U.S. patent application number 11/533034 was filed with the patent office on 2007-01-11 for novel compounds.
This patent application is currently assigned to SmithKline Beecham Corporation. Invention is credited to Jerry L. Adams, Neil W. Johnson, Jeffrey H. Murray.
Application Number | 20070010534 11/533034 |
Document ID | / |
Family ID | 27496721 |
Filed Date | 2007-01-11 |
United States Patent
Application |
20070010534 |
Kind Code |
A1 |
Adams; Jerry L. ; et
al. |
January 11, 2007 |
NOVEL COMPOUNDS
Abstract
The present invention is directed to novel compounds of Formula
(I) for use in the treatment of diseases, in a mammal, in which
inappropriate, excessive or undesirable angiogenesis has occurred
and/or where excessive Tie2 receptor activity has occurred.
Inventors: |
Adams; Jerry L.; (Wayne,
PA) ; Johnson; Neil W.; (Downingtown, PA) ;
Murray; Jeffrey H.; (Norristown, PA) |
Correspondence
Address: |
SMITHKLINE BEECHAM CORPORATION;CORPORATE INTELLECTUAL PROPERTY-US, UW2220
P. O. BOX 1539
KING OF PRUSSIA
PA
19406-0939
US
|
Assignee: |
SmithKline Beecham
Corporation
|
Family ID: |
27496721 |
Appl. No.: |
11/533034 |
Filed: |
September 19, 2006 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10130018 |
May 10, 2002 |
|
|
|
PCT/US00/31791 |
Nov 20, 2000 |
|
|
|
11533034 |
Sep 19, 2006 |
|
|
|
60166886 |
Nov 22, 1999 |
|
|
|
60166885 |
Nov 22, 1999 |
|
|
|
60166814 |
Nov 22, 1999 |
|
|
|
60166895 |
Nov 22, 1999 |
|
|
|
Current U.S.
Class: |
514/256 ;
514/269; 514/341; 544/310; 544/314; 546/272.7 |
Current CPC
Class: |
A61P 9/00 20180101; A61P
43/00 20180101; C07D 417/14 20130101; C07D 409/14 20130101; C07D
401/14 20130101; A61P 9/10 20180101; C07D 401/04 20130101; C07D
403/04 20130101; A61P 27/02 20180101; A61P 25/28 20180101; A61P
25/02 20180101; C07D 405/14 20130101; A61P 35/00 20180101; A61P
35/04 20180101 |
Class at
Publication: |
514/256 ;
514/269; 514/341; 544/310; 544/314; 546/272.7 |
International
Class: |
A61K 31/513 20060101
A61K031/513; A61K 31/506 20060101 A61K031/506; A61K 31/4439
20060101 A61K031/4439; C07D 403/04 20060101 C07D403/04 |
Claims
1. A compound of formula (I): ##STR39## wherein V is CH or N; Ar is
a napth-2-yl, napth-1-yl, a bicyclic or tricyclic heteroaromatic
ring, which ring may be optionally substituted; Y is
NR.sup.10R.sup.11, NR.sup.10C(Z)NR.sup.10R.sup.11,
NR.sup.10C(Z)NR.sup.10C(Z)OR.sup.11, NR.sup.10COOR.sup.11 or
NR.sup.10SO.sub.2R.sup.11; n is 0, 1, 2, 3 or 4; X is O, CH.sub.2,
S or NH; Z is oxygen or sulfur; R.sup.1 is independently hydrogen,
X--R.sup.4, halogen, hydroxy, optionally substituted C.sub.1-6
alkyl, optionally substituted C.sub.1-6alkylsulfinyl,
CH.sub.2OR.sup.5, amino, mono or di-C.sub.1-6alkylamino,
N(R.sup.6)C(O)R.sup.7, N(R.sup.6)S(O).sub.2R.sup.8, or a 5 to
7-membered N-heterocyclyl ring which optionally contains an
additional heteroatom selected from O, S and NR.sup.9; R.sup.2 and
R.sup.3 independently represent optionally substituted
C.sub.1-6alkyl, or R.sup.2 and R.sup.3 together with the carbon
atom to which they are attached form an optionally substituted
C.sub.3-7cycloalkyl or C.sub.5-7 cycloalkenyl ring, or R.sup.2 and
R.sup.3 together with the carbon atom to which they are attached
form an optionally substituted 5 to 7-membered heterocyclyl ring
containing up to 3 heteroatoms selected from N, O and S; R.sup.4 is
independently C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl,
heterocyclyl, heterocyclylC.sub.1-6alkyl, heteroaryl, or a
heteroarylC.sub.1-6alkyl moiety, and wherein any of these moieties
may be optionally substituted; R.sup.5 is hydrogen, C(Z)R.sup.12 or
optionally substituted C.sub.1-6alkyl, optionally substituted aryl,
optionally substituted arylC.sub.1-6alkyl, or S(O).sub.2R.sup.8;
R.sup.6 is hydrogen or C.sub.1-6alkyl; R.sup.7 is hydrogen,
C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl, arylC.sub.1-6alkyl,
heteroaryl, heteroarylC.sub.1-6alkyl, heterocyclyl, or
heterocyclylC.sub.1-6alkyl; R.sup.8 is C.sub.1-6alkyl,
C.sub.3-7cycloalkyl, aryl, arylC.sub.1-6alkyl, heteroaryl,
heteroarylC.sub.1-6alkyl, heterocyclyl, or
heterocyclylC.sub.1-6alkyl; R.sup.9 is hydrogen, C.sub.1-4alkyl,
C.sub.3-7cycloalkyl or aryl; R.sup.10 and R.sup.12 are
independently selected from hydrogen, C.sub.1-6alkyl,
C.sub.3-7cycloalkyl, C.sub.3-7cycloalkyl C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, heterocyclyl, heterocyclylC.sub.1-6alkyl,
heteroaryl and heteroarylC.sub.1-6alkyl, any of which may be
optionally substituted; and R.sup.11 is hydrogen, C.sub.1-6alkyl,
C.sub.3-7cycloalkyl, C.sub.3-7cycloalkyl C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, heterocyclyl, heterocyclylC.sub.1-6alkyl,
heteroaryl, or heteroarylC.sub.1-6alkyl, any of which may be
optionally substituted; or R.sup.10 and R.sup.11 may together with
the nitrogen may form a 5 to 7 membered ring optionally containing
an additional heteroatom selected from O, S, or NR.sup.9; or a
pharmaceutically acceptable salt thereof.
2. The compound according to claim 1 wherein V is CH.
3. The compound according to claim 1 wherein V is N.
4. The compound according to claim 1 wherein R.sup.1 is hydrogen,
or the moiety X--R.sup.4.
5. The compound according to claim 4 wherein X is oxygen or
nitrogen.
6. The compound according to claim 4 wherein R.sup.4 is an
optionally substituted alkyl, aryl or arylC.sub.1-6alkyl.
7. The compound according to claim 1 wherein Ar is an optionally
substituted naphthyl, benzothiophene or benzofuran ring.
8. The compound according to claim 7 wherein the Ar ring is
substituted by up to 3 substituents independently selected from
halo, hydroxy, hydroxy C.sub.1-6alkyl, or C.sub.1-6alkoxy.
9. The compound according to claim 8 wherein Ar is a napth-2-yl,
optionally substituted by a C.sub.1-6alkoxy group.
10. The compound according to claim 1 wherein R.sup.2 and R.sup.3
independently an optionally substituted C.sub.1-6 alkyl.
11. The compound according to claim 1 wherein R.sup.2 and R.sup.3
together with the carbon atom to which they are attached form an
optionally substituted C.sub.3-7cycloalkyl or C.sub.5-7cycloalkenyl
ring.
12. The compound according to claim 1 wherein R.sup.2 and R.sup.3
together with the carbon atom to which they are attached form an
optionally substituted 5 to 7 membered heterocyclyl ring containing
up to 3 heteroatoms selected from N, O and S.
13. The compound according to claim 1 which is:
(2-(4-(6-methoxynapthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-carbamic acid tert-butyl ester;
2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propylamine;
2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-acetamide;
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methy-
l-propyl)-methanesulfonamide;
1,1,1-trifluoro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imid-
azol-2-yl)-2-methyl-propyl)-methanesulfonamide;
2,2,2-trifluoro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imid-
azol-2-yl)-2-methyl-propyl)-ethanesulfonamide;
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methy-
l-propyl)-propanesulfonamide;
3-Chloro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2--
yl)-2-methyl-propyl)-propane sulfonamide;
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methy-
l-propyl)-butanesulfonamide;
1-Ethyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-y-
l)-2-methyl-propyl)-urea;
1-(2-Chloroethyl)-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-im-
idazol-2-yl)-2-methyl-propyl)-urea;
1-n-Propyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol--
2-yl)-2-methyl-propyl)-urea;
1-Isopropyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-
-2-yl)-2-methyl-propyl)-urea;
1-tert-Butyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazo-
l-2-yl)-2-methyl-propyl)-urea;
1-Methylformyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imida-
zol-2-yl)-2-methyl-propyl)-urea;
1-(3,5-Dimethyl-isoxazol-4-yl)-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyrid-
in-4-yl-1H-imidazol-2-yl)-2-methyl-propyl)-urea;
1-n-Propyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol--
2-yl)-2-methyl-propyl)-thiourea;
1-n-Butyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-
-yl)-2-methyl-propyl)-thiourea;
1-Isopropyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-
-2-yl)-2-methyl-propyl)-thiourea;
1-Ethyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-y-
l)-2-methyl-propyl)-thiourea;
1-Methyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2--
yl)-2-methyl-propyl)-thiourea;
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-met-
hyl-propyl)-3-(2-methoxyethyl)-thiourea;
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-met-
hyl-propyl)-3-(2-morpholin-4-yl-ethyl)-thiourea;
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-met-
hyl-propyl)-3-(2-piperidin-4-yl-ethyl)-thiourea;
Cyclohexylmethyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imida-
zol-2-yl)-2-methyl-propyl)-amine;
Bis-n-butyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-
-yl)-2-methyl-propyl)-amine;
Bis-cyclohexylmethyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-i-
midazol-2-yl)-2-methyl-propyl)-amine;
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methy-
l-propyl)-(3-methylsulfanyl-propyl)-amine;
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methy-
l-propyl)-bis-(3-methylsulfanyl-propyl)-amine;
Isobutyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl-
)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine;
Bis-isobutyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol--
2-yl)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine;
(2,2-Dimethylpropyl)-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-i-
midazol-2-yl)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine;
n-Propyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl-
)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine; or a
pharmaceutically acceptable salt thereof.
14. A pharmaceutical composition comprising a compound according to
claim 1 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
15. A method of treating, including prophylaxis, of a TIE2 receptor
mediated disease in a mammal in need thereof, which comprises
administering to said mammal an effective amount of a compound
according to claim 1.
16. The method according to claim 15 wherein the disease is
characterized by excessive, undesired, or inappropriate
angiogenesis.
17. The method according to claim 16 wherein the disease is
diabetic retinopathy, macular degeneration, or other ocular
neovascularizations.
18. The method according to claim 15 wherein the disease is
characterized by excessive or increased proliferation of
vasculature.
19. The method according to claim 18 wherein the disease is tumor
growth and metastasis.
20. The method according to claim 15 wherein the disease is
atherosclerosis.
21. The compound according to claim 1 wherein Ar is a naphthyl-2-yl
ring.
22. The compound according to claim 21 wherein the naphthyl-2-yl
ring is substituted one or more times by halo, hydroxy,
C.sub.1-6alkyl, halosubstituted C.sub.1-6alkyl, hydroxy
C.sub.1-6alkyl, and C.sub.1-6 alkoxy.
23. The compound according to claim 22 wherein the naphthyl-2-yl
ring is substituted in the 6-position.
24. The compound according to claim 23 wherein the substituent is a
C.sub.1-6 alkoxy.
25. The compound according to claim 24 wherein the substituent is
methoxy.
Description
[0001] This application is a divisional of application Ser. No.
10/130,018 (allowed) filed 10 May 2002, which is a .sctn.371
national stage entry of PCT/US00/31791 filed 20 Nov. 2000, which
claims benefit of provisional applications, U.S. Ser. No.
60/166,886 filed 22 Nov. 1999; 60/166,885 filed 22 Nov. 1999;
60/166,814 filed 22 Nov. 1999; and 60/166,895 filed 22 Nov.
1999.
FIELD OF THE INVENTION
[0002] The present invention relates to the treatment of diseases,
in a mammal, in which inappropriate, excessive or undesirable
angiogenesis has occurred and/or where excessive Tie2 receptor
activity has occurred.
BACKGROUND OF THE INVENTION
[0003] Chronic proliferative diseases are often accompanied by
profound angiogenesis, which can contribute to or maintain an
inflammatory and/or proliferative state, or which leads to tissue
destruction through the invasive proliferation of blood vessels.
(Folkman, E X S 79:1-8, 1997; Folkman, Nature Medicine 1:27-31,
1995; Folkman and Shing, J. Biol. Chem. 267:10931, 1992).
[0004] Angiogenesis is generally used to describe the development
of new or replacement blood vessels, or neovascularisation. It is a
necessary and physiological normal process by which the vasculature
is established in the embryo. Angiogenesis does not occur, in
general, in most normal adult tissues, exceptions being sites of
ovulation, menses and wound healing. Many diseases, however, are
characterized by persistent and unregulated angiogenesis. For
instance, in arthritis, new capillary blood vessels invade the
joint and destroy cartilage (Colville-Nash and Scott, Ann. Rheum.
Dis., 51, 919, 1992). In diabetes (and in many different eye
diseases), new vessels invade the macula or retina or other ocular
structures, and may cause blindness (Brooks et al., Cell, 79, 1157,
1994). The process of atherosclerosis has been linked to
angiogenesis (Kahlon et al., Can. J. Cardiol. 8, 60, 1992). Tumor
growth and metastasis have been found to be angiogenesis-dependent
(Folkman, Cancer Biol, 3, 65, 1992; Denekamp, Br. J. Rad. 66, 181,
1993; Fidler and Ellis, Cell, 79, 185, 1994).
[0005] The recognition of the involvement of angiogenesis in major
diseases has been accompanied by research to identify and develop
inhibitors of angiogenesis. These inhibitors are generally
classified in response to discrete targets in the angiogenesis
cascade, such as activation of endothelial cells by an angiogenic
signal; synthesis and release of degradative enzymes; endothelial
cell migration; proliferation of endothelial cells; and formation
of capillary tubules. Therefore, angiogenesis occurs in many stages
and attempts are underway to discover and develop compounds that
work to block angiogenesis at these various stages.
[0006] There are publications that teach that inhibitors of
angiogenesis, working by diverse mechanisms, are beneficial in
diseases such as cancer and metastasis (O'Reilly et al., Cell, 79,
315, 1994; Ingber et al., Nature, 348, 555, 1990), ocular diseases
(Friedlander et al., Science, 270, 1500, 1995), arthritis (Peacock
et al., J. Exp. Med. 175, 1135, 1992; Peacock et al., Cell. Immun.
160, 178, 1995) and hemangioma (Taraboletti et al., J. Natl. Cancer
Inst. 87, 293, 1995).
[0007] Angiogenesis signals result from the interaction of specific
ligands with their receptors. The Tie1 and Tie2 receptors are
single-transmembrane, tyrosine kinase receptors (Tie stands for
lyrosine kinase receptors with immunoglobulin and EGF homology
domains). Both were recently cloned and reported by several groups
(Dumont et al., Oncogene 8:1293-1301, 1993; Partanen et al., Mol.
Cell Biol. 12:1698-1707, 1992; Sato et al., Proc. Natl. Acad. Sci.
USA 90:9355-9358, 1993).
[0008] Based upon the importance of Tie2 receptors in angiogenesis,
inhibition of Tie2 kinase activity is predicted to interrupt
angiogenesis, providing disease-specific therapeutic effects.
Recently, Lin et al. (J. Clin. Invest. 100:2072-2078, 1997) has
shown that exogenously administered soluble Tie2 receptor inhibited
angiogenesis and cancer growth in animal models. Thus inhibition of
Tie2 receptors by other means, such as inhibition of Tie2 receptor
kinase activity, is expected to have therapeutic benefit in
proliferative diseases involving angiogenesis.
[0009] The current application teaches the novel finding that
compounds of specific structure can inhibit the kinase activity of
the Tie2 receptor, block its signal transduction and thus may be
beneficial for proliferative diseases via inhibition of signals for
angiogenesis.
SUMMARY OF THE INVENTION
[0010] The present invention is directed to novel compounds of
Formula (I), and pharmaceutical compositions comprising a compound
of Formula (I) and a pharmaceutically acceptable diluent or
carrier.
[0011] Another aspect of the present invention is the use of
compounds of Formula (I) as Tie2 receptor kinase inhibitors. Tie2
receptor kinase inhibitors may be used in the treatment, including
prophylaxis, of inhibition of angiogenesis, or chronic inflammatory
or proliferative or angiogenic diseases or disorders which are
caused by excessive or inappropriate angiogenesis in a mammal in
need thereof.
[0012] The novel compounds of Formula (I) are represented by the
structure: ##STR1## wherein
[0013] V is CH or N;
[0014] Ar is a napth-2-yl, napth-1-yl, a bicyclic or a tricyclic
heteroaromatic ring, which rings may be optionally substituted;
[0015] Y is NR.sup.10R.sup.11, NR.sup.10C(Z)NR.sup.10R.sup.11,
NR.sup.10C(Z)NR.sup.10C(Z)OR.sup.11, NR.sup.10COOR.sup.11 or
NR.sup.10SO.sub.2R.sup.11;
[0016] X is O, CH.sub.2, S or NH;
[0017] Z is oxygen or sulfur;
[0018] n is 0, 1, 2, 3 or 4;
[0019] R.sup.1 is independently hydrogen, X--R.sup.4, halogen,
hydroxy, optionally substituted C.sub.1-6 alkyl, optionally
substituted C.sub.1-6alkylsulfinyl, CH.sub.2OR.sup.5, amino, mono
or di-C.sub.1-6alkylamino, N(R.sup.6)C(O)R.sup.7,
N(R.sup.6)S(O).sub.2R.sup.8, or a 5 to 7-membered N-heterocyclyl
ring which optionally contains an additional heteroatom selected
from O, Sand NR.sup.9;
[0020] R.sup.2 and R.sup.3 independently represent optionally
substituted C.sub.1-6alkyl, or R.sup.2 and R.sup.3 together with
the carbon atom to which they are attached form an optionally
substituted C.sub.3-7cycloalkyl or C.sub.5-7 cycloalkenyl ring, or
R.sup.2 and R.sup.3 together with the carbon atom to which they are
attached form an optionally substituted 5 to 7-membered
heterocyclyl ring containing up to 3 heteroatoms selected from N, O
and S;
[0021] R.sup.4 is independently C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, heterocyclyl, heterocyclylC.sub.1-6alkyl,
heteroaryl, or a heteroarylC.sub.1-6alkyl moiety, and wherein any
of these moieties may be optionally substituted;
[0022] R.sup.5 is hydrogen, C(Z)R.sup.12 or optionally substituted
C.sub.1-6alkyl, optionally substituted aryl, optionally substituted
arylC.sub.1-6alkyl, or S(O).sub.2R.sup.8;
[0023] R.sup.6 is hydrogen or C.sub.1-6alkyl;
[0024] R.sup.7 is hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl,
aryl, arylC.sub.1-6alkyl, heteroaryl, heteroarylC.sub.1-6alkyl,
heterocyclyl, or heterocyclylC.sub.1-6alkyl;
[0025] R.sup.8 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, aryl,
arylC.sub.1-6alkyl, heteroaryl, heteroarylC.sub.1-6alkyl,
heterocyclyl, or heterocyclylC.sub.1-6alkyl;
[0026] R.sup.9 is hydrogen, C.sub.1-4alkyl, C.sub.3-7cycloalkyl or
aryl;
[0027] R.sup.10 and R.sup.12 are independently selected from
hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, C.sub.3-7cycloalkyl
C.sub.1-16alkyl, aryl, arylC.sub.1-6alkyl, heterocyclyl,
heterocyclylC.sub.1-6alkyl, heteroaryl and
heteroarylC.sub.1-6alkyl, any of which may be optionally
substituted; and
[0028] R.sup.11 is hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl,
C.sub.3-7cycloalkyl C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl,
heterocyclyl, heterocyclylC.sub.1-6alkyl, heteroaryl, or
heteroarylC.sub.1-6alkyl, any of which may be optionally
substituted; or R.sup.10 and R.sup.11 together with the nitrogen
may form a 5 to 7 membered ring optionally containing an additional
heteroatom selected from O, S, or NR.sup.9;
[0029] or a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
[0030] The present invention is the directed to novel compounds
which can inhibit Tie2 kinase, and use of these compounds for
inhibition of angiogenesis in the treatment of chronic inflammatory
or proliferative or angiogenic diseases which are caused by
excessive or inappropriate angiogenesis in a mammal in need
thereof.
[0031] In the compounds of formula (I), V is suitably CH or N,
preferably carbon.
[0032] Suitably, the pyridyl or pyrimidine ring is optionally
substituted, independently, one to two times independently by
R.sup.1.
[0033] Suitably, R.sup.1 is independently hydrogen, X--R.sup.4,
halogen, hydroxy, optionally substituted C.sub.1-6 alkyl,
optionally substituted C.sub.1-6alkylsulfinyl, CH.sub.2OR.sup.5,
amino, mono or di-C.sub.1-6 alkylamino, N(R.sup.6)C(O)R.sup.7,
N(R.sup.6)S(O).sub.2R.sup.8, or a 5 to 7-membered N-heterocyclyl
ring which optionally contains an additional heteroatom selected
from O, S and NR.sup.9. Preferably, the pyridyl or pyrimidine is
substituted in the 2-position. Preferably, R.sup.1 is hydrogen or
X--R.sup.4.
[0034] X is suitably, O, CH.sub.2, S or NH. Preferably X is oxygen
or nitrogen.
[0035] R.sup.4 is independently C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, heterocyclyl, heterocyclylC.sub.1-6alkyl,
heteroaryl, or a heteroarylC.sub.1-6alkyl moiety, and wherein any
of these moieties may be optionally substituted. Preferably R.sup.4
is an optionally substituted alkyl, aryl, or arylalkyl group.
[0036] When R.sup.4 is aryl, it is preferably an optionally
substituted phenyl. When R.sup.4 is aryl alkyl, it is preferably an
optionally substituted benzyl or phenethyl.
[0037] These R.sup.4 moieties may be optionally substituted one or
more times, preferably 1 to 3 times, independently with halogen;
C.sub.1-4 alkyl, such as methyl, ethyl, propyl, isopropyl, or
t-butyl; halosubstituted alkyl, such as CF.sub.3; hydroxy; hydroxy
substituted C.sub.1-4 alkyl; C.sub.1-4 alkoxy, such as methoxy or
ethoxy; S(O).sub.malkyl and S(O)m aryl (wherein m is 0, 1, or 2);
C(O)OR.sup.11, such as C(O)C.sub.1-6 alkyl or C(O)OH moieties;
C(O)R.sup.11; OC(O)R.sup.8; O--(CH.sub.2)s-O--, such as in a ketal
or dioxyalkylene bridge (s is a number having a value of 1 to 5);
amino; mono- and di-C.sub.1-6 alkyl substituted amino;
N(R.sup.10)C(O)R.sup.7; C(O)NR.sup.10R.sup.11; cyano, nitro, or an
N-heterocyclyl ring which ring has from 5 to 7 members and
optionally contains an additional heteroatom selected from oxygen,
sulfur or NR.sup.9; optionally substituted aryl, such as phenyl; an
optionally substituted arylalkyl, such as benzyl or phenethyl;
aryloxy, such as phenoxy; or arylalkyloxy such as benzyloxy; these
aryl and arylalkyl moieties may be substituted with halogen, alkyl,
alkoxy, S(O)m alkyl, amino, or mono- and di-C.sub.1-6 alkyl
substituted amino.
[0038] Preferably the R.sup.4 moieties are substituted with an
amino, mono- or di-C.sub.1-6 alkyl substituted amino, or an
N-heterocyclyl ring which ring has from 5 to 7 members and
optionally contains an additional heteroatom selected from oxygen,
sulfur or NR.sup.9.
[0039] Suitably, R.sup.2 and R.sup.3 independently represent
optionally substituted C.sub.1-6alkyl, or R.sup.2 and R.sup.3
together with the carbon atom to which they are attached form an
optionally substituted C.sub.3-7cycloalkyl or C.sub.5-7
cycloalkenyl ring, or R.sup.2 and R.sup.3 together with the carbon
atom to which they are attached form an optionally substituted 5 to
7-membered heterocyclyl ring containing up to 3 heteroatoms
selected from N, O and S. Preferably R.sup.2 and R.sup.3
independently represent optionally substituted C.sub.1-6 alkyl.
[0040] Suitably, n is 0, 1, 2, 3 or 4. Preferably, n is 1.
[0041] Suitably, Y is NR.sup.10R.sup.11,
NR.sup.10C(Z)NR.sup.10R.sup.11,
NR.sup.10C(Z)NR.sup.10C(Z)OR.sup.11, NR.sup.10COOR.sup.11 or
NR.sup.10SO.sub.2R.sup.11.
[0042] Suitably, R.sup.11 is hydrogen, C.sub.1-6alkyl,
C.sub.3-7cycloalkyl, C.sub.3-7cycloalkyl C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, heterocyclyl, heterocyclylC.sub.1-6alkyl,
heteroaryl, or heteroarylC.sub.1-6alkyl, any of which may be
optionally substituted; or R.sup.10 and R.sup.11 together with the
nitrogen to which they are attached form a 5 to 7 membered
heterocyclic ring optionally containing an additional heteroatom
selected from O, S, or NR.sup.9.
[0043] While R.sup.11 is optionally substituted as defined in the
specification, it is preferably, an unsubstituted or substituted
alkyl, C.sub.3-7cycloalkyl, C.sub.3-7cycloalkyl C.sub.1-6alkyl,
heterocyclyl, heterocyclyl C.sub.1-6alkyl, heteroaryl, or a
heteroarylC.sub.1-6alkyl. The alkyl if substituted is preferably
substituted one or more times by halogen, such as in CF.sub.3,
CH.sub.2CF.sub.3, or (CH.sub.2).sub.2Cl or (CH.sub.2).sub.3Cl, or
by a C.sub.1-6alkoxy, a C.sub.1-6alkylthio, C.sub.1-6alkyl
sulphinyl or C.sub.1-6alkyl sulphonyl group; if R.sup.11 is a
C.sub.3-7cycloalkyl C.sub.1-6alkyl, it is preferably a 5 or 6
membered ring, such as cyclohexylmethyl; if R.sup.11 is a
heterocyclyl C.sub.1-6alkyl, it is preferably a morpholino
C.sub.1-6alkyl, or a piperidine C.sub.1-6alkyl; or if a
heteroarylC.sub.1-6alkyl moiety, an optionally substituted
isoxazolyl.
[0044] Suitably, R.sup.10 and R.sup.12 are independently selected
from hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl,
C.sub.3-7cycloalkyl C.sub.1-6alkyl, heterocyclyl,
heterocyclylC.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl, heteroaryl or
heteroarylC.sub.1-6alkyl, any of which may be optionally
substituted. The R.sup.10 group and R.sup.12 moieties may be
optionally substituted as defined for the alkyl term.
[0045] Suitably, R.sup.5 is hydrogen, C(Z)R.sup.12 or optionally
substituted C.sub.1-6alkyl, optionally substituted aryl, optionally
substituted arylC.sub.1-6alkyl, or S(O).sub.2R.sup.8.
[0046] Suitably, R.sup.6 is hydrogen or C.sub.1-6alkyl.
[0047] Suitably, R.sup.7 is hydrogen, C.sub.1-6alkyl,
C.sub.3-7cycloalkyl, aryl, arylC.sub.1-6alkyl, heteroaryl,
heteroarylC.sub.1-6alkyl, heterocyclyl, or
heterocyclylC.sub.1-6alkyl.
[0048] Suitably, R.sup.8 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl,
aryl, arylC.sub.1-6alkyl, heteroaryl, heteroarylC.sub.1-6alkyl,
heterocyclyl, or heterocyclylC.sub.1-6alkyl.
[0049] Suitably, R.sup.9 is hydrogen, C.sub.1-4alkyl,
C.sub.3-7cycloalkyl or aryl.
[0050] Suitably, R.sup.10 and R.sup.12 are independently selected
from hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, heterocyclyl,
heterocyclylC.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl, heteroaryl or
heteroarylC.sub.1-6alkyl, any of which may be optionally
substituted.
[0051] Suitably, Z is oxygen or sulfur.
[0052] Suitably, Ar is a napth-2-yl, napth-1-yl, a bicyclic or a
tricyclic heteroaromatic ring, which ring may be optionally
substituted in any ring. A bicyclic or tricyclic heteroaromatic
ring system is a fused ring system that may include a carbocyclic
ring. Examples of such ring systems include quinoline,
isoquinoline, benzimidazole, benzothiophene or benzofuran,
benzoxazole, benzthiazole, dibenzofuran, dibenzothiophene,
benzthiodiazole, benztriazole, or indolyl.
[0053] The Ar ring may be optionally substituted one or more times,
preferably 1 to 3 times, independently, in any ring. Suitable
substituents include halogen, C.sub.1-6alkyl, aryl,
arylC.sub.1-6alkyl, cycloalkyl, cycloalkyl C.sub.1-6alkyl,
C.sub.1-6alkoxyC.sub.1-6alkyl, halosubstituted C.sub.1-6alkyl,
arylC.sub.1-6alkoxy, (CR.sup.13R.sup.14).sub.tOR.sup.12, nitro,
cyano, (CR.sup.13R.sup.14).sub.t NR.sup.1OR.sup.1,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(Z)R.sup.12,
(CR.sup.13R.sup.14).sub.tC(Z)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tCOR.sup.12,
(CR.sup.13R.sup.14).sub.tZC(Z)R.sup.12,
(CR.sup.13R.sup.14).sub.tC(Z)OR.sup.12,
(CR.sup.13R.sup.14).sub.tC(O)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(Z)NR.sup.1OR.sup.1,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(.dbd.NH)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tC(.dbd.NH)--NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10S(O).sub.2R.sup.8,
(CR.sup.13R.sup.14).sub.tS(O).sub.2NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tS(O).sub.mR.sup.12, heterocyclyl,
heteroaryl, heterocyclylC.sub.1-6alkyl or heteroaryl
C.sub.1-6alkyl.
[0054] Suitably t is 0, or an integer having a value of 1 to 10.
Preferably t is 0, or 1, more preferably 0.
[0055] Suitably, R.sup.13 and R.sup.14 are independently hydrogen,
or a C.sub.1-6 alkyl.
[0056] Preferably, Ar is substituted one or more times by halo,
cyano, (CR.sup.13R.sup.14).sub.tC(Z)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.t C(Z)OR.sup.12, (CR.sup.13R.sup.14).sub.t
COR.sup.12, (CR.sup.13R.sup.14).sub.tS(O).sub.mR.sup.12,
(CR.sup.13R.sup.14).sub.t OR.sup.12, halo-substituted-C.sub.1-6
alkyl, C.sub.1-6 alkyl,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(Z)R.sup.12,
(CR.sup.13R.sup.14).sub.tNR.sup.10S(O).sub.2R.sup.8,
(CR.sup.13R.sup.14).sub.tS(O).sub.2NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.t ZC(Z)R.sup.12, or
(CR.sup.13R.sup.14).sub.tNR.sup.10R.sup.11.
[0057] Preferably, the Ar ring is substituted one or more times by
halo, hydroxy, C.sub.1-6alkyl, halosubstituted C.sub.1-6alkyl,
hydroxy C.sub.1-6alkyl, and C.sub.1-6alkoxy. More preferred
substitution is C.sub.1-6alkoxy group, such as methoxy; a C.sub.1-6
alkyl, such as methyl, or halogen, such as fluorine or
chlorine.
[0058] Preferably the Ar ring is a naphthyl ring, more preferably a
napth-2-yl ring. If Ar is a bicycloheteroaryl ring it is preferably
a benzothiophene or a benzofuran ring. A preferred ring placement
for the napth-2-yl ring is in the 6-position.
[0059] For use herein, the term "alkyl", and "alkenyl" groups,
individually or as part of a larger group e.g. "alkoxy", may be
straight or branched chain radicals containing up to six carbon
atoms, unless otherwise limited, including, but not limited to,
methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl,
tert-butyl, and the like. The alkyl and alkenyl groups may be
optionally substituted as herein defined.
[0060] For use herein, "cycloalkyl" includes cyclic radicals having
from three to eight ring carbon atoms, including but not limited to
cyclopropyl, cyclopentyl, cyclohexyl, and the like. The cycloalkyl
groups may be optionally substituted as herein defined.
[0061] For use herein, "cycloalkenyl" includes cyclic radicals,
preferably of 5 to 8 carbons, which have at least one bond
including but not limited to cyclopentenyl, cyclohexenyl, and the
like. The cycloalkenyl groups may be optionally substituted as
herein defined.
[0062] For use herein the term "aryl" (on its own or in any
combination, such as "arylalkyl" or "aryloxy") includes a single or
fused ring system, suitably containing from 4 to 7, preferably 5 or
6 ring atoms in each ring, which rings, may each be unsubstituted
or substituted by, independently for example, up to three
substituents. A fused ring system may include an aliphatic ring,
such as a saturated or partially saturated ring, and need include
only one aromatic ring. Suitable aryl groups include phenyl and
naphthyl such as 1-naphthyl or 2-naphthyl. The aryl rings may be
optionally substituted as herein defined unless otherwise
indicated. For use herein the term "heterocyclyl" (on its own or in
any combination, such as "heterocyclyl alkyl" or "heterocyclyl
oxy") suitably includes, unless otherwise defined, non-aromatic,
single and fused rings suitably containing up to four heteroatoms
in each ring, each of which independently selected from O, N and S,
and which rings, may be unsubstituted or substituted independently
by, for example, up to three substituents. Each heterocyclic ring
suitably has from 4 to 7, preferably 5 or 6, ring atoms. A fused
heterocyclic ring system may include carbocyclic rings and need
include only one heterocyclic ring. Examples of heterocyclyl groups
include pyrrolidine, piperidine, piperazine, morpholine,
imidazolidine and pyrazolidine. The heterocyclic and heterocyclic
rings may be optionally substituted as herein defined, unless
otherwise indicated.
[0063] When used herein, the term "heteroaryl" (on its own or in
any combination, such as "heteroaryloxy" or "heteroarylalkyl")
suitably includes, unless otherwise defined, mono- and bicyclic
heteroaromatic ring systems comprising up to four, preferably 1 or
2, heteroatoms each selected from O, N and S. Each ring may have
from 4 to 7, preferably 5 or 6, ring atoms. A bicyclic
heteroaromatic ring system may include a carbocyclic ring. Examples
of heteroaryl groups include pyrrole, quinoline, isoquinoline,
pyridine, pyrimidine, oxazole, thiazole, thiadiazole, triazole,
imidazole, benzimidazole, isoxazole, thiophene, benzothiophene,
furan and benzofuran. The heteroaryl rings may be optionally
substituted as defined herein unless otherwise indicated.
[0064] Suitably the when the term "optionally substituted" is used
herein, such as on the alkyl, alkenyl, cycloalkyl, cycloalkenyl,
aryl, arylalkyl, heterocyclyl, heterocyclicalkyl, heteroaryl, and
heteroarylalkyl groups, unless otherwise defined, shall mean that
the group may be optionally substituted one or more times,
preferably by one to three substituents, each independently
selected from halogen, C.sub.1-6alkyl, aryl, arylC.sub.1-6alkyl,
cycloalkyl, cycloalkyl C.sub.1-6alkyl,
C.sub.1-6alkoxyC.sub.1-6alkyl, halosubstituted C.sub.1-6alkyl,
arylC.sub.1-6alkoxy, (CR.sup.13R.sup.14).sub.tOR.sup.12, nitro,
cyano, (CR.sup.13R.sup.14).sub.tNR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(Z)R.sup.12,
(CR.sup.13R.sup.14).sub.tC(Z)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tCOR.sup.12,
(CR.sup.13R.sup.14).sub.tZC(Z)R.sup.12, (CR.sup.13R.sup.14).sub.t
C(Z)OR.sup.12, (CR.sup.13R.sup.14).sub.tC(O)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(Z)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10C(.dbd.NH)NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tC(.dbd.NH)--NR.sup.10R.sup.11,
(CR.sup.13R.sup.14).sub.tNR.sup.10S(O).sub.2R.sup.8,
(CR.sup.13R.sup.14).sub.tS(O).sub.2NR.sup.10R.sup.11,
S(O).sub.mR.sup.12, heterocyclyl, heteroaryl,
heterocyclylC.sub.1-6alkyl or heteroaryl C.sub.1-6alkyl. In
addition, two adjacent ring carbon atoms may be linked to form a
bicyclic system.
[0065] Particular compounds according to the invention include
those mentioned in the examples and their pharmaceutically
acceptable salts.
[0066] It will be appreciated that for use in medicine the salts of
the compounds of formula (I) should be pharmaceutically acceptable.
Suitable pharmaceutically acceptable salts will be apparent to
those skilled in the art and include those described in J. Pharm.
Sci., 1977, 66, 1-19, such as acid addition salts formed with
inorganic acids e.g. hydrochloric, hydrobromic, sulphuric, nitric
or phosphoric acid; and organic acids e.g. succinic, maleic,
acetic, fumaric, citric, tartaric, benzoic, p-toluenesulfonic,
methanesulfonic or naphthalenesulfonic acid. Other salts e.g.
oxalates, may be used, for example in the isolation of compounds of
formula (I) and are included within the scope of this
invention.
[0067] The compounds of this invention may be in crystalline or
non-crystalline form, and, if crystalline, may optionally be
hydrated or solvated. This invention includes within its scope
stoichiometric hydrates as well as compounds containing variable
amounts of water.
[0068] The invention extends to all isomeric forms including
stereoisomers and geometric isomers of the compounds of formula (I)
including enantiomers and mixtures thereof e.g. racemates. The
different isomeric forms may be separated or resolved one from the
other by conventional methods, or any given isomer may be obtained
by conventional synthetic methods or by stereospecific or
asymmetric syntheses.
[0069] Since the compounds of formula (I) are intended for use in
pharmaceutical compositions it will readily be understood that they
are each preferably provided in substantially pure form, for
example at least 60% pure, more suitably at least 75% pure and
preferably at least 85%, especially at least 98% pure (% are on a
weight for weight basis). Impure preparations of the compounds may
be used for preparing the more pure forms used in the
pharmaceutical compositions.
[0070] Compounds of formula (I) are imidazole derivatives which may
be readily prepared using procedures well-known to those skilled in
the art, and described in, for instance, Comprehensive Heterocyclic
Chemistry, Editors Katritzky and Rees, Pergamon Press, 1984, 5,
457-497, from starting materials which are either commercially
available or can be prepared from such by analogy with well-known
processes. A key step in many such syntheses is the formation of
the central imidazole nucleus, to give compounds of formula (I).
These patents describe the synthesis of .alpha.-ketooximes and
.alpha.-hydroxyketones (benzoins) and their subsequent use in
preparing imidazoles and N-hydroxylimidazoles. Thereafter, further
compounds of formula (I) may be obtained by manipulating
substituents in any of the groups HetAr, Ar and Y.sub.1 using
conventional functional group interconversion procedures.
[0071] In particular, in a first process, compounds of formula (I)
may be prepared by condensing an alpha-diketone of formula (II):
HetArCOCOAr (II) wherein Ar is defined for formula (I), or an
equivalent thereof and HetAr is defined as the ##STR2## fragment of
Formula I, (it should be noted that in the schemes below R.sup.1 is
shown as X--R.sup.4 for representative purposes only); with an
aldehyde of the formula (III): ##STR3## wherein Y.sub.1 is a group
convertible to Y, and R.sup.2 and R.sup.3 is as hereinbefore
defined, or an equivalent thereof, and, if necessary, with ammonia
or a source thereof, under imidazole-ring forming conditions.
[0072] The group Y.sub.1, as used in these schemes, is a group
convertible to Y. The conversions to Y are well known to the
skilled artisan using readily available techniques. For instance,
Y.sub.1 could be a protected amine, such as CBZ or and S--BOC
amine, which is deprotected and converted to Y; alternatively
Y.sub.1 could be a carboxylic acid or a carboxylic acid ester,
which can be converted by well known means to an amine, a bromide
or an azide (i.e., a 1 carbon degradation) and further
functionalized; alternatively Y.sub.1 could be a protected alcohol,
and using standard functional group interconversion converted to an
amine, such as azide and reduced, etc. to the group Y.
[0073] Suitable equivalents of the alpha-diketone are well known to
those skilled in the art and include the corresponding
alpha-keto-oxime and alpha-dioxime. Suitable equivalents of the
aldehyde of formula (III) are well known in the art and include the
corresponding oxime and acetal.
[0074] Ammonia, or a source thereof, is preferably used in excess,
with at least a dimolar amount being used in the case of the
alpha-diketone and at least an equimolar amount in the case of the
alpha-keto-oxime.
[0075] Suitable sources of ammonia include ammonium salts of
organic carboxylic acids, such as ammonium trifluoroacetate or an
ammonium C.sub.1-6 alkanoate, for instance ammonium acetate and
ammonium formate, preferably ammonium acetate, and carboxylic
amides, in particular of formic acid, such as formamide. An
ammonium salt is generally used in large excess and in the presence
of an acid, such as a C.sub.1-6 carboxylic acid which acid may also
be used as a solvent for the reaction. If formamide is used, this
may be used in excess, as the reaction solvent. An alternative
solvent such as ethanol or dimethyl sulphoxide (Lantos et al, J Het
Chem, 19, 1375, 1982) may be used. An additional solvent may also
be employed, for instance, dimethyl formamide may be used with
formamide. The reaction is generally carried out at elevated
temperatures, for instance under reflux conditions, and if desired,
in a sealed vessel optionally under pressure and/or an inert gas
atmosphere, for instance nitrogen. It is also possible to run the
reaction at elevated temperature in a molten salt, which contains a
source of ammonia, for example in ammonium trifluoroacetate.
[0076] A further suitable source of ammonia is hydroxylamine, in
which case the initially formed imidazole is an N-hydroxy-N-oxide
imidazole. This may then be reduced to the corresponding N-hydroxy
imidazole by treating with a suitable reducing agent such as sodium
borohydride, in an appropriate solvent such as methanol, following
the method of Akange and Allan, Chem and Ind, 5 Jan. 1975, 38. The
N-hydroxy imidazole may in turn be converted to an imidazole of
formula (I) by treatment with a conventional deoxygenating agent
such as titanium trichloride, phosphorus trichloride or a
trialkylphosphite such as trimethyl- or triethyl-phosphite.
N-hydroxy-N-oxide imidazoles may be readily obtained by treating an
alpha-diketone of formula (II) with an aldehyde of formula (III)
with about two equivalents of hydroxylamine or the corresponding
aldoxime and about one equivalent of hydroxylamine, under proton
catalysis. Alternatively, the N-oxide may be obtained by the acid
catalyzed condensation of the corresponding alpha-dioxime or
alpha-keto-oxime with an aldoxime of the aldehyde of formula
(III).
[0077] When the compound of formula (II) above is an
alpha-keto-oxime derivative, it will be appreciated that the
product initially obtained will be a compound of formula (I) in
which the imidazole is N-hydroxylated and which may be converted
into a compound of formula (I) as described above.
[0078] It will be appreciated by those skilled in the art that in
some instances, it will not be necessary to provide a separate
source of ammonia as the alpha-diketone or aldehyde equivalent may
already contain such a source. Examples of this include
alpha-dioxime or alpha-keto-oxime and aldoxime.
[0079] Preferred methods for preparing compounds of this invention
are as outlined in the Schemes I-III.
[0080] In Scheme I, the anion prepared from 1 (a specific example
being V=CH, X=S, R.sub.1=Me), by treatment with a strong base such
as lithium di-iso-propylamide, is condensed with an aryl aldehyde,
to give, after removal of the protecting group, the diol 2. This
may then be converted to the alpha-diketone 3 by a Swern oxidation
of which any number of potentially useful variations are known and
may be used. The alpha-diketone 3 is then cyclised to an imidazole
4 by heating 3 with a substituted aldehyde of formula (III) in a
mixture of ammonium acetate, as the source of ammonia, and an
appropriate solvent, for example acetic acid or DMSO. The imidazole
4, with the group Y.sub.1 may be converted into a group Y using
conventional functional group interconversion procedures to afford
imidazole 5, a compound of formula I. Functional group
transformations are well known in the art and are described in, for
instance, Comprehensive Organic Functional Group Transformations,
eds. A. R. Katritzky, O. Meth-Cohn, and C. W. Rees (Elsevier
Science Ltd., Oxford, 1995), Comprehensive Organic Chemistry, eds.
D. Barton and W. D. Ollis (Pergamon Press, Oxford, 1979), and
Comprehensive Organic Transformations, R. C. Larock (VCH Publishers
Inc., New York, 1989). An example of a preferred Y.sub.1 group is
CH.sub.2NH.sub.2 or a protected form thereof e.g. CH.sub.2NHBoc.
Scheme I also illustrates the preparation of a protected
alpha-hydroxyketone 6, by condensing the anion of 1 with an
appropriately activated carbonyl derivative of a substituted
benzamide, such as the N-methoxy-N-methylamide, to yield a
protected a-hydroxyketone. This adduct 6 may then be directly
converted to the imidazole 4, using a combination of a copper (II)
salt, such as copper (II) acetate, as an oxidizing agent and
ammonium acetate as a source of ammonia. The alpha-hydroxyketone 6
may also be deprotected and then oxidized to give an alpha-diketone
3, for instance using Swern oxidation. ##STR4##
[0081] Scheme II illustrates the use of an alpha-keto-oxime for
preparing a compound of formula (I). A heterocyclic ketone 7 (for
example, V=N, X=S, R.sub.1=Me) is prepared by adding the anion of
2-methylthio-6-methylpyrimidine (prepared by treatment thereof with
an alkyl lithium, such as n-butyl lithium) to an
N-alkyl-O-alkoxybenzamide. Alternatively, the anion may be
condensed with a benzaldehyde, to give an alcohol that is then
oxidized to the ketone 7. The alpha-keto-oxime 8 is then prepared
from 7 using standard conditions, such as reaction with sodium
nitrite, and this may then be reacted with an aldehyde of formula
(III) to afford an N-hydroxyimidazole 9. This may converted to 10,
by treating it with a deoxygenating agent such as titanium
trichloride, phosphorus trichloride or a trialkyl phosphite, such
as trimethyl or triethylphosphite. Functional group interconversion
then may be employed to convert Y.sub.1 of 10 to afford 11, a
compound of formula (I) with Y as hereinbefore defined.
Furthermore, oxidation of the sulfur (X.dbd.S) of 5 or 11 to the
sulfoxide or sulfone followed by nucleophilic displacement with
oxygen, nitrogen or sulfur nucleophiles affords compounds of
Formula I (X.dbd.O, N, or S; compounds 5 and 11 in Schemes I and
II). ##STR5##
[0082] Compounds of the general formula (V.dbd.CH, X.dbd.O, N, S)
can be prepared as in Schemes I or II except substituting
4-methyl-2-chloropyridine or 4-methyl-2-fluoropyridine for the
starting 2-methylthio-6-methylpyrimidine or pyridine (1) (Gallagher
et al Bioorg. Med. Chem. 5, 49, 1997). Nucleophilic substitution of
the resulting 2-halopyridinylimidazole can be effected by the
procedure described in U.S. Pat. No. 5,670,527. Alternatively,
oxidation of the sulfur (X.dbd.S) to the sulfoxide or sulfone
followed by nucleophilic displacement with oxygen, nitrogen or
sulfur nucleophiles affords compounds of Formula I (compounds 5 and
11 in Schemes I and II).
[0083] Alternatively compounds may be prepared as in Scheme III
wherein the Ar group is added last. (X, R.sup.2, R.sup.3, and Y are
as defined for formula (I), and Y.sub.1 is a group convertible to
Y). ##STR6##
[0084] Oxidation of a 4-acetyl substituted pyridine derivative
(V.dbd.CH, X.dbd.S, R.sub.1=Me) with sodium nitrite affords the
ketooxime. Heating this product with an alkyl aldehyde and ammonium
acetate in acetic acid allows access to the imidazole nucleus.
Reduction of the hydroxyimidazole may be accomplished with heating
with a trialkyl phosphite or stirring at ambient temperature with
titanium trichloride. Treatment of the imidazole with
N-iodosuccinimide gives the iodoimidazole with then may be reacted
with various boronic acids under palladium catalysis to give the
aryl or heteroaryl imidazoles. Alternative biaryl coupling
reactions may also be used.
[0085] During the synthesis of the compounds of formula (I) labile
functional groups in the intermediate compounds, e.g. hydroxy,
carboxy and amino groups, may be protected. A comprehensive
discussion of the ways in which various labile functional groups
may be protected and methods for cleaving the resulting protected
derivatives is given in for example Protective Groups in Organic
Chemistry, T. W. Greene and P. G. M. Wuts, (Wiley-Interscience, New
York, 2nd edition, 1991).
[0086] The compounds of formula (I) may be prepared singly or as
compound libraries comprising at least 2, for example 5 to 1,000
compounds, and more preferably 10 to 100 compounds of formula (I).
Libraries of compounds of formula (I) may be prepared by a
combinatorial `split and mix` approach or by multiple parallel
synthesis using either solution phase or solid phase chemistry, by
procedures known to those skilled in the art.
[0087] Thus according to a further aspect of the invention there is
provided a compound library comprising at least 2 compounds of
formula (I), or pharmaceutically acceptable salts thereof.
[0088] Pharmaceutically acceptable salts may be prepared
conventionally by reaction with the appropriate acid or acid
derivative.
Methods of Treatment
[0089] The Tie receptors are proteins of approximately 125 kDa,
with a single putative transmembrane region. The extracellular
domain of these receptors is divided into several regions: there
are 3 regions that have a pattern of cysteine expression found in
EGF-like domains; there are 2 regions that have some weak homology
to and structural characteristics of immunoglobulin-like domains;
and there are 3 regions with homology to the fibronectin III repeat
structure. This particular combination of extracellular domains is
unique to the Tie receptors. The intracellular portion of Tie2 is
most closely related (.about.40% identity) to the kinase domains of
FGF-R1, PDGF-R and c-kit. The intracellular portions of Tie2
contain all of the features of tyrosine kinases, including a GXGXXG
ATP binding site consensus sequence and typical tyrosine kinase
motifs (i.e., HRDLAARN and DFGL).
[0090] These receptors have sparked interest because they are the
only receptor tyrosine kinases, other than those receptors for
vascular endothelial cell growth factor (VEGF), that are largely
restricted to endothelial cells in their expression. There are
several lines of evidence showing that Tie2 is important in
angiogenesis, detailed in the following paragraphs.
[0091] a. Tie1 and Tie2 Receptor Location
[0092] i. Embryological Vascular Development
[0093] The location of the Tie receptors in the embryo has been
studied by a number of investigators using in situ hybridization.
Korhonen et al. (Blood 80:2548-2555, 1992) showed that the mRNA for
Tie receptors is located in endothelial cells of all forming blood
vessels and in the endocardium of mouse embryos. During embryonic
development, expression of the Tie receptors is seen in angioblasts
and all developing vasculature. Expression of the Tie receptors
follows expression of the major VEGF receptor, Flk-1, by 12-24
hours during mouse embryogenesis, perhaps suggesting sequential and
different actions of these receptor systems (Schnurch and Risau,
Development 119: 957-968, 1993). Cloning of a 1.2 Kb genomic 5'
flanking region of Tie2, coupled to a lacZ gene and expressed in
transgenic mice, demonstrated a selective pattern of expression in
endothelial cells during embryonic development (Schlaeger et al.,
Development 121:1089-1098, 1995). Thus, the Tie2 promoter acts to
assure endothelial cell-specific expression of Tie2.
[0094] ii. In Adult Tissues
[0095] The similarities between embryonic angiogenesis and
pathologic angiogenesis yields the hypothesis that blocking Tie2
function, in tumors or chronic inflammatory settings, for examples,
may block angiogenesis, thus blocking further cell proliferation
and provide therapeutic benefit. Tie mRNA cannot be observed in
adult skin, except at sites of active wound healing, where the
proliferating capillaries in the granulation tissue contain
abundant Tie mRNA (Korhonen et al., Blood 80:2548-2555, 1992). PCR
amplification of cDNA from normal skin fails to show a signal for
Tie receptor (Kaipainen et al., Cancer Res. 54:6571-6577, 1994). In
contrast, a strong signal is seen with cDNA from metastasizing
melanomas, where in situ studies localize this signal to the
vascular endothelium. While Tie receptor expression is
down-regulated in the established vasculature, it is upregulated in
the angiogenesis that occurs in the ovary during ovulation, in
wounds and in tumor (breast, melanoma and renal cell carcinoma)
vasculature, consistent with prevailing views that angiogenesis in
the adult borrows from embryonic angiogenic mechanisms.
[0096] b. Tie Knockout Animals
[0097] Homozygous mice with a Tie2 knockout, or carrying a
transgene encoding a "dominant-negative" Tie2 receptor, confirmed
that the Tie2 receptor is critical for embryonic development
(Dumont et al., Genes Dev. 8:1897-1909, 1994; Sato et al., Nature
376:70-74, 1995). Embryonic death in these mice occurred due to
vascular insufficiency and there were dramatically reduced numbers
of endothelial cells. Vasculogenesis--that is the differentiation
of endothelial cells and the in situ formation of vessels--appeared
relatively normal in mice lacking Tie2. The subsequent sprouting
and remodelling resulting in formation of vessel branches
(angiogenesis) was drastically reduced in the Tie2 mutant mice
embryos. This lack of sprouting and angiogenesis resulted in
substantial growth retardation, particularly of the brain, neural
tube and heart, resulting in lack of viability. This exemplifies
the critical importance of Tie2 in angiogenesis. This is
significant, as angiogenesis is regulated by a number of growth
factors. Interestingly, Flk1 (VEGF receptor) knockout mice exhibit
embryo lethal defects in vasculogenesis that occur earlier than
those of Tie2 disruption. Disruption of the Tie1 receptor yields a
much different, and later, defective phenotype; the mouse embryo
dies late in development due to haemorrhage resulting from
defective integrity of an otherwise well formed vasculature. Taken
together, these studies suggest that the VEGF/Flk1 and Tie systems
operate in sequential fashion, with Tie2 having a critical role in
angiogenesis.
[0098] c. Tie2 Ligands
[0099] Recently, two ligands for the Tie2 receptor have been
reported. Angiopoietin-1 binds and induces the tyrosine
phosphorylation of Tie2 and its expression in vivo is in close
proximity with developing blood vessels (Davis et al., Cell
87:1161-1169, 1996). Mice engineered to lack Angiopoietin-1 display
angiogenic deficits reminiscent of those previously seen in mice
lacking Tie2 receptors, demonstrating that Angiopoietin-1 is a
primary physiologic ligand for Tie2 and that Tie2 has critical in
vivo angiogenic actions (Suri et al., Cell 87:1171-1180, 1996).
Angiopoietin-2 was identified by homology screening and shown to be
a naturally occurring antagonist for Tie2 receptors. Transgenic
overexpression of Angiopoietin-2 disrupts blood vessel formation in
the mouse embryo (Maisonpierre et al., Science 277:55-60, 1997).
Together, these results support a role for Tie2 receptors in
angiogenesis.
[0100] In order to use the compounds of formula (I) in therapy,
they will normally be formulated into a pharmaceutical composition
in accordance with standard pharmaceutical practice.
[0101] According to a further aspect of the invention there is
provided a pharmaceutical composition comprising a compound of
formula (I) or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
[0102] No toxicological effects are indicated/expected when a
compound of formula (I) is administered in the above mentioned
dosage range.
[0103] Compounds of Formula (I), pharmaceutically acceptable salts
thereof and pharmaceutical compositions incorporating such may
conveniently be administered by any of the routes conventionally
used for drug administration, for instance, orally, topically,
parenterally or by inhalation. The compounds of Formula (I) may be
administered in conventional dosage forms prepared by combining a
compound of Formula (I) with standard pharmaceutical carriers
according to conventional procedures. The compounds of Formula (I)
may also be administered in conventional dosages in combination
with a known, second therapeutically active compound. These
procedures may involve mixing, granulating and compressing or
dissolving the ingredients as appropriate to the desired
preparation. It will be appreciated that the form and character of
the pharmaceutically acceptable character or diluent is dictated by
the amount of active ingredient with which it is to be combined,
the route of administration and other well-known variables. The
carrier(s) must be "acceptable" in the sense of being compatible
with the other ingredients of the formulation and not deleterious
to the recipient thereof.
[0104] The pharmaceutical carrier employed may be, for example,
either a solid or liquid. Exemplary of solid carriers is lactose,
terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium
stearate, stearic acid and the like. Exemplary of liquid carriers
is syrup, peanut oil, olive oil, water and the like. Similarly, the
carrier or diluent may include time delay material well known to
the art, such as glyceryl mono-stearate or glyceryl distearate
alone or with a wax.
[0105] A wide variety of pharmaceutical forms can be employed.
Thus, if a solid carrier is used, the preparation can be tableted,
placed in a hard gelatin capsule in powder or pellet form or in the
form of a troche or lozenge. The amount of solid carrier will vary
widely but preferably will be from about 25 mg. to about 1 g. When
a liquid carrier is used, the preparation will be in the form of a
syrup, emulsion, soft gelatin capsule, sterile injectable liquid
such as an ampule or nonaqueous liquid suspension.
[0106] Compounds of Formula (I) may be administered topically, that
is by non-systemic administration. This includes the application of
a compound of Formula (I) externally to the epidermis or the buccal
cavity and the instillation of such a compound into the ear, eye
and nose, such that the compound does not significantly enter the
blood stream. In contrast, systemic administration refers to oral,
intravenous, intraperitoneal and intramuscular administration.
[0107] Formulations suitable for topical administration include
liquid or semi-liquid preparations suitable for penetration through
the skin to the site of inflammation such as liniments, lotions,
creams, ointments or pastes, and drops suitable for administration
to the eye, ear or nose. The active ingredient may comprise, for
topical administration, from 0.001% to 10% w/w, for instance from
1% to 2% by weight of the formulation. It may however comprise as
much as 10% w/w but preferably will comprise less than 5% w/w, more
preferably from 0.1% to 1% w/w of the formulation.
[0108] Lotions according to the present invention include those
suitable for application to the skin or eye. An eye lotion may
comprise a sterile aqueous solution optionally containing a
bactericide and may be prepared by methods similar to those for the
preparation of drops. Lotions or liniments for application to the
skin may also include an agent to hasten drying and to cool the
skin, such as an alcohol or acetone, and/or a moisturizer such as
glycerol or an oil such as castor oil or arachis oil.
[0109] Creams, ointments or pastes according to the present
invention are semi-solid formulations of the active ingredient for
external application. They may be made by mixing the active
ingredient in finely-divided or powdered form, alone or in solution
or suspension in an aqueous or non-aqueous fluid, with the aid of
suitable machinery, with a greasy or non-greasy base. The base may
comprise hydrocarbons such as hard, soft or liquid paraffin,
glycerol, beeswax, a metallic soap; a mucilage; an oil of natural
origin such as almond, corn, arachis, castor or olive oil; wool fat
or its derivatives or a fatty acid such as steric or oleic acid
together with an alcohol such as propylene glycol or a macrogel.
The formulation may incorporate any suitable surface active agent
such as an anionic, cationic or non-ionic surfactant such as a
sorbitan ester or a polyoxyethylene derivative thereof. Suspending
agents such as natural gums, cellulose derivatives or inorganic
materials such as silicaceous silicas, and other ingredients such
as lanolin, may also be included.
[0110] Drops according to the present invention may comprise
sterile aqueous or oily solutions or suspensions and may be
prepared by dissolving the active ingredient in a suitable aqueous
solution of a bactericidal and/or fungicidal agent and/or any other
suitable preservative, and preferably include a surface active
agent. The resulting solution may then be clarified by filtration,
transferred to a suitable container which is then sealed and
sterilized by autoclaving or maintaining at 98-100.degree. C. for
half an hour. Alternatively, the solution may be sterilized by
filtration and transferred to the container by an aseptic
technique. Examples of bactericidal and fungicidal agents suitable
for inclusion in the drops are phenylmercuric nitrate or acetate
(0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate
(0.01%). Suitable solvents for the preparation of an oily solution
include glycerol, diluted alcohol and propylene glycol.
[0111] Compounds of formula (I) may be administered parenterally,
that is by intravenous, intramuscular, subcutaneous intranasal,
intrarectal, intravaginal or intraperitoneal administration. The
subcutaneous and intramuscular forms of parenteral administration
are generally preferred. Appropriate dosage forms for such
administration may be prepared by conventional techniques.
Compounds of Formula (I) may also be administered by inhalation,
that is by intranasal and oral inhalation administration.
Appropriate dosage forms for such administration, such as an
aerosol formulation or a metered dose inhaler, may be prepared by
conventional techniques.
[0112] The compounds of Formula (I) may also be used topically in
the treatment or prophylaxis of disease states exacerbated by
excessive or inappropriate angiogenesis.
[0113] The compounds of Formula (I) are administered in an amount
sufficient to inhibit Tie2 receptor activity such that it is
regulated down to normal levels, or in some case to subnormal
levels, so as to ameliorate or prevent the disease state.
[0114] Chronic diseases which have an inappropriate angiogenic
component are various ocular neovasularizations, such as diabetic
retinopathy and macular degeneration.
[0115] Other chronic diseases which have an excessive or increased
proliferation of vasculature are tumor growth and metastasis,
athrosclerosis, and certain arthritic conditions. Therefore Tie2
tyrosine kinase receptor inhibitors will be of utility in the
blocking of the angiogenic component of these disease states.
[0116] The term "excessive or increased proliferation of
vasculature inappropriate angiogenesis" as used herein includes,
but not limited to, diseases which are characterized by hemangiomas
and ocular diseases. The term "inappropriate angiogenesis" as used
herein includes, but not limited to, diseases which are
characterized by vessel proliferation with accompanying tissue
proliferation, such as occurs in cancer, metastasis, arthritis,
psoriasis and atherosclerosis.
[0117] For all methods of use disclosed herein for the compounds of
Formula (I), the daily oral dosage regimen will preferably be from
about 0.1 to about 80 mg/kg of total body weight, preferably from
about 0.2 to 30 mg/kg, more preferably from about 0.5 mg to 15 mg.
The daily parenteral dosage regimen will be from about 0.1 to about
80 mg/kg of total body weight, preferably from about 0.2 to about
30 mg/kg, and more preferably from about 0.5 mg to 15 mg/kg. The
daily topical dosage regimen will preferably be from 0.1 mg to 150
mg, administered one to four, preferably two or three times daily.
The daily inhalation dosage regimen will preferably be from about
0.01 mg/kg to about 1 mg/kg per day. It will also be recognized by
one of skill in the art that the optimal quantity and spacing of
individual dosages of a compound of Formula (I) or a
pharmaceutically acceptable salt thereof will be determined by the
nature and extent of the condition being treated, the form, route
and site of administration, and the particular patient being
treated, and that such optimums can be determined by conventional
techniques. It will also be appreciated by one of skill in the art
that the optimal course of treatment, i.e., the number of doses of
a compound of Formula (I) or a pharmaceutically acceptable salt
thereof given per day for a as defined number of days, can be
ascertained by those skilled in the art using conventional course
of treatment determination tests.
Pharmacological Test Methods
A. Measurement of Tie2 Kinase Activity
[0118] A partial cDNA clone for the Tie2 receptor was used to make
protein for Tie kinase studies. In order to generate the primary
screening assay, a baculovirus expressed GST fusion for Tie2 kinase
domain was constructed and expressed using the commercial vector
pAcG1 (Pharmingen).
[0119] This final construct was transfected into Baculovirus and
soluble GST fusion products used in the screening assay. Prior work
had demonstrated the use of a baculovirus expressed GST fusion for
the murine Tie2 kinase domain to screen for candidate
target/signaling molecules (Huang et al., Oncogene 11:2097-2103,
1995).
Tie2 kinase activity assay: The Tie2 kinase activity assay was
typically run in one of 2 ways described as follows. Minor
variations in the assay give similar results.
1. Autophosphorylation Flashplate Assay
Materials:
Kinase buffer (final 20 mM Tris-HCl, pH7.0, 100 mM NaCl, 12 mM
MgCl.sub.2, 1 mM DTT).
Gamma .sup.33p-ATP (usually final amount of 0.5-1 uCi/well)
ATP (final 30 uM, or other desired concentration)
Flashplate (96-well, polystyrene microplate with plastic
scintillator coated wells)
TopCount (microplate scintillation counter)
Procedures:
Turn on incubator shaker and adjust temperature to 30.degree.
C.
Add 20 ul of 3.times. kinase buffer per well to the Flashplate
Add 20 ul of protein per well except for background. Compounds
added, typically in DMSO stocks, at 1-2 ul.
Add 20 ul mixture of gamma .sup.33p-ATP and cold ATP per well.
Total volume is 60 ul.
Cover with transparent polyester film.
Incubate at 30.degree. C. for two hours in shaker, or desired
time.
Take Flashplate out of the shaker, wash five times (for example,
with 300 ul of 10 uM ATP in 1.times.PBS per well).
Read plate on TopCount or other counting instrument. Results are
calculated as % inhibition
and IC50, using normal calculation methods.
[0120] Representative compounds of Formula (I), Example 1 was found
to be active in the assay, having an IC50 of <1 uM.
2. Fluorescence Polarization for Tie 2 Kinase
Final Assay Conditions:
[0121] 50 mM HEPES pH 7.5
[0122] 2% DMSO (when screening compounds)
[0123] 250 uM ATP
[0124] 2 mM MgCl.sub.2
[0125] 1 mM DTT
[0126] 50 uM NaVanidate
[0127] 10 uM peptide substrate
[0128] activated tie 2 kinase * see activation protocol below
Peptide Substrate:
[0129] RFWKYEFWR--OH
[0130] MW (TFA salt)=1873 Da
[0131] Make a 1 mM peptide stock and store at -20.degree. C.
[0132] Dilute to 100 uM just prior to use.
9.times. Kinase Buffer:
[0133] 450 mM HEPES pH 7.5 [0134] 900 mM NaCl [0135] 450 uM
NaVanidate [0136] 18 mM MgCl.sub.2 [0137] 100 mM DTT
[0138] Can be made up ahead of time and stored in aliquots at
-20.degree. C.
ATP Stock:
[0139] Make a 25 mM ATP stock and store in aliquots at -20.degree.
C. until needed.
[0140] Dilute to 2.5 mM prior to use.
Procedure:
[0141] For a 50 ul reaction add the following to each well of a
96-well black half-area plate (Costar, cat# 3694) [0142] 1. 5 ul of
compound in 20% DMSO [0143] 2. 5 ul 9.times. kinase buffer. [0144]
3. 5 ul 2.5 mM ATP. [0145] 4. 5 ul 100 uM peptide substrate. [0146]
5. 25 ul PTK detection mix (Panvera, P-2652, 50 ml--UK distributor
is Cambridge Bioscience) [0147] 6. 5 ul activated tie 2 kinase
(protocol below) diluted in 1.times. buffer to initiate the
reaction. [0148] 7. Read polarization on an FP instrument cycling
for 30-50 minutes in accordance with enzyme activity.
[0149] Representative compounds of Formula (I), Example 1 was found
to be active in this fluorescence assay, having an IC50 of <1
uM.
Activation of Tie 2 Kinase Protocol:
Final Buffer Conditions:
[0150] 20 mM Tris-HCl pH 7.5
[0151] 12 mM MgCl.sub.2
[0152] 100 mM NaCl
[0153] 20 uM NaVanidate
[0154] 1 mM DTT
[0155] 300 uM ATP
Procedure
[0156] 1. Incubate 5 uM tie 2 kinase in the 300 uM ATP and the
buffering conditions described above. [0157] 2. Allow to incubate
at 27.degree. C. for 2 hours. [0158] 3. Add 2.5 ml reaction to a
NAP-25 desalting column (Pharmacia Biotech cat. no. 17-0852-02)
pre-equilibrated in 20 mM Tris-HCl pH 7.5, 100 mM NaCl.sub.2 to
separate the ATP from the enzyme. [0159] 4. Elute the enzyme with
5.0 ml 20 mM Tris-HCl pH 7.5, 100 mM NaCl.sub.2; the protein
concentration should be 2.5 uM at this point. [0160] 5. Aliquot out
the enzyme and store at -80.degree. C. as soon as possible. B.
Measurement of Tie2 Receptor Signal Transduction--a Cellular
Assay
[0161] HEL cells (ATCC # TIB180) are cultured at between 1 and
5.times.10.sup.5 ml in RPMI-1640 medium supplemented with 2 mM
glutamine and 10% FBS as a suspension culture. Sixteen to
thirty-six hours prior to an experiment, the necessary number of
cells are passaged into 0.5% FBS/RPMI medium. On the day of an
experiment, cells are harvested and resuspended at a density of
0.5-1.0.times.10.sup.7 cells ml in 0.5% FBS RPMI and seeded at 2-3
ml/well in six well plates.
[0162] Alternatively, Human Umbilical Vein Endothelial Cells
(HUVECs) (Clonetics--Walkersville, Md.) may be used for the assay.
HUVECs between passages 2 and 12, are plated at 2.times.10.sup.5
and 1.times.10.sup.6 cells per well in a six well plate in
supplemented EGM (Clonetics). After 24 hours the media is changed
to EBM containing 3% BSA (Clonetics), and the cells are cultured
overnight and used for assay the following day.
[0163] Cells are treated with inhibitory compounds at appropriate
concentrations for 30-45 minutes. The contents of the wells are
mixed briefly on a rocker (approx. 30 seconds) and then incubated
at 37.degree. C. The cells are then treated with a source of native
ligand, such as serum or fibroblast conditioned medium for 10
minutes
[0164] At the end of 10 minute incubation period the plate is
placed on ice. The cells are harvested and the media is removed.
The cells are lysed in denaturing sample buffer
(Invitrogen--Carlsbad, Calif.). The suspension is sonicated for 5
pulses at a medium setting and returned to ice. The phosphorylation
state of the Tie2 receptor is determined Western blotting-and
detection by an anti-phospho-Tie2 antibody, as detailed below
(Harlow, E., and Lane, D. P., Antibodies--A Laboratory Manual, Cold
Spring Harbor Laboratory Press: New York, 1988.). Thirty ul of the
lysate are run on a 7.5% SDS/polyacrylamide gel. The gel is then
transferred to a nitrocellulose or PVDF membrane as per the
manufacturer's instructions for Western blotting.
[0165] The blots are washed with PBS 0.05% between -20 and then
blocked with 3% BSA/PBS/Tween for 1 hour at room temperature. The
blots are then incubated with 1 ug/ml anti-phospho-Tie2 antibody
(SmithKline Beecham) in PBS/0.05% Tween for 1 hour. The blot is
then washed 4 times with PBS/Tween for 5 minutes each. The blot is
incubated with an anti-mouse-HRP conjugate secondary antibody at
the dilution recommended by the manufacturer, in PBS/Tween for 1
hour. The blot is washed in PBS/Tween, 4 times for 5 minutes each.
After the last wash, the blot is developed by the ECL method
(Amersham) or some equivalent.
[0166] Using a densitometer or graphics program (e.g.
ImageQuant--Molecular Dynamics), each blot is scanned. The Tie-2
band is isolated and "boxed out" for each lane. The pixel volume or
comparable measure for each sample is analyzed. Also, an
appropriate background region of the same dimensions is determined
for each sample. After adjusting for background, phosphorylation is
expressed as the ratio of phosphotyrosine staining, relative to the
untreated control.
Angiogenesis In Vivo Model
Measurement of Angiogenesis In Vivo--Murine Air Pouch Granuloma
Model:
[0167] Described below a model of inflammatory angiogenesis used to
show that inhibition of Tie2 will stop the tissue destruction of
excessive or inappropriate proliferation of blood vessels. The
murine airpouch granuloma model of chronic inflammation (Kimura et
al., 1985, J. Pharmacobio-Dyn., 8:393-400; Colville-Nash et al.,
1995, J. Pharm. and Exp. Ther., 274:1463-1472) whose disclosure is
incorporated herein by reference in its entirety, is characterized
by inflammatory cell influx, fibrous tissue proliferation and
intense angiogenesis. It is representative of inflammatory
angiogenesis and demonstrates that the angiogenic component can be
pharmacologically modulated independently of granuloma growth and
size. In addition, angiogenesis can be accurately quantitated by a
vascular casting method.
[0168] Day 1, mice are anesthetized using Aerrane (isoflurane) gas
(5%) or other approved methods, after which 3 mls of air is
injected into the dorsal subcutaneous tissue using a 27 g needle.
Mice are allowed to recover.
[0169] Day 0, mice are again anesthetized using Aerrane or other
approved methods, once anesthetized 0.5 ml of Freunds complete
adjuvant with 0.1% v/v croton oil is injected into the air pouch
formed on Day -1. The animals also begin their dosing regime
(number of day's dependent upon study) with the animals typically
receiving compound in 0.2 ml N,N, Dimethyl Acetoacetamide(DMA)
(Sigma, St. Louis, Mo.)/Cremephor El (Sigma, St. Louis, Mo.),
saline (10/10/80) or other appropriate vehicle. The animals are
allowed to recover and all subsequent dosing is performed on the
animals in the absence of anesthetics.
[0170] Days 1-5, animals are dosed according to schedule.
[0171] On Day 6 the animals are again anesthetized after which a
vascular cast is made (Kimura et al., 1986, J. Pharmacobio-Dyn.,
9:442-446); this involves a 1 ml tail vein i.v. injection of a
Carmine Red (10%) (Sigma, St. Louis, Mo.)/gelatin (5%) (Sigma, St.
Louis, Mo.) solution. The animals are then sacrificed by lethal
dose of anesthesia and chilled at 4.degree. C. for 2 hours prior to
the removal of the granuloma tissue.
[0172] When the granuloma is removed it is weighed and then dried
for 3 days at 45.degree. C. and reweighed. The dried tissue is then
digested in 0.9 ml of a 0.05M phosphate buffer pH 7.0 containing 12
U/ml.sup.-1 papain (Sigma, St. Louis, Mo.) and 0.33 g/L.sup.-1
N-acetyl-1-Cysteine (Sigma, St. Louis, Mo.) at 57.degree. C. for 3
days. After 3 days digestion the carmine red is solublized by the
addition of 0.1 ml 5 mM NaOH. Samples are centrifuged and then
filtered using 0.2 um acrodiscs. The carmine content is then
determined against a carmine red standard curve (0.5 to 2 mg/ml)
generated in extracted tissue from non carmine treated animals and
read at 490 nm. Sample and standard values are determined typically
using DeltaSoft Elisa analysis software (Biometallics Inc.,
Princeton, N.J.). The carmine content is then used to determine the
vascular indexes for the various treatments, vascular index being
the mg carmine dye/gm dry tissue.
[0173] The effect of compounds on vascular density was typically
measured for 6 days after induction of the granuloma. This time
point has been determined to be at or near the peak of
angiogenesis. As a positive control medroxyprogesterone, an
angiostatic steroid (Gross et al., 1981, Proc. Natl. Acad. Sci.
USA, 78:1176-1180), whose disclosure is hereby incorporated by
reference in its entirety, was utilized. This control demonstrated
a maximum reduction of 50% in this model. Medroxyprogesterone had
no effect on granuloma size as measured by dry weight.
Angiogenesis Model--In Vivo
Measurement of Angiogenesis In Vivo--Matrigel Model:
[0174] Angiogenesis is modeled in-vivo by placing an extra-cellular
matrix gel, beneath the skin of a mouse for approximately one week,
and then employing several measures to quantitate angiogenic
invasion of the gel (Biancone, L, et. al. Development of
Inflammatory Angiogenesis by Local Stimulation of Fas In Vivo. J.
Exp. Med. 186:147, 1997.). Briefly, reduced growth factor,
endotoxin free Matrigel.RTM. (Becton-Dickinson, Bedford, Mass.) is
a gel at low temperatures. Antibodies or known angiogenic agents
are mixed with the gel, such that they do not constitute more than
2% of the total volume. Eight week old or older, C57 female mice
are administered 0.5 ml of the Matrigel.RTM. by dorsal subcutaneous
injection through chilled syringes. At physiological temperature,
the liquid Matrigel.RTM. rapidly forms a solid and cohesive gel.
During the course of the experiment, mice receive test compounds or
controls administered as described above. After six days, the mice
are sacrificed and the Matrigel.RTM. plugs recovered. Angiogenesis
is quantitated by analyzing the hemoglobin content of the gel by
the method of Drabkin (Drabkin, D L and Austin, J H:
Spectrophotometric Studies II. Preparations from washed blood
cells; nitric oxide hemoglobin and sulflhemoglobin. J Biol Chem
112:51, 1935.)(Sigma, St. Louis, Mo.), or by staining and
quantitating blood vessel with CD31 staining as described
above.
Synthetic Chemistry
[0175] The invention will now be described by reference to the
following examples which are merely illustrative and are not to be
construed as a limitation of the scope of the present invention.
All temperatures are given in degrees centigrade, all solvents are
highest available purity and all reactions run under anhydrous
conditions in an argon atmosphere unless otherwise indicated.
[0176] In the Examples, all temperatures are in degrees Centigrade
(.degree. C.). Mass spectra were performed upon a VG Zab mass
spectrometer using fast atom bombardment, unless otherwise
indicated. .sup.1H-NMR (hereinafter "NMR") spectra were recorded at
250 MHz using a Bruker AM 250 or Am 400 spectrometer.
Multiplicities indicated are: s=singlet, d=doublet, t=triplet,
q=quartet, m=multiplet and br indicates a broad signal. Sat.
indicates a saturated solution, eq indicates the proportion of a
molar equivalent of reagent relative to the principal reactant.
[0177] Flash chromatography is run over Merck Silica gel 60
(230-400 mesh).
EXAMPLE 1
(2-(4-(6-methoxynapthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl--
propyl)-carbamic acid tert-butyl ester
[0178] ##STR7##
a) 6-Methoxy-naphthylene-2-carboxylic
acid-N-methoxy-N-methyl-amide
[0179] A suspension of 6-methoxynapthoic acid (10.95 grams
(hereinafter "g"), 0.05 moles (herein after "mol"), triethylamine
(29.8 milliliters (hereinafter "mL" or "ml")), 0.2 mol) in
dichloromethane (150 ml) was cooled to 0.degree. C. and slowly
thionyl chloride (4.35 ml) was added, upon which the solution
became brown and homogenous. The ice bath was removed and continued
stirring at room temperature for about 1 hour (hereinafter "h") at
which point T methoxymethylamine hydrochloride (7.02 g, 0.06 mol)
was added. The mixture was stirred at room temperature for 3 hours.
The solution was concentrated and the residue was partitioned
between dichloromethane and saturated potassium carbonate solution.
The organic layer was separated, washed with brine dried over
anhydrous magnesium sulphate, filtered and concentrated at reduced
pressure to give the title compound (9.65 g, 73%) as a brown solid;
MS(ES) m/e 246 [M+H].sup.+.
b) 4-t-Butyldimethylsiloxymethylpyridine
[0180] 4-Pyridylcarbinol (8.5 g, 0.08 mol) was dissolved in a 9:1
mixture of N,N-dimethylformamide and dichloromethane (150 mL).
t-Butyldimethylsilylchloride (13 g, 0.09 mol) and imidazole (6.4 g,
0.09 mol) were added and the solution was stirred for 12 hours at
room temperature. The reaction mixture was concentrated at reduced
pressure and the residue was partitioned between ethyl acetate and
water. The organic layer was separated, dried over anhydrous
magnesium sulphate, filtered and concentrated at reduced pressure
to give the title compound (17.4 g, 92%) as a colorless oil; MS(ES)
m/e 244 [M+H].sup.+.
c)
1-(t-Butyl-dimethylsiloxy)-2-(6-methoxy-naphthalen-2-yl)-1-pyridin-4-yl-
-ethan-2-one
[0181] To a stirring solution of diisopropylamine (5.00 mL, 0.035
mol) in tetrahydrofuran (50 mL) at 0.degree. C. was added n-butyl
lithium 15.5 mL, 0.039 mol) to generate lithium diisopropylamide in
situ. To the cooled -78.degree. C. LDA solution was added
4-t-butyldimethylsiloxypyridine (7.84 g, 0.035 mol) and stirring
was continued for 30 minutes at which time
6-methoxynaphthylene-2-carboxylic acid-N-methoxy-N-methyl-amide
(5.51 g, 0.023 mol) was added. The reaction was gradually allowed
to warm to room temperature. The reaction mixture was extracted
with ethyl acetate and the organic layers were combined, dried over
anhydrous magnesium sulphate, filtered and concentrated at reduced
pressure. The title compound was purified by flash chromatography
and isolated as a yellow solid; MS(ES) m/e 377 [M+H].sup.+.
d)
(2-(4-(6-methoxy-naphthalen-2-yl)-5-pyridin-4-yl-1-imidazol-2-yl)-2-met-
hyl-propyl)-carbamic acid tert-butyl ester
[0182] Acetic acid (15 ml) was added into a mixture of
(2,2-dimethyl-3-oxo-propyl)-carbamic acid tert-butyl ester (Y.
Guindon et al., J. Am. Chem. Soc., 1997, 119, 9289) (0.26 g, 1.2
mmol), ammonium acetate (0.98 g, 12.0 mmol), and
2-(t-butyl-dimethylsilanyloxy)-2-(6-methoxy-naphthalen-2-yl)-1-pyridin-4--
yl-ethanone (0.26 g, 0.6 mmol). The resulting solution was heated
at 90.degree. C. overnight, then cooled to 0.degree. C., and
NH.sub.4OH was added to the solution slowly with stirring. The
resulting precipitate was filtered, and dried to yield the title
compound; MS(ES) m/e 472 [M+H].sup.+.
EXAMPLE 2
2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl--
propylamine
[0183] ##STR8##
[0184] The product of Example 1 (200 milligrams (hereinafter "mg"),
0.4 millimoles (hereinafter "mmol")) was dissolved in a 1:1 mixture
of trifluoroacetic acid and dichloromethane (5 mL) and stirred at
room temperature for 4 hours. The solvents were concentrated at
reduced pressure. The crude product was purified by column
chromatography to afford the title compound (50 mg, 32%). MS(ES)
m/e 373 [M+H].sup.+.
EXAMPLE 3
n-Propyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-
-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine
[0185] ##STR9##
[0186] A solution of Example 2 (20 mg, 0.05 mmol) in
N,N-dimethylformamide (2 ml) was treated with potassium carbonate
(10 mg, 0.07 mmol) and 1-bromopropane (9 mg, 0.07 mmol). The
solution was stirred at room temperature for 4 hours. The mixture
was diluted with water and extracted with ethyl acetate. The
aqueous layer was separated and extracted with additional ethyl
acetate, the organic layers were combined, dried over anhydrous
magnesium sulphate, filtered and concentrated at reduced pressure.
The residue was chromatographed on a Gilson HPLC to give the title
compound (3 mg, 13%) as a yellow solid; MS(ES+) m/e. 415
[M+H]+.
EXAMPLE 4
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-methanesulfonamide
[0187] ##STR10##
[0188] A solution of the product of Example 2 (15 mg, 0.04 mmol) in
a 1:1 mixture of tetrahydrofuran and dichloromethane (2 mL)
containing Hunig's base (9 .mu.L, 0.05 mmol) was treated with
methanesulfonyl chloride (4 .mu.L, 0.05 mmol) and stirred at room
temperature for 8 hours. The mixture was diluted with water and
extracted with dichloromethane. The aqueous layer was separated and
extracted with additional dichloromethane, the organic layers were
combined, dried over anhydrous magnesium sulphate, filtered and
concentrated at reduced pressure. The residue was chromatographed
on a Gilson HPLC to give the title compound (8 mg, 44%) as a yellow
solid; MS(ES) m/e. 451 [M+H].sup.+.
EXAMPLE 5
1,1,1-trifluoro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imida-
zol-2-yl)-2-methyl-propyl)-methanesulfonamide
[0189] ##STR11##
[0190] The title compound (9 mg, 33%) was prepared starting from
the product of Example 2 and 1,1,1-trifluoromethanesulfonyl
chloride using the method described in Example 4; MS(LS) m/e 505
[M+H].sup.+.
EXAMPLE 6
2,2,2-trifluoro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imida-
zol-2-yl)-2-methyl-propyl)-ethanesulfonamide
[0191] ##STR12##
[0192] The title compound (12 mg, 43%) was prepared starting from
the product of Example 2 and 2,2,2-trifluoroethanesulfonyl chloride
using the method described in Example 4; MS(ES) m/e 519
[M+H].sup.+.
EXAMPLE 7
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-propanesulfonamide
[0193] ##STR13##
[0194] The title compound (7 mg, 27%) was prepared starting from
the product of Example 2 and propanesulfonyl chloride using the
method described in Example 4; MS(LS) m/e 479 [M+H].sup.+.
EXAMPLE 8
3-Chloro-N-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-y-
l)-2-methyl-propyl)-propane sulfonamide
[0195] ##STR14##
[0196] The title compound (11 mg, 40%) was prepared starting from
the product of Example 2 and 3-chloro-propanesulfonyl chloride
using the method described in Example 4; MS(LS) m/e 514
[M+H].sup.+.
EXAMPLE 9
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-butane sulfonamide
[0197] ##STR15##
[0198] The title compound (8 mg, 30%) was prepared starting from
the product of Example 2 and butanesulfonyl chloride using the
method described in Example 4; MS(ES) m/e 493 [M+H].sup.+.
EXAMPLE 10
1-Ethyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl-
)-2-methyl-propyl)-urea
[0199] ##STR16##
[0200] A solution of the product of Example 2 (20 mg, 0.05 mmol) in
dichloromethane (2 mL) containing Hunig's base (10 .mu.L, 0.07
mmol) was treated with ethyl isocyanate (6 .mu.L, 0.07 mmol) and
stirred at room temperature for 8 hours. The mixture was diluted
with water and extracted with dichloromethane. The aqueous layer
was separated and extracted with additional dichloromethane, the
organic layers were combined, dried over anhydrous magnesium
sulphate, filtered and concentrated at reduced pressure. The
residue was chromatographed on a Gilson HPLC to give the title
compound (12 mg, 50%) as a yellow solid; MS(ES) m/e 444
[M+H].sup.+.
EXAMPLE 11
1-(2-Chloroethyl)-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imi-
dazol-2-yl)-2-methyl-propyl)-urea
[0201] ##STR17##
[0202] The title compound (13 mg, 51%) was prepared starting from
the product of Example 2 and 3-chloroethyl isocyanate using the
method described in Example 10; MS(ES) m/e 478 [M+H].sup.+.
EXAMPLE 12
1-n-Propyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-
-yl)-2-methyl-propyl)-urea
[0203] ##STR18##
[0204] The title compound (12 mg, 49%) was prepared starting from
the product of Example 2 and n-propyl isocyanate using the method
described in Example 10; MS(LS) m/e 458 [M+H].sup.+.
EXAMPLE 13
1-Isopropyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol--
2-yl)-2-methyl-propyl)-urea
[0205] ##STR19##
[0206] The title compound (12 mg, 49%) was prepared starting from
the product of Example 2 and n-propyl isocyanate using the method
described in Example 10; MS(ES) m/e 458 [M+H].sup.+.
EXAMPLE 14
1-tert-Butyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-
-2-yl)-2-methyl-propyl)-urea
[0207] ##STR20##
[0208] The title compound (10 mg, 39%) was prepared starting from
the product of Example 2 and tert-butyl isocyanate using the method
described in Example 10; MS(ES) m/e 472 [M+H].sup.+.
EXAMPLE 15
1-Methylformyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidaz-
ol-2-yl)-2-methyl-prolpyl)-urea
[0209] ##STR21##
[0210] The title compound (13 mg, 51%) was prepared starting from
the product of Example 2 and methylformyl isocyanate using the
method described in Example 4; MS(LS) m/e 474 [M+H].sup.+.
EXAMPLE 16
1-(3,
5-Dimethyl-isoxazol-4-yl)-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyrid-
in-4-yl-1H-imidazol-2-yl)-2-methyl-propyl)-urea
[0211] ##STR22##
[0212] The title compound (11 mg, 40%) was prepared starting from
the product of Example 2 and tert-butyl isocyanate using the method
described in Example 10; MS(ES) m/e 511 [M+H].sup.+.
EXAMPLE 17
1-n-Propyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-
-yl)-2-methyl-propyl)-thiourea
[0213] ##STR23##
[0214] The title compound (7 mg, 28%) was prepared starting from
the product of Example 2 and n-propyl thioisocyanate using the
method described in Example 10; MS(ES) m/e 474 [M+H].sup.+.
EXAMPLE 18
1-n-Butyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2--
yl)-2-methyl-propyl)-thiourea
[0215] ##STR24##
[0216] The title compound (8 mg, 31%) was prepared starting from
the product of Example 2 and n-butyl thioisocyanate using the
method described in Example 10; MS(LS) m/e 488 [M+H].sup.+.
EXAMPLE 19
1-Isopropyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol--
2-yl)-2-methyl-propyl)-thiourea
[0217] ##STR25##
[0218] The title compound (6 mg, 23%) was prepared starting from
the product of Example 2 and sec-butyl thioisocyanate using the
method described in Example 10; MS(LS) m/e 488 [M+H].sup.+.
EXAMPLE 20
1-Ethyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl-
)-2-methyl-propyl)-thiourea
[0219] ##STR26##
[0220] The title compound (12 mg, 49%) was prepared from Example 2
and ethyl thioisocyanate using the method described in Example 10.
MS(ES+) m/e 460 [M+H]+.
EXAMPLE 21
1-Methyl-3-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-y-
l)-2-methyl-propyl)-thiourea
[0221] ##STR27##
[0222] The title compound (8 mg, 30%) was prepared from Example 2
and methyl thioisocyanate using the method described in Example 10.
MS(ES+) m/e 446 [M+H]+.
EXAMPLE 22
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-meth-
yl-propyl)-3-(2-methoxyethyl)-thiourea
[0223] ##STR28##
[0224] The title compound (7 mg, 27%) was prepared from Example 2
and 2-methoxyethyl isothiocyanate using the method described in
Example 10. MS(ES+) m+e 490 [M+H]+.
EXAMPLE 23
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-meth-
yl-propyl)-3-(2-morpholin-4-yl-ethyl)-thiourea
[0225] ##STR29##
[0226] The title compound (10 mg, 34%) was prepared from Example 2
and sec-butyl thioisocyanate using the method described in Example
10. MS(ES+) m/e 545 [M+H]+.
EXAMPLE 24
1-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-meth-
yl-propyl)-3-(2-piperidin-4-yl-ethyl)-thiourea
[0227] ##STR30##
[0228] The title compound (9 mg, 31%) was prepared from Example 2
and sec-butyl thioisocyanate using the method described in Example
10. MS(ES+) m/e 543 [M+H]+.
EXAMPLE 25
Cyclohexylmethyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidaz-
ol-2-yl)-2-methyl-propyl)-amine
[0229] ##STR31##
[0230] A solution of the product of Example 2 (20 mg, 0.05 mmol) in
dichloromethane (2 mL) was treated with cyclohexaldehyde (3 mg,
0.07 mmol) and sodium trimethoxyborohydride (9 mg, 0.07 mmol). The
solution was stirred at room temperature for 4 hours. The mixture
was diluted with water and extracted with ethyl acetate. The
aqueous layer was separated and extracted with additional ethyl
acetate, the organic layers were combined, dried over anhydrous
magnesium sulphate, filtered and concentrated at reduced pressure.
The residue was chromatographed on a Gilson HPLC to give the title
compound (10 mg, 40%) as a yellow solid; MS(ES) m/e 469
[M+H].sup.+.
EXAMPLE 26
Bis-n-butyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2--
yl)-2-methyl-propyl)-amine
[0231] ##STR32##
[0232] The title compound (12 mg, 47%) was prepared starting from
the product of Example 2 and butyraldehyde using the method
described in Example 25; MS(ES) m/e 485 [M+H].sup.+.
EXAMPLE 27
Bis-cyclohexylmethyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-im-
idazol-2-yl)-2-methyl-propyl)-amine
[0233] ##STR33##
[0234] The title compound (4 mg, 13%) was prepared starting from
the product of Example 2 and cyclohexylaldehyde using the method
described in Example 25; MS(ES) m/e 565 [M+H].sup.+.
EXAMPLE 28
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-(3-methylsulfanyl-propyl)-amine
[0235] ##STR34##
[0236] The title compound (4 mg, 16%) was prepared starting from
the product of Example 2 and 1-thiomethylproprionaldehyde using the
method described in Example 25; MS(ES) m/e 461 [M+H].sup.+.
EXAMPLE 29
(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-2-methyl-
-propyl)-bis-(3-methylsulfanyl-propyl)-amine
[0237] ##STR35##
[0238] The title compound (11 mg, 46%) was prepared starting from
the product of Example 2 and 1-thiomethylproprionaldehyde using the
method described in Example 25; MS(ES) m/e 549 [M+H].sup.+.
EXAMPLE 30
Isobutyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-yl)-
-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine
[0239] ##STR36##
[0240] The title compound (11 mg, 48%) was prepared starting from
the product of Example 2 and isoproprionaldehyde using the method
described in Example 25; MS(ES) m/e 429 [M+H].sup.+.
EXAMPLE 31
Bis-isobutyl-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-imidazol-2-
-yl)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine
[0241] ##STR37##
[0242] The title compound (11 mg, 42%) was prepared starting from
the product of Example 2 and i-propionaldehyde using the method
described in Example 25; MS(ES) m/e 485 [M+H].sup.+.
EXAMPLE 32
(2,2-Dimethylpropyl)-(2-(4-(6-methoxy-napthalen-2-yl)-5-pyridin-4-yl-1H-im-
idazol-2-yl)-2-methyl-propyl)-(3-methylsulfanyl-propyl)-amine
[0243] ##STR38##
[0244] The title compound (16 mg, 67%) was prepared starting from
the product of Example 2 and t-butyraldehyde using the method
described in Example 25; MS(ES) m/e 443 [M+H].sup.+.
[0245] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth.
[0246] The above description fully discloses the invention
including preferred embodiments thereof. Modifications and
improvements of the embodiments specifically disclosed herein are
within the scope of the following claims. Without further
elaboration, it is believed that one skilled in the are can, using
the preceding description, utilize the present invention to its
fullest extent. Therefore, the Examples herein are to be construed
as merely illustrative and not a limitation of the scope of the
present invention in any way. The embodiments of the invention in
which an exclusive property or privilege is claimed are defined as
follows.
* * * * *