U.S. patent application number 09/975566 was filed with the patent office on 2002-11-21 for modulation of ccr4 function.
This patent application is currently assigned to Tularik Inc.. Invention is credited to Collins, Tassie, Dairaghi, Daniel J., Mahmud, Hossen, McMaster, Brian E., Medina, Julio C., Schall, Thomas J., Wang, Xuemei, Xu, Feng.
Application Number | 20020173524 09/975566 |
Document ID | / |
Family ID | 26933078 |
Filed Date | 2002-11-21 |
United States Patent
Application |
20020173524 |
Kind Code |
A1 |
Collins, Tassie ; et
al. |
November 21, 2002 |
Modulation of CCR4 function
Abstract
Compounds and compositions are provided that bind to the CCR4
chemokine receptor and which are useful for treating diseases
associated with CCR4 activity, such as contact
hypersensitivity.
Inventors: |
Collins, Tassie; (San Mateo,
CA) ; Dairaghi, Daniel J.; (Palo Alto, CA) ;
Mahmud, Hossen; (San Antonio, TX) ; McMaster, Brian
E.; (Mountain View, CA) ; Medina, Julio C.;
(San Carlos, CA) ; Schall, Thomas J.; (Palo Alto,
CA) ; Xu, Feng; (Palo Alto, CA) ; Wang,
Xuemei; (San Mateo, CA) |
Correspondence
Address: |
TOWNSEND AND TOWNSEND AND CREW, LLP
TWO EMBARCADERO CENTER
EIGHTH FLOOR
SAN FRANCISCO
CA
94111-3834
US
|
Assignee: |
Tularik Inc.
Two Corporate Drive
So. San Francisco
CA
94080
|
Family ID: |
26933078 |
Appl. No.: |
09/975566 |
Filed: |
October 11, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60240022 |
Oct 11, 2000 |
|
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60293781 |
May 23, 2001 |
|
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Current U.S.
Class: |
514/364 ;
514/369; 514/370; 514/438; 514/452; 514/471; 514/473; 514/513;
514/602 |
Current CPC
Class: |
C07D 277/42 20130101;
A61K 31/341 20130101; C07D 277/82 20130101; A61K 31/357 20130101;
A61K 31/4245 20130101; C07D 513/04 20130101; A61K 31/36 20130101;
A61K 31/426 20130101; A61K 31/381 20130101; C07D 417/12 20130101;
A61K 31/00 20130101; A61P 11/00 20180101; A61P 33/00 20180101; A61K
31/265 20130101; A61K 31/351 20130101 |
Class at
Publication: |
514/364 ;
514/369; 514/370; 514/438; 514/452; 514/471; 514/473; 514/513;
514/602 |
International
Class: |
A61K 031/426; A61K
031/4245; A61K 031/381; A61K 031/36; A61K 031/351; A61K 031/265;
A61K 031/18 |
Claims
What is claimed is:
1. A method of treating a CCR4-mediated condition or disease in a
subject, said method comprising administering to a subject in need
of such treatment an effective amount of a compound having the
formula: Ar.sup.1-X-Ar.sup.2 (I) wherein Ar.sup.1 and Ar.sup.2 are
each members independently selected from the group consisting of
substituted or unsubstituted aryl, substituted or unsubstituted
fused aryl-heterolcyclic ring systems and substituted or
unsubstituted heteroaryl; and X is a linking group selected from
the group consisting of --N(R)--, --C(O)S--, --CH.dbd.CHSO.sub.2--
and --SO.sub.2N(R)-- wherein R is a member selected from the group
consisting of H and substituted or unsubstituted
(C.sub.1-C.sub.8)alkyl.
2. A method in accordance with claim 1, wherein X is --NH--.
3. A method in accordance with claim 1, wherein X is
--SO.sub.2NH--.
4. A method in accordance with claim 1, wherein Ar.sup.1 and
Ar.sup.2 are each substituted or unsubstituted members
independently selected from the group consisting of: 34
5. A method in accordance with claim 2, wherein Ar.sup.1 is
substituted heteroaryl and Ar.sup.2 is substituted or unsubstituted
aryl.
6. A method in accordance with claim 5, wherein said Ar.sup.1 is a
substituted heteroaryl selected from the group consisting of
substituted thiazolyl, substituted thienyl, and substituted
furanyl.
7. A method in accordance with claim 5, wherein said Ar.sup.2 is a
substituted or unsubstituted phenyl or a substituted or
unsubstituted naphthyl.
8. A method in accordance with claim 3, wherein Ar.sup.2 is a
phenyl group having from 1 to 4 substituents independently selected
from the group consisting of halogen, hydroxy,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkylthio, (C.sub.1-C.sub.4)haloalkyl,
(C.sub.1-C.sub.4)haloalkoxy, nitro, cyano, (C.sub.1-C.sub.4)acyl,
amino, (C.sub.1-C.sub.4)alkylamino, and
di(C.sub.1-C.sub.4)alkyamino.
9. A method in accordance with claim 8, wherein said phenyl group
has from 1 to 3 substituents independently selected from the group
consisting of halogen, (C.sub.1-C.sub.4)haloalkyl,
(C.sub.1-C.sub.4)haloalkoxy, nitro, cyano, and
(C.sub.1-C.sub.4)acyl.
10. A method in accordance with claim 3, wherein Ar.sup.1 is a
substituted or unsubstituted monocyclic or bicyclic
heterocycle.
11. A method in accordance with claim 10, wherein said heterocycle
is selected from the group consisting of pyrrolyl, pyrazolyl,
imidazolyl, pyrazinyl, oxazolyl, isoxazolyl, thiazolyl, furyl,
thienyl, pyridyl, pyrimidyl, benzothiazolyl, benzoxadiazolyl,
purinyl, benzimidazolyl, indolyl, isoquinolyl, quinoxalinyl and
quinolyl.
12. A method in accordance with claim 11, wherein said heterocycle
is selected from the group consisting of thienyl, thiazolyl and
benzoxadiazolyl.
13. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is selected from the group consisting of
contact hypersensitivity, atopic dermatitis, allergic airway
hypersensitivity, allergic rhinitis, atherosclerosis, septic shock,
angina, myocardial infarction, restenosis, ischemia/reperfusion
injury, multiple sclerosis, rheumatoid arthritis, type I diabetes,
psoriasis, cancer and HIV infection.
14. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is psoriasis, contact hypersensitivity or
atopic dermatitis.
15. A method in accordance with claim 14, wherein said
CCR4-mediated condition or disease is psoriasis.
16. A method in accordance with claim 14, wherein said
CCR4-mediated condition or disease is contact hypersensitivity.
17. A method in accordance with claim 14, wherein said
CCR4-mediated condition or disease is atopic dermatitis.
18. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is a disease of the airway.
19. A method in accordance with claim 18, wherein said disease of
the airway is selected from the group consisting of allergic asthma
and allergic rhinitis.
20. A method in accordance with claim 18, wherein said disease of
the airway is allergic asthma.
21. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is a disease of innate immunity.
22. A method in accordance with claim 21, wherein said disease of
innate immunity is septic shock.
23. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is atherosclerosis.
24. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is a disease or condition characterized by
platelet aggregation or thrombosis.
25. A method in accordance with claim 24, wherein said
CCR4-mediated disease or condition is selected from the group
consisting of angina, myocardial infarction, restenosis, stroke and
ischemia/reperfusion injury.
26. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is an allergic condition and said compound is
used alone or in combination with at least one therapeutic agent
wherein said therapeutic agent is an antihistamine.
27. A method in accordance with claim 1, wherein said CCR4-mediated
disease or condition is psoriasis and said compound is used alone
or in combination with at least one therapeutic agent selected from
a corticosteroid, a lubricant, a keratolytic agent, a vitamin
D.sub.3 derivative, PUVA, or anthralin.
28. A method in accordance with claim 1, wherein said CCR4-mediated
disease or condition is atopic dermatitis and said compound is used
alone or in combination with at least one therapeutic agent
selected from a lubricant and corticosteroid.
29. A method in accordance with claim 1, wherein said CCR4-mediated
condition or disease is asthma and said compound is used alone or
in combination with at least one therapeutic agent selected from a
.beta.2-agonist and a corticosteroid.
30. A method in accordance with claim 1, wherein said compound
interferes with the interaction between CCR4 and a ligand.
31. A method in accordance with claim 1, wherein said
administration is oral or intravenous.
32. A method in accordance with claim 1, wherein said subject is
selected from the group consisting of human, rat, dog, cow, horse,
and mouse.
33. A method in accordance with claim 1, wherein said subject is
human.
34. A method in accordance with claim 1, wherein said compound is
selected from the group consisting of 35
35. A method in accordance with claim 1, wherein said CCR4-mediated
disease or condition is selected from the group consisting of
multiple sclerosis, rheumatoid arthritis, type I diabetes,
psoriasis, cancer and HIV infection; Ar.sup.1 is a substituted
heterocycle; X is --SO.sub.2NH--; and Ar.sup.2 is a substituted
phenyl.
36. A method in accordance with claim 1, wherein said CCR4-mediated
disease or condition is selected from the group consisting of
multiple sclerosis, rheumatoid arthritis, type I diabetes,
psoriasis, cancer and HIV infection; Ar.sup.1 is a substituted
heterocycle; X is --NH--; and Ar.sup.2 is naphthyl.
37. A pharmaceutical composition for the treatment of a
CCR4-mediated disease or condition, said composition comprising a
pharmaceutically acceptable carrier and an effective amount of a
compound which inhibits the binding of MDC or TARC to CCR4, said
compound having the formula: Ar.sup.1-X-Ar.sup.2 (I) Ar.sup.1 and
Ar.sup.2 are each members independently selected from the group
consisting of substituted or unsubstituted aryl, substituted or
unsubstituted fused aryl-heteocyclic ring systems and substituted
or unsubstituted heteroaryl; and X is a linking group selected from
the group consisting of --N(R)--, --C(O)S--, --CH.dbd.CHSO.sub.2--
and --SO.sub.2N(R)-- wherein R is a member selected from the group
consisting of H and substituted or unsubstituted
(C.sub.1-C.sub.8)alkyl.
38. A composition of claim 37, wherein X is --NH--.
39. A composition of claim 37, wherein X is --SO.sub.2NH--.
40. A composition of claim 37, wherein Ar.sup.1 and Ar.sup.2 are
each substituted or unsubstituted members independently selected
from the group consisting of: 36
41. A composition of claim 37, wherein Ar.sup.1 is substituted
heteroaryl and Ar.sup.2 is substituted or unsubstituted aryl.
42. A composition of claim 41, wherein said Ar.sup.1 is a
substituted heteroaryl selected from the group consisting of
substituted thiazolyl, substituted thienyl, and substituted
furanyl.
43. A composition of claim 41, wherein said Ar.sup.2 is a
substituted or unsubstituted phenyl or a substituted or
unsubstituted naphthyl.
44. A composition of claim 41, wherein Ar.sup.2 is a phenyl group
having from 1 to 4 substituents independently selected from the
group consisting of halogen, hydroxy, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, (C.sub.1-C.sub.4)alkylthio,
(C.sub.1-C.sub.4)haloalkyl, (C.sub.1-C.sub.4)haloalkoxy, nitro,
cyano, (C.sub.1-C.sub.4)acyl, amino, (C.sub.1-C.sub.4)alkylamino,
and di(C.sub.1-C.sub.4)alkylamino.
45. A composition of claim 44, wherein said phenyl group has from 1
to 3 substituents independently selected from the group consisting
of halogen, (C.sub.1-C.sub.4)haloalkyl,
(C.sub.1-C.sub.4)haloalkoxy, nitro, cyano, and
(C.sub.1-C.sub.4)acyl.
46. A composition of claim 37, wherein Arl is a substituted or
unsubstituted monocyclic or bicyclic heterocycle.
47. A composition of claim 46, wherein said heterocycle is selected
from the group consisting of pyrrolyl, pyrazolyl, imidazolyl,
pyrazinyl, oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl,
pyridyl, pyrimidyl, benzothiazolyl, benzoxadiazolyl, purinyl,
benzimidazolyl, indolyl, isoquinolyl, quinoxalinyl and
quinolyl.
48. A composition of claim 47, wherein said heterocycle is selected
from the group consisting of thienyl, thiazolyl and
benzoxadiazolyl.
49. A composition of claim 37, wherein said compound is selected
from the group consisting of 37
50. A method for modulating CCR4 function in a cell, comprising
contacting said cell with a CCR4-modulating amount of a composition
of claim 37.
51. A method for modulating CCR4 function, in which said cell is
contacted with a CCR4 protein with a therapeutically effective
amount of the composition of claim 37.
52. A compound of formula (I): 38or a pharmaceutically acceptable
salt thereof, wherein W is selected from aryl, heteroaryl,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, cycloalkyl and
heterocycloalkyl; X is selected from N(R.sup.5), S, O,
C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4) and, optionally, when Z
is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)--(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; Z is selected from N
and C(R.sup.8); R.sup.1 and R.sup.2 are independently selected from
H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl, and when R' and R" are attached to
nitrogen atom, they may be combined with the nitrogen atom to form
a 5-, 6-, or 7-membered ring; R.sup.3, R.sup.4 and R.sup.8are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl; with the provisos that R.sup.2 is other than H when W is
unsubstituted phenyl, X is S, Y is NH, Z is N and R.sup.1 is
(C.sub.1-C.sub.8)alkyl; and R.sup.1 is other than phenyl, when W is
phenyl or unsubstituted naphthyl, X is S, Y is NH, and Z is N.
53. A compound of claim 52, wherein Z is N.
54. A compound of claim 52, wherein X is S.
55. A compound of claim 52, wherein Y is N(R.sup.5).
56. A compound of claim 52, wherein Z is N, X is S and Y is
N(R.sup.5).
57. A compound of claim 52, wherein W is aryl or heteroaryl.
58. A compound of claim 57, wherein W is substituted or
unsubstituted phenyl or naphthyl.
59. A compound of claim 57, wherein W is substituted or
unsubstituted pyridyl or quinolyl.
60. A compound of claim 52, wherein R.sup.1 and R.sup.2 are each
independently selected from H and (C.sub.1-C.sub.8)alkyl.
61. A compound of claim 52, wherein R.sup.1 and R.sup.2 are
combined to form a fused 6-membered aryl or heteroaryl ring.
62. A compound of claim 52, wherein Z is N, X is S, Y is N(R.sup.5)
and R.sup.1 and R.sup.2 are each independently selected from H and
(C.sub.1-C.sub.8)alkyl.
63. A compound of claim 52, wherein Z is N, X is S, Y is N(R.sup.5)
and R.sup.1 and R.sup.2 are combined to form a fused 6-membered
aryl or heteroaryl ring.
64. A compound of claim 52, said compound being selected from the
group consisting of: 39
65. A compound of claim 52, said compound being selected from the
group consisting of: 40
66. A compound of claim 52, wherein W is selected from substituted
phenyl, substituted or unsubstituted naphthyl, pyridyl, quinolyl,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, cycloalkyl and
heterocycloalkyl; X is selected from N(R.sup.5), S, O,
C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4) and, optionally, when Z
is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)--(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; Z is selected from N
and C(R.sup.8); R.sup.1 and R.sup.2 are independently selected from
H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl, and when R' and R" are attached to
a nitrogen atom, they may be combined with the nitrogen atom to
form a 5-, 6-, or 7-membered ring; R.sup.3, R.sup.4 and R.sup.8are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
67. A compound of claim 66, wherein Z is N.
68. A compound of claim 66, wherein X is S.
69. A compound of claim 66, wherein Y is N(R.sup.5).
70. A compound of claim 66, wherein Z is N, X is S and Y is
N(R.sup.5).
71. A compound of claim 66, wherein W is substituted phenyl or
substituted or unsubstituted naphthyl.
72. A compound of claim 66, wherein W is substituted or
unsubstituted pyridyl or substituted or unsubstituted quinolyl.
73. A compound of claim 66, wherein R.sup.1 and R.sup.2 are
independently selected from the group consisting of H and
(C.sub.1-C.sub.8)alkyl.
74. A compound of claim 66, wherein R and R.sup.2 are combined to
form a fused 6-membered aryl or heteroaryl ring.
75. A compound of claim 66, wherein W is substituted phenyl or
substituted or unsubstituted naphthyl, Z is N, X is S, Y is N(R
.sup.5), and R.sup.1 and R.sup.2 are independently selected from
the group consisting of H and (C.sub.1-C.sub.8)alkyl.
76. A compound of claim 66, wherein W is substituted phenyl or
substituted or unsubstituted naphthyl, Z is N, X is S, Y is
N(R.sup.5), and R.sup.1 and R.sup.2 are combined to form a fused
6-membered aryl or heteroaryl ring.
77. A compound of claim 66, wherein W is substituted or
unsubstituted pyridyl or substituted or unsubstituted quinolyl, Z
is N, X is S, Y is N(R.sup.5), and R.sup.1 and R.sup.2 are
independently selected from the group consisting of H and
(C.sub.1-C.sub.8)alkyl.
78. A compound of claim 66, wherein W is substituted or
unsubstituted pyridyl or substituted or unsubstituted quinolyl, Z
is N, X is S, Y is N(R.sup.5), and R.sup.1 and R.sup.2 are combined
to form a fused 6-membered aryl or heteroaryl ring.
79. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound of formula (I): 41or a
pharmaceutically acceptable salt thereof, wherein W is selected
from aryl, heteroaryl, (C.sub.1-C.sub.8)alkyl, heteroalkyl,
cycloalkyl and heterocycloalkyl; X is selected from N(R.sup.5), S,
O, C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4) and, optionally,
when Z is N, X can be C(R.sup.6)(R7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)--(C.sub.1-C.sub.8)alkylen- e, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; Z is selected from N
and C(R.sup.8); R.sup.1 and R.sup.2 are independently selected from
H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl, and when R' and R" are attached to
nitrogen atom, they may be combined with the nitrogen atom to form
a 5-, 6-, or 7-membered ring; R.sup.3, R.sup.4 and R.sup.8are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
80. A method for treating a CCR4-mediated condition in a subject,
said method comprising administering to a subject in need of such
treatment an effective amount of a compound of of formula (I): 42or
a pharmaceutically acceptable salt thereof, wherein W is selected
from aryl, heteroaryl, (C.sub.1-C.sub.8)alkyl, heteroalkyl,
cycloalkyl and heterocycloalkyl; X is selected from N(R.sup.5), S,
O, C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4) and, optionally,
when Z is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a
bond, N(R.sup.5), N(R.sup.5)--(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; Z is selected from N
and C(R.sup.8); R.sup.1 and R.sup.2 are independently selected from
H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl, and when R' and R" are attached to
nitrogen atom, they may be combined with the nitrogen atom to form
a 5-, 6-, or 7-membered ring; R.sup.3, R.sup.4 and R.sup.8are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
81. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a compound selected from the group
consisting of: 43
82. A pharmaceutical composition of claim 81, wherein said compound
is selected from the group consisting of: 44
83. A method for treating a CCR4-mediated condition in a subject,
said method comprising administering to a subject in need of such
treatment an effective amount of a compound selected from the group
consisting of: 45
84. A method in accordance with claim 83, wherein said compound is
selected from the group consisting of: 46
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application claims the benefit of Provisional
Application Ser. No. 60/240,022, filed Oct. 11, 2000, and further
claims the benefit of Provisional Application Ser. No. 60/293,781,
filed May 23, 2001, the disclosures of each being incorporated
herein by reference.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH AND DEVELOPMENT
[0002] Not applicable
BACKGROUND OF THE INVENTION
[0003] Chemokines are chemotactic cytokines that are released by a
wide variety of cells to attract macrophages, T cells, eosinophils,
basophils and neutrophils to sites of inflammation (reviewed in
Schall, Cytokine, 3:165-183 (1991), Schall, et al, Curr. Opin.
Immunol. 6:865-873 (1994) and Murphy, Rev. Immun., 12:593-633
(1994)). In addition to stimulating chemotaxis, other changes can
be selectively induced by chemokines in responsive cells, including
changes in cell shape, transient rises in the concentration of
intracellular free calcium ions ([Ca.sup.2+]).sub.i, granule
exocytosis, integrin upregulation, formation of bioactive lipids
(e.g., leukotrienes) and respiratory burst, associated with
leukocyte activation. Thus, the chemokines are early triggers of
the inflammatory response, causing inflammatory mediator release,
chemotaxis and extravasation to sites of infection or
inflammation.
[0004] There are four classes of chemokines, CXC (.alpha.), CC
(.beta.), C(.gamma.), and CX.sub.3C (.delta.), depending on whether
the first two cysteines are separated by a single amino acid
(C--X--C), are adjacent (C--C), have a missing cysteine pair (C),
or are separated by three amino acids (CX.sub.3C). The
.alpha.-chemokines, such as interleukin-8 (IL-8), melanoma growth
stimulatory activity protein (MGSA), and stromal cell derived
factor 1 (SDF-1) are chemotactic primarily for neutrophils and
lymphocytes, whereas .beta.-chemokines, such as RANTES,
MIP-1.alpha., MIP-1.beta., monocyte chemotactic protein-1 (MCP-1),
MCP-2, MCP-3 and eotaxin are chemotactic for macrophages, T-cells,
eosinophils and basophils (Deng, et al., Nature, 381:661-666
(1996)). The C chemokine lymphotactin shows specificity for
lymphocytes (Kelner, et al., Science, 266:1395-1399 (1994)) while
the CX.sub.3C chemokine fractalkine shows specificity for
lymphocytes and monocytes (Bazan, et al., Nature, 385:640-644
(1997).
[0005] Chemokines bind to specific cell-surface receptors belonging
to the family of G-protein-coupled seven-transmembrane-domain
proteins (reviewed in Horuk, Trends Pharm. Sci. 15:159-165 (1994))
which are termed "chemokine receptors." On binding their cognate
ligands, chemokine receptors transduce an intracellular signal
through the associated heterotrimeric G protein, resulting in a
rapid increase in intracellular calcium concentration. There are at
least twelve human chemokine receptors that bind or respond to
.beta.-chemokines with the following characteristic pattern: CCR1
(or "CKR-1" or "CC-CKR-1") to MIP-1.alpha., MIP-1.beta., MCP-3,
RANTES (Ben-Barruch, et al., J. Biol. Chem., 270:22123-22128
(1995); Neote, et al., Cell, 72:415-425 (1993)); CCR2A and CCR2B
(or "CKR-2A"/"CKR-2A" or "CC-CKR-2A"/"CC-CKR2A") to MCP-1, MCP-3,
MCP-4; CCR3 (or "CKR-3" or "CC-CKR-3") to eotaxin, RANTES, MCP;
(Ponath, et al., J. Exp. Med., 183:2437-2448 (1996)); CCR4 (also
referred to as "CKR-4","CC-CKR-4" or "CMKBR4"), to TARC, MDC (Imai
et al. (1998) J. Biol. Chem. 273:1764-1768); CCR5 (or "CKR-5" or
"CC-CKR-5") to MIP-1.alpha., RANTES, MIP-1.beta. (Sanson, et al.,
Biochemistry, 35:3362-3367 (1996)); CCR6 to MIP-3 alpha (Greaves,
et al., J. Exp. Med., 186:837-844 (1997)); CCR7 to MIP-3 beta and
6Ckine (Campbell, et al., J. Cell. Biol., 141:1053-1059(1998));
CCR8 to I-309, HHV8 vMIP-I, HHV-8 vMIP-II, MCV vMCC-I (Dairaghi, et
al., J. Biol. Chem., 274:21569-21574 (1999)); CCR9 to TECK
(Zaballos, et al., J. Immunol., 162:5671-5675 (1999)), D6 MIP-1
beta, RANTES, and MCP-3 (Nibbs, et al., J. Biol. Chem.,
272:32078-32083 (1997)), the Duffy blood-group antigen to IL-8,
Gro.alpha., RANTES, MCP-1 (Chaudhun et al. (1994) J. Biol. Chem.
269:7835-7838, Murphy et al. (2000) Pharm. Rev. 52:145-176) and
CCR10 to CTACK, CCL28 (Jarmin et al. (2000) J. Immunol.
164:3460-3464, Homey et al. (2000) J. Immunol. 164:3465-3470, Wang
et al. (2000) J. Biol. Chem. 275:22313-22323).
[0006] Chemokine receptors, such as CCR1, CCR2, CCR2A, CCR2B, CCR3,
CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CXCR1, CXCR2, CXCR3, CXCR4,
CXCR5, CX.sub.3CR1 and XCR1 have been implicated as being important
mediators of inflammatory and immunoregulatory disorders and
diseases, including asthma and allergic diseases, as well as
autoimmune pathologies such as rheumatoid arthritis and
atherosclerosis.
[0007] The CCR4 chemokine receptor, first identified by Power et
al. (Power et al. (1995) J. Biol. Chem. 270:19495-19500), is
expressed primarily in peripheral blood T lymphocytes. CCR4 is
involved in T lymphocyte homing to the skin and lungs (see, e.g.,
Campbell et al. (1999) Nature 400:776-780, Gonzalo et al. (1999) J.
Immunol. 163:403-411, Lloyd et al. (2000) J. Exp. Med. 191:265-273,
Kawasaki et al. (2001) J. Immunol. 166:2055-2062).
[0008] The identification of compounds that modulate the function
of CCR4 represents an attractive approach to the development of
therapeutic agents for the treatment of inflammatory conditions and
diseases associated with CCR4 activation, such as psoriasis, asthma
and allergic diseases.
SUMMARY OF THE INVENTION
[0009] The present invention is directed to compounds which are
modulators of CCR4 chemokine receptor function and are useful in
the prevention or treatment of inflammatory conditions and diseases
such as allergic diseases, psoriasis, atopic dermatitis and asthma.
The invention is also directed to pharmaceutical compositions
comprising these compounds and the use of these compounds and
compositions in the prevention or treatment of diseases in which
CCR4 chemokine receptors are involved.
[0010] In one aspect, the present invention provides compounds
having the general formula (I): 1
[0011] wherein W is selected from aryl, heteroaryl,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, cycloalkyl and
heterocycloalkyl; X is selected from N(R.sup.5), S, O,
C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4), and optionally, when Z
is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)-(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; and Z is selected from
N and C(R.sup.8). R.sup.1 and R.sup.2 are independently selected
from H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl. When R' and R" are attached to
nitrogen, they may be combined with the nitrogen atom to form a 5-,
6-, or 7-membered ring. R.sup.3, R.sup.4 and R.sup.8 are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl; with the provisos that R.sup.1 is other than phenyl,
when W is phenyl or unsubstituted naphthyl, X is S, Y is NH, and Z
is N; and R.sup.2 is other than H when W is unsubstituted phenyl, X
is S, Y is NH, Z is N and R.sup.1 is (C.sub.1-C.sub.8)alkyl.
[0012] Unless otherwise indicated, the compounds provided in the
above formula are meant to include pharmaceutically acceptable
salts and prodrugs thereof.
[0013] A number of embodiments are preferred within the above
formula. Among the preferred, but separate, embodiments, are those
wherein Z is N; wherein X is S; wherein Y is N(R.sup.5); wherein Z
is N, X is S and Y is N(R.sup.5); wherein W is aryl or heteroaryl;
wherein W is aryl or heteroaryl and aryl is phenyl or naphthyl;
wherein W is aryl or heteroaryl and heteroaryl is pyridyl or
quinolyl; wherein R.sup.1 and R.sup.2 are independently H or
(C.sub.1-C.sub.8)alkyl; wherein R.sup.1 and R.sup.2 combine to form
a fused 6-membered aryl or heteroaryl ring; wherein W is aryl or
heteroaryl, X is S, Y is N(R.sup.5), Z is N, and R.sup.1 and
R.sup.2 are independently H or (C.sub.1-C.sub.8)alkyl; wherein W is
aryl or heteroaryl, X is S, Y is N(R.sup.5), Z is N, and R.sup.1
and R.sup.2 combine to form a fused 6-membered aryl or heteroaryl
ring.
[0014] In one group of preferred embodiments, the compound is
represented by formula (Ia): 2
[0015] wherein W, Y, R.sup.1 and R.sup.2 have the meanings provided
above.
[0016] In another aspect, the present invention provides a
composition comprising a pharmaceutically acceptable carrier and a
compound of formula I as provided above, or a compound of formula I
wherein R.sup.1 is phenyl, when W is phenyl or unsubstituted
naphthyl, X is S, Y is NH, and Z is N; and R.sup.2 is H when W is
unsubstituted phenyl, X is S, Y is NH, Z is N and R.sup.1 is
(C.sub.1-C.sub.8)alkyl.
[0017] In still another aspect, the present invention provides a
method of treating a CCR4-mediated condition or disease, comprising
administering to a subject in need of such treatment, a
therapeutically effective amount of a composition as described
herein.
[0018] In preferred, but separate, embodiments of this aspect of
the invention the CCR4-mediated condition or disease is selected
from an allergic disease, psoriasis, atopic dermatitis and asthma;
the CCR4-mediated condition or disease is an allergic disease; the
CCR4-mediated condition or disease is an allergic disease and the
allergic disease is selected from systemic anaphylaxis,
hypersensitivity responses, drug allergies, insect sting allergies
and food allergies; the CCR4-mediated condition or disease is
psoriasis; the CCR4-mediated condition or disease is atopic
dermatitis; the CCR4-mediated condition or disease is asthma; the
CCR4-mediated condition or disease is an allergic condition and the
compound is used alone or in combination with at least one
therapeutic agent wherein the therapeutic agent is an
antihistamine; the CCR4-mediated condition or disease is psoriasis
and the compound is used alone or in combination with at least one
therapeutic agent selected from a corticosteroid, a lubricant, a
keratolytic agent, a vitamin D.sub.3 derivative, PUVA and
anthralin; the CCR4-mediated condition or disease is atopic
dermatitis and the compound is used alone or in combination with at
least one therapeutic agent selected from a lubricant and a
corticosteroid; the CCR4-mediated condition or disease is asthma
and the compound is used alone or in combination with at least one
therapeutic agent selected from a .beta.2-agonist and a
corticosteroid; the compound interferes with the interaction
between CCR4 and a ligand; the subject is a human.
[0019] In yet another aspect, the present invention provides a a
method of modulating CCR4 function in a cell, comprising contacting
the cell with a therapeutically effective amount of a compound of
formula I.
[0020] In still another aspect, the present invention provides a
method for modulating CCR4 function, comprising contacting a CCR4
protein with a therapeutically effective amount of a compound of
formula I.
[0021] In another aspect, the present invention provides
compositions and methods using compounds having the general
formula:
Ar.sup.1--X--Ar.sup.2 (II)
[0022] in which the symbols Ar.sup.1 and Ar.sup.2 each
independently represent a substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, or a substituted or
unsubstituted fused aryl-heterocyclic ring system. The letter X
represents a linking group selected from --N(R)--, --C(O)S--,
--CH.dbd.CHSO.sub.2-- and --SO.sub.2N(R)-- wherein R is H or a
substituted or unsubstituted (C.sub.1-C.sub.8)alkyl group. In this
formula, the linking group can be in either orientation relative to
the Ar.sup.1 and Ar.sup.2 moieties. For example, the above general
formula is meant to include both Ar.sup.1--SO.sub.2--NH--Ar.sup.2
and Ar.sup.1--NH--SO.sub.2--Ar.sup.2. These compounds are useful in
compositions and methods for the treatment of CCR4-mediated
conditions or diseases, particularly those recited above with
reference to compounds of formula I.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 is a graph showing the results of a CCR4/TARC
competition assay using compound 1.1.
[0024] FIG. 2 is a graph showing the results of a CCR4/TARC
competition assay using compound 1.2.
[0025] FIG. 3 is a graph showing the results of a CCR4/TARC
competition assay using compound 1.3.
[0026] FIG. 4 illustrates the results achieved with compound 1.2 in
CEM migration assays with CCR4 and TARC.
[0027] FIG. 5 illustrates the effect of compound 1.3 on CEM calcium
response.
[0028] FIG. 6 illustrates the effect of compound 1.2 on CEM calcium
response.
DESCRIPTION OF THE INVENTION
[0029] Abbreviations and Definitions
[0030] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain, or cyclic hydrocarbon radical, or combination thereof, which
may be fully saturated, mono- or polyunsaturated and can include
di- and multivalent radicals, having the number of carbon atoms
designated (i.e. C.sub.1-C.sub.10 means one to ten carbons).
Examples of saturated hydrocarbon radicals include groups such as
methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl,
sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl,
homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl,
n-octyl, and the like. An unsaturated alkyl group is one having one
or more double bonds or triple bonds. Examples of unsaturated alkyl
groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl,
2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1-
and 3-propynyl, 3-butynyl, and the higher homologs and isomers. The
term "alkyl," unless otherwise noted, is also meant to include
those derivatives of alkyl defined in more detail below as
"heteroalkyl." Alkyl groups which are limited to hydrocarbon groups
are termed "homoalkyl".
[0031] The term "alkylene" by itself or as part of another
substituent means a divalent radical derived from an alkane, as
exemplified by --CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, and further
includes those groups described below as "heteroalkylene."
Typically, an alkyl (or alkylene) group will have from 1 to 24
carbon atoms, with those groups having 10 or fewer carbon atoms
being preferred in the present invention. A "lower alkyl" or "lower
alkylene" is a shorter chain alkyl or alkylene group, generally
having eight or fewer carbon atoms.
[0032] The terms "alkoxy," "alkylamino" and "alkylthio" (or
thioalkoxy) are used in their conventional sense, and refer to
those alkyl groups attached to the remainder of the molecule via an
oxygen atom, an amino group, or a sulfur atom, respectively.
[0033] The term "heteroalkyl," by itself or in combination with
another term, means, unless otherwise stated, a stable straight or
branched chain, or cyclic hydrocarbon radical, or combinations
thereof, consisting of the stated number of carbon atoms and from
one to three heteroatoms selected from the group consisting of O,
N, Si and S, and wherein the nitrogen and sulfur atoms may
optionally be oxidized and the nitrogen heteroatom may optionally
be quaternized. The heteroatom(s) O, N and S may be placed at any
interior position of the heteroalkyl group. The heteroatom Si may
be placed at any position of the heteroalkyl group, including the
position at which the alkyl group is attached to the remainder of
the molecule. Examples include --CH.sub.2--CH.sub.2--O--CH.s- ub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3- )--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3, --CH.sub.2--CH.sub.2,
--S(O)--CH.sub.3, --CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH- .sub.3, and
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3. Up to two heteroatoms may be
consecutive, such as, for example, --CH.sub.2--NH--OCH.sub.3 and
--CH.sub.2--O--Si(CH.sub.3).sub.3. Similarly, the term
"heteroalkylene" by itself or as part of another substituent means
a divalent radical derived from heteroalkyl, as exemplified by
--CH.sub.2--CH.sub.2--S--CH.s- ub.2CH.sub.2-- and
--CH.sub.2--S--CH.sub.2--CH.sub.2--NH--CH.sub.2--. For
heteroalkylene groups, heteroatoms can also occupy either or both
of the chain termini (e.g., alkyleneoxy, alkylenedioxy,
alkyleneamino, alkylenediamino, and the like). Still further, for
alkylene and heteroalkylene linking groups, no orientation of the
linking group is implied.
[0034] The terms "cycloalkyl" and "heterocycloalkyl", by themselves
or in combination with other terms, represent, unless otherwise
stated, cyclic versions of "alkyl" and "heteroalkyl", respectively.
Additionally, for heterocycloalkyl, a heteroatom can occupy the
position at which the heterocycle is attached to the remainder of
the molecule. Examples of cycloalkyl include cyclopentyl,
cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the
like. Examples of heterocycloalkyl include
1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,
3-piperidinyl, 4-morpholinyl, 3-moorpholinyl, tetrahydrofuran-2-yl,
tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,
1-piperazinyl, 2-piperazinyl, and the like.
[0035] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "halo(C.sub.1-C.sub.4)alkyl" is mean to
include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,
3-bromopropyl, and the like.
[0036] The term "aryl" means, unless otherwise stated, a
polyunsaturated, typically aromatic, hydrocarbon substituent which
can be a single ring or multiple rings (up to three rings) which
are fused together or linked covalently. The term "heteroaryl"
refers to aryl groups (or rings) that contain from zero to four
heteroatoms selected from N, O, and S, wherein the nitrogen and
sulfur atoms are optionally oxidized, and the nitrogen atom(s) are
optionally quatemized. A heteroaryl group can be attached to the
remainder of the molecule through a heteroatom. Non-limiting
examples of aryl and heteroaryl groups include phenyl, 1-naphthyl,
2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl,
3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl,
4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,
2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl,
4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl,
2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of the above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below.
[0037] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both
aryl and heteroaryl rings as defined above. Thus, the term
"arylalkyl" is meant to include those radicals in which an aryl
group is attached to an alkyl group (e.g., benzyl, phenethyl,
pyridylmethyl and the like) including those alkyl groups in which a
carbon atom (e.g., a methylene group) has been replaced by, for
example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl,
3-(1-naphthyloxy)propyl, and the like).
[0038] Each of the above terms (e.g., "alkyl," "heteroalkyl,"
"aryl" and "heteroaryl") are meant to include both substituted and
unsubstituted forms of the indicated radical. Preferred
substituents for each type of radical are provided below.
[0039] Substituents for the alkyl and heteroalkyl radicals
(including those groups often referred to as alkylene, alkenyl,
heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be a
variety of groups selected from: --OR', .dbd.O, .dbd.NR',
.dbd.N--OR', --NR'R", --SR', -halogen, --SiR'R"R'", --OC(O)R',
--C(O)R', --CO.sub.2R', --CONR'R", --OC(O)NR'R", --NR"C(O)R',
--NR'--C(O)NR"R'", --NR"C(O).sub.2R', --NH--C(NH.sub.2).dbd.NH,
--NR'C(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR', --S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R", --CN and --NO.sub.2 in a number
ranging from zero to (2 m'+1), where m' is the total number of
carbon atoms in such radical. R', R" and R'" each independently
refer to hydrogen, unsubstituted (C.sub.1-C.sub.8)alkyl and
heteroalkyl, unsubstituted aryl, aryl substituted with 1-3
halogens, unsubstituted alkyl, alkoxy or thioalkoxy groups, or
aryl-(C.sub.1-C.sub.4)alkyl groups. When R' and R" are attached to
the same nitrogen atom, they can be combined with the nitrogen atom
to form a 5-, 6-, or 7-membered ring. For example, --NR'R" is meant
to include 1-pyrrolidinyl and 4-morpholinyl. From the above
discussion of substituents, one of skill in the art will understand
that the term "alkyl" is meant to include groups such as haloalkyl
(e.g., --CF.sub.3 and --CH.sub.2CF.sub.3) and acyl (e.g.,
--C(O)CH.sub.3, --C(O)CF.sub.3, --C(O)CH.sub.2OCH.sub.3, and the
like).
[0040] Similarly, substituents for the aryl and heteroaryl groups
are varied and are selected from: -halogen, --OR', --OC(O)R',
--NR'R", --SR', --R', --CN, --NO.sub.2, --CO.sub.2R', --CONR'R",
--C(O)R', --OC(O)NR'R", --NR"C(O)R', --NR"C(O).sub.2R', ,
--NR'--C(O)NR"R'", --NH--C(NH.sub.2).dbd.NH,
--NR'C(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR', --S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R", --N.sub.3, --CH(Ph).sub.2,
perfluoro(C.sub.1-C.sub.4)alkoxy, and
perfluoro(C.sub.1-C.sub.4)alkyl, in a number ranging from zero to
the total number of open valences on the aromatic ring system; and
where R', R" and R'" are independently selected from hydrogen,
(C.sub.1-C.sub.8)alkyl and heteroalkyl, unsubstituted aryl and
heteroaryl, (unsubstituted aryl)-(C.sub.1-C.sub.4)alkyl, and
(unsubstituted aryl)oxy-(C.sub.1-C.sub.4)alkyl.
[0041] Two of the substituents on adjacent atoms of the aryl or
heteroaryl ring may optionally be replaced with a substituent of
the formula --T--C(O)--(CH.sub.2).sub.q--U--, wherein T and U are
independently --NH--, --O--, --CH.sub.2-- or a single bond, and q
is an integer of from 0 to 2. Alternatively, two of the
substituents on adjacent atoms of the aryl or heteroaryl ring may
optionally be replaced with a substituent of the formula
--A--(CH.sub.2).sub.r--B--, wherein A and B are independently
--CH.sub.2--, --O--, --NH--, --S--, --S(O)--, --S(O).sub.2--,
--S(O).sub.2NR'-- or a single bond, and r is an integer of from 1
to 3. One of the single bonds of the new ring so formed may
optionally be replaced with a double bond. Alternatively, two of
the substituents on adjacent atoms of the aryl or heteroaryl ring
may optionally be replaced with a substituent of the formula
--(CH.sub.2).sub.s--X--(CH.sub.2).sub.t- --, where s and t are
independently integers of from 0 to 3, and X is --O--, --NR'--,
--S--, --S(O)--, --S(O).sub.2--, or --S(O).sub.2NR'--. The
substituent R' in --NR'-- and --S(O).sub.2NR'-- is selected from
hydrogen or unsubstituted (C.sub.1-C.sub.6) alkyl.
[0042] As used herein, the term "heteroatom" is meant to include
oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).
[0043] The term "pharmaceutically acceptable salts" is meant to
include salts of the active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituents found on the compounds described herein. When
compounds of the present invention contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired base, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable base addition salts include sodium,
potassium, calcium, ammonium, organic amino, or magnesium salt, or
a similar salt. When compounds of the present invention contain
relatively basic functionalities, acid addition salts can be
obtained by contacting the neutral form of such compounds with a
sufficient amount of the desired acid, either neat or in a suitable
inert solvent. Examples of pharmaceutically acceptable acid
addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,
phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, maleic, malonic, benzoic,
succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic,
p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
Also included are salts of amino acids such as arginate and the
like, and salts of organic acids like glucuronic or galactunoric
acids and the like (see, for example, Berge, S. M., et al,
"Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977,
66, 1-19). Certain specific compounds of the present invention
contain both basic and acidic functionalities that allow the
compounds to be converted into either base or acid addition
salts.
[0044] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents, but otherwise the
salts are equivalent to the parent form of the compound for the
purposes of the present invention.
[0045] In addition to salt forms, the present invention provides
compounds which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0046] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present invention and are intended to be within the scope of the
present invention.
[0047] Certain compounds of the present invention possess
asymmetric carbon atoms (optical centers) or double bonds; the
racemates, diastereomers, geometric isomers and individual isomers
are all intended to be encompassed within the scope of the present
invention.
[0048] The compounds of the present invention may also contain
unnatural proportions of atomic isotopes at one or more of the
atoms that constitute such compounds. For example, the compounds
may be radiolabeled with radioactive isotopes, such as for example
tritium (.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C).
All isotopic variations of the compounds of the present invention,
whether radioactive or not, are intended to be encompassed within
the scope of the present invention.
[0049] General
[0050] Chemokine receptors are attractive targets for the
development of antiinflammatory agents. Small molecule antagonists
of chemokine receptors, e.g., CC chemokine receptors, however, are
not widely known. U.S. Pat. No. 6,207,665 to Hesselgesser et al.
describes piperazine derivatives as CCR1 antagonists and is hereby
incorporated by reference.
[0051] The present invention is directed to compounds, compositions
and methods useful in the modulation of chemokine receptor
function, particularly CCR4 function. Accordingly, the compounds of
the present invention are compounds which inhibit at least one
function or characteristic of a mammalian CCR4 protein, for
example, a human CCR4 protein.
[0052] The full-length human CCR4 protein (GenBank Accession No.
X85740; SWISS-PROT Accession No. P51679) has been described, see,
e.g, Imai et al. (1998) J. Biol. Chem. 273:1764-1768, and has the
sequence shown in SEQ ID NO:1.
[0053] The ability of a compound to inhibit the function of CCR4,
can be demonstrated in a binding assay (e.g., ligand binding or
agonist binding), a signalling assay (e.g., activation of a
mammalian G protein, induction of rapid and transient increase in
the concentration of cytosolic free calcium), and/or cellular
response assay (e.g., stimulation of chemotaxis, exocytosis or
inflammatory mediator release by leukocytes).
[0054] In view of the above, the present invention is directed to
compounds, compositions and methods useful in the modulation of
chemokine receptor activity, particularly CCR4. Accordingly, the
compounds of the present invention are those which inhibit at least
one function or characteristic of a mammalian CCR4 protein, for
example, a human CCR4 protein. The ability of a compound to inhibit
such a function can be demonstrated in a binding assay (e.g.,
ligand binding or promotor binding), a signalling assay (e.g.,
activation of a mammalian G protein, induction of rapid and
transient increase in the concentration of cytosolic free calcium),
and/or cellular response function (e.g., stimulation of chemotaxis,
exocytosis or inflammatory mediator release by leukocytes).
[0055] Compounds that Modulate CCR4 Activity
[0056] CCR4 Antagonists
[0057] The present invention provides compounds having
antiinflammatory or anti-immunoregulatory activity. The compounds
of the invention are thought to interfere with inappropriate T-cell
trafficking by specifically modulating or inhibiting a chemokine
receptor function. Chemokine receptors are integral membrane
proteins which interact with an extracellular ligand, or chemokine,
and mediate a cellular response to the chemokine, e.g., chemotaxis,
increased intracellular calcium ion concentration. Therefore,
inhibition of a chemokine receptor function, e.g., interference
with a chemokine receptor-ligand interaction, will inhibit a
chemokine receptor-mediated response and treat or prevent a
chemokine receptor-mediated condition or disease.
[0058] Without intending to be bound by theory, it is believed that
the compounds provided herein interfere with the interaction
between a chemokine receptor and one or more cognate ligands. In
particular, it is believed that the compounds interfere with the
interaction between CCR4 and a CCR4 ligand, e.g., TARC, MDC, etc.
Compounds contemplated by the invention include, but are not
limited to, the exemplary compounds provided herein.
[0059] The compounds provided herein have the general formula (I):
3
[0060] wherein W is selected from aryl, heteroaryl,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, cycloalkyl and
heterocycloalkyl; X is selected from N(R.sup.5), S, O,
C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4), and optionally, when Z
is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)-(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; and Z is selected from
N and C(R.sup.8). R.sup.1 and R.sup.2 are independently selected
from H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl. When R' and R" are attached to
nitrogen, they may be combined with the nitrogen atom to form a 5-,
6-, or 7-membered ring. R.sup.3, R.sup.4 and R.sup.8 are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl; with the provisos that R.sup.1 is other than phenyl,
when W is phenyl or unsubstituted naphthyl, X is S, Y is NH, and Z
is N; and R.sup.2 is other than H when W is unsubstituted phenyl, X
is S, Y is NH, Z is N and R.sup.1 is (C.sub.1-C.sub.8)alkyl.
[0061] Embodiments represented by formula I can be appreciated by
replacing the ring system containing X and Z with an appropriate
scaffold, wherein the attachment points represent the attachment of
Y, R.sup.1 and R.sup.2 groups: 4
[0062] For example, the ring system or "scaffold" is meant to
include the following (including substituted versions thereof)
wherein the "A" ring is selected from the following embodiments:
5
[0063] In one group of preferred embodiments Z is N. In another
group, X is S. In still another group of preferred embodiments, Y
is N(R.sup.5). Particularly preferred are those embodiments that
combine each of these preferred groups. Accordingly, in one group
of particularly preferred embodiments X is S, Y is N(R.sup.5) and Z
is N.
[0064] In another group of preferred embodiments, W is selected
from aryl and heteroaryl. Particularly preferred are those
embodiments in which aryl is phenyl or naphthyl. Other particularly
preferred embodiments are those in which heteroaryl is pyridyl or
quinolyl. In separate, but preferred embodiments, R.sup.1 and
R.sup.2 are independently H or (C.sub.1-C.sub.8)alkyl. In still
other separate, but preferred embodiments, R.sup.1 and R.sup.2
combine to form a fused 6-membered aryl or heteroaryl ring. In each
of the above groups of preferred embodiments, R.sup.5 is most
preferably H.
[0065] In another group of preferred embodiments, the compounds of
the invention have the formula(I), wherein W is selected from
substituted naphthyl, pyridyl quinolyl, (C.sub.1-C.sub.8)alkyl,
heteroalkyl, cycloalkyl and heterocycloalkyl; X is selected from
N(R.sup.5), S, O, C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4), and
optionally, when Z is N, X can be C(R.sup.6)(R.sup.7); Y is
selected from a bond, N(R.sup.5),
N(R.sup.5)-(C.sub.1-C.sub.8)alkylene, 0, S and S(O).sub.n, wherein
the integer n is 1 or 2; and Z is selected from N and C(R.sup.8).
R.sup.1 and R.sup.2 are independently selected from H, halogen, CN,
CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl,
heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and optionally, R.sup.1
and R.sup.2 combine to form a 5- to 8-membered ring containing from
0 to 3 heteroatoms selected from N, O and S, wherein R' and R" are
independently selected from H, (C.sub.1-C.sub.8)alkyl and aryl.
When R' and R" are attached to nitrogen, they may be combined with
the nitrogen atom to form a 5-, 6-, or 7-membered ring. R.sup.3,
R.sup.4 and R.sup.8 are independently selected from H, halogen, CN,
OH, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
[0066] A number of embodiments are particularly preferred within
this group and are separately provided as those wherein Z is N;
wherein X is S; wherein Y is N(R.sup.5); wherein Z is N, X is S and
Y is N(R.sup.5); wherein W is substituted naphthyl; wherein W is
pyridyl or quinolyl; wherein R.sup.1 is other than phenyl; wherein
R.sup.1 and R.sup.2 are independently H or (C.sub.1-C.sub.8)alkyl;
wherein R.sup.1 and R.sup.2 combine to form a fused 6-membered aryl
or heteroaryl ring; wherein W is substituted naphthyl, X is S, Y is
N(R.sup.5), Z is N, and R.sup.1 and R.sup.2 are independently H or
(C.sub.1-C.sub.8)alkyl; wherein W is substituted naphthyl, X is S,
Y is N(R.sup.5), Z is N, and R.sup.1 and R.sup.2 combine to form a
fused 6-membered aryl or heteroaryl ring; wherein W is pyridyl or
quinolyl, X is S, Y is N(R.sup.5), Z is N, and R.sup.1 and R.sup.2
are independently H or (C.sub.1-C.sub.8)alkyl; and wherein W is
pyridyl or quinolyl, X is S, Y is N(R.sup.5), Z is N, and R.sup.1
and R.sup.2 combine to form a fused 6-membered aryl or heteroaryl
ring.
[0067] In a particularly preferred group of embodiments, the A ring
is a thiazole ring (see formula Ia). 6
[0068] In formula Ia, W, Y, R.sup.1 and R.sup.2 have the meanings
(and preferred groupings) provided above. Exemplary structures
within this preferred group of embodiments are shown below. 7
[0069] The compounds provided above are contemplated for use in
compositions and methods for the treatment of CCR4-mediated
conditions and diseases as described in more detail below. However,
the present invention further includes aspects directed to
pharmaceutical compositions and methods using compounds that are
known, particularly those that are commercially available.
Exemplary commercially available compounds include: 4-(1,1
-dimethylethyl)-N-phenyl-2-thiazolamine,
4-methyl-N-phenyl-2-thiazolamine,
4-(1-methylethyl)-N-phenyl-2-thiazolami- ne,
4-dodecyl-N-phenyl-2-thiazolamine, 2-anilino-4-isobutyl-thiazole,
and 2-anilino-4-methyl-thiazole picrate.
[0070] Still other commercially available compounds that are useful
in the composition and method aspects of this invention include:
8
[0071] Synthesis of CCR4 Antagonists
[0072] Synthesis routes to the compounds provided above are
described in the Examples. One of skill in the art will appreciate
that the substituents (e.g., R', R", R'", etc.) can be altered
before, during or after preparation of the heterocyclic scaffolding
and that suitable adjustments in the exemplary conditions (e.g.,
temperatures, solvents, etc.) can be made. Additionally, one of
skill in the art will recognize that protecting groups may be
necessary for the preparation of certain compounds and will be
aware of those conditions compatible with a selected protecting
group.
[0073] Compositions that Modulate CCR4 Activity
[0074] In another aspect, the present invention provides
compositions that modulate CCR4 activity.
[0075] Generally, the compositions will comprise a pharmaceutically
acceptable excipient and a compound having the formula provided
above as formula I: 9
[0076] wherein W is selected from aryl, heteroaryl,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, cycloalkyl and
heterocycloalkyl; X is selected from N(R.sup.5), S, O,
C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4), and optionally, when Z
is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a bond,
N(R.sup.5), N(R.sup.5)-(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; and Z is selected from
N and C(R.sup.8). R.sup.1 and R.sup.2 are independently selected
from H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl. When R' and R" are attached to
nitrogen, they may be combined with the nitrogen atom to form a 5-,
6-, or 7-membered ring. R.sup.3, R.sup.4 and R.sup.8 are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
[0077] Preferred compounds useful in the present compositions are
those of formula I in which W is selected from substituted
naphthyl, pyridyl quinolyl, (C.sub.1-C.sub.8)alkyl, heteroalkyl,
cycloalkyl and heterocycloalkyl; X is selected from N(R.sup.5), S,
O, C(R.sup.3).dbd.C(R.sup.4), N.dbd.C(R.sup.4), and optionally,
when Z is N, X can be C(R.sup.6)(R.sup.7); Y is selected from a
bond, N(R.sup.5), N(R.sup.5)-(C.sub.1-C.sub.8)alkylene, O, S and
S(O).sub.n, wherein the integer n is 1 or 2; and Z is selected from
N and C(R.sup.8). R.sup.1 and R.sup.2 are independently selected
from H, halogen, CN, CO.sub.2R', CONR'R", (C.sub.1-C.sub.8)alkyl,
heteroalkyl, aryl, heteroaryl, N(R.sup.6)(R.sup.7), OR.sup.9 and
optionally, R.sup.1 and R.sup.2 combine to form a 5- to 8-membered
ring containing from 0 to 3 heteroatoms selected from N, O and S,
wherein R' and R" are independently selected from H,
(C.sub.1-C.sub.8)alkyl and aryl. When R' and R" are attached to
nitrogen, they may be combined with the nitrogen atom to form a 5-,
6-, or 7-membered ring. R.sup.3, R.sup.4 and R.sup.8 are
independently selected from H, halogen, CN, OH,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl, heteroaryl,
O(C.sub.1-C.sub.8)alkyl, N(R.sup.6)(R.sup.7) and OR.sup.9; R.sup.5
is selected from H, (C.sub.1-C.sub.8) alkyl, heteroalkyl, aryl and
heteroaryl; R.sup.6 and R.sup.7 are independently selected from H,
(C.sub.1-C.sub.8)alkyl, heteroalkyl, aryl and heteroaryl; and
R.sup.9 is selected from (C.sub.1-C.sub.8)alkyl, heteroalkyl and
haloalkyl.
[0078] Additional compounds useful in the present invention are
those having the general formula:
Ar.sup.1--X--Ar.sup.2 (II)
[0079] In formula II, the symbols Ar.sup.1 and Ar.sup.2 each
independently represent a substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, or a substituted or
unsubstituted fused aryl-heterocyclic ring system. The letter X
represents a linking group selected from the group consisting of
--N(R)--, --C(O)S--, --CH.dbd.CHSO.sub.2-- and --SO.sub.2N(R)--
wherein R is H or a substituted or unsubstituted
(C.sub.1-C.sub.8)alkyl group. The linking group can be in either
orientation relative to the Ar.sup.1 and Ar.sup.2 moieties. For
example, the above general formula is meant to include both
Ar.sup.1--SO.sub.2--NH--Ar.sup.2 and
Ar.sup.1--NH--SO.sub.2--Ar.sup.2.
[0080] Returning to formula II above, in one group of preferred
embodiments, Ar.sup.1 is a substituted or unsubstituted heteroaryl
group. Preferably Ar.sup.1 is a substituted or unsubstituted
2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,
pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,
2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4
-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl,
2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, or 6-quinolyl group.
More preferably, Ar.sup.1 is a substituted or unsubstituted
thiazolyl, thienyl, benzoxadiazolyl or oxazolyl group.
[0081] In another group of preferred embodiments, Ar.sup.2 is a
substituted or unsubstituted aryl group. More preferably, Ar.sup.2
is a substituted or unsubstituted phenyl or naphthyl group. For
those embodiments in which Ar.sup.2 is a substituted aryl group,
there will preferably be from 1 to 4 substituents, independently
selected from halogen, hydroxy, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, (C.sub.1-C.sub.4)alkylthio,
(C.sub.1-C.sub.4)haloalkyl, (C.sub.1-C.sub.4)haloalkoxy, nitro,
cyano, (C.sub.1-C.sub.4)acyl, amino, (C.sub.1-C.sub.4)alkylamino,
and di(C.sub.1-C.sub.4)alkylamino. More preferably, there will be 1
to 3 substituents independently selected from halogen,
(C.sub.1-C.sub.4)haloalkyl, (C.sub.1-C.sub.4)haloalkoxy, nitro,
cyano and (C.sub.1-C.sub.4)acyl. In the most preferred embodiments,
Ar.sup.2 is a substituted or unsubstituted phenyl or naphthyl
group. The substituted phenyl and napthyl groups will preferably
have from 1 to 3 substituents independently selected from halogen,
(C.sub.1-C.sub.4)haloal- kyl, (C.sub.1-C.sub.4)haloalkoxy, nitro,
cyano and (C.sub.1-C.sub.4)acyl.
[0082] In another group of preferred embodiments, Ar.sup.1 and
Ar.sup.2 are each members independently selected from the group
consisting of: 10
[0083] Further preferred are those embodiments in which Ar.sup.1 is
selected from substituted or unsubstituted phenyl or 2-naphthyl and
Ar.sup.2 is selected from substituted or unsubstituted 11
[0084] In the most preferred embodiments, the compounds used in the
present compositions are selected from: 12
[0085] Preparation of CCR4 Modulators of Formula II
[0086] A number of compounds useful as modulators of CCR4
signalling can be obtained from commercial sources such as
Maybridge Chemical Co. and Aldrich Chemical Co. (Milwaukee, Wis.,
USA).
[0087] Other compounds can be prepared using conventional methods.
For example, Scheme 1 illustrates the preparation of
diarylsulfonamides (certain compounds of formula II). 13
[0088] In this scheme, an aryl (or heteroaryl) sulfonyl chloride i
can be combined with a suitable aryl (or heteroaryl) amine ii to
provide the target sulfonamides iii. The starting sulfonyl
chlorides are typically prepared in one step from a sulfonic acid
using a chlorinating agent such as POCl.sub.3 or SOCl.sub.2.
Similarly, starting arylamines ii are available from commercial
sources or can be prepared in one step from the corresponding
nitroaryl compounds.
[0089] In addition to the compounds provided above (of Formula I
and Formula II), compositions for modulating chemokine receptor
activity in humans and animals will typically contain a
pharmaceutical carrier or diluent.
[0090] "Modulation" of chemokine receptor activity, as used herein
in its various forms, is intended to encompass antagonism, agonism,
partial antagonism and/or partial agonism of the activity
associated with a particular chemokine receptor, preferably the
CCR4 receptor. The term "composition" as used herein is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly
or indirectly, from combination of the specified ingredients in the
specified amounts. By "pharmaceutically acceptable" it is meant the
carrier, diluent or excipient must be compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0091] The pharmaceutical compositions for the administration of
the compounds of this invention may conveniently be presented in
unit dosage form and may be prepared by any of the methods well
known in the art of pharmacy. All methods include the step of
bringing the active ingredient into association with the carrier
which constitutes one or more accessory ingredients. In general,
the pharmaceutical compositions are prepared by uniformly and
intimately bringing the active ingredient into association with a
liquid carrier or a finely divided solid carrier or both, and then,
if necessary, shaping the product into the desired formulation. In
the pharmaceutical composition the active object compound is
included in an amount sufficient to produce the desired effect upon
the process or condition of diseases.
[0092] The pharmaceutical compositions containing the active
ingredient may be in a form suitable for oral use, for example, as
tablets, troches, lozenges, aqueous or oily suspensions,
dispersible powders or granules, emulsions, hard or soft capsules,
or syrups or elixirs. Compositions intended for oral use may be
prepared according to any method known to the art for the
manufacture of pharmaceutical compositions. Such compositions may
contain one or more agents selected from sweetening agents,
flavoring agents, coloring agents and preserving agents in order to
provide pharmaceutically elegant and palatable preparations.
Tablets contain the active ingredient in admixture with other
non-toxic pharmaceutically acceptable excipients which are suitable
for the manufacture of tablets. These excipients may be for
example, inert diluents, such as calcium carbonate, sodium
carbonate, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example, corn starch, or
alginic acid; binding agents, for example starch, gelatin or
acacia, and lubricating agents, for example magnesium stearate,
stearic acid or talc. The tablets may be uncoated or they may be
coated by known techniques to delay disintegration and absorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate may be
employed. They may also be coated by the techniques described in
the U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form
osmotic therapeutic tablets for control release.
[0093] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin, or olive oil.
[0094] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxy-propylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents may be a naturally-occurring phosphatide, for
example lecithin, or condensation products of an alkylene oxide
with fatty acids, for example polyoxy-ethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or 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, for example polyethylene
sorbitan monooleate. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl,
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose or saccharin.
[0095] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil, for example 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, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0096] 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,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0097] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, for example olive oil or arachis oil, or a mineral
oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally-occurring gums, for example gum
acacia or gum tragacanth, naturally-occurring phosphatides, for
example soy bean, lecithin, and esters or partial esters derived
from fatty acids and hexitol anhydrides, for example sorbitan
monooleate, and condensation products of the said partial esters
with ethylene oxide, for example polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening and flavoring
agents.
[0098] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative and
flavoring and coloring agents.
[0099] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous or oleagenous suspension. This
suspension may be formulated according to the known art using those
suitable dispersing or wetting agents and suspending agents which
have been mentioned above. The sterile injectable preparation may
also be a sterile injectable solution or suspension in a non-toxic
parenterally-acceptable diluent or solvent, for example as a
solution in 1,3-butane diol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally 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 injectables.
[0100] The compounds of the present invention may also be
administered in the form of suppositories for rectal administration
of the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient which is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols.
[0101] For topical use, creams, ointments, jellies, solutions or
suspensions, etc., containing the compounds of the present
invention are employed. As used herein, topical application is also
meant to include the use of mouth washes and gargles.
[0102] The pharmaceutical composition and method of the present
invention may further comprise other therapeutically active
compounds as noted herein which are usually applied in the
treatment of the above mentioned pathological conditions.
[0103] Methods of Treating CCR4-Mediated Conditions or Diseases
[0104] In yet another aspect, the present invention provides
methods of treating or preventing a CCR4-mediated condition or
disease by administering to a subject having such a condition or
disease, a therapeutically effective amount of any compound of
formula I above (e.g., without provisos) or a compound of formula
II. Preferred compounds for use in the present methods are those
compounds provided above as preferred embodiments, as well as
compounds specifically exemplified in the Examples below, and
provided with specific structures herein. The "subject" is defined
herein to include animals such as mammals, including, but not
limited to, primates (e.g., humans), cows, sheep, goats, horses,
dogs, cats, rabbits, rats, mice and the like. In preferred
embodiments, the subject is a human.
[0105] As used herein, the phrase "CCR4-mediated condition or
disease" and related phrases and terms refer to a condition or
disease characterized by inappropriate, e.g., less than or greater
than normal, CCR4 functional activity. Inappropriate CCR4
functional activity might arise as the result of CCR4 expression in
cells which normally do not express CCR4, increased CCR4 expression
(leading to, e.g., inflammatory and immunoregulatory disorders and
diseases) or decreased CCR4 expression. Inappropriate CCR4
functional activity might also arise as the result of TARC and/or
MDC secretion by cells which normally do not secrete TARC and/or
MDC, increased TARC and/or MDC expression (leading to, e.g.,
inflammatory and immunoregulatory disorders and diseases) or
decreased TARC and/or MDC expression. A CCR4-mediated condition or
disease may be completely or partially mediated by inappropriate
CCR4 functional activity. However, a CCR4-mediated condition or
disease is one in which modulation of CCR4 results in some effect
on the underlying condition or disease (e.g., a CCR4 antagonist
results in some improvement in patient well-being in at least some
patients).
[0106] The term "therapeutically effective amount" means the amount
of the subject compound that will elicit the biological or medical
response of a tissue, system, animal or human that is being sought
by the researcher, veterinarian, medical doctor or other
clinician.
[0107] Diseases and conditions associated with inflammation,
infection and cancer can be treated or prevented with the present
compounds and compositions. In one group of embodiments, diseases
or conditions, including chronic diseases, of humans or other
species can be treated with inhibitors of CCR4 function. These
diseases or conditions include: (1) allergic diseases such as
systemic anaphylaxis or hypersensitivity responses, drug allergies,
insect sting allergies and food allergies, (2) inflammatory bowel
diseases, such as Crohn's disease, ulcerative colitis, ileitis and
enteritis, (3) vaginitis, (4) psoriasis and inflammatory dermatoses
such as dermatitis, eczema, atopic dermatitis, allergic contact
dermatitis, urticaria and pruritus, (5) vasculitis, (6)
spondyloarthropathies, (7) scleroderma, (8) asthma and respiratory
allergic diseases such as allergic asthma, allergic rhinitis,
hypersensitivity lung diseases and the like, (9) autoimmune
diseases, such as arthritis (including rheumatoid and psoriatic),
multiple sclerosis, systemic lupus erythematosus, type I diabetes,
glomerulonephritis, and the like, (10) graft rejection (including
allograft rejection and graft-v-host disease), and (11) other
diseases in which undesired inflammatory responses are to be
inhibited, such as atherosclerosis, myositis, neurodegenerative
diseases (e.g., Alzheimer's disease), encephalitis, meningitis,
hepatitis, nephritis, sepsis, sarcoidosis, allergic conjunctivitis,
otitis, chronic obstructive pulmonary disease, sinusitis, Behcet's
syndrome and gout.
[0108] In another group of embodiments, diseases or conditions can
be treated with agonists of CCR4 function. Examples of diseases to
be treated with CCR4 agonists include cancers, diseases in which
angiogenesis or neovascularization play a role (neoplastic
diseases, retinopathy and macular degeneration), infectious
diseases (viral infections, e.g., HIV infection, and bacterial
infections) and immunosuppressive diseases such as organ transplant
conditions and skin transplant conditions. The term "organ
transplant conditions" is meant to include bone marrow transplant
conditions and solid organ (e.g., kidney, liver, lung, heart,
pancreas or combination thereof) transplant conditions.
[0109] Preferably, the present methods are directed to the
treatment of diseases or conditions selected from allergic
diseases, psoriasis, atopic dermatitis and asthma.
[0110] Depending on the disease to be treated and the subject's
condition, the compounds of the present invention may be
administered by oral, parenteral (e.g., intramuscular,
intraperitoneal, intravenous, ICV, intracisternal injection or
infusion, subcutaneous injection, or implant), inhalation, nasal,
vaginal, rectal, sublingual, or topical routes of administration
and may be formulated, alone or together, in suitable dosage unit
formulations containing conventional non-toxic pharmaceutically
acceptable carriers, adjuvants and vehicles appropriate for each
route of administration. The present invention also contemplates
administration of the compounds of the present invention in a depot
formulation.
[0111] In the treatment or prevention of conditions which require
chemokine receptor modulation an appropriate dosage level will
generally be about 0.001 to 100 mg per kg patient body weight per
day which can be administered in single or multiple doses.
Preferably, the dosage level will be about 0.01 to about 25 mg/kg
per day; more preferably about 0.05 to about 10 mg/kg per day. A
suitable dosage level may be about 0.01 to 25 mg/kg per day, about
0.05 to 10 mg/kg per day, or about 0.1 to 5 mg/kg per day. Within
this range the dosage may be 0.005 to 0.05, 0.05 to 0.5 or 0.5 to
5.0 mg/kg per day. For oral administration, the compositions are
preferably provided in the form of tablets containing 1.0 to 1000
milligrams of the active ingredient, particularly 1.0, 5.0, 10.0,
15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,
400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of
the active ingredient for the symptomatic adjustment of the dosage
to the patient to be treated. The compounds may be administered on
a regimen of 1 to 4 times per day, preferably once or twice per
day.
[0112] It will be understood, however, that the specific dose level
and frequency of dosage for any particular patient may be varied
and will depend upon a variety of factors including the activity of
the specific compound employed, the metabolic stability and length
of action of that compound, the age, body weight, general health,
sex, diet, mode and time of administration, rate of excretion, drug
combination, the severity of the particular condition, and the host
undergoing therapy.
[0113] The compounds of the present invention can be combined with
other compounds having related utilities to prevent and treat
inflammatory conditions and diseases, including allergic diseases,
psoriasis, atopic dermatitis and asthma, and those pathologies
noted above.
[0114] For example, in the treatment or prevention of inflammation,
the present compounds may be used in conjunction with an
antiinflammatory or analgesic agent such as an opiate agonist, a
lipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, a
cyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, an
interleukin inhibitor, such as an interleukin-1 inhibitor, an NMDA
antagonist, an inhibitor of nitric oxide or an inhibitor of the
synthesis of nitric oxide, a non-steroidal antiinflammatory agent,
or a cytokine-suppressing antiinflammatory agent, for example with
a compound such as acetaminophen, aspirin, codeine, fentanyl,
ibuprofen, indomethacin, ketorolac, morphine, naproxen, phenacetin,
piroxicam, a steroidal analgesic, sufentanyl, sunlindac, tenidap,
and the like. Similarly, the instant compounds may be administered
with an analgesic listed above; a potentiator such as caffeine, an
H2-antagonist (e.g., ranitidine), simethicone, aluminum or
magnesium hydroxide; a decongestant such as phenylephrine,
phenylpropanolamine, pseudophedrine, oxymetazoline, ephinephrine,
naphazoline, xylometazoline, propylhexedrine, or
levo-desoxy-ephedrine; an antiitussive such as codeine,
hydrocodone, caramiphen, carbetapentane, or dextromethorphan; a
diuretic; and a sedating or non-sedating antihistamine.
[0115] Likewise, compounds and compositions of the present
invention may be used in combination with other drugs that are used
in the treatment, prevention, suppression or amelioration of the
diseases or conditions for which compounds of the present invention
are useful. Such other drugs may be administered, by a route and in
an amount commonly used therefor, contemporaneously or sequentially
with a compound of the present invention. When a compound of the
present invention is used contemporaneously with one or more other
drugs, a pharmaceutical composition containing such other drugs in
addition to the compound of the present invention is preferred.
Accordingly, the pharmaceutical compositions of the present
invention include those that also contain one or more other active
ingredients or therapeutic agents, in addition to a compound of the
present invention. Examples of other therapeutic agents that may be
combined with a compound of the present invention, either
administered separately or in the same pharmaceutical compositions,
include, but are not limited to: (a) VLA-4 antagonists, (b)
corticosteroids, such as beclomethasone, methylprednisolone,
betamethasone, prednisone, prenisolone, dexamethasone, fluticasone,
hydrocortisone, budesonide, triamcinolone, salmeterol, salmeterol,
salbutamol, formeterol; (c) immunosuppressants such as cyclosporine
(cyclosporine A, Sandimmune.RTM., Neoral.RTM.), tacrolimus (FK-506,
Prograf.RTM.), rapamycin (sirolimus, Rapamune.RTM.) and other
FK-506 type immunosuppressants, and mycophenolate, e.g.,
mycophenolate mofetil (CellCept.RTM.); (d) antihistamines
(H1-histamine antagonists) such as bromopheniramine,
chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine,
diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine,
methdilazine, promethazine, trimeprazine, azatadine,
cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,
terfenadine, loratadine, cetirizine, fexofenadine,
descarboethoxyloratadine, and the like; (e) non-steroidal
anti-asthmatics such as .beta.2-agonists (e.g., terbutaline,
metaproterenol, fenoterol, isoetharine, albuterol, bitolterol and
pirbuterol), theophylline, cromolyn sodium, atropine, ipratropium
bromide, leukotriene antagonists (e.g., zafirlukast, montelukast,
pranlukast, iralukast, pobilukast and SKB-106,203), leukotriene
biosynthesis inhibitors (zileuton, BAY-1005); (f) non-steroidal
antiinflammatory agents (NSAIDs) such as propionic acid derivatives
(e.g., alminoprofen, benoxaprofen, bucloxic acid, carprofen,
fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,
indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen,
pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic
acid derivatives (e.g., indomethacin, acemetacin, alclofenac,
clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac,
furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac,
tolmetin, zidometacin and zomepirac), fenamic acid derivatives
(e.g., flufenamic acid, meclofenamic acid, mefenamic acid, niflumic
acid and tolfenamic acid), biphenylcarboxylic acid derivatives
(e.g., diflunisal and flufenisal), oxicams (e.g., isoxicam,
piroxicam, sudoxicam and tenoxican), salicylates (e.g., acetyl
salicylic acid and sulfasalazine) and the pyrazolones (e.g.,
apazone, bezpiperylon, feprazone, mofebutazone, oxyphenbutazone and
phenylbutazone); (g) cyclooxygenase-2 (COX-2) inhibitors such as
celecoxib (Celebrex.RTM.) and rofecoxib (Vioxx.RTM.); (h)
inhibitors of phosphodiesterase type IV (PDE-IV); (i) gold
compounds such as auranofin and aurothioglucose, (j) etanercept
(Enbrel.RTM.), (k) antibody therapies such as orthoclone (OKT3),
daclizumab (Zenapax.RTM.), basiliximab (Simulect.RTM.) and
infliximab (Remicade.RTM.), (l) other antagonists of the chemokine
receptors, especially CCR1, CCR2, CCR3, CCR5, CCR6, CCR8 and CCR10;
(m) lubricants or emollients such as petrolatum and lanolin, (n)
keratolytic agents (e.g., tazarotene), (o) vitamin D.sub.3
derivatives, e.g., calcipotriene or calcipotriol (Dovonex.RTM.),
(p) PUVA, (q) anthralin (Drithrocreme.RTM.), (r) etretinate
(Tegison.RTM.) and isotretinoin and (s) multiple sclerosis
therapeutic agents such as interferon .beta.-1.beta.
(Betaseron.RTM.), interferon .beta.-1.alpha. (Avonex.RTM.),
azathioprine (Imurek.RTM., Imuran.RTM.), glatiramer acetate
(Capoxone.RTM.), a glucocorticoid (e.g., prednisolone) and
cyclophosphamide (t) other compounds such as 5-aminosalicylic acid
and prodrugs thereof; hydroxychloroquine; D-penicillamine;
antimetabolites such as azathioprine, 6-mercaptopurine and
methotrexate; DNA synthesis inhibitors such as hydroxyurea and
microtubule disrupters such as colchicine. The weight ratio of the
compound of the present invention to the second active ingredient
may be varied and will depend upon the effective dose of each
ingredient. Generally, an effective dose of each will be used.
Thus, for example, when a compound of the present invention is
combined with an NSAID the weight ratio of the compound of the
present invention to the NSAID will generally range from about
1000:1 to about 1:1000, preferably about 200:1 to about 1:200.
Combinations of a compound of the present invention and other
active ingredients will generally also be within the aforementioned
range, but in each case, an effective dose of each active
ingredient should be used.
[0116] In still other particularly preferred embodiments, the
present methods are directed to the treatment of allergic diseases,
wherein the compound of the invention is administered either alone
or in combination with a second therapeutic agent, wherein said
second therapeutic agent is an antihistamine. When used in
combination, the practitioner can administer a combination of the
therapeutic agents, or administration can be sequential.
[0117] In yet other particularly preferred embodiments, the present
methods are directed to the treatment of psoriasis wherein the
compound of the invention is used alone or in combination with a
second therapeutic agent selected from a corticosteroid, a
lubricant, a keratolytic agent, a vitamin D.sub.3 derivative, PUVA
and anthralin.
[0118] In particularly preferred embodiments, the present methods
are directed to the treatment of atopic dermatitis using a compound
of the invention either alone or in combination with a second
therapeutic agent selected from a lubricant and a
corticosteroid.
[0119] In particularly preferred embodiments, the present methods
are directed to the treatment of asthma using a compound of the
invention either alone or in combination with a second therapeutic
agent selected from a .beta.2-agonist and a corticosteroid.
[0120] Method of Evaluating Putative CCR4 Modulators
[0121] In yet another aspect, the present invention includes
methods to evaluate putative specific agonists or antagonists of
CCR4 function. Accordingly, the present invention is directed to
the use of these compounds in the preparation and execution of
screening assays for compounds which modulate the function of the
CCR4 chemokine receptor. For example, the compounds of this
invention are useful for isolating receptor mutants, which are
excellent screening tools for potent compounds. Furthermore, the
compounds of this invention are useful in establishing or
determining the binding site of other compounds to the CCR4
chemokine receptor, e.g., by competitive inhibition. The compounds
of the instant invention are also useful for the evaluation of
putative specific modulators of the CCR4 chemokine receptor,
relative to other chemokine receptors including CCR1, CCR2, CCR2A,
CCR2B, CCR3, CCR5, CCR6, CCR8, CCR10, CXCR3 and CXCR4. One of skill
in the art will appreciate that thorough evaluation of specific
agonists and antagonists of the above chemokine receptors has been
hampered by the lack of availability of non-peptidyl (metabolically
resistant) compounds with high binding affinity for these
receptors. The compounds provided herein are particularly useful in
this context.
[0122] Combinatorial libraries of putative CCR4 agonists or
antagonists can be screened for pharmacological activity in in
vitro or in vivo assays. Conventionally, new chemical entities with
useful properties are generated by identifying a chemical compound
(called a "lead compound") with some desirable property or
activity, e.g., CCR4 chemokine receptor modulation activity,
creating variants of the lead compound, and evaluating the
properties and activities of those variant compounds. However, the
current trend is to shorten the time scale for all aspects of drug
discovery. Because of the ability to test large numbers of
compounds quickly and efficiently, high throughput screening (HTS)
methods are replacing conventional lead compound identification
methods.
[0123] In one preferred embodiment, high throughput screening
methods involve providing a library containing a large number of
potential therapeutic compounds (candidate compounds). Such
"combinatorial chemical libraries" are then screened in one or more
assays to identify those library members (particular chemical
species or subclasses) that display a desired characteristic
activity. The compounds thus identified can serve as conventional
"lead compounds" or can themselves be used as potential or actual
therapeutics.
[0124] A combinatorial chemical library is a collection of diverse
chemical compounds generated by either chemical synthesis or
biological synthesis by combining a number of chemical "building
blocks", such as reagents. For example, a linear combinatorial
chemical library, such as a polypeptide (e.g., mutein) library, is
formed by combining a set of chemical building blocks called amino
acids in every possible way for a given compound length (i.e., the
number of amino acids in a polypeptide compound). Millions of
chemical compounds can be synthesized through such combinatorial
mixing of chemical building blocks (Gallop et. al. (1994) J. Med.
Chem. 37(9):1233-1251).
[0125] Preparation and screening of combinatorial chemical
libraries is well known to those of skill in the art. Such
combinatorial chemical libraries include, but are not limited to,
peptide libraries (see, e.g., U.S. Pat. No. 5,010,175, Furka (1991)
Int. J. Pept. Prot. Res. 37:487-493, Houghton et. al. (1991) Nature
354: 84-88), peptoid libraries (PCT Publication No WO 91/19735),
encoded peptide libraries (PCT Publication WO 93/20242), random
bio-oligomer libraries (PCT Publication WO 92/00091),
benzodiazepine libraries (U.S. Pat. No. 5,288,514), libraries of
diversomers, such as hydantoins, benzodiazepines and dipeptides
(Hobbs et. al. (1993) Proc. Nat. Acad. Sci. USA 90:6909-6913),
vinylogous polypeptide libraries (Hagihara et al. (1992) J. Amer.
Chem. Soc. 114:6568), libraries of nonpeptidyl peptidomimetics with
a Beta-D-Glucose scaffolding (Hirschmann et al (1992) J. Amer.
Chem. Soc. 114:9217-9218), analogous organic syntheses of small
compound libraries (Chen et. al. (1994) J. Am. Chem. Soc.
116:2661), oligocarbamate libraries (Cho et al. (1993) Science
261:1303) and/or peptidyl phosphonate libraries (Campbell et al.
(1994) J. Org. Chem. 59:658). See, generally, Gordon et al. (1994)
J. Med. Chem. 37:1385-1401, nucleic acid libraries (see, e.g.,
Stratagene Corp.), peptide nucleic acid libraries (see, e.g., U.S.
Pat. No. 5,539,083), antibody libraries (see, e.g., Vaughn et. al.
(1996) Nature Biotechnology 14(3):309-314), and PCT/US96/10287),
carbohydrate libraries (see, e.g., Liang et al. (1996) Science
274:1520-1522, and U.S. Pat. No. 5,593,853), and small organic
molecule libraries (see, e.g., benzodiazepines, Baum (1993)
C&EN January 18, page 33 and U.S. Pat. No. 5,288,514;
isoprenoids, U.S. Pat. No. 5,549,974; pyrrolidines, U.S. Pat. Nos.
5,525,735 and 5,519,134; morpholino compounds, U.S. Pat. No.
5,506,337; and the like).
[0126] Devices for the preparation of combinatorial libraries are
commercially available (see, e.g., 357 MPS, 390 MPS, Advanced Chem
Tech, Louisville Ky.; Symphony, Rainin, Woburn Mass.; 433A Applied
Biosystems, Foster City Calif.; 9050 Plus, Millipore, Bedford,
Mass.).
[0127] A number of well known robotic systems have also been
developed for solution phase chemistries. These systems includes
automated workstations like the automated synthesis apparatus
developed by Takeda Chemical Industries, LTD. (Osaka, Japan) and
many robotic systems utilizing robotic arms (Zymate II, Zymark
Corporation, Hopkinton Mass.; Orca, Hewlett-Packard, Palo Alto
Calif.), which mimic the manual synthetic operations performed by a
chemist. Any of the above devices are suitable for use with the
present invention. The nature and implementation of modifications
to these devices (if any) so that they can operate as discussed
herein will be apparent to persons skilled in the relevant art. In
addition, numerous combinatorial libraries are themselves
commercially available (see e.g., ComGenex, Princeton N.J.; Asinex,
Moscow, Russia; Tripos, Inc., St. Louis Mo.; ChemStar, Ltd, Moscow,
Russia; 3D Pharmaceuticals, Exton Pa.; Martek Biosciences, Columbia
Md.; etc.).
[0128] High throughput assays for the presence, absence,
quantification, or other properties of particular compounds may be
used to test a combinatorial library that contains a large number
of potential therapeutic compounds (potential modulator compounds).
The assays are typically designed to screen large chemical
libraries by automating the assay steps and providing compounds
from any convenient source to the assays, which are typically run
in parallel (e.g., in microtiter formats on microtiter plates in
robotic assays). Preferred assays detect enhancement or inhibition
of CCR4 receptor function.
[0129] High throughput screening systems are commercially available
(see e.g., Zymark Corp., Hopkinton Mass.; Air Technical Industries,
Mentor Ohio; Beckman Instruments, Inc., Fullerton Calif.; Precision
Systems, Inc., Natick Mass.; etc.). These systems typically
automate entire procedures, including all sample and reagent
pipetting, liquid dispensing, timed incubations, and final readings
of the microplate in detector(s) appropriate for the assay. These
configurable systems provide high throughput and rapid start-up as
well as a high degree of flexibility and customization. The
manufacturers of such systems provide detailed protocols for
various high throughput systems. Thus, for example, Zymark Corp.
provides technical bulletins describing screening systems for
detecting the modulation of gene transcription, ligand binding, and
the like.
[0130] Other Evaluation Assays for CCR4 Modulators
[0131] A variety of assays can be used to evaluate the compounds
provided herein, including CCR4 binding assays, CCR4 signalling
assays, chemotaxis assays, and other assays of cellular
response.
[0132] In a suitable assay, a CCR4 protein (whether isolated or
recombinant) is used which has at least one property, activity or
functional charateristic of a mammalian CCR4 protein. The property
can be a binding property (to, for example, a ligand or inhibitor),
a signalling activity (e.g., activation of a mammalian G protein,
induction of rapid and transient increase in the concentration of
cytosolic free calcium [Ca.sup.++].sub.i), cellular response
function (e.g., stimulation of chemotaxis or inflammatory mediator
release by leukocytes), and the like.
[0133] In one embodiment, a composition containing a CCR4 protein
or variant thereof is maintained under conditions suitable for
binding. The CCR4 receptor is contacted with a putative agent (or a
second composition containing at least one putative agent) to be
tested, and binding is detected or measured.
[0134] In one group of preferred embodiments, the assay is a
cell-based assay and cells are used which are stably or transiently
transfected with a vector or expression cassette having a nucleic
acid sequence which encodes the CCR4 receptor. The cells are
maintained under conditions appropriate for expression of the
receptor and are contacted with a putative agent under conditions
appropriate for binding to occur. Binding can be detected using
standard techniques. For example, the extent of binding can be
determined relative to a suitable control (for example, relative to
background in the absence of a putative agent, or relative to a
known ligand). Optionally, a cellular fraction, such as a membrane
fraction, containing the receptor can be used in lieu of whole
cells.
[0135] Detection of binding or complex formation can be detected
directly or indirectly. For example, the putative agent can be
labeled with a suitable label (e.g., fluorescent label,
chemiluminescent label, isotope label, enzyme label, and the like)
and binding can be determined by detection of the label. Specific
and/or competitive binding can be assessed by competition or
displacement studies, using unlabeled agent or a ligand (e.g., TARC
or MDC) as a competitor.
[0136] In other embodiments, binding inhibition assays can be used
to evaluate the present compounds. In these assays, the compounds
are evaluated as inhibitors of ligand binding using, for example,
TARC or MDC. In this embodiment, the CCR4 receptor is contacted
with a ligand such as TARC or MDC and a measure of ligand binding
is made. The receptor is then contacted with a test agent in the
presence of a ligand (e.g., TARC or MDC) and a second measurement
of binding is made. A reduction in the extent of ligand binding is
indicative of inhibition of binding by the test agent. The binding
inhibition assays can be carried out using whole cells which
express CCR4, or a membrane fraction from cells which express
CCR4.
[0137] The binding of a G protein-coupled receptor by, for example,
an agonist, can result in a signalling event by the receptor.
Accordingly, signalling assays can also be used to evaluate the
compounds of the present invention and induction of signalling
function by an agent can be conitored using any suitable method.
For example, G protein activity, such as hydrolysis of GTP to GDP,
or later signalling events triggered by receptor binding can be
assayed by known methods (see, for example, PCT/US97/15915; Neote,
et al., Cell, 72:415-425 (1993); Van Riper, et al., J. Exp. Med.,
177:851-856 (1993) and Dahinden, et al, J. Exp. Med., 179:751-756
(1994)).
[0138] Chemotaxis assays can also be used to assess receptor
function and evaluate the compounds provided herein. These assays
are based on the functional migration of cells in vitro or in vivo
induced by an agent, and can be used to assess the binding and/or
effect on chemotaxis of ligands, inhibitors, or agonists. Suitable
assays are described in PCT/US97/15915; Springer, et al., WO
94/20142; Berman et al., Immunol. Invest., 17:625-677 (1988); and
Kavanaugh et al., J. Immunol., 146:4149-4156 (1991)).
[0139] The compounds provided herein can also be evaluated using
models of inflammation to assess the ability of the compound to
exert an effect in vivo. Suitable models are described as follows:
a sheep model for asthma (see, Weg, et al., J. Exp. Med., 177:561
(1993)); and a rat delayed-type hypersensitivity model (see Rand,
et al., Am. J. Pathol., 148:855-864 (1996)). Another useful model
for evaluating the instant compounds is the experimental autoimmune
encephalomyelitis (EAE) model for multiple sclerosis, which probes
chemokine receptor expression and function (see, Ransohoff, et al.,
Cytokine Growth Factor Rev., 7:35-46 (1996), and Karpus, et al., J.
Immunol. 161:2667-2671 (1998)).
[0140] In addition, leukocyte infiltration assays can also be used
to evaluate a compound (see, Van Damme, et al., J. Exp. Med.
176:59-65 (1992); Zachariae, et al., J. Exp. Med. 171:2177-2182
(1990); and Jose, et al., J. Exp. Med. 179:881-887 (1994)).
[0141] The following examples are offered to illustrate, but not to
limit the claimed invention.
EXAMPLES
[0142] Reagents and solvents used below can be obtained from
commercial sources such as Aldrich Chemical Co. (Milwaukee, Wis.,
USA). .sup.1H-NMR spectra were recorded on a Varian Gemini 400 MHz
NMR spectrometer. Significant peaks are tabulated in the order:
number of protons, multiplicity (s, singlet; d, doublet; t,
triplet; q, quartet; m, multiplet; br s, broad singlet) and
coupling constant(s) in Hertz (Hz). Electron Ionization (El) mass
spectra were recorded on a Hewlett Packard 5989A mass spectrometer.
Mass spectrometry results are reported as the ratio of mass over
charge, followed by the relative abundance of each ion (in
parentheses). In tables, a single m/e value is reported for the M+H
(or, as noted, M-H) ion containing the most common atomic isotopes.
Isotope patterns correspond to the expected formula in all cases.
Electrospray ionization (ESI) mass spectrometry analysis was
conducted on a Hewlett Packard 1100 MSD electrospray mass
spectrometer using the HP1 100 HPLC for sample delivery. Normally
the analyte was dissolved in methanol at 0.1 mg/mL and 1 microliter
was infused with the delivery solvent into the mass spectrometer,
which scanned from 100 to 1500 daltons. All compounds could be
analyzed in the positive ESI mode, using 1:1 acetonitrile/water
with 1% acetic acid as the delivery solvent. The compounds provided
below could also be analyzed in the negative ESI mode, using 2 mM
NH.sub.4OAc in acetonitrile/water as delivery solvent.
General Procedures
[0143] 14
General Procedures
[0144] 15
[0145] General Procedure for Preparing Aryl Thioureas
[0146] To a solution of aryl amine (4.1 mmol) in dry acetone (30
mL) at room temperature was added dropwise benzoylisothiocyanate
(0.56 mL, 4.5 mmol). After 1 h, the reaction mixture was
concentrated in vacuo to afford a solid compound, which was
triturated with 50% aqueous ethanol (50 mL) to dissolve inorganic
materials. The crude product was stirred with 20 mL of 10% NaOH at
98.degree. C. for 10 min. After cooling, the basic solution was
neutralized by adding 10% HCI solution. The precipitate was
filtered and then washed with water.
[0147] General Procedure for Preparing 2-Amino-4-Alkyl-Substituted
Thiazoles 16
[0148] Procedure 1
[0149] To a solution of N-arylthiourea or N-benylthiourea (1 mmol)
in methanol (5 mL) at room temperature was added .alpha.-haloketone
(1.05 mmol). The reaction mixture was refluxed for 3 h and then
concentrated in vacuo. The crude compound was then recrystalized
with ethyl ether and methylene chloride to afford 2-aminothiazole
as the salt form.
[0150] Procedure 2
[0151] To a boiling suspension of N-arylthiourea (1.0 mmol) and
MgSO.sub.4 (60 mg) in acetone (6.0 mL) was added a solution of
.alpha.-haloketone (1.1 mmol) in acetone (1.0 mL) dropwise. The
reaction mixture was refluxed for 2 to 8 h. The mixture was then
cooled to room temperature, poured into a saturated solution of
brine (5 mL). The solution was then basified with concentrated
NH.sub.4OH aqueous solution, diluted with water (5 mL) and then
extracted with EtOAc (2.times.20 mL). The organic layers were
combined, dried over anhydrous MgSO.sub.4 and concentrated in
vacuo. The crude product was purified by silica gel flash column
chromatography (ethyl acetate and hexanes).
[0152] In some cases, the desired thiazole products were
precipitated from reaction solution. The isolation was then carried
out by filtration through a Buckner funnel and washed the solid
with acetone. The final products were isolated as the salt
forms.
Example 1
[0153] 17
[0154] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 7.4-7.5 (m, 1H),
7.3-7.4 (m, 2H), 7.2-7.3 (m, 1H), 6.17 (s, 1H), 4.04 (m, 4H), 2.96
(m, 4H), 1.47 (s, 9H); MS (ES+): 318.2 (M+H).
Example 2
[0155] 18
[0156] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 11.2(s, br, 1H),
7.1-7.2 (m, 3H), 6.04 (s, 1H), 2.35 (s, 3H), 1.57 (s, 3H), 1.44 (s,
9H). MS (ES+): 261.1 (M+H).
Example 3
[0157] 19
[0158] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 10.1 (s, br, 1H),
7.3-7.4 (m, 5H), 6.01 (s, 1H), 4.51 (s, 2H), 1.39 (s, 9H). MS
(ES+): 247.1 (M+H).
Example 4
[0159] 20
[0160] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 12.0 (s, br, 1H),
8.3-8.4 (m, 2H), 7.7-7.8 (m, 2H), 7.63 (d, J=8.1 Hz, 1H), 7.53 (d,
J=8.1 Hz, 1H), 6.13 (s, 1H), 1.44 (s, 9H). MS (ES+): 317.1
(M+H).
Example 5
[0161] 21
[0162] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 7.1-7.2 (m, 3H),
6.14 (s, 1H), 2.82 (t, J=6 Hz, 2H), 2.77 (t, J=6 Hz, 2H), 1.8-1.9
(m, 4H), 1.41 (s, 9H). MS (ES+): 287.1 (M+H).
Example 6
[0163] 22
[0164] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 11.46 (s, br, 1H),
7.60 (d, J=8 Hz, 1H), 7.36 (d, J=8 Hz, 1H), 7.17 (t, J=8 Hz, 1H),
6.11 (s, 1H), 2.54 (s, 3H), 1.44 (s, 9H). MS (ES+): 325.1
(M+H).
Example 7
[0165] 23
[0166] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 9.34 (s, br, 1H),
9.23 (d, J=8.5 Hz, 1H), 8.49 (d, J=8.1 Hz, 1H), 7.9-8.1 (m, 3H),
6.23 (s, 1H), 1.46 (s, 9H). MS (ES+): 284.1 (M+H).
Example 8
[0167] 24
[0168] To a solution of 4,4-dimethyl-2-pentanone (0.25 mL, 1.7
mmol) in methanol (10 mL) was added bromine (0.09 mL, 1.7 mmol) in
methanol (3 mL) dropwise at 0.degree. C. The reaction mixture was
warmed up from 0.degree. C. to room temperature, and kept at room
temperature for 1 h. 1-Naphthylthiourea (0.35 g, 1.7 mmol) was then
added to the reaction mixture and refluxed for 3 h. The reaction
mixture was concentrated in vacuo. The crude compound was
recrystalized with ethyl ether and methylene chloride to afford
2-aminothiazole as the HBr salt. .sup.1H-NMR (400 MHz,
CDCl.sub.3):.delta. 8.12 (d, J=8.1 Hz, 1H), 7.8-8.0 (m, 2H),
7.1-7.7 (m, 4H), 6.04 (s, 1H), 2.76 (s, 2H), 1.44 (s, 9H). MS
(ES+): 297.2 (M+H).
Example 9
[0169] 25
[0170] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 12.6 (s, br, 1H),
8.41 (d, J=8.1 Hz, 1H), 7.91 (dd, J=8.1, 21 Hz, 2H), 7.5-7.7 (m,
4H), 2.73 (t, J=8 Hz, 2H), 1.5-1.6 (m, 2H), 1.55 (s, 9H), 1.0 (t,
J=8 Hz, 3H). MS (ES+): 325.2 (M+H).
Example 10
[0171] 26
[0172] .sup.1H NMR (400 MHz, CDCl.sub.3):.delta. 13.6 (s, br, 1H),
8.12 (d, J=8.1 Hz, 1H), 7.91 (dd, J=8.1, 21 Hz, 2H), 7.4-7.6 (m,
4H), 2.27 (s, 3H), 1.49 (s, 9H) MS (ES+): 297.2 (M+H).
Example 11
[0173] 27
[0174] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 11.02 (s, 1H),
8.09-8.07 (m, 1H), 7.92-7.89 (m, 1H), 7.82 (d, J=7.5 Hz, 1H), 7.69
(d, J=8.3 Hz, 1H), 7.58-7.53 (m, 2H), 7.50 (dd, J =7.8, 7.8 Hz,
1H), 6.21 (s, 1H), 1.37 (s, 9H); MS (ES+): 283.1 (M+H).
Example 12
[0175] 28
[0176] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.10 (d, J=9.4
Hz, 1H), 7.91 (dd, J=9.4, 2.7 Hz, 1H), 7.73 (d, J=7.7 Hz, 2H),
7.55-7.28 (m, 3H), 6.14 (s, 1H), 2.95 (septet, J=6.9 Hz, 1H), 6.9
(d, J=6.9 Hz, 6H); MS (ES+) 269.1 (M+H).
Example 13
[0177] 29
[0178] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.06 (dd, J=6.3,
3.5 Hz, 1H), 7.92-7.89 (m, 1H), 7.81 (d, J=7.5 Hz, 1H), 7.69 (d,
J=8.3 Hz, 1H), 7.53 (dd, J=6.3, 3.2 Hz, 2H), 7.49 (dd, J=7.8, 7.8
Hz, 1H), 6.25 (s, 1H), 4.09 (q, J=7.1 Hz, 2H), 2.72 (s, 2H), 1.46
(s, 6H), 1.21 (t, J=7.1 Hz, 3H); MS (ES+) 355.1 (M+H).
Example 14-15
[0179] 30
[0180] A mixture of 14 (70 mg, 0.22 mmol, prepared from
1,3-dichloroacetone and N-naphthylthiourea following general
procedure 2) and diisopropylamine (129 mg, 1.0 mmol) in ethanol
(1.5 mL) was stirred at reflux for 6 h. After evaporating ethanol
the crude product was purified by HPLC to afford the desired
product (TFA salt) as colorless liquid (33 mg, 40% yield).
.sup.1H-NMR (400 MHz, CDCl.sub.3): .delta.11.1 (s, 1H), 8.05-8.03
(m, 1H), 7.96-7.94 (m, 1H), 7.87 (d, J=8.3, 1H), 7.69 (dd, J=8.3,
0.9 Hz, 1H), 7.65-7.59 (m, 2H), 7.53 (dd, J=7.8, 7.8 Hz, 1H), 7.06
(s, 1H), 4.37 (s, 2H), 3.00 (d, J=6.7 Hz, 4H), 2.20-2.11 (m, 2H),
1.09 (d, J=6.7 Hz, 12H). MS (ES+) 368.3 (M+H).
Examples 16-18
[0181] 31
Example 16
[0182] Compound 16 was prepared from N-naphthylthiourea (606 mg,
3.0 mmol) and 1-bromo-2,3-butanedione (4.0 mmol) according to
general procedure 2. This material was carried to the next step
without further purification.
Example 17
[0183] To a stirred suspension of 4-acetoyl-2-naphthylamino
thiazole (65 mg, 0.18 mmol) and iso-butylamine (53 mg, 0.72 mmol)
in methanol (3 mL) was added a solution of sodium cyanoborohydride
(30 mg) in methanol (1 mL) at 0.degree. C. After stirring the
mixture for 12 h, it was diluted with EtOAc (20 mL) and washed with
water (2.times.5 mL). The organic layer was dried over MgSO4,
filtered and concentrated in vacuo. The residue was purified by
HPLC to give 4-iso-butylamino-2'-ethyl-2-naphthyl- aminothiazole
(17) as white solid (27 mg, 45% yield).
Example 18
[0184] Compound 18 was prepared from
4-iso-butylamino-2'-ethyl-2-naphthyla- mino thiazole and
iso-butylaldehyde according to the general procedure of reductive
amination outlined above. Purification of the crude product by HPLC
afforded the desired product as white solid (61 mg, 50% yield).
.sup.1H-NMR (400 MHz, CDCl.sub.3): .delta. 7.97-7.95 (m, 1H),
7.83-7.81 (m, 1H), 7.68-7.62 (m, 2H), 7.47-7.45 (m, 2H), 7.39 (dd,
J=7.8, 7.8 Hz, 1H), 6.64 (s, 1H), 4.53 (q, J=6.7 Hz, 1H), 2.92 (m,
2H), 2.66 (m, 2H) 1.96 (m, 2H), 1.59 (d, J=6.7 Hz, 3H), 0.94 (d,
J=6.6 Hz, 6H), 0.80 (d, J=6.6 Hz, 6H); MS (ES+) 382.3 (M+H).
Example 19
[0185] 32
[0186] A mixture of 1-aminonaphthalene (2.0 mmol, 268 mg) and
2,6-dichlorobenzenethiazole (1.0 mmol, 203 mg) in DMF (3.0 mL) was
heated at 110.degree. C. for 8 h. The mixture was allowed to cool
to room temperature, diluted with EtOAc (15.0 mL) and then
extracted with H.sub.2O (4.times.10 mL) to remove DMF. The organic
layer was dried over anhydrous MgSO.sub.4, filtered and
concentrated in vacuo. The residue was purified by silica gel flash
column chromatography (hexanes/EtOAc) to afford the desired product
as yellow solid (124 mg, 40% yield). .sup.1H-NMR (400 MHz,
CDCl.sub.3): .delta.7.87-7.83 (m, 4H), 7.51-7.49 (m, 2H), 7.36-7.33
(m, 2H), 6.82 (dd, J=6.8, 1.5Hz, 2H). MS (ES+) 311.0 (M+H).
Example 20
[0187] 33
[0188] This compound was prepared from
1-benzyl-3-bromo-4-piperidone (which is synthesized from
1-benzyl-4-piperidone by regioselective bromination) and
N-naphthylthiourea following a general procedure. Purification of
the crude product by HPLC gave (20) as white solid (178 mg, 48%
yield). .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.89 (d, J=9.1
Hz, 1H), 7.87 (d, J=8.4 Hz, 1H), 7.73-7.68 (m, 2H), 7.58-7.41 (m,
4H), 7.32-7.21 (m, 4H), 3.49 (s, 2H), 3.14 (s, 2H), 2.68 (t, J=5.4
Hz, 2H), 2.60 (t, J=5.4 Hz, 2H); MS (ES+) 372.2 (M+H).
Example 21
[0189] This example illustrates a CCR4 binding assay that can be
used for evaluating the compounds of the present invention.
[0190] Detection of radiolabelled TARC and/or MDC binding to
CCR4
[0191] .sup.125I-labelled TARC and MDC are available from
commercial sources (e.g., Amersham-Pharmacia or Perkin Elmer Life
Sciences). All buffers and materials are available from commercial
sources (e.g., Gibco BRL, Sigma). To measure binding of
.sup.125I-TARC or .sup.125I-MDC to cells expressing CCR4 (e.g., CEM
cells, available from the ATCC), the .sup.125I-TARC or
.sup.125I-MDC is diluted to a concentration of approximately 200 pM
in a buffered saline solution (e.g., RPMI supplemented with 0.5%
bovine serum albumin), and added to an equal volume of a suspension
of cells (e.g., CEM cells at 5.times.10.sup.6 cells/mL). The
resulting mixture is incubated for a period of time (e.g., 2
hours). The unbound .sup.125I-TARC or .sup.125I-MDC is separated
from the cells by filtration, e.g., by passage through GF/B filter
plate (Packard Biosciences) pre-treated with 0.3% polyethyleneimine
(Sigma), using a Packard Filtermate 96 (Packard Biosciences). The
amount of .sup.125I-TARC or .sup.125I-MDC retained with the cells
on the filterplate is measured by adding a small amount of
scintillation fluid (e.g., 50 .mu.L of Microscint-20, obtained from
Packard Biosciences), and reading scintillation on appropriate
detection equipment, e.g., a Packard TopCount 383 (Packard
Biosciences).
[0192] Non-specific binding of .sup.125I-TARC or .sup.125I-MDC can
be estimated by measuring the amount of .sup.125I-TARC or
.sup.125I-MDC retained with the cells on the filterplate when the
assay is performed in the presence of a large excess of unlabelled
TARC or MDC.
[0193] Inhibition of .sup.125I-TARC or .sup.125I-MDC binding to
CCR4 is defined as a decrease in the retention of .sup.125I-TARC or
.sup.125I-MDC to the cells on the filterplate.
[0194] The assay described above varies only moderately from
standardly used procedures, e.g., Imai et al. (1997) J. Biol. Chem.
272:15036-15042, Imai et al. (1998) J. Biol. Chem.
273:1764-1768.
1TABLE 1 CCR4 antagonist activity for compounds of the invention
(inhibition of .sup.125I-TARC binding). Compound IC.sub.50 (nM) 12
+ 2 + 11 + 4 ++ 9 + 10 + 1 ++ 3 ++ 13 + 5 + 19 ++ 8 + 15 + 6 + 18 +
7 ++ 20 + +denotes IC.sub.50 > 1000 nM ++denotes IC.sub.50 <
1000 nM
EXAMPLE 22
[0195] This example illustrates additional screening procedures
used in characterizing the compounds of the present invention.
[0196] Source plates of chemical libraries were obtained from
commercial vendors and contained individual compounds at 5 mg/mL in
DMSO, or in some instances, at 1 mg/nL. From these, multiple
compound plates containing 10 compounds in each well were made, and
these were diluted in 20% DMSO to a concentration of 50 .mu.g/mL
(10 .mu.g/mL for those beginning at 1 mg/mL). An aliquot of 20
.mu.L of each mixture was put into the test plates, which were
stored frozen until use.
[0197] A CCR4 expressing stable transfectant cell line was prepared
using current standard molecular biological methods. The CCR4
transfectants were cultured in IMDM-5% FBS, and harvested when the
concentration was between 0.5-1.0.times.10.sup.6 cells/ml. The
cells were centrifuged and resuspended in assay buffer (20 mM HEPES
pH 7.1, 140 mM NaCl, 1 mM CaCl.sub.2, 5mM MgCl.sub.2, and with 0.2%
bovine serum albumin) to a concentration of 5.6.times.10.sup.6
cells/ml. To set up the screening assays, first 0.09 ml of cells
was added to the assay plates containing the compounds. (For a
final compound concentration of 1-5 .mu.g/ml each [.about.2-10
.mu.M]). Then 0.09 ml of .sup.125I labeled MDC or TARC diluted in
assay buffer (final concentration .about.50 pM, with .about.30,000
cpm per well) was added. The plates were sealed and incubated for
approximately 3 hours at 4.degree. C. on a shaker platform. The
assay plates were harvested using Packard filter plates, pre-soaked
in 0.3% PEI (polyethyleneimine) solution, on a Packard vacuum cell
harvester. Scintillation fluid (50 .mu.l) was added to all wells
and the plates were sealed and counted in a Top Count scintillation
counter. Control wells containing either diluent only (for total
counts) or excess MDC or TARC (1 .mu.g/ml, for non-specific
binding) were used to calculate the percent of total inhibition for
each set of compounds. Further tests on individual compounds were
carried out in the same manner. IC.sub.50 values are those
concentrations required to reduce the binding of labeled MDC or
TARC to the receptor by 50%.
[0198] The calcium mobilization experiments were performed by
labeling the human T-cell line CEM with NDO-1 dye (45 min at room
temperature), washing with PBS, and resuspending into flux buffer
(HBSS with 1% fetal bovine serum). For each test, 1.times.10.sup.6
cells were incubated at 37.degree. C. in the cuvette of a PTI
spectrometer, and the ratio of 410/490 nm emission plotted over
time (typically 2-3 minutes), with compounds added at 5 seconds,
followed by MDC, TARC or other chemokines.
[0199] Chemotaxis assays were performed using 5 .mu. filter plates
(Neuroprobe) with the chemoattractant (MDC, TARC, or SDF) placed in
the lower chamber, and a cell suspension of 100,000 CEM cells in
the upper chamber. The assays were incubated 1-2 hours at
37.degree. C., and the number of cells in the lower chamber
quantified by the CyQuant assay (Molecular Probes).
[0200] Results are provided in Figures.
EXAMPLE 23
[0201] This example describes a procedure to evaluate the efficacy
of CCR4 antagonists for treatment of septic shock.
[0202] An animal model of endotoxic shock can be induced by
injecting rodents with lipopolysaccharide (LPS). Three series of
mouse groups, comprising 15 mice per group, are treated with an
intra-peritoneal injection of an L.D.(lethal dose)-90 of LPS
(precise dose requires titration of the particular batch of
lipopolysaccharide in the actual mouse colony in use to determine a
re-producible dose of LPS that produces 90% mortality in mice). One
series of mice additionally receives phosphate buffered saline
(PBS) and Tween 0.5% i.p. 30 minutes before LPS administration. A
second series consists of groups of mice receiving different doses
of the CCR4 antagonist(s) given either intra-peritoneally,
intra-venously, sub-cutaneously, intramuscularly, orally, or via
any other mode of administration 30 minutes before, or concurrently
with, LPS administration. A third series of mice, serving as
positive control, consists of groups treated with either mouse
IL-10 i.p., or anti-TNF antibodies i.p., 30 minutes before LPS
administration.
[0203] Mice are monitored for death for 72 hours following the LPS
injection.
EXAMPLE 24
[0204] This example describes a procedure to evaluate the efficacy
of CCR4 antagonists for treatment of asthma.
[0205] An animal model of asthma can be induced by sensitizing
rodents to an experimental antigen (e.g. OVA) by standard
immunization, and then subsequently introducing that same antigen
into the rodents lung by aerosolization. Three series of rodent
groups, comprising 10 rodents per goup, are actively sensitized on
Day 0 by a single intraperitoneal injection with 100 ug OVA in
phosphate-buffered saline (PBS), along with an IgE-selective
adjuvant e.g. aluminum hydroxide. At 11 days after sensitization,
at the peak of their IgE response, the animals are placed in a
Plexiglas chamber and challenged with aerosolized OVA (1%) for 30
minutes using the ultrasonic nebulizer (De Vilbliss). One series of
mice additionally receives phosphate buffered saline (PBS)and Tween
0.5% i.p. at the initial sensitization, and at different dosing
schedules thereafter, up until the aerosolized OVA challenge. A
second series consists of groups of mice receiving different doses
of the CCR4 antagonist(s) given either intra-peritoneally,
intra-venously, sub-cutaneously, intra-muscularly, orally, or via
any other mode of administration at the initial sensitization, and
at different dosing schedules thereafter, up until the aerosolized
OVA challenge. A third series of mice, serving as positive control,
consists of groups treated with either mouse IL-10 i.p., anti-IL4
antibodies i.p., or anti-IL5 antibodies i.p. at the initial
sensitization, and at different dosing schedules thereafter, up
until the aerosolized OVA challenge.
[0206] Animals were subsequently analyzed at different time points
after the aerosolized OVA challenge for pulmonary function,
cellular infiltrates in bronchoalveolar lavage (BAL), histological
examination of lungs, and measurement of serum OVA-specific IgE
titers.
EXAMPLE 25
[0207] This example describes a procedure to evaluate the efficacy
of CCR4 antagonists for augmenting protective immunity against
viruses, bacteria and parasites.
[0208] Protective immunity to microbial pathogens is frequently
mediated by Th1 regulatory T cells. Since CCR4 antagonists are
likely inhibitors of Th2 regulatory cells, they may alter the cross
regulation that normally exists between Th1 and Th2 cells, and
potentiate Th1 cells, thereby augmenting protection against
infectious disease. Three series of mouse groups, comprising 15
mice per group, are infected with the intracellular parasite
Leishmania major(L.major) by injecting L.major promastigotes
sub-cutaneously into their left hind footpads. Four weeks after
infection, the animals are challenged with either Leishmania
freeze-thawed antigen, or PBS as a negative control, in the
contra-lateral footpad. One series of mice additionally receives
phosphate buffered saline (PBS)and Tween 0.5% i.p. at the initial
sensitization, and at different dosing schedules thereafter, up
until the Leishmania antigen challenge. A second series consists of
groups of mice receiving different doses of the CCR4 antagonist(s)
given either intra-peritoneally, intra-venously, sub-cutaneously,
intra-muscularly, orally, or via any other mode of administration
at the initial sensitization, and at different dosing schedules
thereafter, up until the Leishmania antigen challenge. A third
series of mice, serving as positive control, consists of groups
treated with either mice IL-12, anti-IL4 antibodies i.p., or
anti-IL5 antibodies i.p. at the initial sensitization, and at
different dosing schedules thereafter, up until the Leishmania
antigen challenge.
[0209] Over the next 48 hours, footpad swelling, caused by a
Delayed-Type Hypersensitivity reaction to the Leishmania antigen
challenge, is monitored with a metric caliper. The response of
draining lymph node T cells to Leishmania antigen stimulation in
vitro is also measured, both at the level of proliferation,
cytokine production, and other phenotypic criteria.
EXAMPLE 26
[0210] This example describes a procedure to evaluate the efficacy
of CCR4 antagonists for treatment of rheumatoid arthritis.
[0211] An animal model of rheumatoid arthritis can be induced in
rodents by injecting them with type II collagen in selected
adjuvants. Three series of rodent groups consisting 15
genetically-susceptible mice or rats per group are injected
sub-cutaneously or intra-dermally with type II collagen emulsified
in Complete Freund's Adjuvant at days 0 and 21. One series of
rodents additionally receives phosphate buffered saline (PBS) and
Tween 0.5% i.p. at the initial sensitization, and at different
dosing schedules thereafter. A second series consists of groups of
rodents receiving different doses of the CCR4 antagonist(s) given
either intra-peritoneally, intravenously, sub-cutaneously,
intra-muscularly, orally, or via any other mode of administration
at the initial sensitization, and at different dosing schedules
thereafter. A third series of rodents, serving as positive control,
consists of groups treated with either mouse IL-10 i.p., or
anti-TNF antibodies i.p.at the initial sensitization, and at
different dosing schedules thereafter.
[0212] Animals are monitored from weeks 3 til 8 for the development
of swollen joints or paws, and graded on a standard disease
severity scale. Disease severity is confirmed by histological
analysis of joints.
EXAMPLE 27
[0213] This example describes a procedure to evaluate efficacy of
CCR4 antagonists for treatment of Systemic Lupus Erythematosus
(SLE).
[0214] Female NZB/W F1 mice spontaneously develop an SLE-like
pathology commencing at 6 months of age that is characterized by
proteinuria, serum autoantibodies, glomerulonephritis, and
eventually death. Three series of NZB/W mouse groups comprising 20
mice per group are tested for efficacy of CCR antagonist(s) as
follows: One series of mice additionally receives phosphate
buffered saline (PBS) and Tween 0.5% i.p. soon after weaning, and
thereafter at varying dosing schedules. A second series consists of
groups of mice receiving different doses of the CCR4 antagonist(s)
given either intra-peritoneally, intra-venously, sub-cutaneously,
intra-muscularly, orally, or via any other mode of administration
soon after weaning, and thereafter at varying dosing schedules. A
third series of mice, serving as positive control, consists of
groups treated with anti-IL10 antibodies given soon after weaning,
and thereafter at varying dosing schedules.
[0215] Disease development is monitored in terms of eventual
mortality, kidney histology, serum autoantibody levels, and
proteinuria.
EXAMPLE 28
[0216] This example describes a procedure to evaluate efficacy of
CCR4 antagonists for treatment of malignancy.
[0217] SCID mice can be transplanted with primary human tumor
cells. Normal mouse strains can be transplanted with a variety of
well-characterized mouse tumor lines, including a mouse thymoma EL4
which has been transfected with OVA to allow easy evaluation of
tumor specific antigen responses following vaccination with OVA.
Three series of mouse groups from any of these tumor models are
tested for CCR4 antagonist efficacy as follows: One series of mice
additionally receives phosphate buffered saline (PBS) and Tween
0.5% i.p. soon after tumor transplant, and thereafter at varying
dosing schedules. A second series consists of groups of mice
receiving different doses of the CCR4 antagonist(s) given either
intra-peritoneally, intra-venously, sub-cutaneously,
intra-muscularly, orally, or via any other mode of administration
soon after tumor transplant, and thereafter at varying dosing
schedules. A third series of mice, serving as positive control,
consists of groups treated with either anti-IL4 antibodies,
anti-IFNg antibodies, IL4, or TNF, given i.p. soon after tumor
transplant, and thereafter at varying dosing schedules.
[0218] Efficacy is monitored via tumor growth versus regression. In
the case of the OVA-transfected EL4 thymoma model, cytolytic
OVA-specific responses can be measured by stimulating draining
lymph node cells with OVA in vitro, and measuring antigen-specific
cytotoxicity at 72 hours.
EXAMPLE 29
[0219] This example describes procedures to evaluate the efficacy
of CCR4 antagonists in psoriasis.
[0220] A rodent model of psoriasis can be obtained by
intra-venously transferring a population of purified T cells
(designated CD45Rbhi T cells) obtained from the spleens of BALB/c
mice into immunodeficient recipient CB.17 scid/scid mice. Mice
develop signs of redness, swelling, and skin lesions resembling
those of human psoriasis in their ear, feet and tail by 8 weeks
after transfer. Three series of mouse groups, comprising 10-15
CB.17 scid/scid mice per group, are injected with purified CD45Rbhi
T cells. One series of mice additionally receives phosphate
buffered saline (PBS) and Tween 0.5% i.p. at the initial cell
transfer, and at different dosing schedules thereafter. A second
series consists of groups of mice receiving different doses of the
CCR4 antagonist(s) given either intra-peritoneally, intravenously,
sub-cutaneously, intramuscularly, orally, or via any other mode of
administration at the initial cell transfer, and at different
dosing schedules thereafter. A third series of mice, serving as
positive control, consists of groups treated with antibodies to
either IL-12, IL-4, IFNg, or TNF, or with cytokine IL-10 at the
initial cell transfer, and at different dosing schedules
thereafter. Animals are monitored for development of psoriatic-like
lesions for 3 months after cell transfer.
EXAMPLE 30
[0221] This example describes a procedure to evaluate the efficacy
of CCR4 antagonists in Inflammatory Bowel Disease (IBD).
[0222] Several mouse models of IBD (including Crohn's Disease and
Ulcerative Colitis) have been developed. Some of these are
spontaneous models occurring in genetically engineered transgenic
mice that have been depleted of certain cytokine genes (e.g. IL-10,
or IL-2) by homologous recombination. Another mouse model of
Inflammatory Bowel Disease is obtained by transferring highly
purified populations of CD4+T lymphocytes bearing a particular
surface marker phenotype (namely CD45 RB hi) into SCID mice. Three
series of mouse groups from any one of these models can be used to
evaluate CCR4 antagonist efficacy as follows. One group of mice
additionally receives phosphate buffered saline (PBS) and Tween
0.5% i.p. soon after weaning in the case of the spontaneous models
in transgenic mice, or at time of cell transfer into SCID mice and
varying dosings thereafter for the cell transfer model. A second
series consists of groups of mice receiving different doses of the
CCR4 antagonist(s) given either intra-peritoneally, intra-venously,
sub-cutaneously, intramuscularly, orally, or via any other mode of
administration soon after weaning in the case of the spontaneous
models in transgenic mice, or at time of cell transfer into SCID
mice and varying dosings thereafter for the cell transfer model. A
third series of mice, serving as positive control, consists of
groups treated with antibodies to either IFNg, or TNF, or with
cytokine IL-10 soon after weaning in the case of the spontaneous
models in transgenic mice, or at time of cell transfer into SCID
mice and varying dosings thereafter for the cell transfer
model.
[0223] Mice are evaluated for 6-8 weeks for disease development,
monitored initially via weight loss and/or prolapsed rectum, and
eventually by histological evaluation of the animals colon and
intestinal tract.
[0224] It is understood that the examples and embodiments described
herein are for illustrative purposes only and that various
modifications or changes in light thereof will be suggested to
persons skilled in the art and are to be included within the spirit
and purview of this application and scope of the appended claims.
All publications, patents, and patent applications cited herein are
hereby incorporated by reference in their entirety for all
purposes.
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