple carbon-carbon
bond, or (ii) substitution with oxo, --COOH, --SO.sub.3H,
--SO.sub.2NH.sub.2, --PO.sub.3H.sub.2, tetrazole or oxadiazolone; A
is absent or CO, O, NR.sub.6, S, SO, SO.sub.2, CONR.sub.6,
NR.sub.6CO, (C.sub.4-C.sub.7cycloalkyl)C.sub.0-C.sub.2alkyl, 4- to
7-membered heterocycloalkyl or 5- or 6-membered heteroaryl; wherein
R.sub.6 is hydrogen or C.sub.1-C.sub.6alkyl; and M is: (i) hydroxy,
halogen, cyano, amino, aminocarbonyl, aminosulfonyl, carboxyalkyl,
or --COOH; or (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy,
(4- to 10-membered carbocycle)C.sub.0-C.sub.4alkyl, (4- to
10-membered heterocycle)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6alkanoyloxy, C.sub.1-C.sub.6alkanoylamino,
C.sub.1-C.sub.6alkylsulfonyl, C.sub.1-C.sub.6alkylsulfonylamino,
C.sub.1-C.sub.6alkylsulfonyloxy, mono- or
di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with 1 to 4 substituents independently chosen from oxo,
amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, alkoxycarbonyl, alkanoyloxy, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle; provided that for a compound of Formula Ia; (a) when U
is CH, then W--X--Y is other than: ##STR00172## wherein Rx is H or
C(.dbd.O)--O-alkyl; (b) when one of Z.sub.1, Z.sub.2, Z.sub.3, and
Z.sub.4 is N, and the others of Z.sub.1, Z.sub.2, Z.sub.3, and
Z.sub.4 are each CH, R.sub.A and Y are each independently alkyl,
aryl aralkyl or heteroaryl, W is --C(.dbd.O)N(H)--, and U is
CR.sub.1A, wherein R.sub.1A is other than OH; (c) when Z.sub.1,
Z.sub.2, Z.sub.3, and Z.sub.4 are each CH, R.sub.A is unsubstituted
phenyl, W is --C(.dbd.O)N(H)--, X is unsubstituted alkylene or
alkylene substituted with one hydroxyl, Y is dialkylamino,
unsubstituted heterocycloalkyl, or heterocycloalkyl substituted
with one alkyl, and U is CR.sub.1A, wherein R.sub.1A is other than
halogen; and (d) when Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are
each CR.sub.1, at least two of R.sub.1 are H and the remaining
R.sub.1 are each independently H, halogen, nitro, hydroxy, or
alkoxy, R.sub.A is cycloalkyl or substituted or unsubstituted
phenyl, W is --C(.dbd.O)N(H)--, X is unsubstituted alkylene, Y is
alkyl or cycloalkyl each optionally substituted with one COOH,
dialkylamino, unsubstituted heteroaryl, unsubstituted
heterocycloalkyl, or heterocycloalkyl substituted with nitrile or
amino, and U is CR.sub.1A, wherein R.sub.1A is other than
alkyl.
2. (canceled)
3. The compound or salt or hydrate thereof according to claim 2,
having formula Ia.
4-6. (canceled)
7. The compound or salt or hydrate thereof according to claim 1,
wherein R.sub.B, R.sub.C, R.sub.D, and R.sub.E are each
independently C.sub.1-C.sub.4alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.2alkyl, or
phenylC.sub.0-C.sub.2alkyl; or any two of R.sub.B, R.sub.C,
R.sub.D, and R.sub.E taken together with the carbon atom or atoms
through which they are connected form a 3- to 7-membered cycloalkyl
or a 4- to 7-membered heterocycloalkyl.
8. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH.
9-10. (canceled)
11. The compound or salt or hydrate thereof according to claim 1,
wherein Y is C.sub.1-C.sub.8alkyl, C.sub.3-C.sub.16cycloalkyl, 6-
to 16-membered aryl or 5- to 16-membered heteroaryl, each
optionally substituted.
12. (canceled)
13. The compound or salt or hydrate thereof according to claim 11,
wherein Y is optionally substituted with from 1 to 3
substituents.
14. The compound or salt or hydrate thereof according to claim 1,
wherein Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are CH.
15. The compound or salt or hydrate thereof according to claim 1,
wherein W is --NR.sub.4C(.dbd.O)--.
16-18. (canceled)
19. The compound or salt or hydrate thereof according to claim 1,
wherein R.sub.A is other than halophenyl, other than haloaryl,
other than cycloalkyl, other than thienyl, and/or other than
carbocyclic.
20-24. (canceled)
25. The compound or salt or hydrate thereof according to claim 1,
wherein when R.sub.A is optionally substituted heteroaryl, said
heteroaryl contains at least one nitrogen ring atom.
26. The compound or salt or hydrate thereof according to claim 1,
wherein when R.sub.A is optionally substituted heteroaryl, said
heteroaryl contains at least two nitrogen ring atoms.
27. (canceled)
28. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is other than unsubstituted
--C(.dbd.O)N(H)aralkyl.
29. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is other than unsubstituted
--C(.dbd.O)N(H)heteroaralkyl.
30. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is other than unsubstituted
--C(.dbd.O)N(H)heterocyclic.
31. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is other than unsubstituted
--C(.dbd.O)N(H)alkoxyalkyl.
32. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is other than unsubstituted
--C(.dbd.O)N(H)R.sub.v, wherein R.sub.v is aminoalkyl,
alkylaminoalkyl, or dialkylaminoalkyl, alkyl, alkenyl, or
alkynyl.
33. The compound or salt or hydrate thereof according to claim 1,
wherein U is CH and W--X--Y is optionally substituted
--C(.dbd.O)N(H)R.sub.v, wherein R.sub.v is arylC.sub.1-C.sub.4alkyl
or heteroarylC.sub.1-C.sub.4alkyl.
34. The compound or salt or hydrate thereof according to claim 1,
wherein W is --C(.dbd.O)N(H)--.
35. The compound or salt or hydrate thereof according to claim 34,
wherein M is optionally substituted heteroaryl.
36. The compound or salt or hydrate thereof according to claim 35,
wherein said heteroaryl contains at least one nitrogen ring
atom.
37. The compound or salt or hydrate thereof according to claim 35,
wherein said heteroaryl contains at least two nitrogen ring
atoms.
38. The compound or salt or hydrate thereof according to claim 37,
wherein M is optionally substituted pyrimidinyl.
39. The compound or salt or hydrate thereof according to claim 38,
wherein M optionally substituted pyrimidin-2-yl.
40. The compound or salt or hydrate thereof according to claim 34,
wherein X is C.sub.1-C.sub.2alkylene, optionally substituted.
41. The compound or salt or hydrate thereof according to claim 40,
wherein X is C.sub.1-C.sub.2alkylene, substituted with
C.sub.1-C.sub.4alkyl.
42. (canceled)
43. The compound or salt or hydrate thereof according to claim 40,
wherein X is substituted with at least 2 substituents selected from
R.sub.B, R.sub.C, R.sub.D, and R.sub.E, wherein: any two of
R.sub.B, R.sub.C, R.sub.D, and R.sub.E taken together with the
carbon atom or atoms through which they are connected form a 5- to
6-membered cycloalkyl.
44. The compound or salt or hydrate thereof according to claim 34,
wherein Y is an optionally substituted carbocycle or an optionally
substituted heterocycle.
45. The compound or salt or hydrate thereof according to claim 44,
wherein Y is adamantyl, phenyl, pyridyl, or morpholinyl, each of
which are optionally substituted.
46. A compound or salt or hydrate thereof according to claim 1,
wherein the compound is:
N-(adamantan-1-ylmethyl)-1-pyrimidin-2-yl-1H-indole-3-carboxamide;
N-{4-methyl-2-[4-(trifluoromethyl)phenyl]pentyl}-1-pyrimidin-2-yl-1H-indo-
le-3-carboxamide;
N-[2-(4-chlorophenyl)pentyl]-1-pyrimidin-2-yl-1H-indole-3-carboxamide;
N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-carboxa-
mide;
1-pyrimidin-2-yl-N-({1-[4-(trifluoromethyl)phenyl]cyclohexyl}methyl)-
-1H-indole-3-carboxamide;
N-{[1-(4-chlorophenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-indole-3-car-
boxamide;
N-{[1-(4-methoxyphenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-in-
dole-3-carboxamide;
N-[(1-morpholin-4-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-carbo-
xamide;
2-{3-[(adamantan-1-ylmethyl)carbamoyl]-1H-indol-1-yl}benzoic acid;
N-[4-methyl-2-(4-methylphenyl)pentyl]-1-pyrimidin-2-yl-1H-indole-3-carbox-
amide;
N-(4-methyl-2-pyridin-3-ylpentyl)-1-pyrimidin-2-yl-1H-indole-3-carb-
oxamide;
N-(4-methyl-2-phenylpentyl)-1-pyrimidin-2-yl-1H-indole-3-carboxam-
ide;
N-{[1-(6-methylpyridin-3-yl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-in-
dole-3-carboxamide;
N-{[1-(4-fluorophenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-indole-3-car-
boxamide;
2-{3-[(adamantan-1-ylmethyl)carbamoyl]-1H-indol-1-yl}pentanoic
acid;
1-(5-fluoropyrimidin-2-yl)-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1H--
indole-3-carboxamide;
1-(3-ethylpyrazin-2-yl)-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1H-indole-3--
carboxamide;
4-chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole--
3-carboxamide;
N-{[1-(4-methoxyphenyl)cyclopentyl]methyl}-1-pyrimidin-2-yl-1H-indole-3-c-
arboxamide;
N-{[1-(4-methylphenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-indole-3-car-
boxamide;
N-{[1-(4-chloro-3-fluorophenyl)cyclohexyl]methyl}-1-pyrimidin-2--
yl-1H-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-yl-1H-indole-3-
-carboxamide; Methyl
2-(3-{[(1-pyridin-3-ylcyclohexyl)methyl]carbamoyl}-1H-indol-1-yl)nicotina-
te;
N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-4-(trifluorometh-
yl)-1H-indole-3-carboxamide;
4-bromo-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-
-carboxamide;
4-cyano-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-
-carboxamide;
N-{[1-(4-methoxyphenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-4-(trifluorome-
thyl)-1H-indole-3-carboxamide;
N-{[1-(4-fluorophenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-4-(trifluoromet-
hyl)-1H-indole-3-carboxamide;
N-{[1-(6-methylpyridin-3-yl)cyclohexyl]methyl}-1-pyrimidin-2-yl-4-(triflu-
oromethyl)-1H-indole-3-carboxamide;
7-chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole--
3-carboxamide;
4-fluoro-N-(4-methyl-2-pyridin-3-ylpentyl)-1-pyrimidin-2-yl-1H-indole-3-c-
arboxamide;
N-[2-(4-chlorophenyl)-4-methylpentyl]-4-fluoro-1-pyrimidin-2-yl-1H-indole-
-3-carboxamide;
4-fluoro-N-[4-methyl-2-(4-methylphenyl)pentyl]-1-pyrimidin-2-yl-1H-indole-
-3-carboxamide;
4-fluoro-N-{4-methyl-2-[4-(trifluoromethyl)phenyl]pentyl}-1-pyrimidin-2-y-
l-1H-indole-3-carboxamide;
N-{[1-(4-chloro-3-fluorophenyl)cyclohexyl]methyl}-4-fluoro-1-pyrimidin-2--
yl-1H-indole-3-carboxamide;
4-fluoro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole--
3-carboxamide;
4-fluoro-N-{[1-(4-methylphenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-ind-
ole-3-carboxamide;
4-fluoro-N-{[1-(4-methoxyphenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-in-
dole-3-carboxamide;
4-fluoro-N-{[1-(4-fluorophenyl)cyclohexyl]methyl}-1-pyrimidin-2-yl-1H-ind-
ole-3-carboxamide;
N-{[1-(4-chlorophenyl)cyclohexyl]methyl}-4-fluoro-1-pyrimidin-2-yl-1H-ind-
ole-3-carboxamide; 1-Pyrimidin-2-yl-1H-indole-3,4-dicarboxylic acid
4-amide 3-[(1-pyridin-3-yl-cyclohexylmethyl)-amide];
1-[(1-methyl-1H-imidazol-2-yl)methyl]-N-[(1-pyridin-3-ylcyclohexyl)methyl-
]-1H-indole-2-carboxamide;
4-methyl-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole--
3-carboxamide;
4-methyl-N-(4-methyl-2-pyridin-3-ylpentyl)-1-pyrimidin-2-yl-1H-indole-3-c-
arboxamide;
N-{[1-(4-methoxyphenyl)cyclohexyl]methyl}-4-methyl-1-pyrimidin-2-yl-1H-in-
dole-3-carboxamide;
N-[2-(4-chlorophenyl)-4-methylpentyl]-4-methyl-1-pyrimidin-2-yl-1H-indole-
-3-carboxamide;
N-{[1-(4-chlorophenyl)cyclohexyl]methyl}-4-methyl-1-pyrimidin-2-yl-1H-ind-
ole-3-carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-1-pyrimidin-2-yl-4-(trifluor-
omethyl)-1H-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-yl-4-(trifluor-
omethyl)-1H-indole-3-carboxamide;
N-(adamantan-1-ylmethyl)-4-chloro-1-[2-(dimethylamino)ethyl]-1H-indole-3--
carboxamide;
N-(adamantan-1-ylmethyl)-4-chloro-1-[3-(dimethylamino)propyl]-1H-indole-3-
-carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-4-methylpentyl]-1-[3-(dimethylamino)propyl-
]-1H-indole-3-carboxamide;
4-chloro-1-[3-(dimethylamino)propyl]-N-(4-methyl-2-pyridin-3-ylpentyl)-1H-
-indole-3-carboxamide;
4-chloro-1-[2-(dimethylamino)ethyl]-N-(4-methyl-2-pyridin-3-ylpentyl)-1H--
indole-3-carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-4-methylpentyl]-1-[2-(dimethylamino)ethyl]-
-1H-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-4-fluoro-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-4-methyl-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-4-fluoro-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
4-fluoro-N-[2-(6-methoxypyridin-3-yl)-2-piperidin-1-ylethyl]-1-pyrimidin--
2-yl-1H-indole-3-carboxamide;
N-[2-(6-methoxypyridin-3-yl)-2-piperidin-1-ylethyl]-4-methyl-1-pyrimidin--
2-yl-1H-indole-3-carboxamide;
N-(adamantan-1-ylmethyl)-4-methyl-1-pyrimidin-2-yl-1H-indole-3-carboxamid-
e;
4-chloro-N-[2-(4-chlorophenyl)-2-piperazin-1-ylethyl]-1-pyrimidin-2-yl--
1H-indole-3-carboxamide;
4-chloro-N-(2-morpholin-4-yl-2-phenylethyl)-1-pyrimidin-2-yl-1H-indole-3--
carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
4-chloro-N-{2-piperidin-1-yl-2-[4-(trifluoromethyl)phenyl]ethyl}-1-pyrimi-
din-2-yl-1H-indole-3-carboxamide;
4-chloro-N-[2-(4-chloro-3-fluorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidi-
n-2-yl-1H-indole-3-carboxamide;
4-chloro-N-[2-(6-methoxypyridin-3-yl)-2-piperidin-1-ylethyl]-1-pyrimidin--
2-yl-1H-indole-3-carboxamide;
4-chloro-N-[2-(3,4-difluorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-y-
l-1H-indole-3-carboxamide;
3-chloro-1-[(1-methyl-1H-imidazol-2-yl)methyl]-N-(4-methyl-2-pyridin-3-yl-
pentyl)-1H-indole-2-carboxamide;
4-chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-pyrrolo-
[2,3-b]pyridine-3-carboxamide;
4-chloro-N-(4-methyl-2-pyridin-3-ylpentyl)-1-pyrimidin-2-yl-1H-pyrrolo[2,-
3-b]pyridine-3-carboxamide;
2-adamantan-1-yl-N-(4-chloro-1-pyrimidin-2-yl-1H-indol-3-yl)acetamide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(adamantan-1-ylmethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(2-adamantan-1-yl-ethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
((R)-6,6-dimethylbicyclo[3.1.1]hept-2-ylmethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(4-methyl-2-p-tolyl-pentyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[2-(4-chloro-phenyl)-4-methyl-pentyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[1-(4-chloro-phenyl)-cyclohexylmethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[1-(4-trifluoromethyl-phenyl)-cyclohexylmethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[4-(4-chloro-phenyl)-tetrahydro-pyran-4-ylmethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[1-(4-methoxy-phenyl)-cyclohexylmethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[4-methyl-2-(4-trifluoromethyl-phenyl)-pentyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[2-(4-chloro-phenyl)-pentyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(4-methyl-2-pyridin-3-yl-pentyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[1-(4-chloro-phenyl)-cyclobutylmethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(2-adamantan-1-yl-2-hydroxy-ethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(3-methyl-butyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(2-phenyl-pentyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(4-methyl-2-phenyl-pentyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(3-cyclopentyl-2-phenyl-propyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(2-cyclohexyl-2-phenyl-ethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(2,3-diphenyl-propyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(3-phenyl-butyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(4-phenyl-butyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[2-(4-bromo-phenyl)-ethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(3,3,5-trimethyl-cyclohexyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
adamantan-2-ylamide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
cycloheptylmethyl-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
[2-(2-bromo-phenyl)-ethyl]-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
((1S,2R)-2-hydroxy-cyclohexylmethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(1-hydroxy-cyclohexylmethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(1-hydroxy-cyclopentylmethyl)-amide;
4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
(4-hydroxy-tetrahydro-thiopyran-4-ylmethyl)-amide;
N-[2-(4-fluorophenyl)-2-piperidin-1-ylethyl]-4-methyl-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
N-[2-(3,4-difluorophenyl)-2-piperidin-1-ylethyl]-4-methyl-1-pyrimidin-2-y-
l-1H-indole-3-carboxamide;
4-methyl-N-(2-morpholin-4-yl-2-phenylethyl)-1-pyrimidin-2-yl-1H-indole-3--
carboxamide;
N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-4-methyl-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
3-chloro-N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-[(1-methyl-1H-imi-
dazol-2-yl)methyl]-1H-indole-2-carboxamide;
N-{[4-(4-chlorophenyl)tetrahydro-2H-pyran-4-yl]methyl}-4-methyl-1-pyrimid-
in-2-yl-1H-indole-3-carboxamide;
3-chloro-N-{[4-(4-chlorophenyl)tetrahydro-2H-pyran-4-yl]methyl}-1-[(1-met-
hyl-1H-imidazol-2-yl)methyl]-1H-indole-2-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-4-methyl-1-pyrazin-2-yl-1H-i-
ndole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-4-methyl-1-pyrazin-2-yl-1H-i-
ndole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-(3-cyanopyridin-2-yl)-4-me-
thyl-1H-indole-3-carboxamide;
1-(3-cyanopyridin-2-yl)-4-methyl-N-(2-morpholin-4-yl-2-phenylethyl)-1H-in-
dole-3-carboxamide;
N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-1-(3-cyanopyridin-2-yl)-4-me-
thyl-1H-indole-3-carboxamide;
4-methyl-N-(4-methyl-2-morpholin-4-ylpentyl)-1-pyrimidin-2-yl-1H-indole-3-
-carboxamide;
4-chloro-N-{2-piperidin-1-yl-2-[6-(trifluoromethyl)pyridin-3-yl]ethyl}-1--
pyrimidin-2-yl-1H-indole-3-carboxamide;
4-chloro-N-[2-(4-fluorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-yl-1H-
-indole-3-carboxamide;
4-chloro-N-[(1-hydroxycycloheptyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-ca-
rboxamide;
4-chloro-N-[2-(3,4-difluorophenyl)-2-piperidin-1-ylethyl]-1-pyr-
imidin-2-yl-1H-indole-3-carboxamide;
4-chloro-N-[2-(2,4-difluorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-y-
l-1H-indole-3-carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-2-morpholin-4-ylethyl]-1-pyrimidin-2-yl-1H-
-pyrrolo[2,3-b]pyridine-3-carboxamide;
4-chloro-N-[2-(4-chlorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-yl-1H-
-pyrrolo[2,3-b]pyridine-3-carboxamide;
4-chloro-N-{[4-(4-chlorophenyl)tetrahydro-2H-pyran-4-yl]methyl}-1-pyrimid-
in-2-yl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide;
4-chloro-N-[(1-hydroxycycloheptyl)methyl]-1-pyrimidin-2-yl-1H-pyrrolo[2,3-
-b]pyridine-3-carboxamide;
4-chloro-N-[(1-hydroxycyclohexyl)methyl]-1-pyrimidin-2-yl-1H-pyrrolo[2,3--
b]pyridine-3-carboxamide;
4-chloro-N-[2-(3,4-difluorophenyl)-2-piperidin-1-ylethyl]-1-pyrimidin-2-y-
l-1H-pyrrolo[2,3-b]pyridine-3-carboxamide;
4-chloro-N-(2-morpholin-4-yl-2-phenylethyl)-1-pyrimidin-2-yl-1H-pyrrolo[2-
,3-b]pyridine-3-carboxamide; or
4-chloro-N-{[1-(4-methylpiperazin-1-yl)cyclohexyl]methyl}-1-pyrimidin-2-y-
l-1H-indole-3-carboxamide.
47-48. (canceled)
49. The pharmaceutical composition, comprising at least one
compound or salt or hydrate thereof according to claim 1 in
combination with a physiologically acceptable carrier or
excipient.
50-53. (canceled)
54. A method for treating a condition responsive to P2X.sub.7
receptor modulation in a patient, comprising administering to the
patient a therapeutically effective amount of at least one compound
or salt or hydrate thereof according to claim 1, and thereby
alleviating the condition in the patient.
55. The method according to claim 54, wherein the condition is
pain.
56. The method according to claim 55, wherein the pain is
neuropathic pain.
57. The method according to claim 55, wherein the pain is
arthritis-associated pain, a neuropathic pain syndrome, visceral
pain, dental pain, headache, stump pain, meralgia paresthetica,
burning-mouth syndrome, pain associated with nerve and root damage,
causalgia, neuritis, neuronitis, neuralgia, surgery-related pain,
musculoskeletal pain, central nervous system pain, spinal pain,
Charcot's pains, ear pain, muscle pain, eye pain, orofacial pain,
carpel tunnel syndrome, acute and chronic back pain, gout, scar
pain, hemorrhoidal pain, dyspeptic pains, angina, nerve root pain,
complex regional pain syndrome, cancer-associated pain, pain
associated with venom exposure, trauma-associated pain, pain
associated with autoimmune diseases or immunodeficiency disorders,
or pain that results from hot flashes, burns, sunburn, or exposure
to heat, cold or external chemical stimuli.
58. The method according to claim 54, wherein the condition is
inflammation, a neurological or neurodegenerative disorder, a
centrally-mediated neuropsychiatric disorder, a cardiovascular
disorder, an ocular disorder or an immune system disorder.
59. The method according to claim 54, wherein the condition is
osteoarthritis, rheumatoid arthritis, arthrosclerosis, glaucoma,
irritable bowel syndrome, inflammatory bowel disease, cirrhosis,
lupus, scleroderma, Alzheimer's disease, traumatic brain injury,
asthma, chronic obstructive pulmonary disease, or interstitial
fibrosis.
60. (canceled)
61. The method according to claim 54, wherein the patient is a
human.
62. The compound or salt or hydrate thereof according to claim 1,
wherein the compound is radiolabeled.
63-67. (canceled)
68. The method according to claim 58, wherein the
centrally-mediated neuropsychiatric disorder is depression,
depression mania, bipolar disease, anxiety, schizophrenia, an
eating disorder, a sleep disorder or a cognition disorder.
69. The method according to claim 58, wherein the neurological
disorder is epilepsy.
Description
FIELD OF THE INVENTION
[0001] This invention relates generally to heteroaryl amide
analogues that have useful pharmacological properties. The
invention further relates to the use of such compounds for treating
conditions related to P2X.sub.7 receptor activation, for
identifying other agents that bind to P2X.sub.7 receptor, and as
probes for the detection and localization of P2X.sub.7
receptors.
BACKGROUND OF THE INVENTION
[0002] Pain perception, or nociception, is mediated by the
peripheral terminals of a group of specialized sensory neurons,
termed "nociceptors." A wide variety of physical and chemical
stimuli induce activation of such neurons in mammals, leading to
recognition of a potentially harmful stimulus. Inappropriate or
excessive activation of nociceptors, however, can result in
debilitating acute or chronic pain.
[0003] Neuropathic pain, which typically results from damage to the
nervous system, involves pain signal transmission in the absence of
stimulus, pain from a normally innocuous stimulus (allodynia) and
increased pain from a normally painful stimulus (hyperalgesia). In
most instances, neuropathic pain is thought to occur because of
sensitization in the peripheral and central nervous systems
following initial damage to the peripheral system (e.g., via direct
injury or systemic disease). Neuropathic pain is typically burning,
shooting and unrelenting in its intensity and can sometimes be more
debilitating than the initial injury or disease process that
induced it.
[0004] Existing treatments for neuropathic pain are generally
suboptimal. Opiates, such as morphine, are potent analgesics, but
their usefulness is limited because of adverse side effects, such
as physical addictiveness and withdrawal properties, as well as
respiratory depression, mood changes, and decreased intestinal
motility with concomitant constipation, nausea, vomiting, and
alterations in the endocrine and autonomic nervous systems. In
addition, neuropathic pain is frequently non-responsive or only
partially responsive to conventional opioid analgesic regimens, or
to treatment with other drugs, such as gabapentin. Treatments
employing the N-methyl-D-aspartate antagonist ketamine or the
alpha(2)-adrenergic agonist clonidine can reduce acute or chronic
pain, and permit a reduction in opioid consumption, but these
agents are often poorly tolerated due to side effects.
[0005] Another common condition for which existing therapies are
insufficient or problematic is inflammation. Transient inflammation
is a beneficial mechanism that protects mammals from invading
pathogens. Uncontrolled inflammation, however, causes tissue damage
and pain and is the underlying cause of many illnesses, including
asthma, as well as other allergic, infectious, autoimmune,
degenerative, and idiopathic diseases. Existing treatments often
exhibit low, delayed or only temporary efficacy, undesirable
side-effects and/or a lack of selectivity. There is a continuing
need for new drugs that overcome one or more of the shortcomings of
drugs currently used for immunosuppression or in the treatment or
prevention of inflammatory disorders, including allergic disorders,
autoimmune disorders, fibrogenic disorders, and neurodegenerative
diseases, such as amyotrophic lateral sclerosis, Alzheimer's
disease, and Huntington's disease.
[0006] The P2X.sub.7 receptor is a ligand-gated ion channel that is
activated by ATP and is present on a variety of cell types,
including microglia in the central nervous system and cells
involved in inflammation and immune system function, such as immune
cells. In particular, P2X.sub.7 is involved in activation of
lymphocytes and monocyte/macrophages leading to the increased
release of pro-inflammatory cytokines (e.g., TNFalpha and IL-1beta)
from these cells. Recent studies indicate that inhibiting P2X.sub.7
receptor activation in situations of inflammation (e.g., rheumatoid
arthritis and other autoimmune diseases, osteoarthritis, uveitis,
asthma, chronic obstructive pulmonary disease and inflammatory
bowel disease) or interstitial fibrosis results in a therapeutic
effect. These and other studies indicate that P2X.sub.7 receptor
antagonists may find use in the treatment and prophylaxis of pain,
including acute, chronic and neuropathic pain, as well as a variety
of other conditions including osteoarthritis, rheumatoid arthritis,
arthrosclerosis, inflammatory bowel disease, Alzheimer's disease,
traumatic brain injury, asthma, chronic obstructive pulmonary
disease, and fibrosis of internal organs (e.g., interstitial
fibrosis).
[0007] Small molecule P2X.sub.7 receptor antagonists are desirable
for such therapies. The present invention fulfills this need, and
provides further related advantages.
SUMMARY OF THE INVENTION
[0008] The present invention provides heteroaryl amide analogues of
Formula Ia or Ib:
##STR00002##
[0009] wherein:
[0010] U is CR.sub.1A or N;
[0011] W is --C(.dbd.O)NR.sub.4--, --NR.sub.4C(.dbd.O)-- or
--NR.sub.4--NR.sub.4--C(.dbd.O)--;
[0012] each R.sub.4 is independently hydrogen,
C.sub.1-C.sub.6alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.2alkyl or taken together
with a substituent of X to form a 4- to 7-membered
heterocycloalkyl;
[0013] X is absent or C.sub.1-C.sub.6alkylene that is optionally
substituted with 1 to 4 substituents selected from R.sub.B,
R.sub.C, R.sub.D, and R.sub.E;
[0014] R.sub.B, R.sub.C, R.sub.D, and R.sub.E are each
independently hydroxy, --COOH, C.sub.1-C.sub.8alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.2-C.sub.8alkyl ether, mono- or
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.0-C.sub.4alkyl, (4- to
7-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl and
phenylC.sub.0-C.sub.2alkyl; or any two of R.sub.B, R.sub.C,
R.sub.D, and R.sub.E taken together with the carbon atom or atoms
through which they are connected form a 3- to 7-membered cycloalkyl
or a 4- to 7-membered heterocycloalkyl; or any one of R.sub.B,
R.sub.C, R.sub.D, and R.sub.E taken together with R.sub.4 and the
atom or atoms through which they are connected form a 4- to
7-membered heterocycloalkyl;
[0015] Y is C.sub.1-C.sub.8alkyl, C.sub.3-C.sub.16cycloalkyl, 4- to
16-membered heterocycloalkyl, 6- to 16-membered aryl or 5- to
16-membered heteroaryl, each of which is optionally substituted
with 1 to 6 substituents independently chosen from hydroxy,
halogen, cyano, amino, nitro, oxo, aminocarbonyl, aminosulfonyl,
COOH, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl and
(C.sub.1-C.sub.6alkyl)sulfonylamino;
[0016] Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are independently
CR.sub.1 or N;
[0017] R.sub.1A is hydrogen, hydroxy, halogen, cyano, amino,
aminocarbonyl, aminosulfonyl, COOH, C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoyl, C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino;
[0018] each R.sub.1 is independently hydrogen, hydroxy, halogen,
cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; and
[0019] R.sub.A is a group of the formula -L-A-M, wherein:
[0020] L is absent or C.sub.1-C.sub.6alkylene that is optionally
modified by (i) the replacement of a carbon-carbon single bond with
a double or triple carbon-carbon bond, or (ii) substitution with
oxo, --SO.sub.3H, --SO.sub.2NH.sub.2, --PO.sub.3H.sub.2, tetrazole
or oxadiazolone;
[0021] A is absent or CO, O, NR.sub.6, S, SO, SO.sub.2, CONR.sub.6,
NR.sub.6CO, (C.sub.4-C.sub.7cycloalkyl)C.sub.0-C.sub.2alkyl, 4- to
7-membered heterocycloalkyl or 5- or 6-membered heteroaryl; wherein
R.sub.6 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0022] M is:
[0023] (i) hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl, carboxyalkyl, or --COOH; or
[0024] (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy, (4- to
10-membered carbocycle)C.sub.0-C.sub.4alkyl, (4- to 10-membered
heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6alkanoyloxy,
C.sub.1-C.sub.6alkanoylamino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, C.sub.1-C.sub.6alkylsulfonyloxy,
mono- or di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with 1 to 4 substituents independently chosen from oxo,
amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, alkoxycarbonyl, alkanoyloxy, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle.
[0025] Within certain aspects, heteroaryl amide analogues of
Formula Ia and/or Ib are P2X.sub.7 receptor antagonists with an
IC.sub.50 value no greater than 20 micromolar, 10 micromolar, 5
micromolar, 1 micromolar, 500 nanomolar or 100 nanomolar in an in
vitro assay for determination of P2X.sub.7 receptor antagonist
activity. In certain embodiments, such P2X.sub.7 receptor
antagonists exhibit no detectable agonist activity in an in vitro
assay of P2X.sub.7 receptor activity (e.g., in an assay provided in
Example 7, herein) at a concentration equal to the IC.sub.50, 10
times the IC.sub.50 or 100 times the IC.sub.50 and/or at a
concentration of 2,500 nM.
[0026] Within certain aspects, heteroaryl amide analogues provided
herein are labeled with a detectable marker (e.g., radiolabeled or
fluorescein conjugated).
[0027] The present invention further provides, within other
aspects, pharmaceutical compositions comprising at least one
heteroaryl amide analogue provided herein in combination with a
physiologically acceptable carrier or excipient.
[0028] Within further aspects, methods are provided for modulating
(e.g., reducing) cellular P2X.sub.7 receptor activation or
activity, comprising contacting a cell (e.g., microglia, astrocyte
or peripheral macrophage or monocyte) that expresses a P2X.sub.7
receptor with at least one P2X.sub.7 receptor modulator as
described herein. Such contact may occur in vivo or in vitro and is
generally performed using a concentration of P2X.sub.7 receptor
modulator that is sufficient to detectably alter P2X.sub.7 receptor
activity in vitro (as determined, for example, using an assay as
described in Example 7).
[0029] The present invention further provides methods for treating
a condition responsive to P2X.sub.7 receptor modulation in a
patient, comprising administering to the patient a therapeutically
effective amount of at least one P2X.sub.7 receptor antagonist as
described herein.
[0030] Within other aspects, methods are provided for treating pain
in a patient, comprising administering to a patient suffering from
(or at risk for) pain a therapeutically effective amount of at
least one P2X.sub.7 receptor antagonist as described herein.
[0031] Within other aspects, methods are provided for treating
inflammation in a patient, comprising administering to a patient
suffering from (or at risk for) inflammation a therapeutically
effective amount of at least one P2X.sub.7 receptor antagonist as
described herein.
[0032] Methods are further provided for treating osteoarthritis,
rheumatoid arthritis, arthrosclerosis, inflammatory bowel disease,
Alzheimer's disease, traumatic brain injury, asthma, chronic
obstructive pulmonary disease, ocular conditions (e.g., glaucoma),
cirrhosis, lupus, scleroderma, or fibrosis of internal organs
(e.g., interstitial fibrosis) in a patient, comprising
administering to a patient suffering from (or at risk for) one or
more of the foregoing conditions a therapeutically effective amount
of at least one P2X.sub.7 receptor antagonist as described
herein.
[0033] Within still further aspects, the present invention provides
methods for inhibiting death of retinal ganglion cells in a
patient, comprising administering to the patient a therapeutically
effective amount of at least one P2X.sub.7 receptor antagonist as
described herein.
[0034] Methods are further provided for identifying an agent that
binds to P2X.sub.7 receptor, comprising: (a) contacting P2X.sub.7
receptor with a labeled compound that is a heteroaryl amide
analogue as described herein under conditions that permit binding
of the compound to P2X.sub.7 receptor, thereby generating bound,
labeled compound; (b) detecting a signal that corresponds to the
amount of bound, labeled compound in the absence of test agent; (c)
contacting the bound, labeled compound with a test agent; (d)
detecting a signal that corresponds to the amount of bound labeled
compound in the presence of test agent; and (e) detecting a
decrease in signal detected in step (d), as compared to the signal
detected in step (b).
[0035] Within further aspects, the present invention provides
methods for determining the presence or absence of P2X.sub.7
receptor in a sample, comprising: (a) contacting a sample with a
compound as described herein under conditions that permit
modulation by the compound of P2X.sub.7 receptor activity; and (b)
detecting a signal indicative of a level of the compound modulating
P2X.sub.7 receptor activity.
[0036] The present invention also provides packaged pharmaceutical
preparations, comprising: (a) a pharmaceutical composition as
described herein in a container; and (b) instructions for using the
composition to (i) treat one or more conditions responsive to
P2X.sub.7 receptor modulation, such as pain, osteoarthritis,
rheumatoid arthritis, arthrosclerosis, inflammatory bowel disease,
Alzheimer's disease, traumatic brain injury, asthma, chronic
obstructive pulmonary disease, ocular conditions (e.g., glaucoma),
cirrhosis, lupus, scleroderma, and/or fibrosis of internal organs
(e.g., interstitial fibrosis) or (ii) provide retinal
neuroprotection (e.g., inhibit death of retinal ganglion
cells).
[0037] In yet another aspect, the present invention provides
methods for preparing the compounds disclosed herein, including the
intermediates.
[0038] These and other aspects of the invention will become
apparent upon reference to the following detailed description.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
Terminology
[0039] Compounds are generally described herein using standard
nomenclature. For compounds having asymmetric centers, it should be
understood that (unless otherwise specified) all of the optical
isomers and mixtures thereof are encompassed. In addition,
compounds with carbon-carbon double bonds may occur in Z- and
E-forms, with all isomeric forms of the compounds being included in
the present invention unless otherwise specified. Where a compound
exists in various tautomeric forms, a recited compound is not
limited to any one specific tautomer, but rather is intended to
encompass all tautomeric forms. Certain compounds are described
herein using a general formula that includes variables (e.g., R1,
A, X). Unless otherwise specified, each variable within such a
formula is defined independently of any other variable, and any
variable that occurs more than one time in a formula is defined
independently at each occurrence.
[0040] The phrase "heteroaryl amide analogue," as used herein,
encompasses all compounds of Formula Ia or Formula Ib, as well as
compounds of other Formulas provided herein (including any
enantiomers, racemates and stereoisomers, and including the various
crystal forms and polymorphs) and pharmaceutically acceptable
salts, solvates (e.g., hydrates, including hydrates of salts),
amides and esters of such compounds.
[0041] A "pharmaceutically acceptable salt" of a compound recited
herein is an acid or base salt that is suitable for use in contact
with the tissues of human beings or animals without excessive
toxicity or carcinogenicity, and preferably without irritation,
allergic response, or other problem or complication. Such salts
include mineral and organic acid salts of basic residues such as
amines, as well as alkali or organic salts of acidic residues such
as carboxylic acids. Specific pharmaceutically acceptable anions
for use in salt formation include, but are not limited to, acetate,
2-acetoxybenzoate, ascorbate, benzoate, bicarbonate, bromide,
calcium edetate, carbonate, chloride, citrate, dihydrochloride,
diphosphate, ditartrate, edetate, estolate (ethylsuccinate),
formate, fumarate, gluceptate, gluconate, glutamate, glycolate,
glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,
hydrochloride, hydroiodide, hydroxymaleate, hydroxynaphthoate,
iodide, isethionate, lactate, lactobionate, malate, maleate,
mandelate, methylbromide, methylnitrate, methylsulfate, mucate,
napsylate, nitrate, pamoate, pantothenate, phenylacetate,
phosphate, polygalacturonate, propionate, salicylate, stearate,
subacetate, succinate, sulfamate, sulfanilate, sulfate, sulfonates
including besylate (benzenesulfonate), camsylate
(camphorsulfonate), edisylate (ethane-1,2-disulfonate), esylate
(ethanesulfonate) 2-hydroxyethylsulfonate, mesylate
(methanesulfonate), triflate (trifluoromethanesulfonate) and
tosylate (p-toluenesulfonate), tannate, tartrate, teoclate and
triethiodide. Similarly, pharmaceutically acceptable cations for
use in salt formation include, but are not limited to ammonium,
benzathine, chloroprocaine, choline, diethanolamine,
ethylenediamine, meglumine, procaine, and metals such as aluminum,
calcium, lithium, magnesium, potassium, sodium and zinc. Those of
ordinary skill in the art will recognize further pharmaceutically
acceptable salts for the compounds provided herein. In general, a
pharmaceutically acceptable acid or base salt can be synthesized
from a parent compound that contains a basic or acidic moiety by
any conventional chemical method. Briefly, such salts can be
prepared by reacting the free acid or base forms of these compounds
with a stoichiometric amount of the appropriate base or acid in
water or in an organic solvent, or in a mixture of the two;
generally, the use of nonaqueous media, such as ether, ethyl
acetate, ethanol, methanol, isopropanol or acetonitrile, is
preferred.
[0042] It will be apparent that each compound provided herein may,
but need not, be formulated as a solvate (e.g., hydrate) or
non-covalent complex. In addition, the various crystal forms and
polymorphs are within the scope of the present invention. Also
provided herein are prodrugs of the compounds of the recited
Formulas. A "prodrug" is a compound that may not fully satisfy the
structural requirements of the compounds provided herein, but is
modified in vivo, following administration to a patient, to produce
a compound of a formula provided herein. For example, a prodrug may
be an acylated derivative of such a compound. Prodrugs include
compounds wherein hydroxy, amine or sulfhydryl groups are bonded to
any group that, when administered to a mammalian subject, cleaves
to form a free hydroxy, amino or sulfhydryl group, respectively.
Examples of prodrugs include, but are not limited to, acetate,
formate, benzoate and peptide derivatives of alcohol and amine
functional groups within a compound provided herein. Prodrugs may
generally be prepared by modifying functional groups present in the
compounds in such a way that the modifications are cleaved in vivo
to yield the parent compounds.
[0043] As used herein, the term "alkyl" refers to a straight or
branched chain saturated aliphatic hydrocarbon. Alkyl groups
include groups having from 1 to 8 carbon atoms (C1-C8alkyl), from 1
to 6 carbon atoms (C1-C6alkyl) and from 1 to 4 carbon atoms
(C1-C4alkyl), such as methyl, ethyl, propyl, isopropyl, n-butyl,
sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl,
hexyl, 2-hexyl, 3-hexyl and 3-methylpentyl. "C0-Cnalkyl" refers to
a single covalent bond (C0) or an alkyl group having from 1 to n
carbon atoms; for example "C0-C4alkyl" refers to a single covalent
bond or a C1-C4alkyl group. In some instances, a substituent of an
alkyl group is specifically indicated. For example,
"C1-C6hydroxyalkyl" is a C1-C6alkyl group substituted with at least
one --OH; "C1-C6-aminoalkyl" is a C1-C6alkyl group substituted with
at least one --NH2; C1-C6cyanoalkyl is a C1-C6alkyl group
substituted with at least one --CN.
[0044] "Alkenyl" refers to straight or branched chain alkene
groups, which comprise at least one unsaturated carbon-carbon
double bond. Alkenyl groups include C2-C8alkenyl, C2-C6alkenyl and
C2-C4alkenyl groups, which have from 2 to 8, 2 to 6 or 2 to 4
carbon atoms, respectively, such as ethenyl, allyl or isopropenyl.
"C2-C6cyanoalkenyl" is a C2-C6alkenyl group substituted with at
least one --CN.
[0045] "Alkynyl" refers to straight or branched chain alkyne
groups, which have one or more unsaturated carbon-carbon bonds, at
least one of which is a triple bond. Alkynyl groups include
C2-C8alkynyl, C2-C6alkynyl and C2-C4alkynyl groups, which have from
2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively.
[0046] "Alkylene" refers to a divalent alkyl group, as defined
above. C1-C2alkylene is methylene or ethylene; C0-C4alkylene is a
single covalent bond or an alkylene group having 1, 2, 3 or carbon
atoms; C0-C2alkylene is a single covalent bond, methylene or
ethylene. A "C1-C6alkylene that is optionally modified by the
replacement of a carbon-carbon single bond with a double or triple
carbon-carbon bond" is a C1-C6alkylene group as described above, or
a divalent C2-C6alkene or C2-C6alkyne.
[0047] A "cycloalkyl" is a group that comprises one or more
saturated and/or partially saturated rings in which all ring
members are carbon, such as cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, myrtanyl and
partially saturated variants of the foregoing, such as
cyclohexenyl. Cycloalkyl groups do not comprise an aromatic ring or
a heterocyclic ring. Certain cycloalkyl groups are C3-C7cycloalkyl,
in which the cycloalkyl group contains a single ring having from 3
to 7 ring members, all of which are carbon. A
"(C3-C7cycloalkyl)C0-C4alkyl" is a C3-C7cycloalkyl group linked via
a single covalent bond or a C1-C4alkylene group.
[0048] A "(C4-C7cycloalkyl)C0-C4alkylene" is a divalent
(C3-C7cycloalkyl)C0-C4alkyl group that is linked via two single
covalent bonds to two specified moieties. In general, one single
covalent bond is located on the cyclic portion and the other is
located on the alkylene portion, if present; alternatively, if no
alkylene group is present, both single covalent bonds are on
different ring members. For example, with respect to the group RA,
if A is (C6cycloalkyl)C2alkylene and M is COOH, one RA moiety so
formed is:
##STR00003##
[0049] By "alkoxy," as used herein, is meant an alkyl group as
described above attached via an oxygen bridge. Alkoxy groups
include C.sub.1-C.sub.6alkoxy and C.sub.1-C.sub.4alkoxy groups,
which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
Methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy,
tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy,
neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy are
representative alkoxy groups.
[0050] The term "oxo" is used herein to refer to an oxygen
substituent of a carbon atom that results in the formation of a
carbonyl group (C.dbd.O). An oxo group that is a substituent of a
nonaromatic carbon atom results in a conversion of --CH.sub.2-- to
--C(.dbd.O)--. An oxo group that is a substituent of an aromatic
carbon atom results in a conversion of --CH-- to --C(.dbd.O)-- and
may result in a loss of aromaticity.
[0051] The term "alkanoyl" refers to an acyl group (e.g.,
--(C.dbd.O)-alkyl), in which carbon atoms are in a linear or
branched alkyl arrangement and where attachment is through the
carbon of the keto group. Alkanoyl groups have the indicated number
of carbon atoms, with the carbon of the keto group being included
in the numbered carbon atoms. For example a C.sub.2alkanoyl group
is --(C.dbd.O)CH.sub.3. Alkanoyl groups include, for example,
C.sub.2-C.sub.8alkanoyl, C.sub.2-C.sub.6alkanoyl and
C.sub.2-C.sub.4alkanoyl groups, which have from 2 to 8, from 2 to 6
or from 2 to 4 carbon atoms, respectively. "C.sub.1alkanoyl" refers
to --(C.dbd.O)H, which (along with C.sub.2-C.sub.8alkanoyl) is
encompassed by the term "C.sub.1-C.sub.8alkanoyl."
[0052] "Alkyl ether" refers to a linear or branched ether
substituent (i.e., an alkyl group that is substituted with an
alkoxy group). Alkyl ether groups include C.sub.2-C.sub.8alkyl
ether, C.sub.2-C.sub.6alkyl ether and C.sub.2-C.sub.4alkyl ether
groups, which have 2 to 8, 6 or 4 carbon atoms, respectively. A
C.sub.2 alkyl ether substituent is --CH.sub.2--O--CH.sub.3.
[0053] The term "alkoxycarbonyl" refers to an alkoxy group attached
through a keto (--(C.dbd.O)--) bridge (i.e., a group having the
general structure --C(.dbd.O)--O-alkyl). Alkoxycarbonyl groups
include C.sub.1-C.sub.8, C.sub.1-C.sub.6 and
C.sub.1-C.sub.4alkoxycarbonyl groups, which have from 1 to 8, 6 or
4 carbon atoms, respectively, in the alkyl portion of the group
(i.e., the carbon of the keto bridge is not included in the
indicated number of carbon atoms). "C.sub.1alkoxycarbonyl" refers
to --C(.dbd.O)--O--CH.sub.3; C.sub.3alkoxycarbonyl indicates
--C(.dbd.O)--O--(CH.sub.2).sub.2CH.sub.3 or
--C(.dbd.O)--O--(CH)(CH.sub.3).sub.2.
[0054] "Alkanoyloxy," as used herein, refers to an alkanoyl group
linked via an oxygen bridge (i.e., a group having the general
structure --O--C(.dbd.O)-alkyl). Alkanoyloxy groups include
C.sub.1-C.sub.8, C.sub.1-C.sub.6 and C.sub.1-C.sub.4alkanoyloxy
groups, which have from 1 to 8, 6 or 4 carbon atoms, respectively,
in the alkyl portion of the group. For example,
"C.sub.1alkanoyloxy" refers to --O--C(.dbd.O)--CH.sub.3.
[0055] Similarly, "alkanoylamino," as used herein, refers to an
alkanoyl group linked via a nitrogen bridge (i.e., a group having
the general structure --N(R)--C(.dbd.O)-alkyl), in which R is
hydrogen or C.sub.1-C.sub.6alkyl. Alkanoylamino groups include
C.sub.1-C.sub.8, C.sub.1-C.sub.6 and C.sub.1-C.sub.4alkanoylamino
groups, which have from 1 to 8, 6 or 4 carbon atoms within the
alkanoyl group, respectively, in the alkyl portion of the
group.
[0056] "Alkylsulfonyl" refers to groups of the formula
--(SO.sub.2)-alkyl, in which the sulfur atom is the point of
attachment. Alkylsulfonyl groups include
C.sub.1-C.sub.6alkylsulfonyl and C.sub.1-C.sub.4alkylsulfonyl
groups, which have from 1 to 6 or from 1 to 4 carbon atoms,
respectively. Methylsulfonyl is one representative alkylsulfonyl
group. "C.sub.1-C.sub.4haloalkylsulfonyl" is an alkylsulfonyl group
that has from 1 to 4 carbon atoms and is substituted with at least
one halogen (e.g., trifluoromethylsulfonyl).
[0057] "Alkylsulfonylamino" refers to groups of the formula
--N(R)--(SO.sub.2)-alkyl, in which R is hydrogen or
C.sub.1-C.sub.6alkyl and the nitrogen atom is the point of
attachment. Alkylsulfonylamino groups include
C.sub.1-C.sub.6alkylsulfonylamino and
C.sub.1-C.sub.4alkylsulfonylamino groups, which have from 1 to 6 or
1 to 4 carbon atoms, respectively. Methylsulfonylamino is a
representative alkylsulfonylamino group.
"C.sub.1-C.sub.6haloalkylsulfonylamino" is an alkylsulfonylamino
group that has from 1 to 6 carbon atoms and is substituted with at
least one halogen (e.g., trifluoromethylsulfonylamino).
[0058] "Aminosulfonyl" refers to groups of the formula
--(SO.sub.2)--NH.sub.2, in which the sulfur atom is the point of
attachment. The term "mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl" refers to groups that
satisfy the formula --(SO.sub.2)--NR.sub.2, in which the sulfur
atom is the point of attachment, and in which one R is
C.sub.1-C.sub.6alkyl and the other R is hydrogen or an
independently chosen C.sub.1-C.sub.6alkyl.
[0059] "Alkylaminoalkyl" refers to an alkylamino group linked via
an alkylene group (i.e., a group having the general structure
-alkylene-NH-alkyl or -alkylene-N(alkyl)(alkyl)) in which each
alkyl is selected independently from alkyl, cycloalkyl and
(cycloalkyl)alkyl groups. Alkylaminoalkyl groups include, for
example, mono- and
di-(C.sub.1-C.sub.8alkyl)aminoC.sub.1-C.sub.8alkyl, mono- and
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.1-C.sub.6alkyl and mono- and
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.1-C.sub.4alkyl. "Mono- or
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.0-C.sub.6alkyl" refers to a
mono- or di-(C.sub.1-C.sub.6alkyl)amino group linked via a single
covalent bond or a C.sub.1-C.sub.6alkylene group. The following are
representative alkylaminoalkyl groups:
##STR00004##
[0060] It will be apparent that the definition of "alkyl" as used
in the teems "alkylamino" and "alkylaminoalkyl" differs from the
definition of "alkyl" used for all other alkyl-containing groups,
in the inclusion of cycloalkyl and (cycloalkyl)alkyl groups (e.g.,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.6alkyl).
[0061] The term "aminocarbonyl" refers to an amide group (i.e.,
--(C.dbd.O)NH.sub.2). "Mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl" refers to groups of the
formula --(C.dbd.O)--N(R).sub.2, in which the carbonyl is the point
of attachment, one R is C.sub.1-C.sub.6alkyl and the other R is
hydrogen or an independently chosen C.sub.1-C.sub.6alkyl.
[0062] "Mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonylC.sub.0-C.sub.4alkyl" is an
aminocarbonyl group in which one or both of the hydrogen atoms is
replaced with C.sub.1-C.sub.6alkyl, and which is linked via a
single covalent bond (i.e., mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl) or a
C.sub.1-C.sub.4alkylene group (i.e.,
--(C.sub.0-C.sub.4alkyl)-(C.dbd.O)N(C.sub.1-C.sub.6alkyl).sub.2).
If both hydrogen atoms are so replaced, the C.sub.1-C.sub.6alkyl
groups may be the same or different.
[0063] The term "aminosulfonyl" refers to a sulfonamide group
(i.e., --(SO.sub.2)NH.sub.2). "Mono- or
di-(C.sub.1-C.sub.8alkyl)aminosulfonyl" refers to groups of the
formula --(SO.sub.2)--N(R).sub.2, in which the sulfur atom is the
point of attachment, one R is C.sub.1-C.sub.8alkyl and the other R
is hydrogen or an independently chosen C.sub.1-C.sub.8alkyl.
[0064] "Mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonylC.sub.0-C.sub.4alkyl" is an
aminosulfonyl group in which one or both of the hydrogen atoms is
replaced with C.sub.1-C.sub.6alkyl, and which is linked via a
single covalent bond (i.e., mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl) or a
C.sub.1-C.sub.4alkylene group (i.e.,
--(C.sub.1-C.sub.4alkyl)-(SO.sub.2)N(C.sub.1-C.sub.6alkyl).sub.2).
If both hydrogen atoms are so replaced, the C.sub.1-C.sub.6alkyl
groups may be the same or different.
[0065] The term "halogen" refers to fluorine, chlorine, bromine or
iodine.
[0066] A "haloalkyl" is an alkyl group that is substituted with 1
or more independently chosen halogens (e.g.,
"C.sub.1-C.sub.6haloalkyl" groups have from 1 to 6 carbon atoms).
Examples of haloalkyl groups include, but are not limited to,
mono-, di- or tri-fluoromethyl; mono-, di- or tri-chloromethyl;
mono-, di-, tri-, tetra- or penta-fluoroethyl; mono-, di-, tri-,
tetra- or penta-chloroethyl; and
1,2,2,2-tetrafluoro-1-trifluoromethyl-ethyl. Typical haloalkyl
groups are trifluoromethyl and difluoromethyl. The term
"haloalkoxy" refers to a haloalkyl group as defined above that is
linked via an oxygen bridge.
[0067] A dash ("-") that is not between two letters or symbols is
used to indicate a point of attachment for a substituent. For
example, --CONH.sub.2 is attached through the carbon atom.
[0068] A "carbocycle" or "carbocyclic group" comprises at least one
ring formed entirely by carbon-carbon bonds (referred to herein as
a carbocyclic ring), and does not contain a heterocycle. Unless
otherwise specified, each ring within a carbocycle may be
independently saturated, partially saturated or aromatic, and is
optionally substituted as indicated. A carbocycle generally has
from 1 to 3 fused, pendant or spiro rings and optionally further
contains one or more alkylene bridges; carbocycles within certain
embodiments have one ring or two fused rings. Typically, each ring
contains from 3 to 8 ring members (i.e., C.sub.3-C.sub.8);
C.sub.5-C.sub.7 rings are recited in certain embodiments.
Carbocycles comprising fused, pendant or spiro rings typically
contain from 9 to 16 ring members. Certain representative
carbocycles are cycloalkyl as described above (e.g., cyclohexyl,
cycloheptyl or adamantly). Other carbocycles are aryl (i.e.,
contain at least one aromatic carbocyclic ring, with or without one
or more additional aromatic and/or cycloalkyl rings). Such aryl
carbocycles include, for example, phenyl, naphthyl (e.g.,
1-naphthyl and 2-naphthyl), fluorenyl, indanyl and
1,2,3,4-tetrahydronaphthyl. The term "haloaryl" refers to an aryl
group that is substituted with at least one halogen.
[0069] Certain carbocycles recited herein are
C.sub.6-C.sub.10arylC.sub.0-C.sub.8alkyl groups (i.e., groups in
which a 6- to 10-membered carbocyclic group comprising at least one
aromatic ring is linked via a single covalent bond or a
C.sub.1-C.sub.8alkylene group). Phenyl groups linked via a single
covalent bond or C.sub.1-C.sub.2alkylene group are designated
phenylC.sub.0-C.sub.2alkyl (e.g., benzyl, 1-phenyl-ethyl and
2-phenyl-ethyl).
[0070] A "heterocycle" or "heterocyclic group" has from 1 to 3
fused, pendant or spiro rings, at least one of which is a
heterocyclic ring (i.e., one or more ring atoms is a heteroatom
independently chosen from O, S and N, with the remaining ring atoms
being carbon). Additional rings, if present, may be heterocyclic or
carbocyclic. Typically, a heterocyclic ring comprises 1, 2, 3 or 4
heteroatoms; within certain embodiments each heterocyclic ring has
1 or 2 heteroatoms per ring. Each heterocyclic ring generally
contains from 3 to 8 ring members (rings having from 4 or 5 to 7
ring members are recited in certain embodiments) and heterocycles
comprising fused, pendant or spiro rings typically contain from 9
to 14 ring members. Certain heterocycles comprise a sulfur atom as
a ring member; in certain embodiments, the sulfur atom is oxidized
to SO or SO.sub.2. Unless otherwise specified, a heterocycle may be
a heterocycloalkyl group (i.e., each ring is saturated or partially
saturated), such as a 4- to 7-membered heterocycloalkyl, which
generally comprises 1, 2, 3 or 4 ring atoms that are independently
chosen from C, O, N and S; or a heteroaryl group (i.e., at least
one ring within the group is aromatic), such as a 5- to 10-membered
heteroaryl (which may be monocyclic or bicyclic) or a 6-membered
heteroaryl (e.g., pyridyl or pyrimidyl). N-linked heterocyclic
groups are linked via a component nitrogen atom.
[0071] As used herein, "aralkyl" refers to a moiety composed of an
alkyl radical bearing an aryl substituent, wherein the aralkyl
moiety has from about 7 to about 50 carbon atoms (and all
combinations and subcombinations of ranges and specific numbers of
carbon atoms therein), and where aryl and alkyl are as previously
defined with from about 7 to about 11 carbon atoms being preferred.
Non-limiting examples include, for example, benzyl, diphenylmethyl,
triphenylmethyl, alpha- or beta-phenylethyl, and diphenylethyl.
[0072] As used herein, "heteroaralkyl" refers to a ring system
composed of a heteroaryl substituted alkyl radical where heteroaryl
and alkyl are as previously defined, and where the heteroaralkyl
group has from about 7 to about 50 carbon atoms (and all
combinations and subcombinations of ranges and specific numbers of
carbon atoms therein). Non-limiting examples include, for example,
2-(1H-pyrrol-3-yl)ethyl, 3-pyridylmethyl, 5-(2H-tetrazolyl)methyl,
and 3-(pyrimidin-2-yl)-2-methylcyclopentanyl.
[0073] A "heterocycleC.sub.0-C.sub.4alkyl" is a heterocyclic group
linked via a single covalent bond or C.sub.1-C.sub.4alkylene group.
A "(4- to 7-membered heterocycloalkyl)C.sub.1-C.sub.4alkyl" is a
heterocycloalkyl ring with from 4 to 7 ring members that is linked
via a C.sub.1-C.sub.4alkylene group.
[0074] A "(4- to 7-membered
heterocycloalkyl)C.sub.0-C.sub.4alkylene" is a divalent (4- to
7-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl group that is
linked via two single covalent bonds to two specified moieties. In
general, one such single covalent bond is located on the cyclic
portion and the other is located on the alkylene portion, if
present; alternatively, if no alkylene group is present, both such
single covalent bonds are located on different ring members. For
example, with respect to the group R.sub.A, if A is a
(piperidinyl)C.sub.2alkylene and M is COOH, one R.sub.A moiety so
formed is:
##STR00005##
[0075] A "substituent," as used herein, refers to a molecular
moiety that is covalently bonded to an atom within a molecule of
interest. For example, a ring substituent may be a moiety such as a
halogen, alkyl group, haloalkyl group or other group that is
covalently bonded to an atom (preferably a carbon or nitrogen atom)
that is a ring member. Substituents of aromatic groups are
generally covalently bonded to a ring carbon atom. The term
"substitution" refers to replacing a hydrogen atom in a molecular
structure with a substituent, such that the valence on the
designated atom is not exceeded, and such that a chemically stable
compound (i.e., a compound that can be isolated, characterized, and
tested for biological activity) results from the substitution.
[0076] Groups that are "optionally substituted" are unsubstituted
or are substituted by other than hydrogen at one or more available
positions, typically 1, 2, 3, 4 or 5 positions, by one or more
suitable groups (which may be the same or different). Optional
substitution is also indicated by the phrase "substituted with from
0 to X substituents," where X is the maximum number of possible
substituents. Certain optionally substituted groups are substituted
with from 0 to 2, 3 or 4 independently selected substituents (i.e.,
are unsubstituted or substituted with up to the recited maximum
number of substituents). Other optionally substituted groups are
substituted with at least one substituent (e.g., substituted with
from 1 to 2, 3 or 4 independently selected substituents).
[0077] The term "P2X.sub.7 receptor" refers to any P2X7 receptor,
preferably a mammalian receptor such as the human or rat P2X7
receptor disclosed in U.S. Pat. No. 6,133,434, as well as
homologues thereof found in other species.
[0078] A "P2X.sub.7 receptor modulator," also referred to herein as
a "modulator," is a compound that increases or decreases P2X.sub.7
receptor activation and/or P2X.sub.7 receptor-mediated activity
(e.g., signal transduction). P2X.sub.7 receptor modulators
specifically provided herein are compounds of Formula I and
pharmaceutically acceptable salts, hydrates and esters thereof. A
modulator may be a P2X.sub.7 receptor agonist or antagonist.
[0079] A modulator is considered an "antagonist" if it detectably
inhibits P2X.sub.7 receptor-mediated signal transduction (using,
for example, a representative assay provided in Example 7); in
general, such an antagonist inhibits P2X.sub.7 receptor activation
with a IC.sub.50 value of less than 20 micromolar, preferably less
than 10 micromolar, more preferably less than 5 micromolar, more
preferably less than 1 micromolar, still more preferably less than
500 nanomolar, and most preferably less than 100 nanomolar within
an assay provided in Example 7. P2X.sub.7 receptor antagonists
include neutral antagonists and inverse agonists.
[0080] An "inverse agonist" of P2X.sub.7 receptor is a compound
that reduces the activity of P2X.sub.7 receptor below its basal
activity level in the absence of added ligand. Inverse agonists of
P2X.sub.7 receptor may also inhibit the activity of ligand at
P2X.sub.7 receptor and/or binding of ligand to P2X.sub.7 receptor.
The basal activity of P2X.sub.7 receptor, as well as a reduction in
P2X.sub.7 receptor activity due to the presence of P2X.sub.7
receptor antagonist, may be determined from a calcium mobilization
assay (e.g., the assay of Example 7).
[0081] A "neutral antagonist" of P2X.sub.7 receptor is a compound
that inhibits the activity of ligand at P2X.sub.7 receptor, but
does not significantly change the basal activity of the receptor
(i.e., within a calcium mobilization assay as described in Example
7 performed in the absence of ligand, P2X.sub.7 receptor activity
is reduced by no more than 10%, preferably by no more than 5%, and
more preferably by no more than 2%; most preferably, there is no
detectable reduction in activity). Neutral antagonists of P2X.sub.7
receptor may inhibit the binding of ligand to P2X.sub.7
receptor.
[0082] As used herein a "P2X.sub.7 receptor agonist" is a compound
that elevates the activity of the P2X.sub.7 receptor above the
basal activity level of the receptor (i.e., enhances P2X.sub.7
receptor activation and/or P2X.sub.7 receptor-mediated activity,
such as signal transduction). P2X.sub.7 receptor agonist activity
may be detected using the representative assay provided in Example
7. P2X.sub.7 receptor agonists include ATP and
2'(3')--O-(4-benzoyl-benzoyl)adenosine 5'-triphosephate
(BzATP).
[0083] A "therapeutically effective amount" (or dose) is an amount
that, upon administration to a patient, results in a discernible
patient benefit (e.g., provides detectable relief from at least one
condition being treated). Such relief may be detected using any
appropriate criteria, including alleviation of one or more symptoms
such as pain. A therapeutically effective amount or dose generally
results in a concentration of compound in a body fluid (such as
blood, plasma, serum, CSF, synovial fluid, lymph, cellular
interstitial fluid, tears or urine) that is sufficient to alter
P2X.sub.7 receptor-mediated signal transduction (using an assay
provided in Example 7). It will be apparent that the discernible
patient benefit may be apparent after administration of a single
dose, or may become apparent following repeated administration of
the therapeutically effective dose according to a predetermined
regimen, depending upon the indication for which the compound is
administered.
[0084] By "statistically significant," as used herein, is meant
results varying from control at the p<0.1 level of significance
as measured using a standard parametric assay of statistical
significance such as a student's T test.
[0085] A "patient" is any individual treated with a compound
provided herein. Patients include humans, as well as other animals
such as companion animals (e.g., dogs and cats) and livestock.
Patients may be experiencing one or more symptoms of a condition
responsive to P2X.sub.7 receptor modulation or may be free of such
symptom(s) (i.e., treatment may be prophylactic in a patient
considered at risk for the development of such symptoms).
Heteroaryl Amide Analogues
[0086] As noted above, the present invention provides heteroaryl
amide analogues. In certain embodiments, the present invention
provides heteroaryl amide analogues of Formula Ia and/or Ib:
##STR00006##
[0087] wherein:
[0088] U is CR.sub.1A or N;
[0089] W is --C(.dbd.O)NR.sub.4--, --NR.sub.4C(.dbd.O)-- or
--NR.sub.4--NR.sub.4--C(.dbd.O)--;
[0090] each R.sub.4 is independently hydrogen,
C.sub.1-C.sub.6alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.2alkyl or taken together
with a substituent of X to faun a 4- to 7-membered
heterocycloalkyl;
[0091] X is absent or C.sub.1-C.sub.6alkylene that is optionally
substituted with 1 to 4 substituents selected from R.sub.R,
R.sub.C, R.sub.D, and R.sub.E;
[0092] R.sub.B, R.sub.C, R.sub.D, and R.sub.E are each
independently hydroxy, --COOH, C.sub.1-C.sub.8alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.2-C.sub.8alkyl ether, mono- or
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.0-C.sub.4alkyl, (4- to
7-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl and
phenylC.sub.0-C.sub.2alkyl; or any two of R.sub.B, R.sub.C,
R.sub.D, and R.sub.E taken together with the carbon atom or atoms
through which they are connected form a 3- to 7-membered cycloalkyl
or a 4- to 7-membered heterocycloalkyl; or any one of R.sub.B,
R.sub.C, R.sub.D, and R.sub.E taken together with R.sub.4 and the
atom or atoms through which they are connected form a 4- to
7-membered heterocycloalkyl;
[0093] Y is C.sub.1-C.sub.8alkyl, C.sub.3-C.sub.16cycloalkyl, 4- to
16-membered heterocycloalkyl, 6- to 16-membered aryl or 5- to
16-membered heteroaryl, each of which is optionally substituted
with 1 to 6 substituents independently chosen from hydroxy,
halogen, cyano, amino, nitro, oxo, aminocarbonyl, aminosulfonyl,
COOH, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl and
(C.sub.1-C.sub.6alkyl)sulfonylamino;
[0094] Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are independently
CR.sub.1 or N;
[0095] R.sub.1A is hydrogen, hydroxy, halogen, cyano, amino,
aminocarbonyl, aminosulfonyl, COOH, C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoyl, C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino;
[0096] each R.sub.1 is independently hydrogen, hydroxy, halogen,
cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; and R.sub.A is a group of the
formula -L-A-M, wherein:
[0097] L is absent or C.sub.1-C.sub.6alkylene that is optionally
modified by (i) the replacement of a carbon-carbon single bond with
a double or triple carbon-carbon bond, or (ii) substitution with
oxo, --COOH, --SO.sub.2NH.sub.2, --PO.sub.3H.sub.2, tetrazole or
oxadiazolone;
[0098] A is absent or CO, O, NR.sub.6, S, SO, SO.sub.2, CONR.sub.6,
NR.sub.6CO, (C.sub.4-C.sub.7cycloalkyl)C.sub.0-C.sub.2alkyl, 4- to
7-membered heterocycloalkyl or 5- or 6-membered heteroaryl; wherein
R.sub.6 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0099] M is:
[0100] (i) hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl, carboxyalkyl, or --COOH; or
[0101] (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy, (4- to
10-membered carbocycle)C.sub.0-C.sub.4alkyl, (4- to 10-membered
heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6alkanoyloxy,
C.sub.1-C.sub.6alkanoylamino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, C.sub.1-C.sub.6alkylsulfonyloxy,
mono- or di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with 1 to 4 substituents independently chosen from oxo,
amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, alkoxycarbonyl, alkanoyloxy, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle.
[0102] In certain embodiments, compounds or salts or hydrates of
formula Ia and/or Ib are provided such that
[0103] (a) when U is N, W is --C(.dbd.O)NH--, and L and A are
absent, then M is other than thienyl or unsubstituted or
halogen-substituted 4- to 10-membered carbocycle;
[0104] (b) when U is N, W is --NHC(.dbd.O)--, and A is absent, then
M is other than thienyl or unsubstituted or halogen-substituted 4-
to 10-membered carbocycle;
[0105] (c) when U is CH, then W--X--Y is other than:
##STR00007##
[0106] wherein Rx is H or C(.dbd.O)--O-alkyl;
[0107] (d) when one of Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 is N,
and the others of Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are each
CH, R.sub.A and Y are each independently alkyl, aryl aralkyl or
heteroaryl, W is --C(.dbd.O)N(H)--, and U is CR.sub.1A, then
R.sub.1A is other than OH;
[0108] (e) when Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are each CH,
R.sub.A is unsubstituted phenyl, W is --C(.dbd.O)N(H)--, X is
unsubstituted alkylene or alkylene substituted with one hydroxyl, Y
is dialkylamino, unsubstituted heterocycloalkyl, or
heterocycloalkyl substituted with one alkyl, and U is CR.sub.1A,
then R.sub.1A is other than halogen; and
[0109] (f) when Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are each
CR.sub.1, at least two of R.sub.1 are H and the remaining R.sub.1
are each independently H, halogen, nitro, hydroxy, or alkoxy,
R.sub.A is cycloalkyl or substituted or unsubstituted phenyl, W is
--C(.dbd.O)N(H)--, X is unsubstituted alkylene, Y is alkyl or
cycloalkyl each optionally substituted with one COOH, dialkylamino,
unsubstituted heteroaryl, unsubstituted heterocycloalkyl, or
heterocycloalkyl substituted with nitrile or amino, and U is
CR.sub.1A, then R.sub.1A is other than alkyl.
[0110] Within certain aspects, such compounds may be used in vitro
or in vivo, as modulators of P2X.sub.7 receptors that may be used
in a variety of contexts, including in the treatment of conditions
responsive to P2X.sub.7 receptor modulation, such as pain.
[0111] Such modulators are also useful as probes for detection and
localization of P2X.sub.7 receptor and as standards in P2X.sub.7
receptor-mediated signal transduction assays.
[0112] In certain embodiments, such compounds exhibit no detectable
agonist activity an in vitro assay of P2X.sub.7 receptor
agonism.
[0113] In certain other embodiments, such compounds are capable of
exhibiting an IC.sub.50 value of 20 micromolar or less in an assay
for P2X.sub.7 receptor antagonism.
[0114] In some embodiments, the compounds of the present invention
or salts or hydrates thereof are compounds of formula Ia.
[0115] In other embodiments, the compounds of the present invention
or salts or hydrates thereof are compounds of formula Ib.
[0116] In certain embodiments of the present invention, the
compounds of Formula Ia and/or Ib are provided as a salt or hydrate
thereof.
[0117] Within Formula Ia and/or Ib, the heteroaryl core
##STR00008##
[0118] comprises at least one nitrogen atom, as indicated, and
optionally comprises additional nitrogen atom(s) at one or more of
U, V, Z.sub.1, Z.sub.2, Z.sub.3, and/or Z.sub.4. In certain
embodiments, U is CR.sub.1A; in further embodiments, U is CH.
Within other embodiments, Z.sub.1, Z.sub.2 and Z.sub.3 are each
CR.sub.1.
[0119] In some other embodiments of compounds of Formula Ia and/or
Ib, X is substituted.
[0120] In other embodiments of compounds of Formula Ia and/or Ib,
R.sub.B, R.sub.C, R.sub.D, and R.sub.E are each independently
--COOH, C.sub.1-C.sub.8alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.2-C.sub.8alkyl ether, mono- or
di-(C.sub.1-C.sub.6alkyl)aminoC.sub.0-C.sub.4alkyl, (4- to
7-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl and
phenylC.sub.0-C.sub.2alkyl; or any two of R.sub.B, R.sub.C,
R.sub.D, and R.sub.E taken together with the carbon atom or atoms
through which they are connected form a 3- to 7-membered cycloalkyl
or a 4- to 7-membered heterocycloalkyl; or any one of R.sub.B,
R.sub.C, R.sub.D, and R.sub.E taken together with R.sub.4 and the
atom or atoms through which they are connected form a 4- to
7-membered heterocycloalkyl
[0121] In certain further embodiments of compounds of Formula Ia
and/or Ib, R.sub.B, R.sub.C, R.sub.D, and R.sub.E are each
independently C.sub.1-C.sub.4alkyl,
(C.sub.3-C.sub.8cycloalkyl)C.sub.0-C.sub.2alkyl, or
phenylC.sub.0-C.sub.2alkyl;
[0122] In further aspects of compounds of Formula Ia and/or Ib, any
two of R.sub.D, R.sub.C, R.sub.D, and R.sub.E taken together with
the carbon atom or atoms through which they are connected form a 3-
to 7-membered cycloalkyl or a 4- to 7-membered
heterocycloalkyl.
[0123] In certain aspects compounds of Formula Ia and/or Ib, any
one of R.sub.B, R.sub.C, R.sub.D, and R.sub.E taken together with
R.sub.4 and the atom or atoms through which they are connected form
a 4- to 7-membered heterocycloalkyl.
[0124] In some embodiments of compounds of Formula Ia and/or Ib, U
is CR.sub.1A (e.g., CH).
[0125] In other embodiments of compounds of Formula Ia and/or Ib, U
is N.
[0126] In certain embodiments of compounds of Formula Ia and/or Ib,
Y is C.sub.1-C.sub.8alkyl, C.sub.3-C.sub.16cycloalkyl, 6- to
16-membered aryl or (5- to 16-membered heteroaryl, each optionally
substituted, preferably C.sub.1-C.sub.8alkyl, 6- to 16-membered
aryl or (5- to 16-membered heteroaryl, each optionally
substituted.
[0127] In other embodiments of compounds of Formula Ia and/or Ib, Y
is optionally substituted with from 1 to 3 substituents. In further
embodiments, Y is optionally substituted with 1 or 2
substituents.
[0128] In still other embodiments of compounds of Formula Ia and/or
Ib, Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 are independently CH. In
other aspects, Z.sub.4 is N. In further aspects, at least one of
Z.sub.1, Z.sub.2, Z.sub.3, and Z.sub.4 is CR.sub.1. In certain
further aspects, Z.sub.1 or Z.sub.4 is CR.sub.1; or Z.sub.1 is
CR.sub.1.
[0129] In certain embodiments of Formula Ia and/or Ib, R.sub.1 is
hydrogen, halogen, cyano, aminocarbonyl, or
C.sub.1-C.sub.6haloalkyl. In certain aspects where R.sub.1 is
halogen, said halogen is fluoro, chloro, or bromo; said halogen is
fluoro. In certain further aspects R.sub.1 is
C.sub.1-C.sub.3haloalkyl; in still other aspects R.sub.1 is
C.sub.1haloalkyl; in yet other aspects, said C.sub.1haloalkyl is
trifluoromethyl. Alternatively, each R.sub.1 is H.
[0130] In certain other embodiments of Formula Ia and/or Ib,
preferably of Formula Ia, R.sub.1A is hydrogen, hydroxy, halogen,
cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; preferably hydrogen, halogen,
cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6hydroxyalkyl, C.sub.1-C.sub.6aminoalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.6alkyl ether, C.sub.1-C.sub.6alkanoyl,
C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino; more preferably hydrogen,
cyano, amino, aminocarbonyl, aminosulfonyl, COOH,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6aminoalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoyl, C.sub.1-C.sub.6alkylsulfonyl,
(C.sub.3-C.sub.7cycloalkyl)C.sub.0-C.sub.4alkyl, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkanoylamino, mono-
or di-(C.sub.1-C.sub.6alkyl)aminocarbonyl, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl or
(C.sub.1-C.sub.6alkyl)sulfonylamino.
[0131] In some embodiments of compounds of Formula Ia and/or Ib, W
is --NR.sub.4C(.dbd.O)-- or --NR.sub.4--NR.sub.4--C(.dbd.O)--. In
other aspects, W is --C(.dbd.O)NR.sub.4--.
[0132] In other embodiments, R.sub.4 is H.
[0133] In certain embodiments of compounds of Formula Ia and/or Ib,
M is:
[0134] (i) hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl or --COOH; or
[0135] (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy, (4- to
10-membered aryl)C.sub.0-C.sub.4alkyl, (4- to 10-membered
heterocycle)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6alkanoyloxy,
C.sub.1-C.sub.6alkanoylamino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, C.sub.1-C.sub.6alkylsulfonyloxy,
mono- or di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with from 1 to 4 substituents independently chosen from
oxo, amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle.
[0136] In certain other embodiments of compounds of Formula Ia
and/or Ib, M is:
[0137] (i) hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl or --COOH; or
[0138] (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy, (4- to
10-membered aryl)C.sub.0-C.sub.4alkyl, (4- to 10-membered
heterocycloalkyl)C.sub.0-C.sub.4alkyl, C.sub.1-C.sub.6alkanoyloxy,
C.sub.1-C.sub.6alkanoylamino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, C.sub.1-C.sub.6alkylsulfonyloxy,
mono- or di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with 1 to 4 substituents independently chosen from oxo,
amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle.
[0139] In still other embodiments of compounds of Formula Ia and/or
Ib, M is arylC.sub.1-C.sub.4alkyl or
heteroarylC.sub.1-C.sub.4alkyl. In some such compounds, M is
substituted; in others, it is unsubstituted.
[0140] In yet other embodiments of compounds of Formula Ia and/or
Ib, M is:
[0141] (i) hydroxy, halogen, cyano, amino, aminocarbonyl,
aminosulfonyl or --COOH; or
[0142] (ii) C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy, (4- to
10-membered heterocycloalkyl)C.sub.0-C.sub.4alkyl,
C.sub.1-C.sub.6alkanoyloxy, C.sub.1-C.sub.6alkanoylamino,
C.sub.1-C.sub.6alkylsulfonyl, C.sub.1-C.sub.6alkylsulfonylamino,
C.sub.1-C.sub.6alkylsulfonyloxy, mono- or
di-C.sub.1-C.sub.6alkylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, or mono- or
di-(C.sub.1-C.sub.6alkyl)aminocarbonyl; each of which is optionally
substituted with 1 to 4 substituents independently chosen from oxo,
amino, halogen, hydroxy, cyano, aminocarbonyl, aminosulfonyl,
--COOH, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6hydroxyalkyl,
C.sub.1-C.sub.6haloalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6haloalkoxy, C.sub.2-C.sub.6alkyl ether,
C.sub.1-C.sub.6alkanoylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)amino, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6alkylsulfonylamino, mono- or
di-(C.sub.1-C.sub.6alkyl)aminosulfonyl, mono- or
di-(C.sub.1-C.sub.6alkylamino)carbonyl, and 4- to 7-membered
heterocycle.
[0143] In certain embodiments of compounds of Formula Ia and/or Ib,
M is optionally substituted with one or two substituents. In
certain aspects these substituents are independently chosen from
halogen, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, and
C.sub.1-C.sub.6alkoxy.
[0144] In some embodiments of compounds of Formula Ia and/or Ib,
R.sub.A is other than halophenyl, other than haloaryl, other than
cycloalkyl, and/or other than thienyl.
[0145] In certain embodiments of compounds of Formula Ia and/or Ib,
when R.sub.A is optionally substituted heteroaryl, said heteroaryl
has at least one oxygen or nitrogen ring atom, preferably at least
one nitrogen ring atom, and more preferably at least two nitrogen
ring atoms.
[0146] In some other embodiments of compounds of Formula Ia and/or
Ib, R.sub.A is other than carbocyclic.
[0147] In still other embodiments of compounds of Formula Ia and/or
Ib, W--X--Y is optionally substituted --C(.dbd.O)N(H)R.sub.v,
Alternatively, in some embodiments, W--X--Y is other than
unsubstituted --C(.dbd.O)N(H)heteroaralkyl; or W--X--Y is other
than unsubstituted --C(.dbd.O)N(H)heterocyclic; or W--X--Y is other
than unsubstituted --C(.dbd.O)N(H)alkoxyalkyl; or W--X--Y is other
than unsubstituted --C(.dbd.O)N(H)aralkyl.
[0148] In some embodiments of compounds of Formula Ia and/or Ib,
R.sub.v is arylC.sub.1-C.sub.4alkyl or
heteroarylC.sub.1-C.sub.4alkyl. In some aspects, R.sub.y is
substituted; in others, it is unsubstituted. Alternatively, in some
embodiments, R.sub.v is other than aminoalkyl, alkylaminoalkyl, or
dialkylaminoalkyl, alkyl, alkenyl, or alkynyl.
[0149] In certain embodiments of compounds of Formula Ia and/or Ib,
W is --C(.dbd.O)N(H)--.
[0150] In other embodiments of compounds of Formula Ia and/or Ib, M
is optionally substituted heteroaryl; preferably said heteroaryl
contains at least one nitrogen ring atom, more preferably said
heteroaryl contains at least two nitrogen ring atoms. In certain
embodiments M is optionally substituted pyrimidinyl, preferably
optionally substituted pyrimidin-2-yl.
[0151] In some embodiments of compounds of Formula Ia and/or Ib, X
is C.sub.1-C.sub.2alkylene, optionally substituted, preferably
substituted with C.sub.1-C.sub.4alkyl. In other embodiments, X is
substituted with at least 2 substituents selected from R.sub.B,
R.sub.C, and R.sub.E, wherein any two of R.sub.B, R.sub.C, R.sub.D,
and R.sub.E taken together with the carbon atom or atoms through
which they are connected form a 3- to 7-membered cycloalkyl.
Alternatively in some embodiments, X is substituted with at least 2
substituents selected from R.sub.B, R.sub.C, R.sub.D, and R.sub.E,
wherein any two of R.sub.B, R.sub.C, R.sub.D, and R.sub.E taken
together with the carbon atom or atoms through which they are
connected form a 3- to 7-membered cycloalkyl, preferably a 5- to
6-membered cycloalkyl.
[0152] In certain embodiments of compounds of Formula Ia and/or Ib,
Y is optionally substituted carbocycle or heterocycle, preferably
adamantyl, phenyl, pyridyl, or morpholinyl, each optionally
substituted.
[0153] In certain other embodiments of compounds of Formula Ia
and/or Ib, the compound is a compound provided in Table A, herein,
or a salt or hydrate thereof.
[0154] Representative heteroaryl amide analogues provided herein
include, but are not limited to, those specifically described in
Examples 1-6 and accompanying Table A. It will be apparent that the
specific compounds recited herein are representative only, and are
not intended to limit the scope of the present invention. Further,
as noted above, all compounds of the present invention may be
present as a free acid or base, or as a pharmaceutically acceptable
salt. In addition, other forms such as hydrates and prodrugs of
such compounds are specifically contemplated by the present
invention.
[0155] Within certain aspects of the present invention, heteroaryl
amide analogues provided herein detectably alter (modulate)
P2X.sub.7 receptor activity, as determined using an assay such as
an assay recited in Example 7, herein. Additional assays that may
be used for this purpose include assays that measure IL-1.beta.
release; assays that measure uptake of a membrane-impermeant
fluorescent dye such as YO-PRO1; assays that measure lucifer yellow
uptake; assays that measure ethidium bromide uptake; and assays
that use calcium imaging to detect P2X.sub.7 activity; all of which
assays are well known in the art. Certain modulators provided
herein detectably modulate P2X.sub.7 receptor activity at
micromolar concentrations, at nanomolar concentrations, or at
subnanomolar concentrations.
[0156] As noted above, compounds that are P2X.sub.7 receptor
antagonists are preferred within certain embodiments. IC.sub.50
values for such compounds may be determined using a standard in
vitro P2X.sub.7 receptor-mediated calcium mobilization assay, as
provided in Example 7. Briefly, cells expressing P2X.sub.7 receptor
are contacted with a compound of interest and with an indicator of
intracellular calcium concentration (e.g., a membrane permeable
calcium sensitivity dye such as Fluo-3, Fluo-4 or Fura-2
(Invitrogen, Carlsbad, Calif.), each of which produce a fluorescent
signal when bound to Ca.sup.++). Such contact is preferably carried
out by one or more incubations of the cells in buffer or culture
medium comprising either or both of the compound and the indicator
in solution. Contact is maintained for an amount of time sufficient
to allow the dye to enter the cells (e.g., 1-2 hours). Cells are
washed or filtered to remove excess dye and are then contacted with
a P2X.sub.7 receptor agonist (e.g., ATP or
2'(3')-O-(4-benzoyl-benzoyl)adenosine 5'-triphosephate at, for
example, a concentration equal to the EC.sub.50 concentration), and
a fluorescence response is measured. When agonist-contacted cells
are contacted with a compound that is a P2X.sub.7 receptor
antagonist, the fluorescence response is generally reduced by at
least 20%, preferably at least 50% and more preferably at least
80%, as compared to cells that are contacted with the agonist in
the absence of test compound. In certain embodiments, P2X.sub.7
receptor antagonists provided herein exhibit no detectable agonist
activity an in vitro assay of P2X.sub.7 receptor agonism at a
concentration of compound equal to the IC.sub.50. Certain such
antagonists exhibit no detectable agonist activity an in vitro
assay of P2X.sub.7 receptor agonism at a concentration of compound
that is 100-fold higher than the IC.sub.50.
[0157] P2X.sub.7 receptor modulating activity may also, or
alternatively, be assessed using an in vivo pain relief assay as
provided in Example 8. Modulators provided herein preferably have a
statistically significant specific effect on P2X.sub.7 receptor
activity within such a functional assay.
[0158] In certain embodiments, preferred modulators are
non-sedating. In other words, a dose of modulator that is twice the
minimum dose sufficient to provide analgesia in an animal model for
determining pain relief (such as a model provided in Example 8,
herein) causes only transient (i.e., lasting for no more than 1/2
the time that pain relief lasts) or preferably no statistically
significant sedation in an animal model assay of sedation (using
the method described by Fitzgerald et al. (1988) Toxicology
49(2-3):433-9). Preferably, a dose that is five times the minimum
dose sufficient to provide analgesia does not produce statistically
significant sedation. More preferably, a modulator provided herein
does not produce sedation at intravenous doses of less than 25
mg/kg (preferably less than 10 mg/kg) or at oral doses of less than
140 mg/kg (preferably less than 50 mg/kg, more preferably less than
30 mg/kg).
[0159] If desired, compounds provided herein may be evaluated for
certain pharmacological properties including, but not limited to,
oral bioavailability (preferred compounds are orally bioavailable
to an extent allowing for therapeutically effective concentrations
of the compound to be achieved at oral doses of less than 140
mg/kg, preferably less than 50 mg/kg, more preferably less than 30
mg/kg, even more preferably less than 10 mg/kg, still more
preferably less than 1 mg/kg and most preferably less than 0.1
mg/kg), toxicity (a preferred compound is nontoxic when a
therapeutically effective amount is administered to a subject),
side effects (a preferred compound produces side effects comparable
to placebo when a therapeutically effective amount of the compound
is administered to a subject), serum protein binding and in vitro
and in vivo half-life (a preferred compound exhibits an in vivo
half-life allowing for Q.I.D. dosing, preferably T.I.D. dosing,
more preferably B.I.D. dosing, and most preferably once-a-day
dosing). In addition, differential penetration of the blood brain
barrier may be desirable for modulators used to treat pain or
neurodegenerative disease by modulating CNS P2X.sub.7 receptor
activity such that total daily oral doses as described above
provide such modulation to a therapeutically effective extent,
while low brain levels of modulators used to treat peripheral nerve
mediated pain or certain inflammatory diseases (e.g. rheumatoid
arthritis) may be preferred (i.e., such doses do not provide brain
(e.g., CSF) levels of the compound sufficient to significantly
modulate P2X.sub.7 receptor activity). Routine assays that are well
known in the art may be used to assess these properties, and
identify superior compounds for a particular use. For example,
assays used to predict bioavailability include transport across
human intestinal cell monolayers, including Caco-2 cell monolayers.
Penetration of the blood brain barrier of a compound in humans may
be predicted from the brain levels of the compound in laboratory
animals given the compound (e.g., intravenously). Serum protein
binding may be predicted from albumin binding assays. Compound
half-life is inversely proportional to the frequency of dosage of a
compound. In vitro half-lives of compounds may be predicted from
assays of microsomal half-life as described, for example, within
Example 7 of U.S. Patent Application Publication Number
2005/0070547.
[0160] As noted above, preferred compounds provided herein are
nontoxic. In general, the term "nontoxic" shall be understood in a
relative sense and is intended to refer to any substance that has
been approved by the United States Food and Drug Administration
("FDA") for administration to mammals (preferably humans) or, in
keeping with established criteria, is susceptible to approval by
the FDA for administration to mammals (preferably humans). In
addition, a highly preferred nontoxic compound generally satisfies
one or more of the following criteria: (1) does not substantially
inhibit cellular ATP production; (2) does not significantly prolong
heart QT intervals; (3) does not cause substantial liver
enlargement, or (4) does not cause substantial release of liver
enzymes.
[0161] As used herein, a compound that does not substantially
inhibit cellular ATP production is a compound that satisfies the
criteria set forth in Example 8 of U.S. Patent Application
Publication Number 2005/0070547. In other words, cells treated as
described therein with 100 .mu.M of such a compound exhibit ATP
levels that are at least 50% of the ATP levels detected in
untreated cells. In more highly preferred embodiments, such cells
exhibit ATP levels that are at least 80% of the ATP levels detected
in untreated cells.
[0162] A compound that does not significantly prolong heart QT
intervals is a compound that does not result in a statistically
significant prolongation of heart QT intervals (as determined by
electrocardiography) in guinea pigs, minipigs or dogs upon
administration of a dose that yields a serum concentration equal to
the EC.sub.50 or IC.sub.50 for the compound. In certain preferred
embodiments, a dose of 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 40 or 50
mg/kg administered parenterally or orally does not result in a
statistically significant prolongation of heart QT intervals.
[0163] A compound does not cause substantial liver enlargement if
daily treatment of laboratory rodents (e.g., mice or rats) for 5-10
days with a dose that yields a serum concentration equal to the
EC.sub.50 or IC.sub.50 for the compound results in an increase in
liver to body weight ratio that is no more than 100% over matched
controls. In more highly preferred embodiments, such doses do not
cause liver enlargement of more than 75% or 50% over matched
controls. If non-rodent mammals (e.g., dogs) are used, such doses
should not result in an increase of liver to body weight ratio of
more than 50%, preferably not more than 25%, and more preferably
not more than 10% over matched untreated controls. Preferred doses
within such assays include 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or 50
mg/kg administered parenterally or orally.
[0164] Similarly, a compound does not promote substantial release
of liver enzymes if administration of twice the minimum dose that
yields a serum concentration equal to the EC.sub.50 or IC.sub.50 at
P2X.sub.7 receptor for the compound does not elevate scrum levels
of ALT, LDH or AST in laboratory animals (e.g., rodents) by more
than 100% over matched mock-treated controls. In more highly
preferred embodiments, such doses do not elevate such serum levels
by more than 75% or 50% over matched controls. Alternatively, a
compound does not promote substantial release of liver enzymes if,
in an in vitro hepatocyte assay, concentrations (in culture media
or other such solutions that are contacted and incubated with
hepatocytes in vitro) that are equal to the EC.sub.50 or IC.sub.50
for the compound do not cause detectable release of any of such
liver enzymes into culture medium above baseline levels seen in
media from matched mock-treated control cells. In more highly
preferred embodiments, there is no detectable release of any of
such liver enzymes into culture medium above baseline levels when
such compound concentrations are five-fold, and preferably ten-fold
the EC.sub.50 or IC.sub.50 for the compound.
[0165] In other embodiments, certain preferred compounds do not
inhibit or induce microsomal cytochrome P450 enzyme activities,
such as CYP1A2 activity, CYP2A6 activity, CYP2C9 activity, CYP2C19
activity, CYP2D6 activity, CYP2E1 activity or CYP3A4 activity at a
concentration equal to the EC.sub.50 or IC.sub.50 at P2X.sub.7
receptor for the compound.
[0166] Certain preferred compounds are not elastogenic (e.g., as
determined using a mouse erythrocyte precursor cell micronucleus
assay, an Ames micronucleus assay, a spiral micronucleus assay or
the like) at a concentration equal the EC.sub.50 or IC.sub.50 for
the compound. In other embodiments, certain preferred compounds do
not induce sister chromatid exchange (e.g., in Chinese hamster
ovary cells) at such concentrations.
[0167] For detection purposes, as discussed in more detail below,
modulators provided herein may be isotopically-labeled or
radiolabeled. For example, compounds may have one or more atoms
replaced by an atom of the same element having an atomic mass or
mass number different from the atomic mass or mass number usually
found in nature. Examples of isotopes that can be present in the
compounds provided herein include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorous, fluorine and chlorine, such as
.sup.2H, .sup.3H, .sup.11C, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
.sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F and .sup.36Cl. In
addition, substitution with heavy isotopes such as deuterium (i.e.,
.sup.2H) can afford certain therapeutic advantages resulting from
greater metabolic stability, for example increased in vivo
half-life or reduced dosage requirements and, hence, may be
preferred in some circumstances.
[0168] Compounds may be radiolabeled by carrying out their
synthesis using precursors comprising at least one atom that is a
radioisotope. Each radioisotope is preferably carbon (e.g., ,
.sup.14C) hydrogen (e.g., .sup.3H), sulfur (e.g., .sup.35S), or
iodine (e.g., .sup.125I). Tritium labeled compounds may also be
prepared catalytically via platinum-catalyzed exchange in tritiated
acetic acid, acid-catalyzed exchange in tritiated trifluoroacetic
acid, or heterogeneous-catalyzed exchange with tritium gas using
the compound as substrate. In addition, certain precursors may be
subjected to tritium-halogen exchange with tritium gas, tritium gas
reduction of unsaturated bonds, or reduction using sodium
borotritide, as appropriate. Preparation of radiolabeled compounds
may be conveniently performed by a radioisotope supplier
specializing in custom synthesis of radiolabeled probe
compounds.
Preparation of Heteroaryl Amide Analogues
[0169] Heteroaryl amide analogues may generally be prepared using
standard synthetic methods. Starting materials are commercially
available from suppliers such as Sigma-Aldrich Corp. (St. Louis,
Mo.), or may be synthesized from commercially available precursors
using established protocols. By way of example, a synthetic route
similar to that shown in any of the following Schemes may be used,
together with synthetic methods known in the art of synthetic
organic chemistry. In some cases, protecting groups may be required
during preparation. Such protecting groups can be removed by
methods well known to those of ordinary skill in the art, such as
methods described in Greene and Wuts, "Protective Groups in Organic
Synthesis" (2.sup.nd Edition, John Wiley & Sons, 1991) or
Philip J. Kocienski, "Protecting Groups", 2.sup.nd ed., John Wiley
& Sons, Inc., New York (2005). In some cases, further organic
transformations may be performed using methods well known to those
of ordinary skill in the art, such as methods described in Richard
C. Larock, "Comprehensive Organic Transformation," (VCH Publisher,
Inc. 1989). Each variable in the following Schemes refers to any
group consistent with the description of the compounds provided
herein. Representative reaction conditions for use within the
following schemes are provided in the Examples.
[0170] Certain abbreviations used in the following Schemes and
elsewhere herein include:
[0171] BOP benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate
[0172] .delta. chemical shift
[0173] DCM dichloromethane
[0174] DMF dimethylformamide
[0175] DMSO dimethylsulfoxide
[0176] Et ethyl
[0177] EtOAc ethyl acetate
[0178] EtOH ethanol
[0179] h hour(s)
[0180] .sup.1H NMR proton nuclear magnetic resonance
[0181] Hz hertz
[0182] iPr isopropyl
[0183] MeOH methanol
[0184] min minute(s)
[0185] Ms methanesulfonyl
[0186] (M+1) mass+1
[0187] Ph.sub.3P triphenylphosphine
[0188] POCl.sub.3 phosphorus oxychloride
[0189] PTLC preparative thin layer chromatography
[0190] rt room temperature
[0191] TEA triethylamine
[0192] TFA trifluoroacetic acid
[0193] (CF.sub.3CO).sub.2O trifluoroacetic anhydride
[0194] Schemes 1-8 illustrate certain embodiments of the present
invention, and are intended to be exemplary only, and nonlimiting.
For example, it will be apparent that each reaction described in a
Scheme may be performed in combination with none, some or all of
the other reactions described therein. In addition, various
modifications to reaction conditions will be apparent, including
the use of different solvents and acids/bases, and changes in
reaction times and temperatures. All processes disclosed in
association with the present invention are contemplated to be
practiced on any scale, including milligram, gram, multigram,
kilogram, multikilogram or commercial industrial scale. It will
further be apparent that starting materials for each step, and each
reaction product, may be the indicated compound or may be a salt
(e.g., a pharmaceutically acceptable salt) or solvate (e.g.,
hydrate) thereof. Unless otherwise specified, each variable in the
following Schemes is as defined above.
##STR00009##
##STR00010##
##STR00011##
##STR00012##
##STR00013##
##STR00014##
[0195] Scheme 6 illustrates a general method of preparing certain
intermediates NH.sub.2--CH.sub.2--CHR.sub.5--Y of Formula X wherein
R.sub.5 is substituted phenyl or heteroaryl and
Y.dbd.N--R.sub.5bR.sub.5b wherein R.sub.5b is independently
hydrogen, C.sub.1-C.sub.6alkyl, or C.sub.3-C.sub.7cycloalkyl; or
both R.sub.5b are taken together to form a heterocycle. Strecker
condensation of the aryl carboxaldehyde and amine either with TMSCN
in a solvent such as acetonitrile, or with NaCN or KCN in a solvent
such MeOH-water or water at pH 3-4 (adjusted by hydrogen chloride)
gives the aminonitrile, which is reduced by LAH in a solvent such
as THF or by hydrogenation with Raney Nickel as a catalyst in a
solvent such as 7N ammonia in methanol to give the amine
intermediate NH.sub.2--CH.sub.2--CHR.sub.5--Y.
##STR00015##
[0196] Scheme 7 illustrates a general method of preparing certain
intermediates NH.sub.2--CH.sub.2--CHR.sub.5--Y or
NH.sub.2--CH.sub.2--CR.sub.5R.sub.5--Y of Formula X wherein Y is
substituted phenyl or heteroaryl. Alkylation of the starting
acetonitrile with one equivalent of X--R.sub.5 (X.dbd.Br or I),
with base such as sodium hydride in a solvent such as THF-DMSO
gives intermediate Y--X(R.sub.5)--CN. Alkylation of the starting
acetonitrile with two equivalents of X--R.sub.5 (X.dbd.Br or I) or
one equivalent of dibromo or diiodo (when R.sub.5 and R.sub.5 are
taken together to form a ring) with base such as sodium hydride in
a solvent such as THF-DMSO gives intermediate
Y--X(R.sub.5)(R.sub.5)--CN. Reduction of either product by LAH in a
solvent such as THF or by hydrogenation with Raney Nickel as a
catalyst in a solvent such as 7N ammonia in methanol provides
NH.sub.2--CH.sub.2--CHR.sub.5--Y or
NH.sub.2--CH.sub.2--CR.sub.5R.sub.5--Y.
##STR00016##
[0197] Scheme 8 illustrates a general method of preparing certain
intermediates NH.sub.2--CH.sub.2--CR.sub.5R.sub.5--Y of Formula X
wherein R.sub.5 is independently hydrogen, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.7cycloalkyl or phenyl; or both R.sub.5 are taken
together to form a C.sub.3-C.sub.8cycloalkyl or heterocycle, and
Y.dbd.N--R.sub.5bR.sub.5b wherein R.sub.5b is C.sub.1-C.sub.6
alkyl, optionally substituted with halogen, hydroxyl,
C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkoxy or CO.sub.2H, or both
R5.sub.b are taken together to form a 5- to 7-membered heterocycle.
Strecker condensation of the ketone and amine either with TMSCN in
a solvent such methanol with a catalyst such as ZnI.sub.2, or with
NaCN or KCN in a solvent such MeOH-water or water at pH 3-4
(adjusted by hydrogen chloride) gives the aminonitrile. Reduction
of the aminonitrile by LAH in a solvent such as THF or by
hydrogenation with Raney Nickel as a catalyst in a solvent such as
7N ammonia in methanol gives
NH.sub.2--CH.sub.2--CR.sub.5R.sub.5--Y.
Pharmaceutical Compositions
[0198] The present invention also provides pharmaceutical
compositions comprising one or more compounds provided herein,
together with at least one physiologically acceptable carrier or
excipient. Pharmaceutical compositions may comprise, for example,
one or more of water, buffers (e.g., sodium bicarbonate, neutral
buffered saline or phosphate buffered saline), ethanol, mineral
oil, vegetable oil, dimethylsulfoxide, carbohydrates (e.g.,
glucose, mannose, sucrose, starch, mannitol or dextrans), proteins,
adjuvants, polypeptides or amino acids such as glycine,
antioxidants, chelating agents such as EDTA or glutathione and/or
preservatives. In addition, other active ingredients may (but need
not) be included in the pharmaceutical compositions provided
herein.
[0199] Pharmaceutical compositions may be formulated for any
appropriate manner of administration, including, for example,
topical, oral, nasal, rectal or parenteral administration. The term
parenteral as used herein includes subcutaneous, intradermal,
intravascular (e.g., intravenous), intramuscular, spinal,
intracranial, intrathecal and intraperitoneal injection, as well as
any similar injection or infusion technique. In certain
embodiments, compositions suitable for oral use are preferred. Such
compositions include, for example, tablets, troches, lozenges,
aqueous or oily suspensions, dispersible powders or granules,
emulsion, hard or soft capsules, or syrups or elixirs. Within yet
other embodiments, pharmaceutical compositions may be formulated as
a lyophilizate. Formulation for topical administration may be
preferred for certain conditions (e.g., in the treatment of skin
conditions such as burns or itch). Formulation for direct
administration into the bladder (intravesicular administration) may
be preferred for treatment of urinary incontinence and overactive
bladder.
[0200] Compositions intended for oral use may further comprise one
or more components such as sweetening agents, flavoring agents,
coloring agents and/or preserving agents in order to provide
appealing and palatable preparations. Tablets contain the active
ingredient in admixture with physiologically acceptable excipients
that are suitable for the manufacture of tablets. Such excipients
include, for example, inert diluents (e.g., calcium carbonate,
sodium carbonate, lactose, calcium phosphate or sodium phosphate),
granulating and disintegrating agents (e.g., corn starch or alginic
acid), binding agents (e.g., starch, gelatin or acacia) and
lubricating agents (e.g., magnesium stearate, stearic acid or
talc). Tablets may be formed using standard techniques, including
dry granulation, direct compression and wet granulation. The
tablets may be uncoated or they may be coated by known
techniques.
[0201] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent (e.g., calcium carbonate, calcium phosphate or
kaolin), or as soft gelatin capsules wherein the active ingredient
is mixed with water or an oil medium (e.g., peanut oil, liquid
paraffin or olive oil).
[0202] Aqueous suspensions contain the active material(s) in
admixture with suitable excipients, such as suspending agents
(e.g., sodium carboxymethylcellulose, methylcellulose,
hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone,
gum tragacanth and gum acacia); and dispersing or wetting agents
(e.g., naturally-occurring phosphatides such as lecithin,
condensation products of an alkylene oxide with fatty acids such as
polyoxyethylene stearate, condensation products of ethylene oxide
with long chain aliphatic alcohols such as
heptadecaethyleneoxycetanol, condensation products of ethylene
oxide with partial esters derived from fatty acids and a hexitol
such as polyoxyethylene sorbitol monooleate, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and hexitol anhydrides such as polyethylene sorbitan
monooleate). Aqueous suspensions may also comprise one or more
preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or
more coloring agents, one or more flavoring agents, and/or one or
more sweetening agents, such as sucrose or saccharin.
[0203] Oily suspensions may be formulated by suspending the active
ingredient(s) in a vegetable oil (e.g., arachis oil, olive oil,
sesame oil or coconut oil) or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent such
as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such
as those set forth above, and/or flavoring agents may be added to
provide palatable oral preparations. Such suspensions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0204] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent, a
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, such as sweetening,
flavoring and coloring agents, may also be present.
[0205] Pharmaceutical compositions may also be formulated as
oil-in-water emulsions. The oily phase may be a vegetable oil
(e.g., olive oil or arachis oil), a mineral oil (e.g., liquid
paraffin) or a mixture thereof. Suitable emulsifying agents include
naturally-occurring gums (e.g., gum acacia or gum tragacanth),
naturally-occurring phosphatides (e.g., soy bean lecithin, and
esters or partial esters derived from fatty acids and hexitol),
anhydrides (e.g., sorbitan monoleate) and condensation products of
partial esters derived from fatty acids and hexitol with ethylene
oxide (e.g., polyoxyethylene sorbitan monoleate). An emulsion may
also comprise one or more sweetening and/or flavoring agents.
[0206] Syrups and elixirs may be formulated with sweetening agents,
such as glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also comprise one or more demulcents,
preservatives, flavoring agents and/or coloring agents.
[0207] Formulations for topical administration typically comprise a
topical vehicle combined with active agent(s), with or without
additional optional components. Suitable topical vehicles and
additional components are well known in the art, and it will be
apparent that the choice of a vehicle will depend on the particular
physical form and mode of delivery. Topical vehicles include water;
organic solvents such as alcohols (e.g., ethanol or isopropyl
alcohol) or glycerin; glycols (e.g., butylene, isoprene or
propylene glycol); aliphatic alcohols (e.g., lanolin); mixtures of
water and organic solvents and mixtures of organic solvents such as
alcohol and glycerin; lipid-based materials such as fatty acids,
acylglycerols (including oils, such as mineral oil, and fats of
natural or synthetic origin), phosphoglycerides, sphingolipids and
waxes; protein-based materials such as collagen and gelatin;
silicone-based materials (both non-volatile and volatile); and
hydrocarbon-based materials such as microsponges and polymer
matrices. A composition may further include one or more components
adapted to improve the stability or effectiveness of the applied
formulation, such as stabilizing agents, suspending agents,
emulsifying agents, viscosity adjusters, gelling agents,
preservatives, antioxidants, skin penetration enhancers,
moisturizers and sustained release materials. Examples of such
components are described in Martindale--The Extra Pharmacopoeia
(Pharmaceutical Press, London 1993) and Remington: The Science and
Practice of Pharmacy, 21.sup.st ed., Lippincott Williams &
Wilkins, Philadelphia, Pa. (2005). Formulations may comprise
microcapsules, such as hydroxymethylcellulose or
gelatin-microcapsules, liposomes, albumin microspheres,
microemulsions, nanoparticles or nanocapsules.
[0208] A topical formulation may be prepared in any of a variety of
physical forms including, for example, solids, pastes, creams,
foams, lotions, gels, powders, aqueous liquids and emulsions. The
physical appearance and viscosity of such pharmaceutically
acceptable forms can be governed by the presence and amount of
emulsifier(s) and viscosity adjuster(s) present in the formulation.
Solids are generally firm and non-pourable and commonly are
formulated as bars or sticks, or in particulate form; solids can be
opaque or transparent, and optionally can contain solvents,
emulsifiers, moisturizers, emollients, fragrances, dyes/colorants,
preservatives and other active ingredients that increase or enhance
the efficacy of the final product. Creams and lotions are often
similar to one another, differing mainly in their viscosity; both
lotions and creams may be opaque, translucent or clear and often
contain emulsifiers, solvents, and viscosity adjusting agents, as
well as moisturizers, emollients, fragrances, dyes/colorants,
preservatives and other active ingredients that increase or enhance
the efficacy of the final product. Gels can be prepared with a
range of viscosities, from thick or high viscosity to thin or low
viscosity. These formulations, like those of lotions and creams,
may also contain solvents, emulsifiers, moisturizers, emollients,
fragrances, dyes/colorants, preservatives and other active
ingredients that increase or enhance the efficacy of the final
product. Liquids are thinner than creams, lotions, or gels and
often do not contain emulsifiers. Liquid topical products often
contain solvents, emulsifiers, moisturizers, emollients,
fragrances, dyes/colorants, preservatives and other active
ingredients that increase or enhance the efficacy of the final
product.
[0209] Suitable emulsifiers for use in topical formulations
include, but are not limited to, ionic emulsifiers, cetearyl
alcohol, non-ionic emulsifiers like polyoxyethylene oleyl ether,
PEG-40 stearate, ceteareth-12, ceteareth-20, ceteareth-30,
ceteareth alcohol, PEG-100 stearate and glyceryl stearate. Suitable
viscosity adjusting agents include, but are not limited to,
protective colloids or non-ionic gums such as
hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate,
silica, microcrystalline wax, beeswax, paraffin, and cetyl
palmitate. A gel composition may be formed by the addition of a
gelling agent such as chitosan, methyl cellulose, ethyl cellulose,
polyvinyl alcohol, polyquaterniums, hydroxyethylcellulose,
hydroxypropylcellulose, hydroxypropylmethylcellulose, carbomer or
ammoniated glycyrrhizinate. Suitable surfactants include, but are
not limited to, nonionic, amphoteric, ionic and anionic
surfactants. For example, one or more of dimethicone copolyol,
polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80,
lauramide DEA, cocamide DEA, and cocamide MEA, oleyl betaine,
cocamidopropyl phosphatidyl PG-dimonium chloride, and ammonium
laureth sulfate may be used within topical formulations. Suitable
preservatives include, but are not limited to, antimicrobials such
as methylparaben, propylparaben, sorbic acid, benzoic acid, and
formaldehyde, as well as physical stabilizers and antioxidants such
as vitamin E, sodium ascorbate/ascorbic acid and propyl gallate.
Suitable moisturizers include, but are not limited to, lactic acid
and other hydroxy acids and their salts, glycerin, propylene
glycol, and butylene glycol. Suitable emollients include lanolin
alcohol, lanolin, lanolin derivatives, cholesterol, petrolatum,
isostearyl neopentanoate and mineral oils. Suitable fragrances and
colors include, but are not limited to, FD&C Red No. 40 and
FD&C Yellow No. 5. Other suitable additional ingredients that
may be included a topical formulation include, but are not limited
to, abrasives, absorbents, anti-caking agents, anti-foaming agents,
anti-static agents, astringents (e.g., witch hazel, alcohol and
herbal extracts such as chamomile extract), binders/excipients,
buffering agents, chelating agents, film forming agents,
conditioning agents, propellants, opacifying agents, pH adjusters
and protectants.
[0210] An example of a suitable topical vehicle for formulation of
a gel is: hydroxypropylcellulose (2.1%); 70/30 isopropyl
alcohol/water (90.9%); propylene glycol (5.1%); and Polysorbate 80
(1.9%). An example of a suitable topical vehicle for formulation as
a foam is: cetyl alcohol (1.1%); stearyl alcohol (0.5%; Quaternium
52 (1.0%); propylene glycol (2.0%); Ethanol 95 PGF3 (61.05%);
deionized water (30.05%); P75 hydrocarbon propellant (4.30%). All
percents are by weight.
[0211] Typical modes of delivery for topical compositions include
application using the fingers; application using a physical
applicator such as a cloth, tissue, swab, stick or brush; spraying
(including mist, aerosol or foam spraying); dropper application;
sprinkling; soaking; and rinsing.
[0212] A pharmaceutical composition may be prepared as a sterile
injectible aqueous or oleaginous suspension. The compound(s)
provided herein, depending on the vehicle and concentration used,
can either be suspended or dissolved in the vehicle. Such a
composition may be formulated according to the known art using
suitable dispersing, wetting agents and/or suspending agents such
as those mentioned above. Among the acceptable vehicles and
solvents that may be employed are water, 1,3-butanediol, Ringer's
solution and isotonic sodium chloride solution. In addition,
sterile, fixed oils may be employed as a solvent or suspending
medium. For this purpose any bland fixed oil may be employed,
including synthetic mono- or diglycerides. In addition, fatty acids
such as oleic acid find use in the preparation of injectible
compositions, and adjuvants such as local anesthetics,
preservatives and/or buffering agents can be dissolved in the
vehicle.
[0213] Pharmaceutical compositions may also be formulated as
suppositories (e.g., for rectal administration). Such compositions
can be prepared by mixing the drug with a suitable non-irritating
excipient that is solid at ordinary temperatures but liquid at the
rectal temperature and will therefore melt in the rectum to release
the drug. Suitable excipients include, for example, cocoa butter
and polyethylene glycols.
[0214] Compositions for inhalation typically can be provided in the
form of a solution, suspension or emulsion that can be administered
as a dry powder or in the form of an aerosol using a conventional
propellant (e.g., dichlorodifluoromethane or
trichlorofluoromethane).
[0215] Pharmaceutical compositions may be formulated for release at
a pre-determined rate. Instantaneous release may be achieved, for
example, via sublingual administration (i.e., administration by
mouth in such a way that the active ingredient(s) are rapidly
absorbed via the blood vessels under the tongue rather than via the
digestive tract). Controlled release formulations (i.e.,
formulations such as a capsule, tablet or coated tablet that slows
and/or delays release of active ingredient(s) following
administration) may be administered by, for example, oral, rectal
or subcutaneous implantation, or by implantation at a target site.
In general, a controlled release formulation comprises a matrix
and/or coating that delays disintegration and absorption in the
gastrointestinal tract (or implantation site) and thereby provides
a delayed action or a sustained action over a longer period. One
type of controlled-release formulation is a sustained-release
formulation, in which at least one active ingredient is
continuously released over a period of time at a constant rate.
Preferably, the therapeutic agent is released at such a rate that
blood (e.g., plasma) concentrations are maintained within the
therapeutic range, but below toxic levels, over a period of time
that is at least 4 hours, preferably at least 8 hours, and more
preferably at least 12 hours. Such formulations may generally be
prepared using well known technology and administered by, for
example, oral, rectal or subcutaneous implantation, or by
implantation at the desired target site. Carriers for use within
such formulations are biocompatible, and may also be biodegradable;
preferably the formulation provides a relatively constant level of
modulator release. The amount of modulator contained within a
sustained release formulation depends upon, for example, the site
of implantation, the rate and expected duration of release and the
nature of the condition to be treated or prevented.
[0216] Controlled release may be achieved by combining the active
ingredient(s) with a matrix material that itself alters release
rate and/or through the use of a controlled-release coating. The
release rate can be varied using methods well known in the art,
including (a) varying the thickness or composition of coating, (b)
altering the amount or manner of addition of plasticizer in a
coating, (c) including additional ingredients, such as
release-modifying agents, (d) altering the composition, particle
size or particle shape of the matrix, and (e) providing one or more
passageways through the coating. The amount of modulator contained
within a sustained release formulation depends upon, for example,
the method of administration (e.g., the site of implantation), the
rate and expected duration of release and the nature of the
condition to be treated or prevented.
[0217] The matrix material, which itself may or may not serve a
controlled-release function, is generally any material that
supports the active ingredient(s). For example, a time delay
material such as glyceryl monosterate or glyceryl distearate may be
employed. Active ingredient(s) may be combined with matrix material
prior to formation of the dosage form (e.g., a tablet).
Alternatively, or in addition, active ingredient(s) may be coated
on the surface of a particle, granule, sphere, microsphere, bead or
pellet that comprises the matrix material. Such coating may be
achieved by conventional means, such as by dissolving the active
ingredient(s) in water or other suitable solvent and spraying.
Optionally, additional ingredients are added prior to coating
(e.g., to assist binding of the active ingredient(s) to the matrix
material or to color the solution). The matrix may then be coated
with a barrier agent prior to application of controlled-release
coating. Multiple coated matrix units may, if desired, be
encapsulated to generate the final dosage form.
[0218] In certain embodiments, a controlled release is achieved
through the use of a controlled release coating (i.e., a coating
that permits release of active ingredient(s) at a controlled rate
in aqueous medium). The controlled release coating should be a
strong, continuous film that is smooth, capable of supporting
pigments and other additives, non-toxic, inert and tack-free.
Coatings that regulate release of the modulator include
pH-independent coatings, pH-dependent coatings (which may be used
to release modulator in the stomach) and enteric coatings (which
allow the formulation to pass intact through the stomach and into
the small intestine, where the coating dissolves and the contents
are absorbed by the body). It will be apparent that multiple
coatings may be employed (e.g., to allow release of a portion of
the dose in the stomach and a portion further along the
gastrointestinal tract). For example, a portion of active
ingredient(s) may be coated over an enteric coating, and thereby
released in the stomach, while the remainder of active
ingredient(s) in the matrix core is protected by the enteric
coating and released further down the GI tract. pH dependent
coatings include, for example, shellac, cellulose acetate
phthalate, polyvinyl acetate phthalate,
hydroxypropylmethylcellulose phthalate, methacrylic acid ester
copolymers and zein.
[0219] In certain embodiments, the coating is a hydrophobic
material, preferably used in an amount effective to slow the
hydration of the gelling agent following administration. Suitable
hydrophobic materials include alkyl celluloses (e.g.,
ethylcellulose or carboxymethylcellulose), cellulose ethers,
cellulose esters, acrylic polymers (e.g., poly(acrylic acid),
poly(methacrylic acid), acrylic acid and methacrylic acid
copolymers, methyl methacrylate copolymers, ethoxy ethyl
methacrylates, cyanoethyl methacrylate, methacrylic acid alkamide
copolymer, poly(methyl methacrylate), polyacrylamide, ammonio
methacrylate copolymers, aminoalkyl methacrylate copolymer,
poly(methacrylic acid anhydride) and glycidyl methacrylate
copolymers) and mixtures of the foregoing. Representative aqueous
dispersions of ethylcellulose include, for example, AQUACOAT.RTM.
(FMC Corp., Philadelphia, Pa.) and SURELEASE.RTM. (Colorcon, Inc.,
West Point, Pa.), both of which can be applied to the substrate
according to the manufacturer's instructions. Representative
acrylic polymers include, for example, the various EUDRAGIT.RTM.
(Rohm America, Piscataway, N.J.) polymers, which may be used singly
or in combination depending on the desired release profile,
according to the manufacturer's instructions.
[0220] The physical properties of coatings that comprise an aqueous
dispersion of a hydrophobic material may be improved by the
addition or one or more plasticizers. Suitable plasticizers for
alkyl celluloses include, for example, dibutyl sebacate, diethyl
phthalate, triethyl citrate, tributyl citrate and triacetin.
Suitable plasticizers for acrylic polymers include, for example,
citric acid esters such as triethyl citrate and tributyl citrate,
dibutyl phthalate, polyethylene glycols, propylene glycol, diethyl
phthalate, castor oil and triacetin.
[0221] Controlled-release coatings are generally applied using
conventional techniques, such as by spraying in the form of an
aqueous dispersion. If desired, the coating may comprise pores or
channels or to facilitate release of active ingredient. Pores and
channels may be generated by well known methods, including the
addition of organic or inorganic material that is dissolved,
extracted or leached from the coating in the environment of use.
Certain such pore-forming materials include hydrophilic polymers,
such as hydroxyalkylcelluloses (e.g.,
hydroxypropylmethylcellulose), cellulose ethers, synthetic
water-soluble polymers (e.g., polyvinylpyrrolidone, cross-linked
polyvinylpyrrolidone and polyethylene oxide), water-soluble
polydextrose, saccharides and polysaccharides and alkali metal
salts. Alternatively, or in addition, a controlled release coating
may include one or more orifices, which may be formed my methods
such as those described in U.S. Pat. Nos. 3,845,770; 4,034,758;
4,077,407; 4,088,864; 4,783,337 and 5,071,607. Controlled-release
may also be achieved through the use of transdermal patches, using
conventional technology (see, e.g., U.S. Pat. No. 4,668,232).
[0222] Further examples of controlled release formulations, and
components thereof, may be found, for example, in U.S. Pat. Nos.
4,572,833; 4,587,117; 4,606,909; 4,610,870; 4,684,516; 4,777,049;
4,994,276; 4,996,058; 5,128,143; 5,202,128; 5,376,384; 5,384,133;
5,445,829; 5,510,119; 5,618,560; 5,643,604; 5,891,474; 5,958,456;
6,039,980; 6,143,353; 6,126,969; 6,156,342; 6,197,347; 6,387,394;
6,399,096; 6,437,000; 6,447,796; 6,475,493; 6,491,950; 6,524,615;
6,838,094; 6,905,709; 6,923,984; 6,923,988; and 6,911,217; each of
which is hereby incorporated by reference for its teaching of the
preparation of controlled release dosage forms.
[0223] In addition to or together with the above modes of
administration, a compound provided herein may be conveniently
added to food or drinking water (e.g., for administration to
non-human animals including companion animals (such as dogs and
cats) and livestock). Animal feed and drinking water compositions
may be formulated so that the animal takes in an appropriate
quantity of the composition along with its diet. It may also be
convenient to present the composition as a premix for addition to
feed or drinking water.
[0224] Compounds are generally administered in a therapeutically
effective amount. Preferred systemic doses are no higher than 50 mg
per kilogram of body weight per day (e.g., ranging from about 0.001
mg to about 50 mg per kilogram of body weight per day), with oral
doses generally being about 5-20 fold higher than intravenous doses
(e.g., ranging from 0.01 to 40 mg per kilogram of body weight per
day).
[0225] The amount of active ingredient that may be combined with
the carrier materials to produce a single dosage unit will vary
depending, for example, upon the patient being treated, the
particular mode of administration and any other co-administered
drugs. Dosage units generally contain between from about 10 .mu.g
to about 500 mg of active ingredient. Optimal dosages may be
established using routine testing, and procedures that are well
known in the art.
[0226] Pharmaceutical compositions may be packaged for treating
conditions responsive to P2X.sub.7 receptor modulation (e.g., pain,
inflammation, neurodegeneration or other condition described
herein). Packaged pharmaceutical compositions generally include (i)
a container holding a pharmaceutical composition that comprises at
least one modulator as described herein and (ii) instructions
(e.g., labeling or a package insert) indicating that the contained
composition is to be used for treating a condition responsive to
P2X.sub.7 receptor modulation in the patient.
Methods of Use
[0227] P2X.sub.7 receptor modulators provided herein may be used to
alter activity and/or activation of P2X.sub.7 receptors in a
variety of contexts, both in vitro and in vivo. Within certain
aspects, P2X.sub.7 receptor antagonists may be used to inhibit the
binding of ligand agonist to P2X.sub.7 receptor in vitro or in
vivo. In general, such methods comprise the step of contacting a
P2X.sub.7 receptor with one or more P2X.sub.7 receptor modulators
provided herein, in the presence of ligand in aqueous solution and
under conditions otherwise suitable for binding of the ligand to
P2X.sub.7 receptor. The modulator(s) are generally present at a
concentration that is sufficient to alter P2X.sub.7
receptor-mediated signal transduction (using an assay provided in
Example 7). The P2X.sub.7 receptor may be present in solution or
suspension (e.g., in an isolated membrane or cell preparation), or
in a cultured or isolated cell. Within certain embodiments, the
P2X.sub.7 receptor is expressed by a cell that is present in a
patient, and the aqueous solution is a body fluid. Preferably, one
or more modulators are administered to an animal in an amount such
that the modulator is present in at least one body fluid of the
animal at a therapeutically effective concentration that is 20
micromolar or less, 10 micromolar or less, 5 micromolar or less, or
1 micromolar or less. For example, such compounds may be
administered at a therapeutically effective dose that is less than
20 mg/kg body weight, preferably less than 5 mg/kg and, in some
instances, less than 1 mg/kg.
[0228] Also provided herein are methods for modulating, preferably
reducing, cellular P2X.sub.7 receptor activation and/or activity,
such as signal-transducing activity (e.g., calcium conductance).
Such modulation may be achieved by contacting a P2X.sub.7 receptor
(either in vitro or in vivo) with one or more modulators provided
herein under conditions suitable for binding of the modulator(s) to
the receptor. The modulator(s) are generally present at a
concentration that is sufficient to alter P2X.sub.7
receptor-mediated signal transduction as described herein. The
receptor may be present in solution or suspension, in a cultured or
isolated cell preparation or in a cell within a patient. For
example, the cell may be contacted in vivo in an animal. Modulation
of signal transducing activity may be assessed by detecting an
effect on calcium ion conductance (also referred to as calcium
mobilization or flux). Modulation of signal transducing activity
may alternatively be assessed by detecting an alteration of a
symptom (e.g., pain or inflammation) of a patient being treated
with one or more modulators provided herein.
[0229] P2X.sub.7 receptor modulator(s) provided herein are
preferably administered to a patient (e.g., a human) orally or
topically, and are present within at least one body fluid of the
animal while modulating P2X.sub.7 receptor signal-transducing
activity.
[0230] The present invention further provides methods for treating
conditions responsive to P2X.sub.7 receptor modulation. Within the
context of the present invention, the term "treatment" encompasses
both disease-modifying treatment and symptomatic treatment, either
of which may be prophylactic (i.e., before the onset of symptoms,
in order to prevent, delay or reduce the severity of symptoms) or
therapeutic (i.e., after the onset of symptoms, in order to reduce
the severity and/or duration of symptoms). A condition is
"responsive to P2X.sub.7 receptor modulation" if it is
characterized by inappropriate activity of a P2X.sub.7 receptor,
regardless of the amount of P2X.sub.7 agonist present locally,
and/or if modulation of P2X.sub.7 receptor activity results in
alleviation of the condition or a symptom thereof. Such conditions
include, for example, pain, inflammation, cardiovascular disorders,
ocular disorders, neurodegenerative disorders and respiratory
disorders (such as cough, asthma, chronic obstructive pulmonary
disease, chronic bronchitis, cystic fibrosis and rhinitis,
including allergic rhinitis, such as seasonal an perennial
rhinitis, and non-allergic rhinitis), fibrosis as well as other
conditions described in more detail below. Such conditions may be
diagnosed and monitored using criteria that have been established
in the art. Patients may include humans, domesticated companion
animals and livestock, with dosages as described above.
[0231] Treatment regimens may vary depending on the compound used
and the particular condition to be treated; however, for treatment
of most disorders, a frequency of administration of 4 times daily
or less is preferred. In general, a dosage regimen of 2 times daily
is more preferred, with once a day dosing particularly preferred.
For the treatment of acute pain, a single dose that rapidly reaches
effective concentrations is desirable. It will be understood,
however, that the specific dose level and treatment regimen for any
particular patient will depend upon a variety of factors including
the activity of the specific compound employed, the age, body
weight, general health, sex, diet, time of administration, route of
administration, and rate of excretion, drug combination and the
severity of the particular disease undergoing therapy. In general,
the use of the minimum dose sufficient to provide effective therapy
is preferred. Patients may generally be monitored for therapeutic
effectiveness using medical or veterinary criteria suitable for the
condition being treated or prevented.
[0232] Pain that may be treated using the modulators provided
herein includes, for example, acute, chronic, inflammatory, and
neuropathic pain. Specific pain indications that may be treated as
described herein include, but are not limited to, pain associated
with osteoarthritis or rheumatoid arthritis; various neuropathic
pain syndromes (such as post-herpetic neuralgia, trigeminal
neuralgia, reflex sympathetic dystrophy, diabetic neuropathy,
Guillian Barre syndrome, fibromyalgia, oral neuropathic pain,
phantom limb pain, post-mastectomy pain, peripheral neuropathy,
myofascial pain syndromes, MS-related neuropathy, HIV or
AIDS-related neuropathy, and chemotherapy-induced and other
iatrogenic neuropathies); visceral pain, (such as that associated
with gastroesophageal reflux disease (GERD), irritable bowel
syndrome, inflammatory bowel disease, pancreatitis, intestinal gas,
gynecological disorders (e.g., menstrual pain, dysmenorrhoea, pain
associated with cystitis, labor pain, chronic pelvic pain, chronic
prostitis, endometriosis, heart pain and abdominal pain), and
urological disorders); dental pain (e.g., toothache, denture pain,
nerve root pain, pain resulting from periodontal disease, and pain
due to dental surgery including operative and post-operative pain);
headache (e.g., headaches involving peripheral nerve activity,
sinus headache, cluster headache (i.e., migranous neuralgia)
tension headache, migraine, temporomandibular pain and maxillary
sinus pain); stump pain; meralgia paresthetica; burning-mouth
syndrome; pain associated with nerve and root damage, including as
pain associated with peripheral nerve disorders (e.g., nerve
entrapment and brachial plexus avulsions, amputation, peripheral
neuropathies including bilateral peripheral neuropathy, tic
douloureux, atypical facial pain, nerve root damage, and
arachnoiditis), causalgia, neuritis (including, for example,
sciatic neuritis, peripheral neuritis, polyneuritis, optic
neuritis, postfebrile neuritis, migrating neuritis, segmental
neuritis and Gombault's neuritis), neuronitis, neuralgias (e.g.,
those mentioned above, cervicobrachial neuralgia, cranial
neuralgia, geniculate neuralgia, glossopharyngial neuralgia,
migranous neuralgia, idiopathic neuralgia, intercostals neuralgia,
mammary neuralgia, mandibular joint neuralgia, Morton's neuralgia,
nasociliary neuralgia, occipital neuralgia, red neuralgia, Sluder's
neuralgia, splenopalatine neuralgia, supraorbital neuralgia and
vidian neuralgia); surgery-related pain; musculoskeletal pain;
central nervous system pain (e.g., pain due to brain stem damage,
sciatica, and ankylosing spondylitis); and spinal pain, including
spinal cord injury-related pain.
[0233] Further pain conditions that can be treated as described
herein include Charcot's pains, ear pain, muscle pain, eye pain,
orofacial pain (e.g., odontalgia), carpel tunnel syndrome, acute
and chronic back pain (e.g., lower back pain), gout, scar pain,
hemorrhoidal pain, dyspeptic pains, angina, nerve root pain,
"non-painful" neuropathies, complex regional pain syndrome,
homotopic pain and heterotopic pain--including pain associated with
carcinoma, often referred to as cancer-associated pain (e.g., in
patients with bone cancer), pain (and inflammation) associated with
venom exposure (e.g., due to snake bite, spider bite, or insect
sting) and trauma-associated pain (e.g., post-surgical pain,
episiotomy pain, pain from cuts, musculoskeletal pain, bruises and
broken bones, and burn pain, especially primary hyperalgesia
associated therewith). Additional pain conditions that may be
treated as described herein include pain associated with autoimmune
diseases or immunodeficiency disorders, hot flashes, burns,
sunburn, and pain that results from exposure to heat, cold or
external chemical stimuli.
[0234] Conditions associated with inflammation and/or immune system
disorders that may be treated using the modulators provided herein
include, but are not limited to, arthritis (including
osteoarthritis, rheumatoid arthritis, psoriatic arthritis, Reiter's
syndrome, gout, traumatic arthritis, rubella arthritis, rheumatoid
spondylitis, gouty arthritis and juvenile arthritis); cystic
fibrosis; uveitis; systemic lupus erythematosus (and associated
glomerulonephritis); spondyloarthropathies; psoriasis; scleritis;
allergic conditions (including allergic reactions, allergic
rhinitis, allergic contact hypersensitivity, allergic dermatitis,
eczema and contact dermatitis), reperfusion injury (e.g., cardiac
and renal reperfusion injury), respiratory system disorders
(including hyper-responsiveness of the airway, cough, asthma (e.g.,
to prevent or decrease the severity of both acute early phase
asthma attack and the late phase reactions that follow such an
asthma attack; including bronchial, allergic, intrinsic, extrinsic,
exercise-induced, drug-induced (e.g., aspirin or NSAID-induced) and
dust-induced asthma), reactive airway disease, emphysema, acute
(adult) respiratory distress syndrome (ARDS), bronchitis (e.g.,
infectious and eosinophilic bronchitis), bronchiectasis, chronic
pulmonary obstructive disorder (COPD), chronic pulmonary
inflammatory disease, silicosis, pulmonary sarcoidosis, farmer's
lung, hypersensitivity pneumonitis and lung fibrosis), viral
infection, fungal infection, bacterial infection, Crohn's disease,
glomerulornephritis, HIV infection and AIDS, irritable bowel
syndrome, inflammatory bowel disease, dermatomyositis, multiple
sclerosis, pemphigus, pemphigoid, scleroderma, myasthenia gravis,
autoimmune hemolytic and thrombocytopenic states, Goodpasture's
syndrome (and associated glomerulonephritis and pulmonary
hemorrhage), tissue graft rejection, hyperacute rejection of
transplanted organs, allograft rejection, organ transplant
toxicity, neutropenia, sepsis, septic shock, endotoxic shock,
conjunctivitis shock, toxic shock syndrome, Alzheimer's disease,
inflammation associated with severe burns, lung injury, systemic
inflammatory response syndrome (SIRS), neonatal-onset multisystem
inflammatory disease (NOMID), Hashimoto's thyroiditis, Grave's
disease, Addison's disease, idiopathic thrombocytopaenic purprua,
eosinophilic fascitis, hyper-IgE syndrome, antiphospholipid
syndrome, leprosy, Sezary syndrome, paraneoplastic syndromes,
Muckle-Wells syndrome, lichen planus, familial cold
autoinflammatory syndrome (FCAS), colitis, ruptured abdominal
aortic aneurysm and multiple organ dysfunction syndrome (MODS).
Also included are pathologic sequellae associated with
insulin-dependent diabetes mellitus (including diabetic
retinopathy), lupus nephropathy, Heyman nephritis, membranous
nephritis and other forms of glomerulonephritis, macular
degeneration, contact sensitivity responses, and inflammation
resulting from contact of blood with artificial surfaces as occurs,
for example, during extracorporeal circulation of blood (e.g.,
during hemodialysis or via a heart-lung machine, for example, in
association with vascular surgery such as coronary artery bypass
grafting or heart valve replacement) such as extracorporeal
post-dialysis syndrome, or in association with contact with other
artificial vessel or container surfaces (e.g., ventricular assist
devices, artificial heart machines, transfusion tubing, blood
storage bags, plasmapheresis, plateletpheresis, and the like).
[0235] Still further conditions that may be treated using the
modulators provided herein include:
[0236] Cardiovascular disorders, such as cardiovascular disease,
stroke, cerebral ischemia, myocardial infarction, atherosclerosis,
ischemic heart disease, ischemia-reperfusion injury, aortic
aneurysm, and congestive heart failure;
[0237] Ocular disorders such as glaucoma;
[0238] Neurological disorders (e.g., neurodegeneration), such as
neurodegenerative conditions associated with progressive CNS
disorders, including, but not limited to, Alzheimer's disease,
Parkinson's disease, amyotrophic lateral sclerosis, Huntington's
disease, Creutzfeldt-Jakob disease, dementia with Lewy bodies,
traumatic brain injury, spinal cord injury, neurotrauma, cerebral
amyloid angiopathy, and encephalitis; epilepsy and seizure
disorders; multiple sclerosis and other demyelinating syndromes;
cerebral atherosclerosis; vasculitis; temporal arteritis;
myasthenia gravis; neurosarcoidosis; and central and peripheral
nervous system complications of malignant, infectious or autoimmune
processes; the modulators provided herein may also be used to
promote neuroregeneration;
[0239] Centrally-mediated neuropsychiatric disorders, such as
depression, depression mania, bipolar disease, anxiety,
schizophrenia, eating disorders, sleep disorders and cognition
disorders; and
[0240] Other disorders, such as cirrhosis, interstitial fibrosis,
prostate, bladder and bowel dysfunction (e.g., urinary
incontinence, urinary hesitancy, rectal hypersensitivity, fecal
incontinence and benign prostatic hypertrophy); itch/pruritus;
obesity; lipid disorders; cancer; hypertension; renal disorders;
abnormal wound healing; myoblastic leukemia; diabetes; meningitis;
varicose veins; muscle degeneration; cachexia; restenosis;
thrombosis; cerebral malaria; disorders of bones and joints (e.g.,
osteoporosis, bone resorption disease, loosening of artificial
joint implants, and others listed above); epidermolysis bullosa;
ocular angiogenesis; corneal injury; corneal scarring; and tissue
ulceration.
[0241] Modulators provided herein may also be used for
neuroprotection of the optic nerve (e.g., to inhibit the death of
retinal ganglion cells in a patient).
[0242] Within other aspects, modulators provided herein may be used
within combination therapy for the treatment of conditions
responsive to P2X.sub.7 receptor modulation (e.g., conditions
involving pain and/or inflammatory components). Such conditions
include, for example, autoimmune disorders and pathologic
autoimmune responses known to have an inflammatory component
including, but not limited to, arthritis (especially rheumatoid
arthritis), psoriasis, Crohn's disease, lupus erythematosus,
irritable bowel syndrome, tissue graft rejection, and hyperacute
rejection of transplanted organs. Other such conditions include
trauma (e.g., injury to the head or spinal cord), cardio- and
cerebro-vascular disease and certain infectious diseases.
[0243] Within such combination therapy, a modulator is administered
to a patient along with a second therapeutic agent (e.g., an
analgesic and/or anti-inflammatory agent). The modulator and second
therapeutic agent may be present in the same pharmaceutical
composition, or may be administered separately in either order.
Anti-inflammatory agents include, for example, non-steroidal
anti-inflammatory drugs (NSAIDs), non-specific and cyclooxygenase-2
(COX-2) specific cyclooxygenase enzyme inhibitors, gold compounds,
corticosteroids, methotrexate, leflunomide, cyclosporine A, IM
gold, minocycline, azathioprine, tumor necrosis factor (TNF)
receptor antagonists, soluble TNF alpha receptor (etanercept),
anti-TNF alpha antibodies (e.g., infliximab and adalimumab),
anti-05 antibodies, interleukin-1 (IL-1) receptor antagonists
(e.g., anakinra or IL-1 trap), IL-18 binding protein, CTLA4-Ig
(e.g., abatacept), anti-human IL-6 receptor monoclonal antibody
(e.g., tocilizumab), LFA-3-Ig fusion proteins (e.g., alefacept),
LFA-1 antagonists, anti-VLA4 monoantibody (e.g., natalizumab),
anti-CD11a monoclonal antibody, anti-CD20 monoclonal antibody
(e.g., rituximab), anti-IL-12 monoclonal antibody, anti-IL-15
monoclonal antibody, CDP 484, CDP 870, chemokine receptor
antagonists, selective iNOS inhibitors, p38 kinase inhibitors,
integrin antagonists, angiogenesis inhibitors, and TMI-1 dual
inhibitors. Further anti-inflammatory agents include meloxicam,
rofecoxib, celecoxib, etoricoxib, parecoxib, valdecoxib and
tilicoxib.
[0244] NSAIDs include, but are not limited to, ibuprofen,
flurbiprofen, naproxen or naproxen sodium, diclofenac, combinations
of diclofenac sodium and misoprostol, sulindac, oxaprozin,
diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium,
ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and
hydroxychloroquine. One class of NSAIDs consists of compounds that
inhibit cyclooxygenase (COX) enzymes; such compounds include
celecoxib and rofecoxib. NSAIDs further include salicylates such as
acetylsalicylic acid or aspirin, sodium salicylate, choline and
magnesium salicylates, and salsalate, as well as corticosteroids
such as cortisone, dexamethasone, methylprednisolone, prednisolone,
prednisolone sodium phosphate, and prednisone.
[0245] Suitable dosages for P2X.sub.7 receptor modulator within
such combination therapy are generally as described above. Dosages
and methods of administration of anti-inflammatory agents can be
found, for example, in the manufacturer's instructions in the
Physician's Desk Reference. In certain embodiments, the combination
administration of a modulator with an anti-inflammatory agent
results in a reduction of the dosage of the anti-inflammatory agent
required to produce a therapeutic effect (i.e., a decrease in the
minimum therapeutically effective amount). Thus, preferably, the
dosage of anti-inflammatory agent in a combination or combination
treatment method is less than the maximum dose advised by the
manufacturer for administration of the anti-inflammatory agent
without combination administration of a modulator. More preferably
this dosage is less than 3/4, even more preferably less than 1/2,
and highly preferably, less than 1/4 of the maximum dose, while
most preferably the dose is less than 10% of the maximum dose
advised by the manufacturer for administration of the
anti-inflammatory agent(s) when administered without combination
administration of a modulator. It will be apparent that the dosage
amount of modulator component of the combination needed to achieve
the desired effect may similarly be reduced by the
co-administration of the anti-inflammatory agent.
[0246] In certain preferred embodiments, the combination
administration of a modulator with an anti-inflammatory agent is
accomplished by packaging one or more modulators and one or more
anti-inflammatory agents in the same package, either in separate
containers within the package or in the same contained as a mixture
of one or more modulators and one or more anti-inflammatory agents.
Preferred mixtures are formulated for oral administration (e.g., as
pills, capsules, tablets or the like). In certain embodiments, the
package comprises a label bearing indicia indicating that the one
or more modulators and one or more anti-inflammatory agents are to
be taken together for the treatment of an inflammatory pain
condition.
[0247] Within further aspects, modulators provided herein may be
used in combination with one or more additional pain relief
medications. Certain such medications are also anti-inflammatory
agents, and are listed above. Other such medications are analgesic
agents, including narcotic agents which typically act at one or
more opioid receptor subtypes (e.g., .mu., .kappa. and/or .delta.),
preferably as agonists or partial agonists. Such agents include
opiates, opiate derivatives and opioids, as well as
pharmaceutically acceptable salts and hydrates thereof. Specific
examples of narcotic analgesics include, within preferred
embodiments, alfentanil, alphaprodine, anileridine, bezitramide,
buprenorphine, butorphanol, codeine, diacetyldihydromorphine,
diacetylmorphine, dihydrocodeine, diphenoxylate, ethylmorphine,
fentanyl, heroin, hydrocodone, hydromorphone, isomethadone,
levomethorphan, levorphane, levorphanol, meperidine, metazocine,
methadone, methorphan, metopon, morphine, nalbuphine, opium
extracts, opium fluid extracts, powdered opium, granulated opium,
raw opium, tincture of opium, oxycodone, oxymorphone, paregoric,
pentazocine, pethidine, phenazocine, piminodine, propoxyphene,
racemethorphan, racemorphan, sulfentanyl, thebaine and
pharmaceutically acceptable salts and hydrates of the foregoing
agents.
[0248] Other examples of narcotic analgesic agents include
acetorphine, acetyldihydrocodeine, acetylmethadol, allylprodine,
alphracetylmethadol, alphameprodine, alphamethadol, benzethidine,
benzylmorphine, betacetylmethadol, betameprodine, betamethadol,
betaprodine, clonitazene, codeine methylbromide, codeine-N-oxide,
cyprenorphine, desomorphine, dextromoramide, diampromide,
diethylthiambutene, dihydromorphine, dimenoxadol, dimepheptanol,
dimethylthiamubutene, dioxaphetyl butyrate, dipipanone, drotebanol,
ethanol, ethylmethylthiambutene, etonitazene, etorphine,
etoxeridine, furethidine, hydromorphinol, hydroxypethidine,
ketobemidone, levomoramide, levophenacylmorphan, methyldesorphine,
methyldihydromorphine, morpheridine, morphine, methylpromide,
morphine methylsulfonate, morphine-N-oxide, myrophin, naloxone,
naltyhexone, nicocodeine, nicomorphine, noracymethadol,
norlevorphanol, normethadone, normorphine, norpipanone,
pentazocaine, phenadoxone, phenampromide, phenomorphan,
phenoperidine, piritramide, pholcodine, proheptazoine, properidine,
propiran, racemoramide, thebacon, trimeperidine and the
pharmaceutically acceptable salts and hydrates thereof.
[0249] Further specific representative analgesic agents include,
for example acetaminophen (paracetamol); aspirin and other NSAIDs
described above; NR2B antagonists; bradykinin antagonists;
anti-migraine agents; anticonvulsants such as oxcarbazepine and
carbamazepine; antidepressants (such as TCAs, SSRIs, SNRIs,
substance P antagonists, etc.); spinal blocks;
pentazocine/naloxone; meperidine; levorphanol; buprenorphine;
hydromorphone; fentanyl; sufentanyl; oxycodone;
oxycodone/acetaminophen, nalbuphine and oxymorphone. Still further
analgesic agents include CB2-receptor agonists, such as AM1241,
capsaicin receptor antagonists and compounds that bind to the
.alpha.2.delta. subunit of voltage-gated calcium channels, such as
gabapentin and pregabalin.
[0250] Representative anti-migraine agents for use in combination
with a modulator provided herein include CGRP antagonists,
capsaicin receptor antagonists, ergotamines and 5-HT.sub.1
agonists, such as sumatripan, naratriptan, zolmatriptan and
rizatriptan.
[0251] Within still further aspects, modulators provided herein may
be used, for example, in the treatment of pulmonary disorders such
as asthma, in combination with one or more beta(2)-adrenergic
receptor agonists or leukotriene receptor antagonists (e.g., agents
that inhibits the cysteinyl leukotriene CysLT.sub.1 receptor).
CysLT.sub.1 antagonists include montelukast, zafirlukast, and
pranlukast.
[0252] For retinal neuroprotection and treatment of ocular
disorders, P2X.sub.7 receptor modulators may be administered to the
eye in combination with, for example, one or more of an agent that
inhibits ATP release, an agent that enhances conversion of ATP to
adenosine and/or an agent that inhibits Ca.sup.+2 influx into
retinal ganglion cells. Such agents include, for example, adenosine
A.sub.3 receptor agonists, adenosine A.sub.1 receptor agonists,
ectonucleotidase agonists, Ca.sup.+2 chelating agents and NMDA
receptor antagonists.
[0253] Suitable dosages for P2X.sub.7 receptor modulator within
such combination therapy are generally as described above. Dosages
and methods of administration of other pain relief medications can
be found, for example, in the manufacturer's instructions in the
Physician's Desk Reference. In certain embodiments, the combination
administration of a modulator with one or more additional pain
medications results in a reduction of the dosage of each
therapeutic agent required to produce a therapeutic effect (e.g.,
the dosage or one or both agent may less than 3/4, less than 1/2,
less than 1/4 or less than 10% of the maximum dose listed above or
advised by the manufacturer).
[0254] For use in combination therapy, pharmaceutical compositions
as described above may further comprise one or more additional
medications as described above. In certain such compositions, the
additional medication is an analgesic. Also provided herein are
packaged pharmaceutical preparations comprising one or more
modulators and one or more additional medications (e.g.,
analgesics) in the same package. Such packaged pharmaceutical
preparations generally include (i) a container holding a
pharmaceutical composition that comprises at least one modulator as
described herein; (ii) a container holding a pharmaceutical
composition that comprises at least one additional medication (such
as a pain relief and/or anti-inflammatory medication) as described
above and (iii) instructions (e.g., labeling or a package insert)
indicating that the compositions are to be used simultaneously,
separately or sequentially for treating or preventing a condition
responsive to P2X.sub.7 receptor modulation in the patient (such as
a condition in which pain and/or inflammation predominates).
[0255] Within separate aspects, the present invention provides a
variety of non-pharmaceutical in vitro and in vivo uses for the
modulator compounds provided herein. For example, such compounds
may be labeled and used as probes for the detection and
localization of P2X.sub.7 receptor (in samples such as cell
preparations or tissue sections, preparations or fractions
thereof). In addition, modulators provided herein that comprise a
suitable reactive group (such as an aryl carbonyl, nitro or azide
group) may be used in photoaffinity labeling studies of receptor
binding sites. In addition, modulators provided herein may be used
as positive controls in assays for receptor activity or as
radiotracers (e.g., in receptor mapping procedures). For example, a
modulator compound may be labeled using any of a variety of well
known techniques (e.g., radiolabeled with a radionuclide such as
tritium, as described herein), and used as a probe for receptor
autoradiography (receptor mapping) of P2X.sub.7 receptor in
cultured cells or tissue samples, which may be performed as
described by Kuhar in sections 8.1.1 to 8.1.9 of Current Protocols
in Pharmacology (1998) John Wiley & Sons, New York, which
sections are incorporated herein by reference. Such receptor
mapping procedures also include methods that can be used to
characterize P2X.sub.7 receptor in living subjects, such as
positron emission tomography (PET) imaging or single photon
emission computerized tomography (SPECT).
[0256] The following Examples are offered by way of illustration
and not by way of limitation. Unless otherwise specified all
reagents and solvent are of standard commercial grade and are used
without further purification. Using routine modifications, the
starting materials may be varied and additional steps employed to
produce other compounds provided herein.
[0257] Mass spectroscopy data provided herein is Electrospray MS,
obtained in positive ion mode. Unless otherwise specified, such
data is obtained using a Micromass Time-of-Flight LCT (Waters
Corp.; Milford, Mass.), equipped with a Waters 600 pump (Waters
Corp.), Waters 996 photodiode array detector (Waters Corp.), and a
Gilson 215 autosampler (Gilson, Inc.; Middleton, Wis.).
MassLynx.TM. (Waters Corp.) version 4.0 software with OpenLynx
Global Server.TM., OpenLynx.TM. and AutoLynx.TM. processing is used
for data collection and analysis. MS conditions are as follows:
capillary voltage=3.5 kV; cone voltage=30 V, desolvation and source
temperature=350.degree. C. and 120.degree. C., respectively; mass
range=181-750 with a scan time of 0.22 seconds and an interscan
delay of 0.05 seconds.
[0258] For data marked with a ".sctn.," mass spectroscopy data is
obtained using a Waters ZMD II Mass Spectrometer (Waters Corp.),
equipped with a Waters 600 pump (Waters Corp.), Waters 996
photodiode array detector (Waters Corp.), and a Gilson 215
autosampler (Gilson, Inc.; Middleton, Wis.). MassLynx.TM. (Waters
Corp.) version 4.0 software with OpenLynx Global Server.TM.,
OpenLynx.TM. and AutoLynx.TM. processing is used for data
collection and analysis. MS conditions are as follows: capillary
voltage=3.5 kV; cone voltage=30 V, desolvation and source
temperature=250.degree. C. and 100.degree. C., respectively; mass
range=100-800 with a scan time of 0.5 seconds and an interscan
delay of 0.1 seconds.
[0259] For either method, sample volume of 1 microliter is injected
onto a 50.times.4.6 mm Chromolith SpeedROD RP-18e column (Merck
KGaA, Darmstadt, Germany), and eluted using a 2-phase linear
gradient at a flow rate of 6 ml/min. Sample is detected using total
absorbance count over the 220-340 nm UV range. The elution
conditions are: Mobile Phase A--95% water, 5% MeOH with 0.05% TFA;
Mobile Phase B--5% water, 95% MeOH with 0.025% TFA. The following
gradient is used: 0-0.5 min 10-100% B, hold at 100% B to 1.2 min,
return to 10% B at 1.21 min. Inject to inject cycle is 2.15
min.
[0260] Where indicated, LC retention times (R.sub.T) are provided
in minutes.
Example 1
N-(Adamantan-1-ylmethyl)-1-pyrimidin-2-yl-1H-indole-3-carboxamide
Step 1. Methyl 1-(pyrimidin-2-yl)-1H-indole-3-carboxylate
##STR00017##
[0262] Potassium t-butoxide (2.52 g, 0.022 mol) is added to a
mixture of methyl 1H-indole-3-carboxylate (3.5 g, 0.02 mol) and
2-chloropyrimidine (2.28 g, 0.02 mol) in 50 mL of dioxane. The
reaction mixture is heated to 110.degree. C. and stirred for 20 h.
The dioxane is removed in vacuo, and the residue is diluted with
water (100 mL). The solid is filtered and purified by silica gel
column chromatography (15% EtOAc/DCM) to afford the title compound
as a white solid.
Step 2. 1-(Pyrimidin-2-yl)-1H-indole-3-carboxylic acid
##STR00018##
[0264] 1.0 N aqueous NaOH (10 mL) is added to a mixture of methyl
1-(pyrimidin-2-yl)-1H-indole-3-carboxylate (1.4 g, 0.0055 moles) in
50 mL of EtOH and heated at 70.degree. C. for 4 h. The reaction
mixture is concentrated in vacuo, diluted with water (50 mL),
acidified with concentrated HCl to pH 2.0. The white solid
separated is filtered, washed with water (2.times.25 mL) and dried
to afford the title compound.
Step 3.
N-(Adamantan-1-ylmethyl)-1-pyrimidin-2-yl-1H-indole-3-carboxamide
##STR00019##
[0266] To a mixture of 1-(pyrimidin-2-yl)-1H-indole-3-carboxylic
acid (72 mg, 0.3 mmol) in 2.0 mL of DMF is added sequentially
diisopropylethylamine (0.2 mL), 1-adamantane methylamine (49.5 mg,
0.3 mmol), and BOP, (150 mg). The resulting mixture is stirred at
rt for 20 h. Water (3 mL) is added, and the solid is filtered and
purified by silica gel column chromatography (25% EtOAc/hexane) to
give the title compound as a white solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 8.81 (1H, d), 8.71 (3H, dd,), 8.13 (1H, d),
7.33 (2H, m), 7.11 (1H, d), 6.13 (1H, s), 3.21 (2H, d), 1.61 (12H,
m). Mass spec. (387.24, M+H).
Example 2
N-[(1-Pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-4-(trifluoromethyl)--
1H-indole-3-carboxamide
Step 1. 4-Bromo-1H-indole-3-carbaldehyde
##STR00020##
[0268] A round bottom flask charged with 50 mL of DMF is cooled to
0.degree. C., and phosphorous oxychloride (8.1 mL, 88 mmol) is
added dropwise. After stirring for approx. 5 min, a solution of
4-bromoindole (5.0 mL, 40 mmol) in 50 mL of DMF is added dropwise.
The ice bath is removed, and the reaction mixture is stirred for 1
h at rt. The reaction becomes a very thick suspension. The mixture
is cooled back to 0.degree. C. and carefully quenched with 22g of
KOH in 80 mL of water. The resulting mixture is partitioned between
EtOAc (200 mL) and sat. NaHCO.sub.3 (100 mL). The EtOAc layer is
washed with brine (100 mL) and water (100 mL), dried
(Na.sub.2SO.sub.4), filtered, and evaporated in vacuo to give a
brown solid, which is triturated with ether to afford the title
compound as an off-white solid.
Step 2. 4-(Trifluoromethyl)-1H-indole-3-carbaldehyde
##STR00021##
[0270] Methyl 2-(fluorosulfonyl)difluoroacetate (3.5 mL, 27.6 mmol)
and copper (I) iodide (5.3 g, 27.6 mmol) are added to a solution of
4-bromo-1H-indole-3-carbaldehyde (3.1 g, 13.8 mmol) in 65 mL of
DMF. The reaction is heated to 85.degree. C. for 18 h. After
cooling to rt, the mixture is diluted with EtOAc and filtered
through celite. The celite is washed well with EtOAc. The filtrate
is washed with water (3 times) and brine, dried (Na.sub.2SO.sub.4),
filtered, and evaporated to give a brown oil. Purification by
silica gel column chromatography (gradient from 10% EtOAc/hexane to
40% EtOAc/hexane) affords the title compound as a brown solid.
Step 3.
1-Pyrimidin-2-yl-4-(trifluoromethyl)-1H-indole-3-carbaldehyde
##STR00022##
[0272] Potassium t-butoxide (753 mg, 6.71 mmol) is added to a
mixture of 4-(trifluoromethyl)-1H-indole-3-carbaldehyde (1.3 g,
6.10 mmol) and 2-chloropyrimidine (699 mg, 6.10 mmol) in 20 mL of
dioxane. The reaction mixture is heated to 100.degree. C. and
stirred for 15 h. The dioxane is removed in vacuo, and the residue
is partitioned between EtOAc and water (30 mL each). The aqueous
phase is extracted twice more with EtOAc (20 mL each), and the
combined EtOAc extracts are dried (Na.sub.2SO.sub.4), filtered, and
evaporated in vacuo to give a brown solid. Purification by silica
gel column chromatography (2% EtOAc/DCM) affords the title compound
as a light brown solid.
Step 4. 1-Pyrimidin-2-yl-4-(trifluoromethyl)-1H-indole-3-carboxylic
acid
##STR00023##
[0274] To a solution of
1-pyrimidin-2-yl-4-(trifluoromethyl)-1H-indole-3-carbaldehyde (450
mg, 1.55 mmol) in 15 mL of dioxane and 5 mL of water is added
sodium chlorite (183 mg, 2.02 mmol) and sulfamic acid (858 mg,
8.,84 mmol). After stirring at rt for 10 min., the reaction is
carefully quenched with sat. aqueous NaHCO.sub.3 and concentrated
in vacuo. The residue is taken up in 10% MeOH/DCM and washed with
10% HCl and water. The combined aqueous phases are back extracted
with 10% MeOH/DCM (4 times), and the combined organic extracts are
dried (Na.sub.2SO.sub.4), filtered, and evaporated in vacuo to give
a pale yellow solid. Trituration with ether affords the title
compound as a white solid.
Step 5. (1-Pyridin-3-yl-cyclohexyl)-methylamine
##STR00024##
[0276] To a mixture of pyridin-3-yl-acetonitrile (14.65 g) and
1,5-dibromo-pentane (28.52 g) in THF (450 mL) and DMSO (450 mL) is
added NaH (60% in mineral oil, 10.42 g) portionwise at 0.degree. C.
over 1 h. The mixture is allowed to warm to RT and stirred
overnight. The reaction mixture is poured into water and extracted
twice with EtOAc. The extract is concentrated and the residue is
purified by silica gel chromatography to give
1-pyridin-3-yl-cyclohexanecarbonitrile.
[0277] To a solution of the above product (18.5 g) in 140 mL of 7.0
N NH.sub.3 in MeOH is added carefully slurry of Raney Nickel (16g).
The mixture is hydrogenated at 50 psi overnight. The mixture is
filtered through Celite and concentrated in vacuo to give the title
compound
Step 6.
N-[(1-Pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-4-(trifluoro-
methyl)-1H-indole-3-carboxamide
##STR00025##
[0279] To a mixture of
1-((1-methyl-1H-imidazol-2-yl)methyl)-1H-indole-2-carboxylic acid
(21 mg, 0.05 mmol) in 1.0 mL of DMF is added sequentially
diisopropylethylamine (0.02 mL, 0.10 mmol),
1-(1-pyridin-3-ylcyclohexyl)methanamine (9.5 mg, 0.05 mmol), and
BOP (27 mg, 0.06 mmol). The resulting mixture is stirred at rt for
20 h. Water (3 mL) is added, and the mixture is extracted with
EtOAc (5 mL). The EtOAc layer is dried (Na.sub.2SO.sub.4),
filtered, and evaporated in vacuo. The residue is purified by
preparative chromatography (4% MeOH/DCM) to give the title compound
as a light brown solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
9.10 (1H, d, J 8.4), 8.95 (2H, d, J 4.8), 8.62 (1H, d, J 2), 8.40
(2H, m), 8.16 (1H, t, J 6), 7.83 (1H, d, J 8), 7.63 (1H, d, J 7.6),
7.51 (2H, m), 7.36 (1H, m), 3.39 (2H, d, J 6.4), 2.20 (2H, d, J
13.6), 1.70 (t, 2H, J 11.2), 1.58 (m, 2H), 1.35 (m, 4H). Mass spec.
(480.24, M+H).
Example 3
4-Chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-indole-3-
-carboxamide
Step 1. 1-(4-Chloro-1H-indol-3-yl)-2,2,2-trifluoroethanone
##STR00026##
[0281] Trifluoroacetic anhydride (27.5 mL, 200 mmol) is added to a
solution of 4-chloroindole (25.0 g, 165 mmol) and DMF (170 mL)
under N.sub.2 over 30 min. The reaction vessel is sealed. After 20
h, the solution is poured into water (700 mL) and extracted with
EtOAc (300 mL). The organics are dried over Na.sub.2SO.sub.4,
filtered, and concentrated. Purification by flash silica gel column
chromatography (4:1 hexane/EtOAc to 1:1 hexane/EtOAc) affords the
title compound as a red-brown solid.
Step 2.
1-(4-Chloro-1-pyrimidin-2-yl-1H-indol-3-yl)-2,2,2-trifluoroethanon-
e
##STR00027##
[0283] A mixture of
1-(4-chloro-1H-indol-3-yl)-2,2,2-trifluoroethanone (10.1 g, 40.8
mmol), 2-chloropyrimidine (9.3 g, 81 mmol), cesium carbonate (26.6
g, 81.6 mmol), and 1,4-dioxane (80 mL) under N.sub.2 is warmed to
100.degree. C. for 3 h. After cooling to rt, water (200 mL) is
added. The precipitate is collected by filtration to afford the
title compound as a tan powder.
Step 3. 4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid
##STR00028##
[0285] A mixture of 1-(4-chloro-1-pyrimidin-2-yl
-1H-indol-3-yl)-2,2,2-trifluoroethanone (8.33 g, 25.6 mmol), 10 M
aqueous NaOH (60 mL), water (20 mL), and ethanol (20 mL) under air
is warmed to 65.degree. C. for 20 h. After cooling to rt, the
volatiles are removed under reduced pressure. The aqueous mixture
is cooled to 0.degree. C. and then acidified with 5 M aqueous HCl
(110 mL). The mixture is filtered. The filtrate is washed with
H.sub.2O (50 mL) and then with Et.sub.2O (100 mL) to afford the
title compound as a tan powder.
Step 4.
4-Chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H--
indole-3-carboxamide
##STR00029##
[0287] BOP (58 mg, 130 mmol) is added to a slurry of
4-chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid (27 mg, 98
mmol), (1-pyridin-3-ylcyclohexyl)methylamine (21 mg, 110 mmol),
iPr.sub.2NEt (50 .mu.L, 290 mmol), and DMF (0.5 mL) under N.sub.2.
The reaction vessel is sealed and the solution is left to stir for
64 h. The solution is poured into 50% sat. aqueous NaHCO.sub.3 (10
mL) and then extracted with EtOAc (2.times.10 mL). The combined
organics are dried over Na.sub.2SO.sub.4, filtered, and
concentrated. Purification by PTLC (95:5 DCM/MeOH) affords the
title compound as a white powder. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta.: 8.61-8.77 (m, 4H), 8.55 (s, 1H), 8.41 (s, 1H), 7.75 (d,
1H), 7.08-7.31 (m, 4H), 6.14 (s, 1H), 3.68 (d, 2H), 2.11-2.22 (m,
2H), 1.28-1.86 (m, 8H). LC-MS m/z (M+H.sup.+): 446.18.
Example 4
N
1-((1-methyl-1H-imidazol-2-yl)methyl)-N-((1-(pyridin-3-yl)cyclohexyl)met-
hyl)-1H-indole-2-carboxamide
Step 1. Ethyl
1-((1-methyl-1H-imidazol-2-yl)methyl)-1H-indole-2-carboxylate
##STR00030##
[0289] 60% NaH (527 mg, 0.0132 mol) is added to a mixture of ethyl
1H-indole-2-carboxylate (1.13 g, 0.0059 mol) in DMF (20 mL) at rt
and stirred for 10 mins. 2-Chloromethyl-1-methylimidazole
hydrochloride is then added to the mixture and stirred for 20 h.
The mixture is diluted with ice water (100 mL) and the white solid
is filtered to afford the title compound.
Step 2.
1-((1-Methyl-1H-imidazol-2-yl)methyl)-1H-indole-2-carboxylic
acid
##STR00031##
[0291] 1.0 N aqueous NaOH (10 mL) is added to a mixture of ethyl
1-((1-methyl-1H-imidazol-2-yl)methyl)-1H-indole-2-carboxylate (1.5
g, 0.0053 moles) in 25 mL of EtOH and stirred at 25.degree. C. for
16 h. The reaction mixture is concentrated in vacuo, diluted with
water (50 mL), and acidified with concentrated HCl to pH 2.0. The
white solid separated is filtered, washed with water (2.times.25
mL) and dried to afford the title compound.
Step 3. N
1-((1-methyl-1H-imidazol-2-yl)methyl)-N-((1-(pyridin-3-yl)cycloh-
exyl)methyl)-1H-indole-2-carboxamide
##STR00032##
[0293] To a mixture of
1-((1-methyl-1H-imidazol-2-yl)methyl)-1H-indole-2-carboxylic acid
(51 mg, 0.2 mmol)) in 2.0 mL of DMF is added sequentially
diisopropylethylamine (0.2 mL),
1-(1-pyridin-3-ylcyclohexyl)methanamine (38 mg, 0.25 mmol), and BOP
reagent (100 mg, 0.22 mmol). The resulting mixture is stirred at rt
for 20 h. Water (3 mL) is added, and the mixture is extracted with
EtOAc (5 mL). The EtOAc layer is dried (Na.sub.2SO.sub.4),
filtered, and evaporated in vacuo. The residue is purified by
silica gel column chromatography (4% MeOH/DCM) to give the title
compound as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.56 (1H, s), 8.34 (2H, m), 7.70 (1H, d), 7.58 (1H, d),
7.53 (1H, d), 7.25 (1H, m), 7.17 (1H, t), 7.00 (3H, m), 6.67 (1H,
s), 5.71 (2H, s), 3.46 (3H, s), 3.32 (2H, d,), 2.14 (d, 2H),
1.14-1.63 (8H, m). Mass spec. (428.28, M+H).
Example 5
2-Adamantan-1-yl-N-(4-chloro-1-pyrimidin-2-yl-1H-indol-3-yl)-acetamide
Step 1. 4-Chloro-1-pyrimidin-2-yl-1H-indole-3-carbonyl azide
##STR00033##
[0295] Ethyl chloroformate (230 .mu.L, 2 4 mmol) is added dropwise
over 5 min to a slurry of
4-chloro-1-pyrimidin-2-yl-1H-indole-3-carboxylic acid (629 mg, 2.30
mmol) in iPr.sub.2NEt (420 .mu.L, 2.4 mmol) and acetone (7 0 mL)
under N.sub.2 at -10.degree. C. After 1 h, sodium azide (450 mg,
6.9 mmol) in water (4 mL) is added in one portion and the mixture
is left to stir under air for 15 h. The volatiles are removed under
reduced pressure. The resulting aqueous mixture is diluted with
water (10 mL) and then filtered. The solids are collected and dried
under reduced pressure to afford the title compound as a tan
powder.
Step 2. 4-Chloro-1-pyrimidin-2-yl-1H-indol-3-ylamine
##STR00034##
[0297] A slurry of 4-chloro-1-pyrimidin-2-yl-1H-indole-3-carbonyl
azide (442 mg, 1.48 mmol) in toluene (5 mL) under air is warmed to
100.degree. C. for 1 h. Concentrated HCl (1 mL) is added dropwise.
The mixture is warmed to 110.degree. C. for 2 h. The volatiles are
removed under reduced pressure. TEA (1 mL) and water (10 mL) are
added. The mixture is extracted with CH.sub.2Cl.sub.2 (4.times.30
mL) and the combined organics are dried over Na.sub.2SO.sub.4,
filtered, and concentrated. Purification by flash column
chromatography (5:1 hexanes:EtOAc to 3:1 hexanes:EtOAc) affords the
title compound as a tan solid. LC-MS m/z (M+H.sup.+): 244.99.
Step 3.
2-Adamantan-1-yl-N-(4-chloro-1-pyrimidin-2-yl-1H-indol-3-yl)-aceta-
mide
##STR00035##
[0299] BOP (135 mg, 305 mmol) is added to a slurry of
4-chloro-1-pyrimidin-2-yl-1H-indol-3-ylamine (50 mg, 200 mmol),
1-adamantaneacetic acid (52 mg, 270 .mu.mol), iPr.sub.2NEt (70
.mu.L, 400 .mu.mol), and DMF (1.0 mL) under N.sub.2. The reaction
vessel is sealed and the solution is left to stir for 4.5 days.
Water (5 mL) is added and the crude product is collected by
filtration. The solid is dissolved in EtOAc (40 mL) and then washed
with a 1:1 solution of water and brine (20 mL). The organics are
dried over Na.sub.2SO.sub.4, filtered, and concentrated.
Purification by preparative layer chromatography (7.5:1
hexanes:EtOAc) affords the title compound as an ivory powder.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta.: 9.10 (s, 1H), 8.81 (d,
1H), 8.67 (d, 2H), 8.64 (bs, 1H), 7.24 (t, 1H), 7.17 (d, 1H), 7.04
(t, 1H), 2.20 (s, 2H), 1.94-2.03 (m, 3H), 1.58-1.81 (m, 12H). LC-MS
m/z (M+H.sup.+): 421.11.
Example 6
4-Chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H-pyrrolo[-
2,3-b]pyridine-3-carboxamide
Step 1. 4-Chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde
##STR00036##
[0301] A mixture of 4-chloro-1H-pyrrolo[2,3-b]pyridine (1.2 g, 7.6
mmol, prepared essentially as described in Wang et al., J. Org.
Chem. 2006, 71, 4021-4023) and hexamethylenetetramine (1.6 g, 11.4
mmol) in 10 ml of 33% aqueous acetic acid is heated to reflux and
stirred for 14 h. The clear solution is cooled to rt and diluted
with water. The mixture becomes cloudy, and a solid begins to
precipitate out of solution. The mixture is cooled in an ice bath
and stirred for 30 min. The precipitated solid is then collected by
vacuum filtration and dried in vacuo to afford the title compound
as a light brown solid. Mass spec. (180.92, M+H).
Step 2.
4-Chloro-1-pyrimidin-2-yl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde
##STR00037##
[0303] A mixture of
4-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde (220 mg, 1.22
mmol), 2-chloropyrimidine (210 mg, 1.83 mmol), and cesium carbonate
(476 mg, 1.46 mmol) in 5-10 ml of dioxane is heated to 100.degree.
C. After stirring for 15 h, the reaction mixture is cooled and the
dioxane is removed in vacuo. The residue is slurried in water and
filtered. The collected solid is washed with more water and dried
in vacuo to afford the title compound as a light brown solid. Mass
spec. (258.95, M+H).
Step 3.
4-Chloro-1-pyrimidin-2-yl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic
acid
##STR00038##
[0305] To a mixture of
4-chloro-1-pyrimidin-2-yl-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde
(88 mg, 0.34 mmol) in 3 ml of dioxane and 1 ml of water is added
sodium chlorite (40 mg, 0.44 mmol) and sulfamic acid (188 mg, 1.94
mmol). After stirring at rt for 20 min, the solvents are removed in
vacuo. The residue is taken up in 10% MeOH/CH.sub.2Cl.sub.2, and
the resulting mixture is filtered. The filtrate is concentrated to
give the title compound as a brown solid. Mass spec. (274.95,
M+H).
Step 4.
4-Chloro-N-[(1-pyridin-3-ylcyclohexyl)methyl]-1-pyrimidin-2-yl-1H--
pyrrolo[2,3-b]pyridine-3-carboxamide
##STR00039##
[0307] To a mixture of
4-chloro-1-pyrimidin-2-yl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic
acid (45 mg, 0.16 mmol) in 1.0 ml of DMF is added sequentially
diisopropylethylamine (0.06 ml, 0.32 mmol),
1-(1-pyridin-3-ylcyclohexyl)methanamine (30 mg, 0.16 mmol), and BOP
(84 mg, 0.19 mmol). The resulting mixture is stirred at rt for 20
h. Water (3 ml) is added, and the precipitated solid is collected
by vacuum filtration and dried in vacuo to afford the title
compound as a brown solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6)
.delta. 8.97 (2H, d, J 4.8), 8.61 (1H, s), 8.35 (3H, m), 8.23 (1H,
s), 7.79 (1H, d, J 6.8), 7.54 (1H, t, J 4.4), 7.39 (1H, d, J 5.2),
7.32 (1H, bs), 3.41 (2H, d, J 5.6), 2.17 (2H, m), 1.2-1.8 (8H, m).
Mass spec. (447.15, M+H).
[0308] Additional heteroaryl amide analogues prepared using the
general methodologies hereindisclosed are listed in Table A. In the
column of Table A labeled "IC.sub.50," a "*" indicates that the
IC.sub.50 determined as described in Example 7A is 2 micromolar or
less (i.e., the concentration of such compounds that is required to
provide a 50% decrease in the fluorescence response of cells
exposed to 80 .mu.M of (2'(3')--O-(4-benzoyl-benzoyl)adenosine
5'-triphosephate is 2 micromolar or less).
[0309] Mass spectroscopy data is provided in Table I as (M+1) in
the column headed "M+H." The LC retention time, in minutes, is
provided in the column headed R.sub.T.
TABLE-US-00001 TABLE A Structure Name M + H .sub.T C.sub.50 1
##STR00040## N-(adamantan-1- ylmethyl)-1-pyrimidin-2-yl-
1H-indole-3-carboxamide 387.24 .43 ##STR00041## N-{4-methyl-2-[4-
(trifluoromethyl)phenyl]pentyl}- 1-pyrimidin-2-yl-1H-indole-
3-carboxamide 467.20 .43 ##STR00042## N-[2-(4-
chlorophenyl)pentyl]-1- pyrimidin-2-yl-1H-indole-3- carboxamide
419.18 .42 ##STR00043## N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 412.22 .18 ##STR00044##
1-pyrimidin-2-yl-N-({1-[4- (trifluoromethyl)phenyl]
cyclohexyl}methyl)-1H-indole-3- carboxamide 479.21 .46 ##STR00045##
N-{[1-(4-chlorophenyl) cyclohexyl]methyl}- 1-pyrimidin-2-yl-1H-
indole-3-carboxamide 445.18 .48 ##STR00046##
N-{[1-(4-methoxyphenyl)cyclohexyl] methyl}-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 441.24 .43 ##STR00047## N-[(1-morpholin-4-
ylcyclohexyl)methyl]-1- pyrimidin-2-yl-1H-indole-3- carboxamide
420.24 .17 ##STR00048## 2-{3-[(adamantan-1- ylmethyl)carbamoyl]-1H-
indo1-1-yl}benzoic acid 429.21 .37 ##STR00049## N-[4-methyl-2-(4-
methylphenyl)pentyl]-1- pyrimidin-2-yl-1H-indole-3- carboxamide
413.22 .44 ##STR00050## N-(4-methyl-2- pyridin-3-ylpentyl)-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 400.21 .21 ##STR00051##
N-(4-methyl-2- phenylpentyl)-1-pyrimidin-2-
yl-1H-indole-3-carboxamide 399.21 .41 ##STR00052##
N-{[1-(6-methylpyridin-3- yl)cyclohexyl]methyl}-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 426.21 .20 ##STR00053##
N-{[1-(4-fluorophenyl) cyclohexyl]methyl)-1- pyrimidin-2-yl-1H-
indole-3-carboxamide 429.19 .44 ##STR00054## 2-{3-[(adamantan-1-
ylmethyl)carbamoyl]-1H- indol-1-yl}pentanoic acid 409.30 .38
##STR00055## 1-(5-fluoropyrimidin- 2-yl)-N-[(1-pyridin-3-
ylcyclohexyl)methyl]-1H- indole-3-carboxamide 430.25 .20
##STR00056## 1-(3-ethylpyrazin-2- yl)-N-[(1-pyridin-3-
ylcyclohexyl)methyl]-1H- indole-3-carboxamide 440.30 .17
##STR00057## 4-chloro-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 446.18 .17 ##STR00058##
N-{[1-(4- methoxyphenyl)cyclopentyl] methyl}-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 427.28 .41 ##STR00059## N-{[l-(4-methyl-
phenyl)cyclohexyl]methyl}-1- pyrimidin-2-yl-1H-
indole-3-carboxamide 425.26 .48 ##STR00060##
N-{[1-(4-chloro-3-fluoro- phenyl)cyclohexyl]methyl}-1-
pyrimidin-2-yl-1H- indole-3-carboxamide 463.20 .46 ##STR00061##
N-[2-(4-chlorophenyl)-2-piperidin- 1-ylethyl]-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 460.24 .22 ##STR00062## methyl
2-(3-{[(1-pyridin-3- ylcyclohexyl)methyl]
carbamoyl}-1H-indo1-1-yl)nicotinate 469.24 .15 32 ##STR00063##
N-[(1-pyridin-3-ylcyclohexyl) methyl]-1-pyrimidin-2-yl-4-
(trifluoromethyl)-1H-indole- 3-carboxamide 480.24 .19 ##STR00064##
4-bromo-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 490.06 .40 ##STR00065##
4-cyano-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 437.17 .31 ##STR00066##
N-{[1-(4-methoxyphenyl)cyclohexyl] methyl}-1-pyrimidin-2-yl-4-
(trifluoromethyl)-1H-indole- 3-carboxamide 509.13 .68 ##STR00067##
N-{[1-(4-fluorophenyl)cyclohexyl] methyl}-1-pyrimidin-2-yl-4-
(trifluoromethyl)-1H-indole- 3-carboxamide 497.12 .70 ##STR00068##
N-{[1-(6-methylpyridin-3- yl)cyclohexyl]nethyl}-1-pyrimidin-
2-yl-4-(trifluoromethyl)-1H-indole- 3-carboxamide 494.13 .38
##STR00069## 7-chloro-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 446.07 .35 ##STR00070##
4-fluoro-N-(4- methyl-2-pyridin-3-ylpentyl)-
1-pyrimidin-2-yl-1H-indole-3- carboxamide 418.16 .65 ##STR00071##
N-[2-(4-chlorophenyl)-4- methylpentyl]-4-fluoro-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 451.10 .85 ##STR00072##
4-fluoro-N-[4-methyl-2-(4- methylphenyl)pentyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 431.12 .86 ##STR00073##
4-fluoro-N-{4-methyl-2-[4- (trifluoromethyl)phenyl]pentyl}-
1-pyrimidin-2-yl-1H-indole- 3-carboxamide 485.15 .26 ##STR00074##
N-{[1-(4-chloro-3- fluorophenyl)cyclohexyl]
methyl}-4-fluoro-1-pyrimidin-2- yl-1H-indole-3-carboxamide 481.09
.89 ##STR00075## 4-fluoro-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 430.16 .55 ##STR00076##
4-fluoro-N-{[1-(4- methylphenyl)cyclohexyl]
methyl}-1-pyrimidin-2-yl-1H- indole-3-carboxamide 443.11 .93
##STR00077## 4-fluoro-N-{[1-(4- methoxyphenyl)cyclohexyl]
methyl}-1-pyrimidin-2-yl-1H- indole-3-carboxamide 459.10 .84
##STR00078## 4-fluoro-N-{[1-(4- fluorophenyl)cyclohexyl]
methyl}-1-pyrimidin-2-yl-1H- indole-3-carboxamide 447.10 .87
##STR00079## N-{[1-(4-chlorophenyl)cyclohexyl]
methyl}-4-fluoro-1-pyrimidin-2- yl-1H-indole-3-carboxamide 463.06
.90 ##STR00080## 1-Pyrimidin-2-yl-1H- indole-3,4-dicarboxylic acid
4-amide 3-[(1-pyridin-3-yl- cyclohexylmethyl)-amide] 455.17 .17 40
##STR00081## 1-[(1-methyl-1H-imidazol-2-
yl)methyl]-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1H-
indole-2-carboxamide 428.28 .04 ##STR00082##
4-methyl-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 426.24 .12 . ##STR00083##
4-methyl-N-(4- methyl-2-pyridin-3-ylpentyl)-
1-pyrimidin-2-yl-1H-indole-3- carboxamide 414.23 .14 . ##STR00084##
N-{[1-(4-methoxyphenyl) cyclohexyl]methyl}-4-methyl-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 455.26 .35 . ##STR00085##
N-[2-(4-chlorophenyl)-4- methylpentyl]-4-methyl-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 447.32 .37 . ##STR00086##
N-{[1-(4-chlorophenyl) cyclohexyl]methyl}-4-
methyl-1-pyrimidin-2-yl-1H- indole-3-carboxamide 459.32 .40 .
##STR00087## 4-chloro-N-[2-(4- chlorophenyl)-2-morpholin-
4-ylethyl]-1-pyrimidin-2-yl- 1H-indole-3-carboxamide 496.14 .14 .
##STR00088## N-[2-(4-chlorophenyl)-2-
morpholin-4-ylethyl]-1-pyrimidin-
2-yl-4-(trifluoromethyl)-1H-indole- 3-carboxamide 530.16 .18 .
##STR00089## N-[2-(4-chlorophenyl)-2-
piperidin-1-ylethyl]-1-pyrimidin-2- yl-4-(trifluoromethyl)-1H-
indole-3-carboxamide 528.20 .15 . ##STR00090## N-(adamantan-1-
ylmethyl)-4-chloro-1-[2- (dimethylamino)ethyl]-1H-
indole-3-carboxamide 414.23 .21 . ##STR00091## N-(adamantan-1-
ylmethyl)-4-chloro-1-[3- (dimethylamino)propyl]-1H-
indole-3-carboxamide 428.24 .22 . ##STR00092##
4-chloro-N-[2-(4-chlorophenyl)- 4-methylpentyl]-1-[3-
(dimethylamino)propyl]-1H- indole-3-carboxamide 474.20 .24 .
##STR00093## 4-chloro-1-[3- (dimethylamino)propyl]-N-(4-
methyl-2-pyridin-3-ylpentyl)- 1H-indole-3-carboxamide 441.24 .02 .
##STR00094## 4-chloro-1-[2- (dimethylamino)ethyl]-N-(4-
methyl-2-pyridin-3-ylpentyl)- 1H-indole-3-carboxamide 427.23 .02 .
##STR00095## 4-chloro-N-[2-(4-chlorophenyl)- 4-methylpentyl]-1-[2-
(dimethylamino)ethyl]-1H- indole-3-carboxamide 460.19 .24 .
##STR00096## N-[2-(4-chlorophenyl)-2-
piperidin-1-ylethyl]-4-fluoro-1- pyrimidin-2-yl-1H-indole-3-
carboxamide 478.19 .15 . ##STR00097## N-[2-(4-chlorophenyl)-2-
piperidin-1-ylethyl]-4- methyl-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 474.21 .16 . ##STR00098##
N-[2-(4-chlorophenyl)-2- morpholin-4-ylethyl]-4-fluoro-
1-pyrimidin-2-yl-1H-indole-3- carboxamide 480.15 .14 . ##STR00099##
4-fluoro-N-[2-(6-methoxypyridin- 3-yl)-2-piperidin-1-ylethyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 475.21 .10 . ##STR00100##
N-[2-(6-methoxypyridin-3-yl)- 2-piperidin-1-ylethyl]-4-methyl-
1-pyrimidin-2-yl-1H-indole-3- carboxamide 471.25 .13 . ##STR00101##
N-(adamantan-1- ylmethyl)-4-methyl-1- pyrimidin-2-yl-1H-indole-3-
carboxamide 401.29 .43 . ##STR00102## 4-chloro-N-[2-(4-
chlorophenyl)-2-piperazin-1- ylethyl]-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 495.19 .23 . ##STR00103## 4-chloro-N-(2-
morpholin-4-yl-2- phenylethyl)-1-pyrimidin-2-
yl-1H-indole-3-carboxamide 462.21 .18 . ##STR00104##
4-chloro-N-[2-(4- chlorophenyl)-2-piperidin-1-
ylethyl]-1-pyrimidin-2-yl-1H- indole-3-carboxamide 494.19 .22 .
##STR00105## 4-chloro-N-{2-piperidin-1-yl-
2-[4-(trifluoromethyl)phenyl] ethyl}-1-pyrimidin-2-yl-1H-indole-
3-carboxamide 528.21 .23 . ##STR00106## 4-chloro-N-[2-(4-chloro-3-
fluorophenyl)-2-piperidin-1-ylethyl]- 1-pyrimidin-2-yl-1H-indole-3-
carboxamide 512.17 .23 . ##STR00107##
4-chloro-N-[2-(6-methoxypyridin-3- yl)-2-piperidin-1-ylethyl]-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 491.21 .18 . ##STR00108##
4-chloro-N-[2-(3,4- difluorophenyl)-2-piperidin-
1-ylethyl]-1-pyrimidin-2-yl- 1H-indole-3-carboxamide 496.21 .21 .
##STR00109## 3-chloro-1-[(1-methyl-1H- imidazol-2-yl)methyl]-N-(4-
methyl-2-pyridin-3-ylpentyl)- 1H-indole-2-carboxamide 450.20 .04 .
##STR00110## 4-chloro-N-[(1-pyridin-3- ylcyclohexyl)methyl]-1-
pyrimidin-2-yl-1H- pyrrolo[2,3-b]pyridine-3- carboxamide 447.19 .03
. ##STR00111## 4-chloro-N-(4-methyl-2-
pyridin-3-ylpentyl)-1-pyrimidin- 2-yl-1H-pyrrolo[2,3-b]pyridine-3-
carboxamide 435.19 .04 . ##STR00112## 2-adamantan-1-yl-
N-(4-chloro-1-pyrimidin-2-yl- 1H-indo1-3-yl)acetamide 421.28 .54 .
##STR00113## 4-Chloro-1-pyrimidin-2-yl- 1H-indole-3-carboxylic acid
(adamantan-1-ylmethyl)-amide 421.16 .41 . ##STR00114##
4-Chloro-1-pyrimidin-2-yl- 1H-indole-3-carboxylic acid (2-
adamantan-1-yl-ethyl)-amide 435.17 .45 . ##STR00115##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid ((R)-6,6-
dimethylbicyclo[3.1.1]hept-2- ylmethyl)-amide 409.17 .41 .
##STR00116## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
(4-methyl- 2-p-tolyl-pentyl)-amide 447.17 .42 . ##STR00117##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid [2-(4-
chloro-phenyl) 4-methyl- pentyl]-amide 467.10 .42 .
##STR00118## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
[1-(4-chloro- phenyl)-cyclohexylmethyl]- amide 479.11 .46 .
##STR00119## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
[1-(4- trifluoromethyl-phenyl)- cyclohexylmethyl]-amide 513.13 .43
. ##STR00120## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic
acid [4-(4- chloro-phenyl)-tetrahydro-pyran- 4-ylmethyl]-amide
481.09 .34 . ##STR00121## 4-Chloro-1-pyrimidin-2-yl-1H-
indole-3-carboxylic acid [1-(4-methoxy-phenyl)-
cyclohexylmethyl]-amide 475.16 .42 . ##STR00122##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid [4-
methyl-2-(4-trifluoromethyl- phenyl)-pentyl]-amide 501.13 .41 .
##STR00123## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
[2-(4- chloro-phenyl)-pentyl]-amide 453.10 .40 . ##STR00124##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (4-methyl-
2-pyridin-3-yl-pentyl)-amide 434.14 .20 . ##STR00125##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid [1-(4-
chloro-phenyl)-cyclobutylmethyl]- amide 451.08 .41 . ##STR00126##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (2-
adamantan-1-yl-2-hydroxy- ethyl)-amide 451.15 .37 . ##STR00127##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (3-methyl-
butyl)-amide 343.13 .31 . ##STR00128##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
(2-phenyl-pentyl)-amide 419.14 .37 . ##STR00129##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (4-methyl-2-
phenyl-pentyl)-amide 433.16 .39 . ##STR00130##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (3-
cyclopentyl-2-phenyl- propyl)-amide 459.17 .44 . ##STR00131##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (2-
cyclohexyl-2-phenyl-ethyl)- amide 459.15 .44 . ##STR00132##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (2,3-
diphenyl-propyl)-amide 467.14 .37 . ##STR00133##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (3-
phenyl-butyl)-amide 405.13 .34 . ##STR00134##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (4-
phenyl-butyl)-amide 405.13 .35 . ##STR00135##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid [2-(4-bromo-
phenyl)-ethyl]-amide 456.99 .34 . ##STR00136##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (3,3,5-
trimethyl-cyclohexyl)-amide 397.20 .39 . ##STR00137##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid adamantan-
2-ylamide 256.06 .65 . ##STR00138## 4-Chloro-1-pyrimidin-2-yl-1H-
indole-3-carboxylic acid cycloheptylmethyl-amide 383.16 .38 .
##STR00139## 4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid
[2-(2- bromo-phenyl)-ethyl]-amide 456.98 .34 . ##STR00140##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid ((1S,2R)-2-
hydroxy-cyclohexylmethyl)- amide 385.14 .28 . ##STR00141##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (1-hydroxy-
cyclohexylmethyl)-amide 385.14 .28 . ##STR00142##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (1-hydroxy-
cyclopentylmethyl)-amide 371.11 .25 . ##STR00143##
4-Chloro-1-pyrimidin-2-yl-1H- indole-3-carboxylic acid (4-hydroxy-
tetrahydro-thiopyran-4- ylmethyl)-amide 403.09 .24 . ##STR00144##
N-[2-(4-fluorophenyl)-2- piperidin-1-ylethyl]-4-methyl-
1-pyrimidin-2-yl-1H-indole-3- carboxamide 458.25 .21 . ##STR00145##
N-[2-(3,4-difluorophenyl)-2- piperidin-1-ylethyl]-4-methyl-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 476.25 .22 . ##STR00146##
4-methyl-N-(2-morpholin-4-yl- 2-phenylethyl)-1-pyrimidin-2-
yl-1H-indole-3-carboxamide 442.25 .18 . ##STR00147##
N-[2-(4-chlorophenyl)-2- morpholin-4-ylethyl]-4-methyl-1-
pyrimidin-2-yl-1H-indole-3- carboxamide 476.22 .23 . ##STR00148##
3-chloro-N-[2-(4-chlorophenyl)- 2-piperidin-1-ylethyl]-1-[(1-
methyl-1H-imidazol-2-yl)methyl]- 1H-indole-2-carboxamide 510.19 .13
. ##STR00149## N-{[4-(4-chlorophenyl)tetrahydro-
2H-pyran-4-yl]methyl}-4- methyl-1-pyrimidin-2-yl-1H-
indole-3-carboxamide 461.20 .36 . ##STR00150## 3-chloro-N-{[4-(4-
chlorophenyl)tetrahydro-2H- pyran-4-yl]methyl}-1-[(1-
methyl-1H-imidazol-2- yl)methyl]-1H-indole-2- carboxamide 497.18
.24 . ##STR00151## N-[2-(4-chlorophenyl)-2-
piperidin-1-ylethyl]-4-methyl-1-
pyrazin-2-yl-1H-indole-3-carboxamide 474.25 .21 . ##STR00152##
N-[2-(4-chlorophenyl)-2-morpholin-
4-ylethyl]-4-methyl-1-pyrazin-2-yl- 1H-indole-3-carboxamide 476.23
.20 . ##STR00153## N-[2-(4-chlorophenyl)-2-piperidin-
1-ylethyl]-1-(3-cyanopyridin-2- yl)-4-methyl-1H-indole-3-
carboxamide 498.22 .20 . ##STR00154## 1-(3-cyanopyridin-2-
yl)-4-methyl-N-(2-morpholin- 4-yl-2-phenylethyl)-1H-
indole-3-carboxamide 466.24 .15 . ##STR00155##
N-[2-(4-chlorophenyl)-2-morpholin- 4-ylethyl]-1-(3-cyanopyridin-
2-yl)-4-methyl-1H-indole-3- carboxamide 500.20 .19 . ##STR00156##
4-methyl-N-(4- methyl-2-morpholin-4- ylpentyl)-1-pyrimidin-2-yl-
1H-indole-3-carboxamide . ##STR00157##
4-chloro-N-{2-piperidin-1-yl- 2-[6-(trifluoromethyl)pyridin-3-
yl]ethyl}-1-pyrimidin-2-yl-1H- indole-3-carboxamide 529.21 .20 .
##STR00158## 4-chloro-N-[2-(4-fluorophenyl)-2-
piperidin-1-ylethyl]-1-pyrimidin- 2-yl-1H-indole-3-carboxamide
478.23 .19 . ##STR00159## 4-chloro-N-[(1-hydroxycyclo-
heptyl)methyl]-1-pyrimidin-2-yl-1H- indole-3-carboxamide 399.22 .31
. ##STR00160## 4-chloro-N-[2-(3,4-difluorophenyl)-
2-piperidin-1-ylethyl]-1-pyrimidin- 2-yl-1H-indole-3-carboxamide
496.22 .21 . ##STR00161## 4-chloro-N-[2-(2,4-difluorophenyl)-
2-piperidin-1-ylethyl]-1-pyrimidin-2- yl-1H-indole-3-carboxamide
496.21 .20 . ##STR00162## 4-chloro-N-[2-(4-chlorophenyl)-
2-morpholin-4-ylethyl]-1-pyrimidin-
2-yl-1H-pyrrolo[2,3-b]pyridine-3- carboxamide 497.17 .11 .
##STR00163## 4-chloro-N-[2-(4-chlorophenyl)-
2-piperidin-1-ylethyl]-1-pyrimidin-
2-yl-1H-pyrrolo[2,3-b]pyridine-3- carboxamide 377.17 .19 .
##STR00164## 4-chloro-N-{[4-(4-chlorophenyl)
tetrahydro-2H-pyran-4-yl]methyl}-
1-pyrimidin-2-yl-1H-pyrrolo[2,3-b] pyridine-3-carboxamide 482.16
.26 . ##STR00165## 4-chloro-N-[(1-hydroxycycloheptyl)
methyl]-1-pyrimidin-2-yl-1H- pyrrolo[2,3-b]pyridine-3- carboxamide
400.21 .22 . ##STR00166## 4-chloro-N-[(1-hydroxycyclohexyl)
methyl]-1-pyrimidin-2-yl-1H- pyrrolo[2,3-b]pyridine-3- carboxamide
386.19 .18 . ##STR00167## 4-chloro-N-[2-(3,4-difluorophenyl)-
2-piperidin-1-ylethyl]-1-pyrimidin-
2-yl-1H-pyrrolo[2,3-b]pyridine-3- carboxamide 497.22 .12 .
##STR00168## 4-chloro-N-(2-morpholin-4-yl-2-
phenylethyl)-1-pyrimidin-2- yl-1H-pyrrolo[2,3-b]pyridine-
3-carboxamide 463.21 .06 . ##STR00169## 4-chloro-N-{[1-(4-methyl-
piperazin-1-yl)cyclohexyl]methyl}-1- pyrimidin-2-yl-1H-indole-3-
carboxamide 467.25 .44 ##STR00170## indicates data missing or
illegible when filed
[0310] Example 7
P2X7 Calcium Mobilization Assay
[0311] This Example illustrates representative calcium mobilization
assays for use in evaluating test compounds for agonist and
antagonist activity.
[0312] A. High Throughput Assay of P2X7 Receptors
[0313] SH-SY5Y cells, ATCC Number CRL-2266, (American Type Culture
Collection, Manassas, Va.) are cultured under DMEM/High medium
supplemented with 10% FBS, and 10 mM HEPES (Invitrogen Corp.,
Carlsbad, Calif.) in 5% CO.sub.2 and at 37.degree. C. One day prior
to the experiment, cells are plated at a density of 100,000
cells/well in a 96 well black/clear TC plate (Corning.RTM.
Costar.RTM., Sigma-Aldrich Co., St. Louis, Mo.). At the beginning
of the experiment, the culture medium is removed and cells are
incubated with 50 .mu.L of 2.3 .mu.M Fluo-4 AM dye (Invitrogen
Corp.) in the assay solution (5 mM KCl, 9.6 mM
NaH.sub.2PO.sub.4H.sub.2O, 25 mM HEPES, 280 mM Sucrose, 5 mM
Glucose, and 0.5 mM CaCl.sub.2; pH is adjusted to 7.4 with NaOH)
for an hour at 37.degree. C. After one hour dye incubation, wells
are rinsed once with 50 .mu.L assay solution, and are then
incubated for an hour at rt with 100 .mu.L assay solution
containing the test compound. The final concentration of test
compound generally ranges from 1 to 2500 nM; for positive control
cells, no test compound is added. After the one hour incubation,
plates are transferred to a FLIPR.sup.TETRA instrument (Molecular
Devices, Sunnyvale, Calif.) for calcium mobilization analysis.
[0314] For determination of antagonist activity, 50 .mu.L of
P2X.sub.7 agonist (2'(3')--O-(4-benzoyl-benzoyl)adenosine
5'-triphosephate (BzATP; Sigma-Aldrich) in the assay solution is
transferred using the FLIPR into the plate, such that the final
agonist concentration is 80 .mu.M (about EC.sub.50). In negative
control cells, 50 .mu.L of assay solution without agonist is added
at this stage. The peak fluorescence signal over a 2 minute period
is then measured.
[0315] The data is analyzed as follows. First, the average maximum
relative fluorescent unit (RFU) response from the negative control
wells (no agonist) is subtracted from the maximum response detected
for each of the other experimental wells. Second, average maximum
RFU response is calculated for the positive control wells (agonist
wells). Then, percent inhibition for each compound tested is
calculated using the equation:
Percent Inhibition=100-100.times.(Peak Signal in Test Cells/Peak
Signal in Control Cells)
[0316] The % inhibition data is plotted as a function of test
compound concentration and test compound IC.sub.50 is determined
using, for example, KALEIDAGRAPH software (Synergy Software,
Reading, Pa.) best fit of the data to the equation:
y=m.sub.1*(1/(1+(m.sub.2/m.sub.0).sup.m3))
[0317] where y is the percent inhibition, m.sub.0 is the
concentration of the agonist, m.sub.1 is the maximum RFU, m.sub.2
corresponds to the test compound IC.sub.50 (the concentration
required to provide a 50% decrease, relative to the response
observed in the presence of agonist and without antagonist) and
m.sub.3 is the Hill coefficient. Alternatively, test compound
IC.sub.50 is determined using a linear regression in which x is
ln(concentration of test compound) and y is ln(percent
inhibition/(100-percent inhibition). Data with a percent inhibition
that is greater than 90% or less than 15% are rejected and are not
used in the regression. The IC.sub.50 calculated in this fashion is
e.sup.(-intercept/slope). For antagonists of the P2X.sub.7
receptor, the calculated IC.sub.50 is preferably below 20
micromolar, more preferably below 10 micromolar, even more
preferably below 5 micromolar and most preferably below 1
micromolar.
[0318] Similar assays are performed in the absence of added agonist
for the determination of agonist activity of the test compounds.
Within such assays, the ability of a test compound to act as an
agonist of P2X.sub.7 receptor is determined by measuring the
fluorescence response elicited by the test compound as a function
of compound concentration. P2X.sub.7 receptor antagonists that
exhibit no detectable agonist activity elicit no detectable
fluorescence response at a concentration of 2,500 nM.
[0319] B. Electrophysiology Assay for P2X7 Receptors
[0320] SH-SY5Y cells are cultured under DMEM/High medium
supplemented with 10% FBS, and 10 mM HEPES (Invitrogen Corp.,
Carlsbad, Calif.) in 5% CO.sub.2 and at 37.degree. C., and are
split onto 12 mm round Poly-D-Lysine (PDL) coated coverslips (BD
Biosciences, San Jose, Calif.) in a 35 mm dish with a density of
130K cells/dish a day prior to the experiment. Whole cell voltage
clamp recordings are made with the Axopatch-200B amplifier (Axon
Instruments, Foster City, Calif.). The recording electrodes are
pulled from borosilicate pipettes (World Precision Instruments,
Sarasota, Fla.) on a horizontal puller (Sutter Instrument Model
P-87) and have resistances ranging from 2 to 3 M.OMEGA. when
backfilled with internal solution. All voltage protocols are
generated using pClamp 8 (Axon Instruments) software. Data are
digitized at 1 or 5 kHz and recorded onto a PC for further
analysis. Data are analyzed using Clampfit (Axon Instruments),
Excel (Microsoft, Redmond, Wash.), and Origin software (MicroCal,
LLC; Northampton, Mass.). All whole-cell recordings are conducted
at rt. Internal solution contains (in mM): 100 KF, 40 KCl, 5 NaCl,
10 EGTA and 10 HEPES (pH=7.4 adjusted with KOH). The external
solution contains 70 mM NaCl, 0.3 mM CaCl.sub.2, 5 mM KCl, 20 mM
HEPES, 10 mM glucose, and 134 mM sucrose (pH=7.4 adjusted with
NaOH). All chemicals are from Sigma, unless otherwise stated.
[0321] P2X.sub.7 receptor is activated by 200 .mu.M of P2X.sub.7
agonist, BzATP. At a holding potential of -80 mV, the activated
inward current is recorded in the presence and absence of the test
compound. Then, percent inhibition for each compound tested is
calculated using the equation:
% Inhibition=100-100.times.(Current Amplitude in Compound/Current
Amplitude in Control).
[0322] To determine a test compound's IC.sub.50 for P2X.sub.7
receptor electrophysiologically, several concentrations of the
compound are tested and their inhibitions on P2X.sub.7 currents are
calculated as above. This dose-response curve is best fitted using
Origin software (Microcal, MA) with the following equation:
Percent Inhibition=100/(1+(IC.sub.50/C).sup.N)
[0323] where C is the concentration of the antagonist, N is the
Hill coefficient, and IC.sub.50 represents the compound IC.sub.50
value against P2X.sub.7 receptors.
Example 8
Carrageenan-Induced Mechanical Hyperalgesia (Paw Pressure) Assay
for Determining Pain Relief
[0324] This Example illustrates a representative method for
assessing the degree of pain relief provided by a test
compound.
[0325] Adult male Sprague Dawley rats (200-300g; obtained from
Harlan Sprague Dawley, Inc., Indianapolis, Ind.) are housed under a
12 h light/dark cycle with access to food and water ad libitum. For
the assay, all animals are habituated once, baselined twice and
tested once, with each procedure being conducted on a separate day.
Prior to each day's procedure, animals are allowed to acclimate for
at least 1 hour in the testing room before the start of the
procedure. For habituation, each animal is gently restrained with
each hindpaw consecutively extended in front of the animal as is
necessary for testing. This procedure is performed by alternating
hindpaws and repeated three times for each hindpaw. Animals are
then subjected to the first baseline, second baseline and testing
on consecutive days. For each baseline, the animal is restrained as
in the habituation session and the paw tested using the paw
pressure testing apparatus (Digital Randall Selitto, IITC Inc.,
Woodland Hills, Calif.). Animals are baselined and tested in groups
of ten, each animal being tested once on the left and right
hindpaws, followed by the next consecutive animal. This procedure
is repeated three times for a total of three measurements on each
hindpaw. If any individual read is drastically different (varies by
more than about 100 g) from the other two on a given hindpaw, the
hindpaw is retested a 4.sup.th time, and the average of the three
most consistent scores is used. On test day, all animals are
injected with 0.1 mL intraplantar 0.5%-1.5% carrageenan (dissolved
in saline) 3 hours prior to testing. Test compounds or vehicle may
be administered by various routes at various timepoints prior to
testing, but for any particular assay, the routes and timepoints
are the same for animals in each treatment group administered test
compound (a different dosage of test compound may be administered
to each such group) and those in the treatment group administered
vehicle control. If a compound is orally administered, the animals
are food-deprived the evening before testing. As with the baseline,
each hindpaw is tested three times and the results recorded for
analysis.
[0326] Hypersensitivity of nociception values are calculated for
each treatment group as the mean of the left foot gram force scores
on test day (left foot only or LFO score). Statistical significance
between treatment groups is determined by running an ANOVA on LFO
scores followed with a least significant difference (LSD) post hoc
test. A p<0.05 is considered to be a statistically significant
difference.
[0327] Compounds are said to relieve pain in this model if they
result in a statistically significant reduction in hypersensitivity
of nociception values compared to vehicle controls, determined as
described above, when administered (0.01-50 mg/kg, orally,
parenterally or topically) immediately prior to testing as a single
bolus, or for several days: once or twice or three times daily
prior to testing.
[0328] Those skilled in the art will appreciate that numerous
changes and modifications can be made to the preferred embodiments
of the invention and that such changes and modifications can be
made without departing from the spirit of the invention. It is,
therefore, intended that the appended claims cover all such
equivalent variations as fall within the true spirit and scope of
the invention.
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