U.S. patent application number 14/261754 was filed with the patent office on 2014-08-21 for chromene modulators of chemokine receptor activity.
The applicant listed for this patent is Bristol-Myers Squibb Company. Invention is credited to Douglas G. Batt.
Application Number | 20140235672 14/261754 |
Document ID | / |
Family ID | 41258645 |
Filed Date | 2014-08-21 |
United States Patent
Application |
20140235672 |
Kind Code |
A1 |
Batt; Douglas G. |
August 21, 2014 |
CHROMENE MODULATORS OF CHEMOKINE RECEPTOR ACTIVITY
Abstract
The present application describes modulators of MCP-1 or CCR-2
of formula ##STR00001## or stereoisomers or prodrugs or
pharmaceutically acceptable salts thereof, wherein W.sup.1,
W.sup.2, W.sup.3, Y, Z, R.sup.2, R.sup.3, R.sup.3' and R.sup.4, are
defined in the specification. In addition, methods of treating and
preventing inflammatory diseases such as asthma and allergic
diseases, as well as autoimmune pathologies such as rheumatoid
arthritis and transplant rejection using modulators of formula (I)
are disclosed.
Inventors: |
Batt; Douglas G.;
(Wilmington, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Bristol-Myers Squibb Company |
Princeton |
NJ |
US |
|
|
Family ID: |
41258645 |
Appl. No.: |
14/261754 |
Filed: |
April 25, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13003777 |
Jan 12, 2011 |
8759390 |
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PCT/US2009/050450 |
Jul 14, 2009 |
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14261754 |
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61081194 |
Jul 16, 2008 |
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Current U.S.
Class: |
514/314 ;
514/337; 514/456; 546/167; 546/282.7; 549/405 |
Current CPC
Class: |
A61P 37/04 20180101;
A61P 37/08 20180101; A61P 17/06 20180101; A61P 11/06 20180101; A61P
37/00 20180101; A61P 35/00 20180101; A61P 37/02 20180101; A61P
19/02 20180101; A61P 1/04 20180101; C07D 311/32 20130101; A61P
25/00 20180101; A61P 1/00 20180101; A61P 9/00 20180101; A61P 9/10
20180101; C07D 311/58 20130101; A61P 19/10 20180101; A61P 43/00
20180101; C07D 405/04 20130101; C07D 407/04 20130101; C07D 311/66
20130101; A61P 31/18 20180101; C07D 311/60 20130101; A61P 29/00
20180101; A61P 9/04 20180101; A61P 13/12 20180101; A61P 11/00
20180101 |
Class at
Publication: |
514/314 ;
514/337; 514/456; 546/167; 546/282.7; 549/405 |
International
Class: |
C07D 407/04 20060101
C07D407/04; C07D 311/58 20060101 C07D311/58; C07D 405/04 20060101
C07D405/04 |
Claims
1. A compound of formula (I): ##STR00007## or a stereoisomer or a
pharmaceutically acceptable salt thereof, wherein: R.sup.2 is H, or
C.sub.1-4 alkyl; R.sup.3 and R.sup.3' are independently H,
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl; or
R.sup.3 and R.sup.3', when attached to neighboring carbons, taken
together form --CH.dbd.CH--CH.dbd.CH--; R.sup.4 is H or C.sub.1-4
alkyl; W.sup.1 is N or C--Z'; W.sup.2 is N or C--Y'; W.sup.3 is N
or C--X; X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl; Y and Y' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, haloC.sub.1-4 alkyl or NO.sub.2; Z and Z'
are independently H or halo; or Y and Z, Y' and Z', Y and X, or Y'
and X are taken together to form --CH.dbd.CH--CH.dbd.CH--; or Y and
X or Y' and X are taken together to form --O--CH.sub.2--O--;
provided that: (1) one and only one of W.sup.1, W.sup.2 or W.sup.3
is N; (2) if W.sup.2 is N, then X, Y, Z and Z' are not all H; (3)
if W.sup.3 is N, then W.sup.1 and W.sup.2 are both CH and Y and Z
are H; (4) if Y is C.sub.1-4alkoxy then X is not H; and (5) if X is
F then one of Y or Y' is not H.
2. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl; R.sup.4 is H or
C.sub.1-4 alkyl; W.sup.1 is N or C--Z'; W.sup.2 is N or C--Y';
W.sup.3 is N or C--X; X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
halo or haloC.sub.1-4 alkyl; Y and Y' are independently H,
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo, haloC.sub.1-4 alkyl or
NO.sub.2; Z and Z' are independently H or halo; or Y and Z, Y' and
Z', Y and X, or Y' and X are taken together to form
--CH.dbd.CH--CH.dbd.CH--; or Y and X or Y' and X are taken together
to form --O--CH.sub.2--O--.
3. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl; R.sup.4 is H or
C.sub.1-4 alkyl; W.sup.1 is N or C--Z'; W.sup.2 is N or C--Y';
W.sup.3 is N or C--X; X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
halo or haloC.sub.1-4 alkyl; Y and Y' are independently H,
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo, haloC.sub.1-4 alkyl or
NO.sub.2; Z and Z' are independently H or halo; or Y and Z, Y' and
Z', Y and X, or Y' and X are taken together to form
--CH.dbd.CH--CH.dbd.CH--.
4. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl; R.sup.4 is H or
C.sub.1-4 alkyl; W.sup.1 is N or C--Z'; W.sup.2 is N or C--Y';
W.sup.3 is N or C--X; X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
halo or haloC.sub.1-4 alkyl; Y and Y' are independently H,
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo, haloC.sub.1-4 alkyl or
NO.sub.2; and Z and Z' are independently H or halo.
5. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, or halo; R.sup.4 is H or C.sub.1-4 alkyl; W.sup.1
is N or C--Z'; W.sup.2 is N or C--Y'; W.sup.3 is N or C--X; X is H,
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or haloC.sub.1-4 alkyl; Y
and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
halo, or haloC.sub.1-4 alkyl; and Z and Z' are independently H or
halo.
6. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkoxy,
or halo; R.sup.4 is H or C.sub.1-4 alkyl; W.sup.1 is N or C--Z';
W.sup.2 is N or C--Y'; W.sup.3 is N or C--X; X is H, C.sub.1-4
alkyl, halo or haloC.sub.1-4 alkyl; Y and Y' are independently H,
C.sub.1-4 alkyl, halo, or haloC.sub.1-4 alkyl; and Z and Z' are
independently H or halo.
7. The compound of claim 1, wherein: R.sup.2 is H, or C.sub.1-4
alkyl; R.sup.3 and R.sup.3' are independently H or halo; R.sup.4 is
H or C.sub.1-4 alkyl; W.sup.1 is N or C--Z'; W.sup.2 is N or C--Y';
W.sup.3 is N or C--X; X is H, halo or haloC.sub.1-4 alkyl; Y and Y'
are independently H, halo, or haloC.sub.1-4 alkyl; and Z and Z' are
independently H or halo.
8. The compound of claim 1, wherein: R.sup.2 is H, methyl or ethyl;
R.sup.3 and R.sup.3' are H; R.sup.4 is H; W.sup.1 is N or C--Z';
W.sup.2 is N or C--Y'; W.sup.3 is N or C--X; X is H, Br, Cl, F or
CF.sub.3; Y and Y' are independently H, Br, Cl, F or CF.sub.3; and
Z and Z' are independently H, Br, Cl or F.
9. The compound of claim 1, or a stereoisomer or pharmaceutically
acceptable salt from thereof, wherein the compound is selected from
the compounds exemplified in the examples.
10. A pharmaceutical composition comprised of a pharmaceutically
acceptable carrier and a therapeutically effective amount of a
compound of claim 1.
11. A method for treating a disorder comprising administering to a
patient in need thereof a therapeutically effective amount of at
least one compound of claim 1; wherein said disorder is selected
from osteoarthritis, aneurysm, fever, cardiovascular effects,
Crohn's disease, congestive heart failure, autoimmune diseases,
HIV-infection, HIV-associated dementia, psoriasis, idiopathic
pulmonary fibrosis, transplant arteriosclerosis, physically- or
chemically-induced brain trauma, inflammatory bowel disease,
alveolitis, colitis, systemic lupus erythematosus, nephrotoxic
serum nephritis, glomerularnephritis, asthma, multiple sclerosis,
atherosclerosis, rheumatoid arthritis, restinosis, organ
transplantation, and cancer.
12. A method for preparing a medicament for the treatment of
osteoarthritis, aneurysm, fever, cardiovascular effects, Crohn's
disease, congestive heart failure, autoimmune diseases,
HIV-infection, HIV-associated dementia, psoriasis, idiopathic
pulmonary fibrosis, transplant arteriosclerosis, physically- or
chemically-induced brain trauma, inflammatory bowel disease,
alveolitis, colitis, systemic lupus erythematosus, nephrotoxic
serum nephritis, glomerularnephritis, asthma, multiple sclerosis,
atherosclerosis, and rheumatoid arthritis comprising formulating at
least one compound of claim 1.
13. A method for treating a disorder comprising administering to a
patient in need thereof a therapeutically effective amount of a
composition of claim 10, wherein said disorder is selected from
osteoarthritis, aneurysm, fever, cardiovascular effects, Crohn's
disease, congestive heart failure, autoimmune diseases,
HIV-infection, HIV-associated dementia, psoriasis, idiopathic
pulmonary fibrosis, transplant arteriosclerosis, physically- or
chemically-induced brain trauma, inflammatory bowel disease,
alveolitis, colitis, systemic lupus erythematosus, nephrotoxic
serum nephritis, glomerularnephritis, asthma, multiple sclerosis,
atherosclerosis, rheumatoid arthritis, restinosis, organ
transplantation, and cancer.
14. A method for preparing a medicament for the treatment of
osteoarthritis, aneurysm, fever, cardiovascular effects, Crohn's
disease, congestive heart failure, autoimmune diseases,
HIV-infection, HIV-associated dementia, psoriasis, idiopathic
pulmonary fibrosis, transplant arteriosclerosis, physically- or
chemically-induced brain trauma, inflammatory bowel disease,
alveolitis, colitis, systemic lupus erythematosus, nephrotoxic
serum nephritis, glomerularnephritis, asthma, multiple sclerosis,
atherosclerosis, and rheumatoid arthritis comprising formulating a
composition of claim 10 into a useful pharmaceutical
dosage-form.
15. A method for treating a patient in need of therapy comprising
administering to said patient in need thereof a therapeutically
effective amount of a composition of claim 10.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a divisional of U.S. Ser. No.
13/003,777, filed Jan. 12, 2011, now allowed, which claims benefit
to International Application No. PCT/US2009/050450, filed on Jul.
14, 2009, which claims priority benefit to U.S. Provisional
Application Ser. No. 61/081,194, filed on Jul. 16, 2008, the
contents of which are herein incorporated by reference in its
entirety.
FIELD OF THE INVENTION
[0002] This invention relates generally to modulators of chemokine
receptor activity, pharmaceutical compositions containing the same,
and methods of using the same as agents for treatment and
prevention of inflammatory diseases, allergic and autoimmune
diseases, and in particular, rheumatoid arthritis and transplant
rejection.
BACKGROUND OF THE INVENTION
[0003] Chemokines are chemotactic cytokines, of molecular weight
6-15 kDa, that are released by a wide variety of cells to attract
and activate, among other cell types, monocytes, macrophages, T and
B lymphocytes, eosinophils, basophils and neutrophils (reviewed in:
Luster, New Eng. J. Med. 1998, 338, 436-445 and Rollins, Blood
1997, 90, 909-928). There are two major classes of chemokines, CXC
and CC, depending on whether the first two cysteines in the amino
acid sequence are separated by a single amino acid (CXC) or are
adjacent (CC). The CXC chemokines, such as interleukin-8 (IL-8),
neutrophil-activating protein-2 (NAP-2) and melanoma growth
stimulatory activity protein (MGSA) are chemotactic primarily for
neutrophils and T lymphocytes, whereas the CC chemokines, such as
RANTES, MIP-1.alpha., MIP-1.beta., the monocyte chemotactic
proteins (MCP-1, MCP-2, MCP-3, MCP-4, and MCP-5) and the eotaxins
(-1 and -2) are chemotactic for, among other cell types,
macrophages, T lymphocytes, eosinophils, dendritic cells, and
basophils. There also exist the chemokines lymphotactin-1,
lymphotactin-2 (both C chemokines), and fractalkine (a CX.sub.3C
chemokine) that do not fall into either of the major chemokine
subfamilies.
[0004] The chemokines bind to specific cell-surface receptors
belonging to the family of G-protein-coupled
seven-transmembrane-domain proteins (reviewed in: Horuk, Trends
Pharm. Sci. 1994, 15, 159-165) which are termed "chemokine
receptors." On binding their cognate ligands, chemokine receptors
transduce an intracellular signal though the associated trimeric G
proteins, resulting in, among other responses, a rapid increase in
intracellular calcium concentration, changes in cell shape,
increased expression of cellular adhesion molecules, degranulation,
and promotion of cell migration. There are at least ten human
chemokine receptors that bind or respond to CC chemokines with the
following characteristic patterns (reviewed in Zlotnik et al.,
Immunity 2000, 12, 121): CCR-1 (or "CKR-1" or "CC-CKR-1")
[MIP-1.alpha., MCP-3, MCP-4, RANTES] (Neote et al., Cell 1993, 72,
415-425, and Luster, New Eng. J. Med. 1998, 338, 436-445); CCR-2A
and CCR-2B (or "CKR-2A"/"CKR-2B" or "CC-CKR-2A"/"CC-CKR-2B")
[MCP-1, MCP-2, MCP-3, MCP-4, MCP-5] (Charo et al., Proc. Natl.
Acad. Sci. USA 1994, 91, 2752-2756, and Luster, New Eng. J. Med.
1998, 338, 436-445); CCR-3 (or "CKR-3" or "CC-CKR-3") [eotaxin-1,
eotaxin-2, RANTES, MCP-3, MCP-4] (Combadiere et al., J. Biol. Chem.
1995, 270, 16491-16494, and Luster, New Eng. J. Med. 1998, 338,
436-445); CCR-4 (or "CKR-4" or "CC-CKR-4") [TARC, MDC] (Power et
al., J. Biol. Chem. 1995, 270, 19495-19500, and Luster, New Eng. J.
Med. 1998, 338, 436-445); CCR-5 (or "CKR-5" OR "CC-CKR-5")
[MIP-1.alpha., RANTES, MIP-1.beta.] (Samson et al., Biochemistry
1996, 35, 3362-3367); CCR-6 (or "CKR-6" or "CC-CKR-6") [LARC] (Baba
et al., J. Biol. Chem. 1997, 272, 14893-14898); CCR-7 (or "CKR-7"
or "CC-CKR-7") [ELC] (Yoshie et al., J. Leukoc. Biol. 1997, 62,
634-644); CCR-8 (or "CKR-8" or "CC-CKR-8") [1-309] (Napolitano et
al., J. Immunol., 1996, 157, 2759-2763); CCR-10 (or "CKR-10" or
"CC-CKR-10") [MCP-1, MCP-3] (Bonini et al., DNA Cell Biol. 1997,
16, 1249-1256); and CCR-11 [MCP-1, MCP-2, and MCP-4] (Schweickart
et al., J. Biol. Chem. 2000, 275, 9550).
[0005] In addition to the mammalian chemokine receptors, mammalian
cytomegaloviruses, herpesviruses and poxviruses have been shown to
express, in infected cells, proteins with the binding properties of
chemokine receptors (reviewed in: Wells et al., Curr. Opin.
Biotech. 1997, 8, 741-748). Human CC chemokines, such as RANTES and
MCP-3, can cause rapid mobilization of calcium via these virally
encoded receptors. Receptor expression may be permissive for
infection by allowing for the subversion of normal immune system
surveillance and response to infection. Additionally, human
chemokine receptors, such as CXCR4, CCR2, CCR3, CCR5 and CCR8, can
act as co-receptors for the infection of mammalian cells by
microbes as with, for example, the human immunodeficiency viruses
(HIV).
[0006] The chemokines and their cognate receptors have been
implicated as being important mediators of inflammatory,
infectious, and immunoregulatory disorders and diseases, including
asthma and allergic diseases, as well as autoimmune pathologies
such as rheumatoid arthritis and atherosclerosis (reviewed in:
Carter, P. H., Curr. Opin. Chem. Biol. 2002, 6, 510; Trivedi et
al., Ann. Reports Med. Chem. 2000, 35, 191; Saunders et al., Drug
Disc. Today 1999, 4, 80; Premack et al., Nature Medicine 1996, 2,
1174). For example, the chemokine macrophage inflammatory protein-1
(MIP-1.alpha.) and its receptor CC Chemokine Receptor 1 (CCR-1)
play a pivotal role in attracting leukocytes to sites of
inflammation and in subsequently activating these cells. When the
chemokine MIP-1.alpha. binds to CCR-1, it induces a rapid increase
in intracellular calcium concentration, increased expression of
cellular adhesion molecules, cellular degranulation, and the
promotion of leukocyte migration.
[0007] In addition, demonstration of the chemotactic properties of
MIP-1.alpha. in humans has been provided experimentally. Human
subjects, when injected intradermally with MIP-1.alpha.,
experienced a rapid and significant influx of leukocytes to the
site of injection (Brummet, M. E., J. Immun. 2000, 164,
3392-3401).
[0008] The chemokines and their cognate receptors have been
implicated as being important mediators of inflammatory,
infectious, and immunoregulatory disorders and diseases, including
asthma and allergic diseases; as well as autoimmune pathologies,
such as rheumatoid arthritis and multiple sclerosis; and metabolic
diseases, such as atherosclerosis and diabetes (reviewed in: Charo
et al., New Eng. J. Med. 2006, 354, 610-621; Gao, Z. et al., Chem.
Rev. 2003, 103, 3733; Carter, P. H., Curr. Opin. Chem. Biol. 2002,
6, 510; Trivedi et al., Ann. Reports Med. Chem. 2000, 35, 191;
Saunders et al., Drug Disc. Today 1999, 4, 80; Premack et al.,
Nature Medicine 1996, 2, 1174). For example, the chemokine monocyte
chemoattractant-1 (MCP-1) and its receptor CC Chemokine Receptor 2
(CCR-2) play a pivotal role in attracting leukocytes to sites of
inflammation and in subsequently activating these cells. When the
chemokine MCP-1 binds to CCR-2, it induces a rapid increase in
intracellular calcium concentration, increased expression of
cellular adhesion molecules, and the promotion of leukocyte
migration. Demonstration of the importance of the MCP-1/CCR-2
interaction has been provided by experiments with genetically
modified mice. MCP-1 -/- mice were unable to recruit monocytes into
sites of inflammation after several different types of immune
challenge (Lu, B. et al., J. Exp. Med. 1998, 187, 601). Likewise,
CCR-2 -/- mice were unable to recruit monocytes or produce
interferon-.gamma. when challenged with various exogenous agents;
moreover, the leukocytes of CCR-2 null mice did not migrate in
response to MCP-1 (Boring, L. et al., J. Clin. Invest. 1997, 100,
2552), thereby demonstrating the specificity of the MCP-1/CCR-2
interaction. Two other groups have independently reported
equivalent results with different strains of CCR-2 -/- mice
(Kuziel, W. A. et al., Proc. Natl. Acad. Sci. USA 1997, 94, 12053,
and Kurihara, T. et al., J. Exp. Med. 1997, 186, 1757). The
viability and generally normal health of the MCP-1 -/- and CCR-2
-/- animals is noteworthy, in that disruption of the MCP-1/CCR-2
interaction does not induce physiological crisis. Taken together,
these data lead one to the conclusion that molecules that block the
actions of MCP-1/CCR2 would be useful in treating a number of
inflammatory and autoimmune disorders (reviewed in: Feria, M. et
al., Exp. Opin. Ther. Patents 2006, 16, 49; and Dawson, J. et al.,
Exp. Opin. Ther. Targets 2003, 7, 35). This hypothesis has now been
validated in a number of different animal disease models, as
described below.
[0009] It is known that MIP-1.alpha. is elevated in the synovial
fluid and blood of patients with rheumatoid arthritis (Koch, A. et
al., J. Clin. Invest. 1994, 93, 921-928). Moreover, several studies
have demonstrated the potential therapeutic value of antagonism of
the MIP-1.alpha./CCR1 interaction in treating rheumatoid arthritis
(Pease, J. E. et al., Expert Opin. Invest. Drugs 2005, 14,
785-796).
[0010] An antibody to MIP-1.alpha. was shown to ameliorate
experimental autoimmune encepahlomytis (EAE), a model of multiple
sclerosis, in mice (Karpus, W. J. et al., J. Immun. 1995,
5003-5010). Likewise, inflammatory disease symptoms could be
controlled via direct administration of antibodies for MIP-1.alpha.
to mice with collagen-induced arthritis (Lukacs, N. W. et al., J.
Clin. Invest. 1995, 95, 2868-2876).
[0011] It is known that MCP-1 is upregulated in patients who
develop bronchiolitis obliterans syndrome after lung
transplantation (Reynaud-Gaubert, M. et al., J. Heart Lung
Transplant., 2002, 21, 721-730; Belperio, J. et al., J. Clin.
Invest. 2001, 108, 547-556). In a murine model of bronchiolitis
obliterans syndrome, administration of an antibody to MCP-1 led to
attenuation of airway obliteration; likewise, CCR2 -/- mice were
resistant to airway obliteration in this same model (Belperio, J.
et al., J. Clin. Invest. 2001, 108, 547-556). These data suggest
that antagonism of MCP-1/CCR2 may be beneficial in treating
rejection of organs following transplantation. In addition, studies
have shown that disruption of MCP-1/CCR2 axis was able to prolong
the survival of islet transplant (Lee, I. et al. J. Immunol. 2003,
171, 6929; Abdi, R. et al., J. Immunol. 2004, 172, 767). In rat
graft models, CCR2 and MCP-1 was shown to be upregulated in grafts
that develop graft vasculopathy (Horiguchi, K. et al., J. Heart
Lung Transplant. 2002, 21, 1090). In another study, anti-MCP-1 gene
therapy attenuated graft vasculopathy (Saiura, A. et al.,
Arterioscler. Thromb. Vasc. Biol. 2004, 24, 1886). One study
described inhibition of experimental vein graft neoinitimal
formation by blockage of MCP-1 (Tatewaki, H. et al., J. Vasc. Surg.
2007, 45, 1236).
[0012] Other studies have demonstrated the potential therapeutic
value of antagonism of the MCP-1/CCR2 interaction in treating
asthma. Sequestration of MCP-1 with a neutralizing antibody in
ovalbumin-challenged mice resulted in marked decrease in bronchial
hyperresponsiveness and inflammation (Gonzalo, J-A., et al., J.
Exp. Med. 1998, 188, 157). It proved possible to reduce allergic
airway inflammation in Schistosoma mansoni egg-challenged mice
through the administration of antibodies for MCP-1 (Lukacs, N. W.
et al., J. Immunol. 1997, 158, 4398). Consistent with this, MCP-1
-/- mice displayed a reduced response to challenge with Schistosoma
mansoni egg (Lu, B. et al., J. Exp. Med. 1998, 187, 601).
[0013] Other studies have demonstrated the potential therapeutic
value of antagonism of the MCP-1/CCR2 interaction in treating
kidney disease. Administration of antibodies for MCP-1 in a murine
model of glomerularnephritis resulted in a marked decrease in
glomerular crescent formation and deposition of type I collagen
(Lloyd, C. M. et al., J. Exp. Med. 1997, 185, 1371). In addition,
MCP-1 -/- mice with induced nephrotoxic serum nephritis showed
significantly less tubular damage than their MCP-1 +/+ counterparts
(Tesch, G. H. et al., J. Clin. Invest. 1999, 103, 73).
[0014] Several studies have demonstrated the potential therapeutic
value of antagonism of the MCP-1/CCR2 interaction in treating
systemic lupus erythematosus. CCR2 -/- mice exhibited prolonged
survival and reduced renal disease relative to their WT
counterparts in a murine model of systemic lupus erythematosus
(Perez de Lema, G. et al., J. Am. Soc. Neph. 2005, 16, 3592). These
data are consistent with the disease-modifying activity found in
recent studies on genetic deletion of MCP-1 (Shimizu, S. et al.,
Rheumatology (Oxford) 2004, 43, 1121; Tesch, G. H. et al., J. Exp.
Med. 1999, 190, 1813) or administration of a peptide antagonist of
CCR2 (Hasegawa, H. et al., Arthritis & Rheumatism 2003, 48,
2555) in rodent models of lupus.
[0015] A remarkable 30-fold increase in CCR2.sup.+ lamina propria
lymphocytes was observed in the small bowels from Crohn's patients
relative to non-diseased ileum (Connor, S. J. et al., Gut 2004, 53,
1287). Also of note, there was an expansion in the subset of
circulating CCR2.sup.+/CD14.sup.+/CD56.sup.+ monocytes in patients
with active Crohn's disease relative to controls. Several rodent
studies have demonstrated the potential therapeutic value of
antagonism of the MCP-1/CCR2 interaction in treating Crohn's
disease/colitis. CCR-2 -/- mice were protected from the effects of
dextran sodium sulfate-induced colitis (Andres, P. G. et al., J.
Immunol. 2000, 164, 6303). Administration of a small molecule
antagonist of CCR2, CCR5, and CXCR3 (murine binding affinities=24,
236, and 369 nM, respectively) also protected against dextran
sodium sulfate-induced colitis (Tokuyama, H. et al., Int. Immunol.
2005, 17, 1023). Finally, MCP-1 -/- mice showed substantially
reduced colonic damage (both macroscopic and histological) in a
hapten-induced model of colitis (Khan, W. I. et al., Am. J.
Physiol. Gastrointest. Liver Physiol. 2006, 291, G803).
[0016] Two reports described the overexpression of MCP-1 in the
intestinal epithelial cells and bowel mucosa of patients with
inflammatory bowel disease (Reinecker, H. C. et al.,
Gastroenterology 1995, 108, 40, and Grimm, M. C. et al., J. Leukoc.
Biol. 1996, 59, 804).
[0017] One study described the association of promoter polymorphism
in the MCP-1 gene with scleroderma (systemic sclerosis) (Karrer, S.
et al., J. Invest. Dermatol. 2005, 124, 92). In related models of
tissue fibrosis, inhibition of CCR2/MCP-1 axis reduced fibrosis in
skin (Yamamoto, T. et al., J. Invest. Dermatol. 2003, 121, 510;
Ferreira, A. M. et al., J. Invest. Dermatol. 2006, 126, 1900), lung
(Okuma, T. et al., J. Pathol. 2004, 204, 594; Gharaee-Kermani, M.
et al., Cytokine 2003, 24, 266), kidney (Kitagawa, K. et al., Am.
J. Pathol. 2004, 165, 237; Wada, T. et al., J. Am. Soc. Nephrol.
2004, 15, 940), heart (Hayashidani, S. et al., Circulation 2003,
108, 2134), and liver (Tsuruta, S. et al., Int. J. Mol. Med. 2004,
14, 837).
[0018] One study has demonstrated the potential therapeutic value
of antagonism of the MCP-1/CCR2 interaction in treating alveolitis.
When rats with IgA immune complex lung injury were treated
intravenously with antibodies raised against rat MCP-1 (JE), the
symptoms of alveolitis were partially alleviated (Jones, M. L. et
al., J. Immunol. 1992, 149, 2147).
[0019] Several studies have shown the potential therapeutic value
of antagonism of the MCP-1/CCR2 interaction in treating cancer
(reviewed in: Craig, M. J. et al., Cancer Metastasis Rev. 2006, 25,
611; Conti, I. et al., Seminars in Cancer Biology 2004, 14, 149;
Giles, R. et al., Curr. Cancer Drug Targets 2006, 6, 659). When
immunodeficient mice bearing human breast carcinoma cells were
treated with an anti-MCP-1 antibody, inhibition of lung
micrometastases and increases in survival were observed (Salcedo,
R. et al., Blood 2000, 96, 34-40). Using human clinical tumor
specimens, CCR2 expression was associated with prostrate cancer
progression (Lu, Y. et al., J. Cell. Biochem. 2007, 101, 676). In
vitro, MCP-1 expression has been shown to mediate prostrate cancer
cell growth and invasion (Lu, Y. et al., Prostate 2006, 66, 1311);
furthermore, MCP-1 expressed by prostate cancer cells induced human
bone marrow progenitors for bone resorption (Lu, Y. et al., Cancer
Res. 2007, 67, 3646).
[0020] Multiple studies have described the potential therapeutic
value of antagonism of the MCP-1/CCR2 interaction in treating
restenosis. In humans, MCP-1 levels correlate directly with risk
for restenosis (Cipollone, F. et al. Arterioscler. Thromb. Vasc.
Biol. 2001, 21, 327). Mice deficient in CCR2 or in MCP-1 showed
reductions in the intimal area and in the intima/media ratio
(relative to wildtype littermates) after arterial injury (Roque, M.
et al. Arterioscler. Thromb. Vasc. Biol. 2002, 22, 554; Schober, A.
et al. Circ. Res. 2004, 95, 1125; Kim, W. J. et al., Biochem.
Biophys. Res. Commun. 2003, 310, 936). In mice, transfection of a
dominant negative inhibitor of MCP-1 in the skeletal muscle
(Egashira, K. et al., Circ. Res. 2002, 90, 1167) also reduced
intimal hyperplasia after arterial injury. Blockade of CCR2 using a
neutralizing antibody reduced neointimal hyperplasia after stenting
in primates (Horvath, C. et al., Circ. Res. 2002, 90, 488).
[0021] Two reports describe the overexpression of MCP-1 rats with
induced brain trauma (King, J. S. et al., J. Neuroimmunol. 1994,
56, 127, and Berman, J. W. et al., J. Immunol. 1996, 156, 3017). In
addition, studies have shown that both CCR2 -/- (Dimitrijevic, O.
B. et al., Stroke 2007, 38, 1345) and MCP-1 -/- mice (Hughes, P. M.
et al., J. Cereb. Blood Flow Metab. 2002, 22, 308) are partially
protected from ischemia/reperfusion injury.
[0022] It is known that monocytes/macrophages play an important
role in the development of neuropathic pain (Liu, T. et al., Pain
2000, 86, 25). Consistent with this notion, a potential role for
CCR2 in the treatment of both inflammatory and neuropathic pain has
been described recently. CCR2 -/- mice showed altered responses to
inflammatory pain relative to their WT counterparts, including
reduced pain behavior after intraplantar formalin injection and
slightly reduced mechanical allodynia after intraplantar CFA
injection (Abbadie, C. et al., Proc. Natl. Acad. Sci., USA 2003,
100, 7947). In addition, CCR2 -/- mice did not display significant
mechanical allodynia after sciatic nerve injury. Likewise, a small
molecule CCR2 antagonist reduced mechanical allodynia to .about.80%
of pre-injury levels after oral administration (Abbadie, C. et al.,
WO 2004/110376).
[0023] One study described the critical role of MCP-1 in ischemic
cardiomyopathy (Frangogiannis, N. G. et al., Circulation 2007, 115,
584). Another study described the attenuation of experimental heart
failure following inhibition of MCP-1 (Hayashidani, S. et al.,
Circulation 2003, 108, 2134).
[0024] Other studies have provided evidence that MCP-1 is
overexpressed in various disease states not mentioned above. These
reports provide correlative evidence that MCP-1 antagonists could
be useful therapeutics for such diseases. Another study has
demonstrated the overexpression of MCP-1 in rodent cardiac
allografts, suggesting a role for MCP-1 in the pathogenesis of
transplant arteriosclerosis (Russell, M. E. et al., Proc. Natl.
Acad. Sci. USA 1993, 90, 6086). The overexpression of MCP-1 has
been noted in the lung endothelial cells of patients with
idiopathic pulmonary fibrosis (Antoniades, H. N. et al., Proc.
Natl. Acad. Sci. USA 1992, 89, 5371). Similarly, the overexpression
of MCP-1 has been noted in the skin from patients with psoriasis
(Deleuran, M. et al., J. Dermatol. Sci. 1996, 13, 228, and
Gillitzer, R. et al., J. Invest. Dermatol. 1993, 101, 127);
correlative findings with predominance of CCR2+ cells have also
been reported (Vestergaard, C. et al., Acta Derm. Venerol. 2004,
84, 353). Finally, a recent report has shown that MCP-1 is
overexpressed in the brains and cerebrospinal fluid of patients
with HIV-1-associated dementia (Garzino-Demo, A., WO 99/46991).
[0025] In addition, CCR2 polymorphism has been shown to be
associated with sarcoidosis at least in one subset of patients
(Spagnolo, P. et al., Am. J. Respir. Grit. Care Med. 2003, 168,
1162).
[0026] It should also be noted that CCR-2 has been implicated as a
co-receptor for some strains of HIV (Doranz, B. J. et al., Cell
1996, 85, 1149). It has also been determined that the use of CCR-2
as an HIV co-receptor can be correlated with disease progression
(Connor, R. I. et al., J. Exp. Med. 1997, 185, 621). This finding
is consistent with the recent finding that the presence of a CCR-2
mutant, CCR2-64I, is positively correlated with delayed onset of
HIV in the human population (Smith, M. W. et al., Science 1997,
277, 959). Although MCP-1 has not been implicated in these
processes, it may be that MCP-1 antagonists that act via binding to
CCR-2 may have beneficial therapeutic effects in delaying the
disease progression to AIDS in HIV-infected patients.
[0027] It should be noted that CCR2 is also the receptor for the
human chemokines MCP-2, MCP-3, and MCP-4 (Luster, New Eng. J. Med.
1998, 338, 436-445). Since the new compounds of formula (I)
described herein antagonize MCP-1 by binding to the CCR-2 receptor,
it may be that these compounds of formula (I) are also effective
antagonists of the actions of MCP-2, MCP-3, and MCP-4 that are
mediated by CCR-2. Accordingly, when reference is made herein to
"antagonism of MCP-1," it is to be assumed that this is equivalent
to "antagonism of chemokine stimulation of CCR-2."
SUMMARY OF THE INVENTION
[0028] Accordingly, the present invention provides novel
antagonists or partial agonists/antagonists of MCP-1 or CCR-2
receptor activity, or pharmaceutically acceptable salts or prodrugs
thereof.
[0029] The present invention provides pharmaceutical compositions
comprising a pharmaceutically acceptable carrier and a
therapeutically effective amount of at least one of the compounds
of the present invention or a pharmaceutically acceptable salt or
prodrug form thereof.
[0030] The present invention provides a method for treating
rheumatoid arthritis and transplant rejection, comprising
administering to a host in need of such treatment a therapeutically
effective amount of at least one of the compounds of the present
invention or a pharmaceutically acceptable salt or prodrug form
thereof.
[0031] The present invention provides a method for treating
inflammatory diseases, comprising administering to a host in need
of such treatment a therapeutically effective amount of at least
one of the compounds of the present invention or a pharmaceutically
acceptable salt or prodrug form thereof.
[0032] The present invention provides novel chromene derivatives
for use in therapy.
[0033] The present invention provides the use of novel chromene
derivatives for the manufacture of a medicament for the treatment
of inflammatory diseases.
[0034] These and other features of the invention, which will become
apparent during the following detailed description, have been
achieved by the inventors' discovery that compounds of formula
(I):
##STR00002##
or stereoisomers or prodrugs or pharmaceutically acceptable salts
thereof, wherein W.sup.1, W.sup.2, W.sup.3, Y, Z, R.sup.2, R.sup.3,
R.sup.3' and R.sup.4, are defined below, are effective modulators
of MCP-1 and chemokine activity.
DETAILED DESCRIPTION OF THE PRESENT INVENTION
[0035] In one embodiment, the present invention provides novel
compounds of formula (I):
##STR00003##
or a stereoisomer or a pharmaceutically acceptable salt thereof,
wherein:
[0036] R.sup.2 is H, or C.sub.1-4 alkyl;
[0037] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl;
[0038] or R.sup.3 and R.sup.3', when attached to neighboring
carbons, taken together form --CH.dbd.CH--CH.dbd.CH--;
[0039] R.sup.4 is H or C.sub.1-4 alkyl;
[0040] W.sup.1 is N or C--Z';
[0041] W.sup.2 is N or C--Y';
[0042] W.sup.3 is N or C--X;
[0043] X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl;
[0044] Y and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4
alkoxy, halo, haloC.sub.1-4 alkyl or NO.sub.2;
[0045] Z and Z' are independently H or halo;
[0046] or Y and Z, Y' and Z', Y and X, or Y' and X are taken
together to form --CH.dbd.CH--CH.dbd.CH--;
[0047] or Y and X or Y' and X are taken together to form
--O--CH.sub.2--O--; provided that:
[0048] (1) only one of W.sup.1, W.sup.2 or W.sup.3 may be N;
[0049] (2) if W.sup.2 is N, then X, Y, Z and Z' are not all H;
[0050] (3) if W.sup.3 is N, then W.sup.1 and W.sup.2 are both CH
and Y and Z are H;
[0051] (4) if W.sup.1, W.sup.2 and W.sup.3 are all CH, then Y
cannot be H;
[0052] (5) if Y is C.sub.1-4alkoxy then X is not H;
[0053] (6) if W.sup.1, W.sup.2 and W.sup.3 are all CH and Z is Cl,
then Y is Cl; and
[0054] (7) if X is F then one of Y or Y' is not H.
[0055] In yet another embodiment, compounds of the present
invention, or a stereoisomer or pharmaceutically acceptable salt
from thereof, are those in which:
[0056] R.sup.2 is H, or C.sub.1-4 alkyl;
[0057] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl;
[0058] R.sup.4 is H or C.sub.1-4 alkyl;
[0059] W.sup.1 is N or C--Z';
[0060] W.sup.2 is N or C--Y';
[0061] W.sup.3 is N or C--X;
[0062] X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl;
[0063] Y and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4
alkoxy, halo, haloC.sub.1-4 alkyl or NO.sub.2;
[0064] Z and Z' are independently H or halo;
[0065] or Y and Z, Y' and Z', Y and X, or Y' and X are taken
together to form --CH.dbd.CH--CH.dbd.CH--;
[0066] or Y and X or Y' and X are taken together to form
--O--CH.sub.2--O--.
[0067] In still yet another embodiment, compounds of the present
invention, or a stereoisomer or pharmaceutically acceptable salt
from thereof, are those in which:
[0068] R.sup.2 is H, or C.sub.1-4 alkyl;
[0069] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl;
[0070] R.sup.4 is H or C.sub.1-4 alkyl;
[0071] W.sup.1 is N or C--Z';
[0072] W.sup.2 is N or C--Y';
[0073] W.sup.3 is N or C--X;
[0074] X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl;
[0075] Y and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4
alkoxy, halo, haloC.sub.1-4 alkyl or NO.sub.2;
[0076] Z and Z' are independently H or halo;
[0077] or Y and Z, Y' and Z', Y and X, or Y' and X are taken
together to form --CH.dbd.CH--CH.dbd.CH--.
[0078] In one embodiment, compounds of the present invention, or a
stereoisomer or pharmaceutically acceptable salt from thereof, are
those in which:
[0079] R.sup.2 is H, or C.sub.1-4 alkyl;
[0080] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, halo, or haloC.sub.1-4 alkyl;
[0081] R.sup.4 is H or C.sub.1-4 alkyl;
[0082] W.sup.1 is N or C--Z';
[0083] W.sup.2 is N or C--Y';
[0084] W.sup.3 is N or C--X;
[0085] X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl;
[0086] Y and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4
alkoxy, halo, haloC.sub.1-4 alkyl or NO.sub.2; and
[0087] Z and Z' are independently H or halo.
[0088] In another embodiment, compounds of the present invention,
or a stereoisomer or pharmaceutically acceptable salt from thereof,
are those in which:
[0089] R.sup.2 is H, or C.sub.1-4 alkyl;
[0090] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, or halo;
[0091] R.sup.4 is H or C.sub.1-4 alkyl;
[0092] W.sup.1 is N or C--Z';
[0093] W.sup.2 is N or C--Y';
[0094] W.sup.3 is N or C--X;
[0095] X is H, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, halo or
haloC.sub.1-4 alkyl;
[0096] Y and Y' are independently H, C.sub.1-4 alkyl, C.sub.1-4
alkoxy, halo, or haloC.sub.1-4 alkyl; and
[0097] Z and Z' are independently H or halo.
[0098] In yet another embodiment, compounds of the present
invention, or a stereoisomer or pharmaceutically acceptable salt
from thereof, are those in which:
[0099] R.sup.2 is H, or C.sub.1-4 alkyl;
[0100] R.sup.3 and R.sup.3' are independently H, C.sub.1-4 alkoxy,
or halo;
[0101] R.sup.4 is H or C.sub.1-4 alkyl;
[0102] W.sup.1 is N or C--Z';
[0103] W.sup.2 is N or C--Y';
[0104] W.sup.3 is N or C--X;
[0105] X is H, C.sub.1-4 alkyl, halo or haloC.sub.1-4 alkyl;
[0106] Y and Y' are independently H, C.sub.1-4 alkyl, halo, or
haloC.sub.1-4 alkyl; and
[0107] Z and Z' are independently H or halo.
[0108] In still yet another embodiment, compounds of the present
invention, or a stereoisomer or pharmaceutically acceptable salt
from thereof, are those in which:
[0109] R.sup.2 is H, or C.sub.1-4 alkyl;
[0110] R.sup.3 and R.sup.3' are independently H or halo;
[0111] R.sup.4 is H or C.sub.1-4 alkyl;
[0112] W.sup.1 is N or C--Z';
[0113] W.sup.2 is N or C--Y';
[0114] W.sup.3 is N or C--X;
[0115] X is H, halo or haloC.sub.1-4 alkyl;
[0116] Y and Y' are independently H, halo, or haloC.sub.1-4 alkyl;
and
[0117] Z and Z' are independently H or halo.
[0118] In one embodiment, compounds of the present invention, or a
stereoisomer or pharmaceutically acceptable salt from thereof, are
those in which:
[0119] R.sup.2 is H, methyl or ethyl;
[0120] R.sup.3 and R.sup.3' are H;
[0121] R.sup.4 is H;
[0122] W.sup.1 is N or C--Z';
[0123] W.sup.2 is N or C--Y';
[0124] W.sup.3 is N or C--X;
[0125] X is H, Br, Cl, F or CF.sub.3;
[0126] Y and Y' are independently H, Br, Cl, F or CF.sub.3; and
[0127] Z and Z' are independently H, Br, Cl or F.
[0128] In one embodiment, compounds of Formula (I), or a
stereoisomer or pharmaceutically acceptable salt from thereof, are
those compounds exemplified in the examples.
[0129] In another embodiment, the present invention is directed to
a pharmaceutical composition, comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of a
compound of the present invention.
[0130] In another embodiment, the present invention is directed to
a method for modulation of chemokine or chemokine receptor activity
comprising administering to a patient in need thereof a
therapeutically effective amount of a compound of the present
invention.
[0131] In another embodiment, the present invention is directed to
a method for modulation of CCR-2 receptor activity comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0132] In another embodiment, the present invention is directed to
a method for modulation of MCP-1 activity that is mediated by the
CCR-2 receptor comprising administering to a patient in need
thereof a therapeutically effective amount of a compound of the
present invention.
[0133] In another embodiment, the present invention is directed to
a method for treating disorders, comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of the present invention, said wherein said disorder is
selected from osteoarthritis, aneurysm, fever, cardiovascular
effects, Crohn's disease, congestive heart failure, autoimmune
diseases, HIV-infection, HIV-associated dementia, psoriasis,
idiopathic pulmonary fibrosis, transplant arteriosclerosis,
physically- or chemically-induced brain trauma, inflammatory bowel
disease, alveolitis, colitis, systemic lupus erythematosus,
nephrotoxic serum nephritis, glomerularnephritis, asthma, multiple
sclerosis, atherosclerosis, rheumatoid arthritis, restinosis, organ
transplantation, psoriatic arthritis, multiple myeloma, allergies,
for example, skin and mast cell degranulation in eye conjunctiva,
hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,
preferably, Crohn's disease, psoriasis, inflammatory bowel disease,
systemic lupus erythematosus, multiple sclerosis, rheumatoid
arthritis, multiple myeloma, allergies, for example, skin and mast
cell degranulation in eye conjunctiva, hepatocellular carcinoma,
osteoporosis and renal fibrosis.
[0134] In another embodiment, the present invention is directed to
a method for treating inflammatory diseases, comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0135] In another embodiment, the present invention is directed to
a method for treating inflammatory bowel disease, comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0136] In another embodiment, the present invention is directed to
a method for treating Crohn's disease, comprising administering to
a patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0137] In another embodiment, the present invention is directed to
a method for treating psoriasis, comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0138] In another embodiment, the present invention is directed to
a method for treating systemic lupus erythematosus, comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0139] In another embodiment, the present invention is directed to
a method for treating multiple sclerosis, comprising administering
to a patient in need thereof a therapeutically effective amount of
a compound of the present invention.
[0140] In another embodiment, the present invention is directed to
a method for treating rheumatoid arthritis, comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0141] In another embodiment, the present invention is directed to
a method for treating psoriatic arthritis, comprising administering
to a patient in need thereof a therapeutically effective amount of
a compound of the present invention.
[0142] In another embodiment, the present invention is directed to
a method for treating multiple myeloma, comprising administering to
a patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0143] In another embodiment, the present invention is directed to
a method for treating allergies, for example, skin and mast cell
degranulation in eye conjunctiva, comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0144] In another embodiment, the present invention is directed to
a method for treating hepatocellular carcinoma, comprising
administering to a patient in need thereof a therapeutically
effective amount of a compound of the present invention.
[0145] In another embodiment, the present invention is directed to
a method for treating osteoporosis, comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0146] In another embodiment, the present invention is directed to
a method for treating renal fibrosis, comprising administering to a
patient in need thereof a therapeutically effective amount of a
compound of the present invention.
[0147] In another embodiment, the present invention is directed to
a method for treating inflammatory diseases, for example,
inflammatory diseases which are at least partially mediated by
CCR-2, comprising administering to a patient in need thereof a
therapeutically effective amount of a compound of the present
invention.
[0148] In another embodiment, the present invention is directed to
a method for modulation of CCR2 activity comprising administering
to a patient in need thereof a therapeutically effective amount of
a compound of the present invention.
[0149] In another embodiment, the present invention is directed the
use of a compound of the present invention in the preparation of a
medicament for the treatment of a disorder, said disorder is
selected from osteoarthritis, aneurysm, fever, cardiovascular
effects, Crohn's disease, congestive heart failure, autoimmune
diseases, HIV-infection, HIV-associated dementia, psoriasis,
idiopathic pulmonary fibrosis, transplant arteriosclerosis,
physically- or chemically-induced brain trauma, inflammatory bowel
disease, alveolitis, colitis, systemic lupus erythematosus,
nephrotoxic serum nephritis, glomerularnephritis, asthma, multiple
sclerosis, atherosclerosis, rheumatoid arthritis, restinosis, organ
transplantation, psoriatic arthritis, multiple myeloma, allergies,
for example, skin and mast cell degranulation in eye conjunctiva,
hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,
preferably, Crohn's disease, psoriasis, inflammatory bowel disease,
systemic lupus erythematosus, multiple sclerosis, rheumatoid
arthritis, multiple myeloma, allergies, for example, skin and mast
cell degranulation in eye conjunctiva, hepatocellular carcinoma,
osteoporosis and renal fibrosis.
[0150] In another embodiment, the present invention is directed to
a compound of the present invention for use in therapy.
[0151] In another embodiment, the present invention is directed to
a pharmaceutical composition comprising a compound of the present
invention and one or more active ingredients.
[0152] In another embodiment, the present invention is directed to
a method for modulation of chemokine or chemokine receptor activity
comprising administering to a patient in need thereof a
therapeutically effective amount of a pharmaceutical composition
comprised of a compound of the present invention and one or more
active ingredients.
[0153] In another embodiment, the present invention is directed to
a method for modulation of CCR-2 receptor activity comprising
administering to a patient in need thereof a therapeutically
effective amount of a pharmaceutical composition comprised of a
compound of the present invention and one or more active
ingredients.
[0154] In yet another embodiment, the present invention is directed
to a method for modulation of MCP-1 activity that is mediated by
the CCR-2 receptor comprising administering to a patient in need
thereof a therapeutically effective amount of a pharmaceutical
composition comprised of a compound of the present invention and
one or more active ingredients.
[0155] In another embodiment, the present invention is directed to
a method for treating a disorder, comprising administering to a
patient in need thereof a therapeutically effective amount of a
pharmaceutical composition comprised of a compound of the present
invention and one or more active ingredients, wherein said disorder
is selected from osteoarthritis, aneurysm, fever, cardiovascular
effects, Crohn's disease, congestive heart failure, autoimmune
diseases, HIV-infection, HIV-associated dementia, psoriasis,
idiopathic pulmonary fibrosis, transplant arteriosclerosis,
physically- or chemically-induced brain trauma, inflammatory bowel
disease, alveolitis, colitis, systemic lupus erythematosus,
nephrotoxic serum nephritis, glomerularnephritis, asthma, multiple
sclerosis, atherosclerosis, rheumatoid arthritis, restinosis, organ
transplantation, psoriatic arthritis, multiple myeloma, allergies,
for example, skin and mast cell degranulation in eye conjunctiva,
hepatocellular carcinoma, osteoporosis, renal fibrosis and cancer,
preferably, Crohn's disease, psoriasis, inflammatory bowel disease,
systemic lupus erythematosus, multiple sclerosis, rheumatoid
arthritis, multiple myeloma, allergies, for example, skin and mast
cell degranulation in eye conjunctiva, hepatocellular carcinoma,
osteoporosis and renal fibrosis.
[0156] In yet another embodiment, the present invention, is
directed to a method for treating inflammatory diseases,
preferably, inflammatory diseases which are at least partially
mediated by CCR-2, comprising administering to a patient in need
thereof a therapeutically effective amount of a pharmaceutical
composition comprised of a compound of present invention and one or
more active ingredients.
[0157] In another embodiment, the present invention is directed to
a method for modulation of CCR-2 activity comprising administering
to a patient in need thereof a therapeutically effective amount of
a pharmaceutical composition comprised of a compound of the present
invention and one or more active ingredients.
[0158] In another embodiment, the present invention is directed to
the use of a pharmaceutical composition comprised of a compound of
the present invention and one or more active ingredients in the
preparation of a medicament for the treatment of a disorder, said
disorder is selected from osteoarthritis, aneurysm, fever,
cardiovascular effects, Crohn's disease, congestive heart failure,
autoimmune diseases, HIV-infection, HIV-associated dementia,
psoriasis, idiopathic pulmonary fibrosis, transplant
arteriosclerosis, physically- or chemically-induced brain trauma,
inflammatory bowel disease, alveolitis, colitis, systemic lupus
erythematosus, nephrotoxic serum nephritis, glomerularnephritis,
asthma, multiple sclerosis, atherosclerosis, rheumatoid arthritis,
restinosis, organ transplantation, psoriatic arthritis, multiple
myeloma, allergies, for example, skin and mast cell degranulation
in eye conjunctiva, hepatocellular carcinoma, osteoporosis, renal
fibrosis and cancer, preferably, Crohn's disease, psoriasis,
inflammatory bowel disease, systemic lupus erythematosus, multiple
sclerosis, rheumatoid arthritis, multiple myeloma, allergies, for
example, skin and mast cell degranulation in eye conjunctiva,
hepatocellular carcinoma, osteoporosis and renal fibrosis.
[0159] In still yet another embodiment, the present invention is
directed to the use of a pharmaceutical composition comprised of a
compound of the present invention and one or more active
ingredients in therapy.
[0160] In another embodiment, the present invention provides a
compound of the present invention for use in therapy for treating
an inflammatory disease.
[0161] In another embodiment, the present invention provides a
combined preparation of a compound of the present invention and
additional therapeutic agent(s) for simultaneous, separate or
sequential use in therapy.
[0162] In another embodiment, the present invention provides a
combined preparation of a compound of the present invention and
additional therapeutic agent(s) for simultaneous, separate or
sequential use in treatment of an inflammatory disease.
[0163] In another embodiment, the present invention provides a
novel article of manufacture, comprising: (a) a first container;
(b) a pharmaceutical composition located within the first
container, wherein the composition, comprises: a first therapeutic
agent, comprising: a compound of the present invention; and (c) a
package insert stating that the pharmaceutical composition can be
used for the treatment of an inflammatory disease.
[0164] In another preferred embodiment, the present invention
provides a novel article of manufacture, further comprising: (d) a
second container; wherein components (a) and (b) are located within
the second container and component (c) is located within or outside
of the second container.
[0165] In another embodiment, the present invention provides a
novel article of manufacture, comprising: (a) a first container;
(b) a pharmaceutical composition located within the first
container, wherein the composition, comprises: a first therapeutic
agent, comprising: a compound of the present invention; and (c) a
package insert stating that the pharmaceutical composition can be
used in combination with a second therapeutic agent to treat an
inflammatory disease.
[0166] The invention may be embodied in other specific forms
without departing from the spirit or essential attributes thereof.
This invention also encompasses all combinations of alternative
aspects of the invention noted herein. It is understood that any
and all embodiments of the present invention may be taken in
conjunction with any other embodiment to describe additional
embodiments of the present invention. Furthermore, any elements of
an embodiment may be combined with any and all other elements from
any of the embodiments to describe additional embodiments.
DEFINITIONS
[0167] The compounds herein described may have asymmetric centers.
Compounds of the present invention containing an asymmetrically
substituted atom may be isolated in optically active or racemic
forms. It is well known in the art how to prepare optically active
forms, such as by resolution of racemic forms or by synthesis from
optically active starting materials. Many geometric isomers of
olefins, C.dbd.N double bonds, and the like can also be present in
the compounds described herein, and all such stable isomers are
contemplated in the present invention. Cis and trans geometric
isomers of the compounds of the present invention are described and
may be isolated as a mixture of isomers or as separated isomeric
forms. All chiral, diastereomeric, racemic forms and all geometric
isomeric forms of a structure are intended, unless the specific
stereochemistry or isomeric form is specifically indicated.
[0168] One enantiomer of a compound of Formula I may display
superior activity compared with the other. Thus, all of the
stereochemistries are considered to be a part of the present
invention. When required, separation of the racemic material can be
achieved by HPLC using a chiral column or by a resolution using a
resolving agent such as camphonic chloride as in Young, S. D. et
al., Antimicrobial Agents and Chemotherapy 1995, 2602-2605.
[0169] The term "substituted," as used herein, means that any one
or more hydrogens on the designated atom or ring is replaced with a
selection from the indicated group, provided that the designated
atom's or ring atom's normal valency is not exceeded, and that the
substitution results in a stable compound. When a substituent is
keto (i.e., .dbd.O), then 2 hydrogens on the atom are replaced.
[0170] When any variable (e.g., R.sub.4) occurs more than one time
in any constituent or formula for a compound, its definition at
each occurrence is independent of its definition at every other
occurrence. Thus, for example, if a group is shown to be
substituted with (R.sub.4).sub.m and m is 0-3, then said group may
optionally be substituted with up to three R.sub.4 groups and
R.sub.4 at each occurrence is selected independently from the
definition of R.sub.4. Also, combinations of substituents and/or
variables are permissible only if such combinations result in
stable compounds.
[0171] When a bond to a substituent is shown to cross a bond
connecting two atoms in a ring, then such substituent may be bonded
to any atom on the ring. When a substituent is listed without
indicating the atom via which such substituent is bonded to the
rest of the compound of a given formula, then such substituent may
be bonded via any atom in such substituent. Combinations of
substituents and/or variables are permissible only if such
combinations result in stable compounds.
[0172] As used herein, "alkyl" is intended to include both branched
and straight-chain saturated aliphatic hydrocarbon groups
containing 1 to 20 carbons, preferably 1 to 10 carbons, more
preferably 1 to 8 carbons, in the normal chain, such as methyl,
ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl,
isohexyl, heptyl, 4,4-dimethylpentyl, octyl,
2,2,4-trimethyl-pentyl, nonyl, decyl, undecyl, dodecyl, the various
branched chain isomers thereof, and the like as well as such groups
may optionally include 1 to 4 substituents such as halo, for
example F, Br, Cl, or I, or CF.sub.3, alkyl, alkoxy, aryl, aryloxy,
aryl(aryl) or diaryl, arylalkyl, arylalkyloxy, alkenyl, cycloalkyl,
cycloalkylalkyl, cycloalkylalkyloxy, amino, hydroxy, hydroxyalkyl,
acyl, heteroaryl, heteroaryloxy, heteroarylalkyl, heteroarylalkoxy,
aryloxyalkyl, alkylthio, arylalkylthio, aryloxyaryl, alkylamido,
alkanoylamino, arylcarbonylamino, nitro, cyano, thiol, haloalkyl,
trihaloalkyl, and/or alkylthio.
[0173] Unless otherwise indicated, the term "alkenyl" as used
herein by itself or as part of another group refers to straight or
branched chain radicals of 2 to 20 carbons, preferably 2 to 12
carbons, and more preferably 1 to 8 carbons in the normal chain,
which include one to six double bonds in the normal chain, such as
vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl,
2-hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl,
3-octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl, 4-dodecenyl,
4,8,12-tetradecatrienyl, and the like, and which may be optionally
substituted with 1 to 4 substituents, namely, halogen, haloalkyl,
alkyl, alkoxy, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl,
amino, hydroxy, heteroaryl, cycloheteroalkyl, alkanoylamino,
alkylamido, arylcarbonyl-amino, nitro, cyano, thiol, alkylthio,
and/or any of the alkyl substituents set out herein.
[0174] Unless otherwise indicated, the term "alkynyl" as used
herein by itself or as part of another group refers to straight or
branched chain radicals of 2 to 20 carbons, preferably 2 to 12
carbons and more preferably 2 to 8 carbons in the normal chain,
which include one triple bond in the normal chain, such as
2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl,
2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl,
3-octynyl, 3-nonynyl, 4-decynyl, 3-undecynyl, 4-dodecynyl, and the
like, and which may be optionally substituted with 1 to 4
substituents, namely, halogen, haloalkyl, alkyl, alkoxy, alkenyl,
alkynyl, aryl, arylalkyl, cycloalkyl, amino, heteroaryl,
cycloheteroalkyl, hydroxy, alkanoylamino, alkylamido,
arylcarbonylamino, nitro, cyano, thiol, and/or alkylthio, and/or
any of the alkyl substituents set out herein.
[0175] Unless otherwise indicated, the term "cycloalkyl" as
employed herein alone or as part of another group includes
saturated or partially unsaturated (containing 1 or 2 double bonds)
cyclic hydrocarbon groups containing 1 to 3 rings, including
monocyclic alkyl, bicyclic alkyl (or bicycloalkyl) and tricyclic
alkyl, containing a total of 3 to 20 carbons forming the ring,
preferably 3 to 10 carbons, forming the ring and which may be fused
to 1 or 2 aromatic rings as described for aryl, which includes
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl,
##STR00004##
any of which groups may be optionally substituted with 1 to 4
substituents such as halogen, alkyl, alkoxy, hydroxy, aryl,
aryloxy, arylalkyl, cycloalkyl, alkylamido, alkanoylamino, oxo,
acyl, arylcarbonylamino, amino, nitro, cyano, thiol, and/or
alkylthio, and/or any of the substituents for alkyl.
[0176] Where alkyl groups as defined above have single bonds for
attachment to other groups at two different carbon atoms, they are
termed "alkylene" groups and may optionally be substituted as
defined above for "alkyl".
[0177] Where alkenyl groups as defined above and alkynyl groups as
defined above, respectively, have single bonds for attachment at
two different carbon atoms, they are termed "alkenylene groups" and
"alkynylene groups", respectively, and may optionally be
substituted as defined above for "alkenyl" and "alkynyl".
[0178] "Halo" or "halogen" as used herein refers to fluoro, chloro,
bromo, and iodo; and "haloalkyl" is intended to include both
branched and straight-chain saturated aliphatic hydrocarbon groups,
for example CF.sub.3, having the specified number of carbon atoms,
substituted with 1 or more halogen (for example --C.sub.vF.sub.w
where v=1 to 3 and w=1 to (2v+1)).
[0179] Unless otherwise indicated, the term "aryl" as employed
herein alone or as part of another group refers to monocyclic and
bicyclic aromatic groups containing 6 to 10 carbons in the ring
portion (such as phenyl or naphthyl, including 1-naphthyl and
2-naphthyl) and may optionally include 1 to 3 additional rings
fused to a carbocyclic ring or a heterocyclic ring (such as aryl,
cycloalkyl, heteroaryl, or cycloheteroalkyl rings for example
##STR00005##
and may be optionally substituted through available carbon atoms
with 1, 2, or 3 substituents, for example, hydrogen, halo,
haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl,
trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkyl-alkyl,
cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl,
arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio, arylazo,
heteroarylalkyl, heteroarylalkenyl, heteroarylheteroaryl,
heteroaryloxy, hydroxy, nitro, cyano, amino, substituted amino
wherein the amino includes 1 or 2 substituents (which are alkyl,
aryl, or any of the other aryl compounds mentioned in the
definitions), thiol, alkylthio, arylthio, heteroarylthio,
arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl,
alkyl-aminocarbonyl, arylaminocarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy,
alkylcarbonylamino, arylcarbonylamino, arylsulfinyl,
arylsulfinylalkyl, arylsulfonylamino, or arylsulfonaminocarbonyl,
and/or any of the alkyl substituents set out herein.
[0180] Unless otherwise indicated, the term "lower alkoxy",
"alkoxy", "aryloxy" or "aralkoxy" as employed herein alone or as
part of another group includes any of the above alkyl, aralkyl, or
aryl groups linked to an oxygen atom.
[0181] Unless otherwise indicated, the term "amino" as employed
herein alone or as part of another group refers to amino that may
be substituted with one or two substituents, which may be the same
or different, such as alkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl,
cycloalkyl, cycloalkylalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl,
or thioalkyl. These substituents may be further substituted with a
carboxylic acid and/or any of the R.sup.1 groups or substituents
for R.sup.1 as set out above. In addition, the amino substituents
may be taken together with the nitrogen atom to which they are
attached to form 1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl,
4-morpholinyl, 4-thiamorpholinyl, 1-piperazinyl,
4-alkyl-1-piperazinyl, 4-arylalkyl-1-piperazinyl, or
4-diarylalkyl-1-piperazinyl, all of which may be optionally
substituted with alkyl, alkoxy, alkylthio, halo, trifluoromethyl,
or hydroxy.
[0182] Unless otherwise indicated, the term "lower alkylthio,"
"alkylthio," "arylthio," or "aralkylthio" as employed herein alone
or as part of another group includes any of the above alkyl,
aralkyl, or aryl groups linked to a sulfur atom.
[0183] Unless otherwise indicated, the term "lower alkylamino,"
"alkylamino," "arylamino," or "arylalkylamino" as employed herein
alone or as part of another group includes any of the above alkyl,
aryl, or arylalkyl groups linked to a nitrogen atom.
[0184] As used herein, the term "heterocyclyl" or "heterocyclic
system" is intended to mean a stable 5, 6, or 7-membered monocyclic
or bicyclic or 7, 8, 9, or 10-membered bicyclic heterocyclic ring
which is saturated, partially unsaturated or unsaturated
(aromatic), and which consists of carbon atoms and 1, 2, 3, or 4
heteroatoms independently selected from the group consisting of N,
NH, O and S and including any bicyclic group in which any of the
above-defined heterocyclic rings is fused to a benzene ring. The
nitrogen and sulfur heteroatoms may optionally be oxidized. The
heterocyclic ring may be attached to its pendant group at any
heteroatom or carbon atom, which results in a stable structure. The
heterocyclic rings described herein may be substituted on carbon or
on a nitrogen atom if the resulting compound is stable. If
specifically noted, a nitrogen in the heterocycle may optionally be
quaternized. It is preferred that when the total number of S and O
atoms in the heterocycle exceeds 1, then these heteroatoms are not
adjacent to one another. As used herein, the term "aromatic
heterocyclic system" or "heteroaryl" is intended to mean a stable
5- to 7-membered monocyclic or bicyclic or 7- to 10-membered
bicyclic heterocyclic aromatic ring which consists of carbon atoms
and from 1 to 4 heteroatoms independently selected from the group
consisting of N, O and S and is aromatic in nature.
[0185] Examples of heterocycles include, but are not limited to,
1H-indazole, 2-pyrrolidonyl, 2H,6H-1,5,2-dithiazinyl, 2H-pyrrolyl,
1H-indolyl, 4-piperidonyl, 4aH-carbazole, 4H-quinolizinyl,
6H-1,2,5-thiadiazinyl, acridinyl, azocinyl, benzimidazolyl,
benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazalonyl, carbazolyl, 4aH-carbazolyl,
.beta.-carbolinyl, chromanyl, chromenyl, cinnolinyl,
decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, isobenzofuranyl, isochromanyl,
isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl,
benzimidazolyl, isothiazolyl, isoxazolyl, morpholinyl,
naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,
1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinylperimidinyl,
phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, pteridinyl, piperidonyl, 4-piperidonyl,
pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,
pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl,
4H-quinolizinyl, quinoxalinyl, quinuclidinyl, carbolinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,
1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl,
thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl,
thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl,
1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, and xanthenyl. In
another aspect of the invention, the heterocycles include, but are
not limited to, pyridinyl, thiophenyl, furanyl, indazolyl,
benzothiazolyl, benzimidazolyl, benzothiaphenyl, benzofuranyl,
benzoxazolyl, benzisoxazolyl, quinolinyl, isoquinolinyl,
imidazolyl, indolyl, isoidolyl, piperidinyl, piperidonyl,
4-piperidonyl, piperonyl, pyrazolyl, 1,2,4-triazolyl,
1,2,3-triazolyl, tetrazolyl, thiazolyl, oxazolyl, pyrazinyl, and
pyrimidinyl. Also included are fused ring and spiro compounds
containing, for example, the above heterocycles.
[0186] Examples of heteroaryls are 1H-indazole,
2H,6H-1,5,2-dithiazinyl, indolyl, 4aH-carbazole, 4H-quinolizinyl,
6H-1,2,5-thiadiazinyl, acridinyl, azocinyl, benzimidazolyl,
benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazalonyl, carbazolyl, 4aH-carbazolyl,
.beta.-carbolinyl, chromanyl, chromenyl, cinnolinyl,
decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, isobenzofuranyl, isochromanyl,
isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl
(benzimidazolyl), isothiazolyl, isoxazolyl, morpholinyl,
naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,
1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinylperimidinyl,
phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, pteridinyl, piperidonyl, 4-piperidonyl,
pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,
pyrazolinyl, pyrazolyl, pyrazolotriazinyl, pyridazinyl,
pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl,
pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
carbolinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl,
tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,
1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, and
xanthenyl. In another aspect of the invention, examples of
heteroaryls are indolyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl, benzoxazolyl, benzthiazolyl, benztriazolyl,
benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalonyl,
cinnolinyl, furanyl, imidazolyl, indazolyl, indolyl, isoquinolinyl
isothiazolyl, isoxazolyl, oxazolyl, pyrazinyl, pyrazolyl,
pyrazolotriazinyl, pyridazinyl, pyridyl, pyridinyl, pyrimidinyl,
pyrrolyl, quinazolinyl, quinolinyl, thiazolyl, thienyl, and
tetrazolyl.
[0187] The term "heterocyclylalkyl" or "heterocyclyl" as used
herein alone or as part of another group refers to heterocyclyl
groups as defined above linked through a C atom or heteroatom to an
alkyl chain.
[0188] The term "heteroarylalkyl" or "heteroarylalkenyl" as used
herein alone or as part of another group refers to a heteroaryl
group as defined above linked through a C atom or heteroatom to an
alkyl chain, alkylene, or alkenylene as defined above.
[0189] The term "cyano" as used herein, refers to a --CN group.
[0190] The term "nitro" as used herein, refers to an --NO.sub.2
group.
[0191] The term "hydroxy" as used herein, refers to an OH
group.
[0192] The phrase "pharmaceutically acceptable" is employed herein
to refer to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio.
[0193] As used herein, "pharmaceutically acceptable salts" refer to
derivatives of the disclosed compounds wherein the parent compound
is modified by making acid or base salts thereof. Examples of
pharmaceutically acceptable salts include, but are not limited to,
mineral or organic acid salts of basic residues such as amines;
alkali or organic salts of acidic residues such as carboxylic
acids; and the like. The pharmaceutically acceptable salts include
the conventional non-toxic salts or the quaternary ammonium salts
of the parent compound formed, for example, from non-toxic
inorganic or organic acids. For example, such conventional
non-toxic salts include those derived from inorganic acids such as
hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric
and the like; and the salts prepared from organic acids such as
acetic, propionic, succinic, glycolic, stearic, lactic, malic,
tartaric, citric, ascorbic, palmidic, maleic, hydroxymaleic,
phenylacetic, glutamic, benzoic, salicylic, sulfanilic,
2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane
disulfonic, oxalic, isethionic, and the like.
[0194] The pharmaceutically acceptable salts of the present
invention can be synthesized from the parent compound which
contains a basic or acidic moiety by conventional chemical methods.
Generally, such salts can be prepared by reacting the free acid or
base forms of these compounds with a stoichiometric amount of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two; generally, nonaqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists
of suitable salts are found in Remington's Pharmaceutical Sciences,
17th ed., p. 1418 (Mack Publishing Company, Easton, Pa., 1985), the
disclosure of which is hereby incorporated by reference.
[0195] Any compound that can be converted in vivo to provide the
bioactive agent (i.e., the compound of formula I) is a prodrug
within the scope and spirit of the invention.
[0196] The term "prodrugs" as employed herein includes esters,
carbamates and carbonates formed by reacting one or more hydroxyls
of compounds of formula I with alkyl, alkoxy, or aryl substituted
acylating agents employing procedures known to those skilled in the
art to generate acetates, pivalates, methylcarbonates, benzoates,
and the like.
[0197] Various forms of prodrugs are well known in the art and are
described in: [0198] a) Wermuth, C. G. et al., The Practice of
Medicinal Chemistry, Ch. 31 (Academic Press, 1996); [0199] b)
Bundgaard, H., ed., Design of Prodrugs (Elsevier, 1985); [0200] c)
Krause, B. R. et al., Chapter 6: "ACAT Inhibitors: Physiologic
Mechanisms for Hypolipidemic and Anti-Atherosclerotic Activities in
Experimental Animals", Inflammation: Mediators and Pathways,
Ruffolo, Jr., R. R. et al., eds., pp. 173-198 (CRC Press, Inc.,
1995); and [0201] d) Testa, B. et al., Hydrolysis in Drug and
Prodrug Metabolism (Wiley-VCH, 2003). Said references are
incorporated herein by reference.
[0202] In addition, compounds of the formula I are, subsequent to
their preparation, preferably isolated and purified to obtain a
composition containing an amount by weight equal to or greater than
99% formula I compound ("substantially pure" compound I), which is
then used or formulated as described herein. Such "substantially
pure" compounds of the formula I are also contemplated herein as
part of the present invention.
[0203] All stereoisomers of the compounds of the instant invention
are contemplated, either in admixture or in pure or substantially
pure form. The compounds of the present invention can have
asymmetric centers at any of the carbon atoms including any one of
the R substituents and/or exhibit polymorphism. Consequently,
compounds of formula I can exist in enantiomeric, or diastereomeric
forms, or in mixtures thereof. The processes for preparation can
utilize racemates, enantiomers, or diastereomers as starting
materials. When diastereomeric or enantiomeric products are
prepared, they can be separated by conventional methods for
example, chromatographic or fractional crystallization.
[0204] "Stable compound" and "stable structure" are meant to
indicate a compound that is sufficiently robust to survive
isolation to a useful degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent. The present
invention is intended to embody stable compounds.
[0205] "Therapeutically effective amount" is intended to include an
amount of a compound of the present invention alone or an amount of
the combination of compounds claimed or an amount of a compound of
the present invention in combination with other active ingredients
effective to inhibit MCP-1 or effective to treat or prevent
inflammatory disorders.
[0206] As used herein, "treating" or "treatment" cover the
treatment of a disease-state in a mammal, particularly in a human,
and include: (a) preventing the disease-state from occurring in a
mammal, in particular, when such mammal is predisposed to the
disease-state but has not yet been diagnosed as having it; (b)
inhibiting the disease-state, i.e., arresting it development;
and/or (c) relieving the disease-state, i.e., causing regression of
the disease state.
Synthesis
[0207] The compounds of the present invention can be prepared in a
number of ways well known to one skilled in the art of organic
synthesis. The compounds of the present invention can be
synthesized using the methods described below, together with
synthetic methods known in the art of synthetic organic chemistry,
or variations thereon as appreciated by those skilled in the art.
Preferred methods include, but are not limited to, those described
below. All references cited herein are hereby incorporated in their
entirety herein by reference.
[0208] The novel compounds of this invention may be prepared using
the reactions and techniques described in this section. The
reactions are performed in solvents appropriate to the reagents and
materials employed and are suitable for the transformations being
effected. Also, in the description of the synthetic methods
described below, it is to be understood that all proposed reaction
conditions, including choice of solvent, reaction atmosphere,
reaction temperature, duration of the experiment and work up
procedures, are chosen to be the conditions standard for that
reaction, which should be readily recognized by one skilled in the
art. It is understood by one skilled in the art of organic
synthesis that the functionality present on various portions of the
molecule must be compatible with the reagents and reactions
proposed. Such restrictions to the substituents that are compatible
with the reaction conditions will be readily apparent to one
skilled in the art and alternate methods must then be used. This
will sometimes require a judgment to modify the order of the
synthetic steps or to select one particular process scheme over
another in order to obtain a desired compound of the invention. It
will also be recognized that another major consideration in the
planning of any synthetic route in this field is the judicious
choice of the protecting group used for protection of the reactive
functional groups present in the compounds described in this
invention. An authoritative account describing the many
alternatives to the trained practitioner is Greene et al.
(Protective Groups In Organic Synthesis, Third Edition (Wiley and
Sons, 1999)).
[0209] Compounds of Formula (I) can be prepared by the route
outlined in Scheme 1, wherein A represents the pendent ring of
Formula (I). A phenolic .beta.-ketoester 1-1 (where R is, for
example, an alkyl group such as methyl, ethyl or ten-butyl) can be
reacted with an aromatic aldehyde A-CHO in the presence of
catalytic amounts of an acid such as acetic acid and a base such as
piperidine in a solvent such as isopropanol to provide an
intermediate 1-2 (R.sup.2=H), for example as reported by Cheng, J.
et al., Bioorg. Med. Chem. Lett. 2003, 13, 3647. Alternatively, an
appropriate chromenone 1-3 (where R is, for example, an alkyl group
such as methyl, ethyl or tert-butyl and R.sup.2=H or alkyl) can be
converted to 1-2 by treatment with a Grignard reagent A-MgBr or
A-MgCl in the presence of a copper (I) compound such as copper(I)
iodide, copper(I) bromide, or copper (I) cyanide, or by treatment
with a lithium organocuprate A.sub.2CuLi, for example as reported
by N. Ishizuka et al., J. Med. Chem. 2002, 45, 2041, or by
Saengchantara, S. et al., Tetrahedron 1990, 46, 3029.
##STR00006##
[0210] Intermediates 1-2 can be treated with a reducing agent such
as sodium borohydride in an appropriate solvent such as methanol or
a mixture of solvents such as methanol and tetrahydrofuran to
provide intermediates 1-4. These intermediates can then be
converted to intermediates 1-5 (where R is, for example, alkyl) by
a number of methods known to one skilled in the art, for example by
treatment with an acid, optionally in an appropriate solvent, for
example trifluoroacetic acid or p-toluenesulfonic acid in toluene.
Alternatively, the hydroxyl group of 1-4 can be converted into a
more reactive leaving group such as a methanesulfonate ester,
trifluoromethanesulfonate ester, arylsulfonate ester or halogen,
using methods well known to one skilled in the art. This
intermediate can then be treated with a base such as, for example,
1,8-diazabicyclo[5.4.0]undec-7-ene to induce elimination to 1-5.
Intermediates 1-5 (R=alkyl) can then be converted to compounds of
Formula (I) represented by 1-5 (R=H) by hydrolysis under basic or
acidic conditions, or (for R=tert-butyl) by treatment with a strong
acid such as trifluoroacetic acid, hydrochloric acid or
p-toluenesulfonic acid, optionally in an appropriate solvent. Such
conversions of esters to acids are well known by those skilled in
the art of organic synthesis.
[0211] Intermediates 1-4 can also be converted directly to
compounds of Formula (I) represented by 1-5 (R=H) by treatment with
an aqueous solution of a base such as sodium hydroxide.
[0212] Intermediates 1-2 can also be converted to intermediates 1-7
wherein R.sup.4 is alkyl using procedures well known in the
chemical literature. Intermediates 1-2 can be converted into 1-6
wherein L is a group suitable for replacement with an alkyl moiety.
For example, treatment of 1-2 with a base such as sodium hydride in
a suitable solvent such as diethyl ether or tetrahydrofuran,
followed by treatment of the resulting anion with a chlorophosphate
such as dimethyl chlorophosphate or diethyl chlorophosphate (as
reported by Sum et al., Can. J. Chem. 1979, 57, 1431), can provide
the intermediates 1-6 where L is a dialkylphosphoryloxy group.
Treatment of these intermediates with a lithium dialkylcuprate can
provide the intermediates 1-7 (where R.sup.4 is an alkyl group),
which can then be converted to the compounds of Formula I
represented by 1-7 (where R.sup.4 is H), for example by hydrolysis
with aqueous sodium or potassium hydroxide as described above.
[0213] The starting intermediates 1-1 and 1-3 in Scheme 1 can be
prepared using methods reported in the chemical literature. For
example, phenolic .beta.-ketoesters 1-1 can be prepared by the
base-catalyzed condensation of a 2'-hydroxyacetophenone with a
dialkyl carbonate as reported by Cushman, M. et al., J. Med. Chem.
1991, 34, 798, or by condensation of a salicylic acid derivative
with an acetate ester (using a procedure such as that reported by
Appendino, G. et al., J. Nat. Prod. 1999, 62, 1627). Chromenone
derivatives 1-3 can be prepared, for example, by the base-catalyzed
condensation of a .beta.-ketoester such as methyl acetoacetate with
a 2-fluorobenzoyl chloride to provide 1-3 (R.sup.2=R=methyl), as
reported by G. Coppola et al., Synthesis 1981, 523, or by the
treatment of a phenolic .beta.-ketoester 1-1 with triethyl
orthoformate and acetic anhydride or with acetic formic anhydride
and sodium formate to provide 1-3 (R.sup.2=H) as reported by
Okumura, K. et al., Chem. Pharm. Bull. 1974, 22, 331.
EXAMPLES
[0214] Abbreviations used in the Examples are defined as "2.times."
for twice, ".degree. C." for degrees Celsius, "g" for gram or
grams, "mmol" for millimolar, "mL" for milliliter or milliliters,
".mu.L" for microliter or microliters, "M" for molar, "min" for
minute or minutes, "mg" for milligram or milligrams, "h" for hour
or hours, "LC" for liquid chromatography, "HPLC" for high
performance liquid chromatography, "TLC" for thin layer
chromatography, "MS" for mass spectroscopy, "rt" or "RT" for room
temperature, "calc." for calculated, "THF" for tetrahydrofuran,
"CDCl.sub.3" for chloroform, "sat." for saturated, "N" for normal,
and "HCl" for hydrochloric acid. "D", "L", "R" and "S" are
stereochemical designations familiar to those skilled in the art.
Chemical names were derived using ChemDraw Ultra Version 9.0.5
(CambridgeSoft). When this program failed to provide a name for the
exact structure in question, an appropriate name was assigned using
the same methodology utilized by the program.
Example 1
Preparation of racemic 2-(3-chlorophenyl)-2H-chromene-3-carboxylic
acid
[0215] Step 1.
[0216] A mixture of methyl 3-(2-hydroxyphenyl)-3-oxopropanoate
(prepared according to Cushman, M. et al., J. Med. Chem. 1991, 34,
798; 250 mg, 1.29 mmol) and 3-chlorobenzaldehyde (146 .mu.L, 1.29
mmol) in isopropanol (5.0 mL) was treated with piperidine (13
.mu.L, 129 .mu.mol) and acetic acid (7.4 .mu.L, 129 .mu.mol) and
heated to reflux for 17 h. After this time, the mixture was cooled
to rt. The cooled solution was concentrated under vacuum and the
resulting residue was purified by rotary TLC, eluting with 9:1
hexane-ethyl acetate, to provide a colorless gum. The colorless gum
was crystallized from a hot isopropanol/water solution to provide
methyl 2-(3-chlorophenyl)-4-oxochroman-3-carboxylate as an
off-white solid (173 mg, 42%). The NMR indicated predominantly the
enolic form. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.78 (s,
3H) 6.21 (s, 1H) 6.82 (d, J=8.1 Hz, 1H) 6.97 (t, J=7.9 Hz, 1H)
7.18-7.32 (m, 4H) 7.34 (m, 1H) 7.69 (dd, J=7.9, 1.8 Hz, 1H) 12.28
(br. s, 1H). High resolution mass spec (M+H).sup.+ calc. 317.0581,
found 317.0597.
[0217] Step 2.
[0218] A solution of methyl
2-(3-chlorophenyl)-4-oxochroman-3-carboxylate (153 mg, 483 .mu.mol)
in tetrahydrofuran (2.0 mL) was diluted with methanol (4.0 mL) and
treated at rt with sodium borohydride (18 mg, 483 .mu.mol). After
30 min, additional sodium borohydride (18 mg, 483 .mu.mol) was
added and stirring at rt was continued. After 3.5 h, additional
sodium borohydride (18 mg, 483 .mu.mol) was added. After 2 h, the
mixture was concentrated under vacuum and stirred with water
containing a few drops of 1.0 M NaOH (pH>10). The resulting
mixture was extracted three times with ethyl acetate, and the
combined organic phases were dried over sodium sulfate and
concentrated under vacuum to yield a residue. The residue was taken
up in methanol (2.0 mL) and 1.0 M aqueous sodium hydroxide (2.0 mL)
and then heated to reflux for 4 h. The solution was cooled to rt
and acidified with 3 mL of 1.0 M aqueous HCl. Methanol (about 10
mL) was added with heating until the gum all dissolved, then the
solution was refrigerated. The resulting white crystals of Example
1 were isolated by filtration (39 mg, 28%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 6.25 (s, 1H) 6.81 (d, J=7.6 Hz, 1H) 6.91
(dt, J=7.6, 1.0 Hz, 1H) 7.16-7.25 (m, 4H) 7.29 (dt, J=4.3, 2.0 Hz,
1H) 7.37 (m, 1H) 7.69 (s, 1H). High resolution mass spectrum
(M+H).sup.+ calc. 287.0475, found 287.0487.
Example 2-1
Preparation of racemic
2-(4-chloro-3-(trifluoromethyl)phenyl)-2H-chromene-3-carboxylic
acid
[0219] Step 1.
[0220] A mixture of methyl 3-(2-hydroxyphenyl)-3-oxopropanoate
(prepared according to Cushman, M. et al., J. Med. Chem. 1991, 200
mg, 1.03 mmol) and 4-chloro-3-(trifluoromethyl)benzaldehyde (148
.mu.L, 1.03 mmol) in isopropanol (4.0 mL) was treated with
piperidine (10 .mu.L, 103 .mu.mol) and acetic acid (5.9 .mu.L, 103
.mu.mol) and heated to reflux for 16.5 h. After this time, water
(2.0 mL) was added and the solution was slowly cooled to rt,
whereupon crystals formed. After standing at rt, the mixture was
cooled on ice, and the resulting crystals were collected by
filtration and dried to provide methyl
2-(4-chloro-3-(trifluoromethyl)phenyl)-4-oxochroman-3-carboxylate
as a white crystalline solid (204 mg, 51%). A second crop (47 mg)
was obtained from the mother liquor, raising the total yield to
63%. The NMR indicated predominantly the enolic form. .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. ppm 3.79 (s, 3H) 6.24 (s, 1H) 6.82
(d, J=8.2 Hz, 1H) 7.00 (t, J=7.7 Hz, 1H) 7.31 (t, J=6.9 Hz, 1H)
7.42 (m, 2H) 7.70 (m, 2H) 12.27 (br. s, 1H). Mass spectrum m/z
407.19, 409.18 (M+Na).sup.+.
[0221] Step 2.
[0222] A solution of methyl
2-(4-chloro-3-(trifluoromethyl)phenyl)-4-oxochroman-3-carboxylate
(300 mg, 780 .mu.mol) in tetrahydrofuran (2.0 mL) was diluted with
methanol (4.0 mL, 780 .mu.mol) and treated at rt with sodium
borohydride (30 mg, 780 .mu.mol). After 2.25 h, the solution was
poured into water (about 25 mL), treated with 1.0 M aqueous HCl
(about 1 mL) and then extracted three times with ethyl acetate. The
combined organic phases were dried over sodium sulfate and
concentrated under vacuum to provide crude racemic methyl
2-(4-chloro-3-(trifluoromethyl)phenyl)-4-hydroxychroman-3-carboxyl-
ate as a white solid (292 mg). Without further purification, this
material was dissolved in dichloromethane (9.5 mL) and treated with
trifluoroacetic acid (0.5 mL). After standing at rt for 1.5 h, the
solution was washed with saturated aqueous sodium bicarbonate,
dried over sodium sulfate and concentrated under vacuum to yield a
residue. The residue was purified by column chromatography (silica
gel, 2-10% ethyl acetate/hexane) to provide crude racemic methyl
2-(4-chloro-3-(trifluoromethyl)phenyl)-2H-chromene-3-carboxylate as
a colorless gum (65 mg). Without further purification, a solution
of a portion of this gum (60 mg) in methanol (2.0 mL) was treated
with 1.0 M aqueous sodium hydroxide (2.0 mL) and heated at reflux
for 4.5 h. After this time, the solution was cooled to rt. Once at
the prescribed temperature, a white solid formed, which was removed
by filtration and washed with water. The combined filtrate and wash
were acidified with 1.0 M aqueous HCl, giving a gummy material. A
small amount of methanol was added to the gummy material and the
resulting mixture was heated briefly, then stirred while cooling to
rt, thereby forming a white solid, which was collected by
filtration, washed with water and dried under vacuum to provide
Example 2-1 (29 mg). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
6.26 (s, 1H) 6.83 (d, J=8.1 Hz, 1H) 6.96 (t, J=7.1 Hz, 1H) 7.23
(dd, J=7.6, 1.0 Hz, 1H) 7.29 (m, 1H) 7.41 (d, J=8.1 Hz, 1H) 7.46
(dd, J=8.6, 2.0 Hz, 1H) 7.72 (d, J=1.5 Hz, 1H) 7.83 (s, 1H. Mass
spectrum m/z 355.24 (M+H).sup.+, 377.16 (M+Na).sup.+.
Examples 2-2 to 2-4
[0223] Examples 2-2 to 2-4 set forth in Table 1 below were prepared
using the methods described in Example 2-1:
TABLE-US-00001 TABLE 1 Example Compound name Mass spectrum 2-2
racemic 2-(4-chlorophenyl)-2H-chromene-3- 287.0 (M + H).sup.+
carboxylic acid 2-3 racemic 2-(4-methoxyphenyl)-2H-chromene- 283.1
(M + H).sup.+ 3-carboxylic acid 2-4 racemic
2-(3,5-dichlorophenyl)-2H- 319.0(M - H).sup.- chromene-3-carboxylic
acid
Example 3
Preparation of racemic
2-(benzo[d][1,3]dioxol-5-yl)-2H-chromene-3-carboxylic acid
[0224] Step 1.
[0225] A mixture of methyl 3-(2-hydroxyphenyl)-3-oxopropanoate
(prepared according to Cushman, M. et al., J. Med. Chem. 1991, 34,
798; 200 mg, 1.03 mmol) and piperonal (155 mg, 1.03 mmol) in
isopropanol (5.0 mL) was treated with piperidine (10 .mu.L, 103
.mu.mol) and acetic acid (5.9 .mu.L, 103 .mu.mol) and then heated
at reflux for 15.5 h. At the conclusion of this period, the
solution was treated with water (1.0 mL) and cooled to rt with
stirring. Once at the prescribed temperature, the mixture was
concentrated and purified by column chromatography (silica gel,
2-25% ethyl acetate/hexane) to provide a cloudy oil (172 mg). The
cloudy oil was crystallized from methanol to provide methyl
2-(benzo[d][1,3]dioxol-5-yl)-4-oxochroman-3-carboxylate as a white
solid (63 mg, 19%). The NMR indicated predominantly the enolic
form. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.76 (s, 3H)
5.91 (m, 2H) 6.15 (s, 1H) 6.70 (d, J=8.1 Hz, 1H) 6.78 (d, J=8.1 Hz,
1H) 6.82 (dd, J=8.1, 1.5 Hz, 1H) 6.86 (d, J=2.0 Hz, 1H) 6.96 (dt,
J=7.6, 1.0 Hz, 1H) 7.28 (m, 1H) 7.68 (dd, J=7.6, 1.5 Hz, 1H) 12.26
(br. s, 1H). Mass spectrum m/z 349.18 (M+Na).sup.+.
[0226] Step 2.
[0227] A solution of methyl
2-(benzo[d][1,3]dioxol-5-yl)-4-oxochroman-3-carboxylate (58 mg, 178
.mu.mol) in tetrahydrofuran (1.0 mL) was diluted with methanol (2.0
mL) and stirred on an ice/acetone bath. Sodium borohydride (7 mg,
178 .mu.mol) was added and the resulting mixture was stirred for 40
min. After this time, water (about 10 mL) was added and the mixture
was stirred while warming to rt. The resulting white solid was
collected by filtration, washed with water and dried under vacuum
to provide racemic methyl
2-(benzo[d][1,3]dioxol-5-yl)-4-hydroxychroman-3-carboxylate (51 mg,
87%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 2.69 (s, 1H)
3.47 (dd, J=6.6, 2.5 Hz, 1H) 3.51 (s, 3H) 5.25 (d, J=5.6 Hz, 1H)
5.31 (d, J=2.5 Hz, 1H) 5.99 (s, 2H) 6.82 (d, J=8.1 Hz, 1H) 6.89 (d,
J=8.1 Hz, 1H) 6.92 (m, 2H) 7.03 (t, J=7.4 Hz, 1H) 7.23 (td, J=7.1,
1.0 Hz, 1H) 7.57 (d, J=7.6 Hz, 1H). Mass spectrum m/z 351.20
(M+Na).sup.+.
[0228] Step 3.
[0229] A suspension of racemic methyl
2-(benzo[d][1,3]dioxol-5-yl)-4-hydroxychroman-3-carboxylate (49 mg,
149 .mu.mol) in dichloromethane (2.0 mL) was stirred on an ice bath
and treated with triethylamine (31 .mu.L, 224 .mu.mol), then
dropwise with methanesulfonyl chloride (13 .mu.L, 164 .mu.mol). The
cooling bath was removed after 90 min and the solution was stirred
at rt for 23 h. At the conclusion of this period, the solution was
diluted with additional dichloromethane, washed with 1.0 M aqueous
HCl, dried over sodium sulfate and concentrated under vacuum to
yield a residue. The residue was dissolved in benzene (2.0 mL),
treated with 1,8-diazabicyclo[5.4.0]undec-7-ene (30 .mu.L, 224
.mu.mol) and stirred at 60.degree. C. for 3.75 h. After this time,
the mixture was cooled to rt and then diluted with ethyl acetate,
washed with 1.0 M aqueous HCl, water, and saturated aqueous sodium
chloride, dried over sodium sulfate and concentrated under vacuum.
The resulting residue was purified by column chromatography (silica
gel, 2-10% ethyl acetate/hexane) to provide racemic methyl
2-(benzo[d][1,3]dioxol-5-yl)-2H-chromene-3-carboxylate as a white
solid (27 mg, 58%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
3.77 (s, 3H) 5.91 (m, 2H) 6.18 (s, 1H) 6.71 (d, J=8.6 Hz, 1H) 6.78
(d, J=8.1 Hz, 1H) 6.85 (m, 2H) 6.90 (t, J=7.4 Hz, 1H) 7.19 (m, 2H)
7.67 (s, 1H).
[0230] Step 4.
[0231] A solution of racemic methyl
2-(benzo[d][1,3]dioxol-5-yl)-2H-chromene-3-carboxylate (26 mg, 84
.mu.mol) in tetrahydrofuran (1.0 mL) was treated with 1.0 M aqueous
sodium hydroxide (1.0 mL) and the mixture was stirred vigorously at
rt for 23.5 h. After this time, the THF was removed under a
nitrogen stream, and the resulting aqueous residue was acidified
with 1.0 M aqueous HCl. The resulting solid was collected by
filtration, washed with water and dried under vacuum to provide
Example 3 as a white powder (19 mg, 77%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 5.91 (m, 2H) 6.16 (s, 1H) 6.71 (d, J=8.6
Hz, 1H) 6.79 (d, J=8.1 Hz, 1H) 6.86 (m, 2H) 6.91 (t, J=7.4 Hz, 1H)
7.21 (m, 2H) 7.76 (s, 1H). Mass spectrum m/z 297.29
(M+H).sup.+.
Example 4-1
Preparation of racemic
2-(3,4-dichlorophenyl)-2-methyl-2H-chromene-3-carboxylic acid
[0232] Step 1.
[0233] In a dried flask, a solution of methyl acetoacetate (2.16
mL, 20.0 mmol) in toluene (100 mL) was treated with sodium hydride
(60% mineral oil dispersion, 880 mg, 22.0 mmol). The resulting
mixture was stirred at rt for 10 min, and then a solution of
2-fluorobenzoyl chloride (2.23 mL, 20.0 mmol) in toluene (50 mL)
was added over about a 1 min. period. The resulting mixture was
heated at reflux for 24.5 h, and then poured into water (200 mL).
The mixture was extracted twice with ether (50 mL each). The
combined organic phases were dried over sodium sulfate and
concentrated under vacuum to provide a viscous yellow oil, which
crystallized. The solid was recrystallized from a ethyl
acetate/hexane solution to provide methyl
2-methyl-4-oxo-4H-chromene-3-carboxylate as off-white crystals in
two crops (2.10 g, 48%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 2.53 (s, 3H) 3.94 (s, 3H) 7.42 (m, 2H) 7.67 (ddd, J=8.5, 7.0,
1.8 Hz, 1H) 8.21 (dd, J=7.9, 1.8 Hz, 1H).
[0234] Step 2.
[0235] A suspension of copper(I) chloride (454 mg, 4.58 mmol) in
tetrahydrofuran (5.0 mL) was stirred in a dry flask under nitrogen
on ice. A solution of (3,4-dichlorophenyl)magnesium bromide (0.5 M)
in THF (9.17 mL, 4.58 mmol) was added over about a 2 min. period.
The resulting mixture was stirred on ice for 5 min, and then
treated with a solution of methyl
2-methyl-4-oxo-4H-chromene-3-carboxylate (500 mg, 2.29 mmol) in THF
(6.0 mL) over about a 2 min. period. The mixture was stirred on ice
for 10 min, and then allowed to warm to rt. After 4.5 h at rt, the
reaction mixture was quenched by adding 9:1 saturated aqueous
ammonium chloride/aqueous ammonia (20 mL). Upon completion of
addition, the reaction mixture was stirred at rt for 10 min. and
then ethyl acetate and additional ammonium chloride were added. The
aqueous and organic layers were mixed well and then separated. The
organic layer was washed twice with saturated ammonium chloride,
dried over sodium sulfate and then concentrated under vacuum to
yield a residue. The residue was purified by column chromatography
(silica gel, 9:1 hexane-ethyl acetate) to provide methyl
2-(3,4-dichlorophenyl)-2-methyl-4-oxochroman-3-carboxylate as a
colorless gum (518 mg, 62%). By NMR, this material was mixture of
the enol form and 2 diastereomeric keto forms (ratio about 1:1:2).
Mass spectrum m/z 387.01, 389.03 (M+Na).sup.+.
[0236] Step 3.
[0237] A solution of methyl
2-(3,4-dichlorophenyl)-2-methyl-4-oxochroman-3-carboxylate (490 mg,
1.34 mmol) in THF (2.0 mL) was diluted with methanol (4.0 mL) and
treated at rt with sodium borohydride (51 mg, 1.34 mmol).
Additional sodium borohydride was added after 4 h. After 2.5 h more
the solution was concentrated under vacuum. The residue was
partitioned between water (containing a few drops of 1.0 M aqueous
sodium hydroxide) and ethyl acetate. The organic phase was dried
over sodium sulfate and concentrated under vacuum to provide crude
methyl
2-(3,4-dichlorophenyl)-4-hydroxy-2-methylchroman-3-carboxylate as a
white solid (485 mg, 98%), which was used without further
purification. Mass spectrum m/z 389, 391 (M+Na).sup.+.
[0238] Step 4.
[0239] A solution of crude methyl
2-(3,4-dichlorophenyl)-4-hydroxy-2-methylchroman-3-carboxylate (100
mg, 272 .mu.mol) in dichloromethane (2.0 mL) was stirred on an ice
bath and treated with triethylamine (76 .mu.L, 545 .mu.mol), then
dropwise with methanesulfonyl chloride (23 .mu.L, 300 .mu.mol).
After stirring for 1.5 h, the solution was warmed to rt. After
stirring at rt for 6 h, the solution was diluted with
dichloromethane, washed with 1.0 M aqueous HCl, dried over sodium
sulfate and concentrated under vacuum. The resulting residue was
dissolved in benzene (2.0 mL), treated with
1,8-diazabicyclo[5.4.0]undec-7-ene (55 .mu.L, 408 .mu.mol) and
stirred at rt for 25 min. After this time, the resulting mixture
was heated to 60.degree. C. where it stirred for 5.25 h. After this
time, the solution was cooled to rt, diluted with ethyl acetate,
washed with water, 1.0 M aqueous HCl, saturated aqueous sodium
bicarbonate and saturated aqueous sodium chloride, dried over
sodium sulfate and then concentrated under vacuum to yield a
residue. The residue was purified by rotary TLC (silica gel, 92:8
hexane-ethyl acetate) to provide racemic methyl
2-(3,4-dichlorophenyl)-2-methyl-2H-chromene-3-carboxylate as a
colorless gum (44 mg, 46%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 2.02 (s, 3H) 3.73 (s, 3H) 6.82 (d, J=8.1 Hz, 1H) 6.92
(dt, J=7.6, 1.0 Hz, 1H) 7.15 (dd, J=7.6, 1.5 Hz, 1H) 7.25 (dt,
J=7.9, 1.5 Hz, 1H) 7.37 (m, 2H) 7.58 (s, 1H) 7.61 (t, J=1.5 Hz,
1H).
[0240] Step 5.
[0241] A solution of racemic methyl
2-(3,4-dichlorophenyl)-2-methyl-2H-chromene-3-carboxylate (44 mg,
126 .mu.mol) in methanol (1.0 mL) was treated with 1.0 M aqueous
sodium hydroxide (1.0 mL) and heated at reflux for 4.25 h. After
this time, the solution was cooled to rt, and then acidified with
1.0 M aqueous HCl, forming a dense white precipitate. The reaction
mixture was diluted with water (2 mL) and the resulting solid was
collected by filtration, washed with water and dried to provide
Example 4-1 as a white solid (33 mg, 78%). .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 2.04 (s, 3H) 6.83 (d, J=8.1 Hz, 1H) 6.93
(dt, J=7.6, 1.0 Hz, 1H) 7.17 (dd, J=7.6, 1.5 Hz, 1H) 7.28 (m, 1H)
7.37 (m, 2H) 7.60 (s, 1H) 7.74 (s, 1H).
Example 4-2
Preparation of racemic
2-(3,4-dichlorophenyl)-2-ethyl-2H-chromene-3-carboxylic acid
[0242] Using the procedures of Example 4-1, methyl 3-oxopentanoate
was converted to Example 4-2 as a white solid (16%). .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. ppm 1.07 (t, J=7.1 Hz, 3H) 2.22 (dq,
J=14.3, 7.4 Hz, 1H) 2.68 (dq, J=14.3, 7.4 Hz, 1H) 6.87 (d, J=8.1
Hz, 1H) 6.90 (t, J=7.6 Hz, 1H) 7.14 (dd, J=7.6, 1.5 Hz, 1H) 7.28
(m, 1H) 7.35 (m, 2H) 7.59 (t, J=1.3 Hz, 1H) 7.80 (s, 1H). High
resolution mass spectrum m/z 347.0247 (M-H).sup.-, calc.
347.0247.
Example 5-1
Preparation of racemic
2-(3,4-dichlorophenyl)-2H-chromene-3-carboxylic acid
[0243] Step 1.
[0244] A mixture of methyl 3-(2-hydroxyphenyl)-3-oxopropanoate
(prepared according to Cushman, M. et al., J. Med. Chem. 1991, 34,
798; 500 mg, 2.56 mmol) and 3,4-dichlorobenzaldehyde (496 mg, 2.83
mmol) in isopropanol (5.0 mL) was treated with piperidine (25
.mu.l, 257 .mu.mol) and acetic acid (15 .mu.l, 257 .mu.mol) and
then heated to reflux for 20 h. At the conclusion of this period,
the mixture was cooled on ice, causing a gum to separate from the
solution. The mixture was treated with a small amount of water and
heated until homogeneous. On slow cooling to rt, a solid formed,
which was collected by filtration and dried under vacuum to provide
racemic methyl 2-(3,4-dichlorophenyl)-4-oxochroman-3-carboxylate as
an off-white powder (496 mg, 55%). An additional sample (57 mg, 6%)
was obtained by partial concentration of the filtrate. The NMR
indicated predominantly the enolic form. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. ppm 3.78 (s, 3H) 6.18 (s, 1H) 6.81 (dd, J=8.1,
1.0 Hz, 1H) 6.99 (td, J=7.4, 1.0 Hz, 1H) 7.19 (dd, J=8.4, 1.8 Hz,
1H) 7.31 (m, 1H) 7.34 (d, J=8.1 Hz, 1H) 7.43 (d, J=2.0 Hz, 1H) 7.69
(dd, J=7.9, 1.8 Hz, 1H) 12.28 (br. s, 1H). High resolution mass
spectrum m/z found 373.0016, calcd 373.0010 (M+Na).sup.+.
[0245] Step 2a.
[0246] A solution of racemic methyl
2-(3,4-dichlorophenyl)-4-oxochroman-3-carboxylate (100 mg, 285
.mu.mol) in THF (2.0 mL) was diluted with methanol (4.0 mL) and
stirred on an ice/acetone bath. Sodium borohydride (11 mg, 285
.mu.mol) was added and the resulting mixture was stirred for 40
min. After this time, the mixture was diluted with water and
stirred while warming to rt. The resulting white solid was
collected by filtration, washed with water and dried under vacuum
to provide methyl
2-(3,4-dichlorophenyl)-4-hydroxychroman-3-carboxylate (90 mg).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.49 (s, 3H) 3.51 (m,
1H) 5.26 (d, J=6.1 Hz, 1H) 5.35 (d, J=2.5 Hz, 1H) 6.94 (dd, J=8.1,
1.0 Hz, 1H) 7.06 (td, J=7.6, 1.0 Hz, 1H) 7.22-7.29 (m, 2H) 7.47 (d,
J=8.6 Hz, 1H) 7.56 (d, J=1.5 Hz, 1H) 7.58 (d, J=7.6 Hz, 1H).
[0247] Step 2b.
[0248] Methyl 2-(3,4-dichlorophenyl)-4-hydroxychroman-3-carboxylate
(90 mg) and p-toluenesulfonic acid hydrate (24 mg, 128 .mu.mol)
were dissolved in toluene (4.0 mL) and heated to reflux. After 40
min, the solution was cooled to rt, diluted with ethyl acetate,
washed with saturated aqueous sodium bicarbonate, dried over sodium
sulfate and concentrated under vacuum to provide racemic methyl
2-(3,4-dichlorophenyl)-2H-chromene-3-carboxylate as a white solid
(90 mg, 94%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.79
(s, 3H) 6.21 (s, 1H) 6.81 (d, J=8.1 Hz, 1H) 6.93 (t, J=7.6 Hz, 1H)
7.17-7.25 (m, 3H) 7.35 (d, J=8.6 Hz, 1H) 7.44 (d, J=2.0 Hz, 1H)
7.70 (s, 1H).
[0249] Step 3.
[0250] Racemic methyl
2-(3,4-dichlorophenyl)-2H-chromene-3-carboxylate (90 mg, 268
.mu.mol) was suspended in methanol (1.5 mL) and then treated with
1.0 M aqueous sodium hydroxide (1.5 mL). The resulting suspension
was heated to reflux. After 2 h at reflux, the solution was
acidified with 1.0 M aqueous HCl (1.5 mL) and cooled to rt with
stirring. The resulting solid was collected by filtration, washed
with water and dried under vacuum to provide Example 5-1 as a pale
yellowish solid (73 mg, 85%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 6.20 (s, 1H) 6.82 (d, J=8.1 Hz, 1H) 6.95 (td, J=7.4,
1.0 Hz, 1H) 7.20-7.29 (m, 3H) 7.36 (d, J=8.1 Hz, 1H) 7.45 (d, J=2.0
Hz, 1H) 7.81 (s, 1H). Mass spectrum m/z 321.24 (M+H).sup.+.
Example 5-2
Preparation of racemic
2-(4-chloronaphthalen-1-yl)-2H-chromene-3-carboxylic acid
[0251] Step 1.
[0252] Following the procedure of Step 1, Example 5-1,
4-chloro-1-naphthaldehyde (prepared according to the procedure
reported in PCT Patent Application WO 02/17712, 196 mg, 1.03 mmol)
was converted to racemic methyl
2-(4-chloronaphthalen-1-yl)-4-oxochroman-3-carboxylate (127 mg,
34%).
[0253] Step 2.
[0254] Following the procedure of Step 2, Example 5-1, racemic
methyl 2-(4-chloronaphthalen-1-yl)-4-oxochroman-3-carboxylate (124
mg, 338 .mu.mol) was converted to racemic methyl
2-(4-chloronaphthalen-1-yl)-2H-chromene-3-carboxylate. In contrast
to the procedure of Step 2, Example 5-1, the crude product was
purified by column chromatography (silica gel, ethyl
acetate-hexane, 2-10%) to provide a white glassy foam (98 mg, 83%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.73 (s, 3H) 6.88
(td, J=7.6, 1.0 Hz, 1H) 7.06 (s, 1H) 7.10 (td, J=7.8, 1.8 Hz, 1H)
7.22-7.27 (m, 2H) 7.38 (d, J=7.6 Hz, 1H) 7.64-7.76 (m, 2H) 7.90 (s,
1H) 8.32 (dd, J=8.6, 1.0 Hz, 1H) 8.62 (d, J=8.1 Hz, 1H).
[0255] Step 3.
[0256] Following the procedure of Step 3, Example 5-1, racemic
methyl 2-(4-chloronaphthalen-1-yl)-2H-chromene-3-carboxylate (98
mg, 279 .mu.mol) was converted to Example 5-2, isolated as a white
powder (85 mg, 90%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
6.58 (d, J=8.1 Hz, 1H) 6.93 (t, J=7.4 Hz, 1H) 7.06 (s, 1H) 7.14 (t,
J=7.6 Hz, 1H) 7.28 (d, J=7.6 Hz, 1H) 7.46 (d, J=7.6 Hz, 1H) 7.61
(d, J=8.1 Hz, 1H) 7.80 (t, J=8.1 Hz, 1H) 7.84 (t, J=7.1 Hz, 1H)
7.96 (s, 1H) 8.27 (d, J=8.1 Hz, 1H) 8.69 (d, J=8.1 Hz, 1H) 12.95
(s, 1H). High resolution mass spectrum m/z 695.0984 (2M+Na).sup.+,
calc. 695.0999.
Example 5-3
Preparation of racemic 2-(pyridin-4-yl)-2H-chromene-3-carboxylic
acid trifluoroacetic acid salt
[0257] Step 1.
[0258] Following the procedure of Step 1, Example 5-1,
picolinaldehyde (120 .mu.l, 1.29 mmol) was converted to racemic
methyl 2-(pyridin-4-yl)-4-oxochroman-3-carboxylate. In contrast to
Step 1, Example 5-1, the product failed to crystallize from the
reaction mixture, which was concentrated and the resulting residue
was purified by column chromatography (silica gel, 1-100% ethyl
acetate-hexane) to provide an oil (100 mg, 27%). By NMR, this oil
was a mixture of the enol and keto forms (ratio about 6:4). .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.70 (s, keto 3H) 3.82 (s,
enol 3H) 4.01 (d, J=11.7 Hz, keto 1H) 5.74 (d, J=11.7 Hz, keto 1H)
6.23 (s, enol 1H) 6.88 (d, J=8.1 Hz, enol 1H) 6.99 (t, J=7.9 Hz,
enol 1H) 7.07 (d, J=8.1 Hz, keto 1H) 7.12 (t, J=8.1 Hz, keto 1H)
7.26 (d, J=6.1 Hz, enol 2H) 7.32 (t, enol 1H) 7.41 (d, J=6.1 Hz,
keto 2H) 7.58 (t, J=8.6 Hz, keto 1H) 7.68 (dd, J=7.6, 1.5 Hz, enol
1H) 7.95 (dd, J=7.9, 1.8 Hz, keto 1H) 8.52 (d, J=6.1 Hz, enol 2H)
8.68 (d, J=6.1 Hz, keto 2H) 12.25 (br. s, enol 1H). Mass spectrum
m/z 284.2 (M+H).sup.+.
[0259] Step 2.
[0260] A solution of racemic methyl
2-(pyridin-4-yl)-4-oxochroman-3-carboxylate (100 mg, 353 .mu.mol)
in methanol (5.0 mL) was stirred on an ice/acetone bath. Sodium
borohydride (27 mg, 707 .mu.mol) was added and the mixture was
stirred for 2 h while warming to rt. Once at the prescribed
temperature, the mixture was diluted with saturated aqueous sodium
bicarbonate and extracted three times with ethyl acetate. The
combined organic phases were dried over sodium sulfate and
concentrated under vacuum. The resulting residue was dissolved in
toluene (5.0 mL), treated with p-toluenesulfonic acid hydrate (135
mg, 709 .mu.mol). The resulting mixture was heated at reflux for 2
h. After this time, the mixture was cooled to rt, diluted with
water and ethyl acetate, and then treated with 1.0 M aqueous sodium
hydroxide (5.0 mL). The aqueous and organic layers were separated
and the aqueous phase was extracted twice more with ethyl acetate.
The combined organic phases were dried over sodium sulfate and
concentrated under vacuum to yield a residue. The residue (100 mg)
was used without further purification. Mass spectrum m/z 268.2
(M+H).sup.+.
[0261] Step 3.
[0262] Following the procedure of Step 3, Example 5-1, crude
racemic methyl 2-(pyridin-4-yl)-2H-chromene-3-carboxylate (100 mg,
375 .mu.mol) was converted to racemic
2-(pyridin-4-yl)-2H-chromene-3-carboxylic acid. In contrast to Step
3, Example 5-1, the product was purified by preparative HPLC
[(C.sub.18, 5-95% acetonitrile-water (containing 0.05%
trifluoroacetic acid)] and lyophilization to provide Example 5-3 as
a yellow solid (30 mg, 22%). .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. ppm 6.47 (s, 1H) 7.00 (d, J=8.1 Hz, 1H) 7.03 (t, J=7.6 Hz,
1H) 7.24 (dd, J=7.4, 1.3 Hz, 1H) 7.37 (t, J=7.1 Hz, 1H) 7.78 (s,
1H) 7.94 (d, J=6.1 Hz, 2H) 8.85 (d, J=6.6 Hz, 2H). High resolution
mass spectrum m/z 254.0817 (M+H).sup.+, calc. 254.0817.
Examples 5-4 to 5-26
[0263] Examples 5-4 to 5-26 set forth in Table 2 below were
prepared using the methods described in Examples 5-1, 5-2 and
5-3:
TABLE-US-00002 TABLE 2 Example Compound name Mass spectrum 5-4
racemic 2-(naphthalen-2-yl)-2H-chromene-3- 325.2 (M + Na).sup.+
carboxylic acid 5-5 racemic 2-(naphthalen-1-yl)-2H-chromene-3-
301.2(M - H).sup.- carboxylic acid 5-6 racemic
2-(3-fluorophenyl)-2H-chromene-3-carboxylic 293.2 (M + Na).sup.+
acid 5-7 racemic 2-(3-fluoro-4-chlorophenyl)-2H-chromene-3- 305.3
(M + H).sup.+ carboxylic acid 5-8 racemic
2-(3-chloro-4-fluorophenyl)-2H-chromene-3- 305.3 (M + H).sup.+
carboxylic acid 5-9 racemic
2-(3-trifluoromethylphenyl)-2H-chromene-3- 321.3 (M + H).sup.+
carboxylic acid 5-10 racemic
2-(4-trifluoromethylphenyl)-2H-chromene-3- 319.1(M - H).sup.-
carboxylic acid 5-11 racemic 2-(3,4-difluorophenyl)-2H-chromene-3-
287.1(M - H).sup.- carboxylic acid 5-12 racemic
2-(3-nitrophenyl)-2H-chromene-3-carboxylic 296.1(M - H).sup.- acid
5-13 racemic 2-(3-nitro-4-chlorophenyl)-2H-chromene-3- 354.2 (M +
Na).sup.+ carboxylic acid 5-14 racemic
2-(3-methyl-4-chlorophenyl)-2H-chromene-3- 299.0(M - H).sup.-
carboxylic acid 5-15 racemic
2-(4-bromophenyl)-2H-chromene-3-carboxylic 329.0(M - H).sup.- acid
5-16 racemic 2-(3-bromophenyl)-2H-chromene-3-carboxylic 329.0(M -
H).sup.- acid 5-17 racemic 2-(3,4-dibromophenyl)-2H-chromene-3-
406.9(M - H).sup.- carboxylic acid 5-18 racemic
2-(3-fluoro-4-trifluoromethylphenyl)-2H- 337.0(M - H).sup.-
chromene-3-carboxylic acid 5-19 racemic
2-(4-methylphenyl)-2H-chromene-3- 265.1(M - H).sup.- carboxylic
acid 5-20 racemic 2-(3-methylphenyl)-2H-chromene-3- 265.1(M -
H).sup.- carboxylic acid 5-21 racemic
2-(pyridin-2-yl)-2H-chromene-3-carboxylic 254.2 (M + H).sup.+ acid
5-22 racemic 2-(quinolin-2-yl)-2H-chromene-3-carboxylic 304.2 (M +
H).sup.+ acid 5-23 racemic
2-(quinolin-3-yl)-2H-chromene-3-carboxylic 304.2 (M + H).sup.+ acid
5-24 racemic 2-(3-trifluoromethyl-4-fluoro-phenyl)-2H- 337.0(M -
H).sup.- chromene-3-carboxylic acid 5-25 racemic
2-(6-trifluoromethylpyridin-2-yl)-2H- 322.3(M + H).sup.+
chromene-3-carboxylic acid 5-26 racemic
2-(2,3-dichlorophenyl)-2H-chromene-3- 319.2(M - H).sup.- carboxylic
acid
Examples 6a and 6b
Preparation of (-)-2-(3,4-dichlorophenyl)-2H-chromene-3-carboxylic
acid and (+)-2-(3,4-dichlorophenyl)-2H-chromene-3-carboxylic acid,
respectively
[0264] A sample of Example 5-26 (31 mg) was resolved using
supercritical fluid chromatography (Chiralcel OJ, carbon
dioxide-methanol 70-30, 100 bar, 35 C..degree.). The (-) isomer
eluted first and was obtained in >99% enantiomeric purity (14
mg). [.alpha.].sub.D=-168.2.degree. (MeOH, c=2.5 mg/mL). The (+)
isomer eluted second and was obtained in >98% enantiomeric
purity (14 mg). [.alpha.].sub.D=+176.8.degree. (MeOH, c=2.5 mg/mL).
NMR spectra of both isomers were identical to that of the racemic
material.
Example 7-1
Preparation of racemic
2-(3-methoxy-4-chlorophenyl)-2H-chromene-3-carboxylic acid
[0265] Step 1.
[0266] A solution of 4-chloro-3-methoxybenzoic acid (1.12 g, 5.98
mmol) in THF (30 mL) was stirred on an ice bath and treated with a
1.0 M tetrahydrofuran solution of lithium aluminum hydride (7.18
mL, 7.18 mmol) over about a 5 min. period. The solution was stirred
on ice for 2 h, and then quenched with rapid stirring by the slow
dropwise addition of water (0.27 mL), followed by 15% aqueous
sodium hydroxide (0.27 mL), then water (0.82 mL) again. Upon
completion of addition, the resulting suspension was filtered and
the collected solid was washed with ethyl acetate. The filtrate was
concentrated under vacuum to provide
(4-chloro-3-methoxyphenyl)methanol as a colorless oil (737 mg,
71%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.92 (s, 3H)
4.67 (s, 2H) 6.87 (d, J=7.6 Hz, 1H) 6.98 (s, 1H) 7.33 (d, J=7.6 Hz,
1H).
[0267] Step 2.
[0268] A suspension of pyridinium chlorochromate (1.37 g, 6.34
mmol) in dichloromethane (35 mL) was treated with Celite (2.0 g)
and stirred at rt. A solution of (4-chloro-3-methoxyphenyl)methanol
(730 mg, 4.23 mmol) in dichloromethane (5 mL) was added in one
portion, and the resulting mixture was stirred at rt for 3 h. After
this time, the mixture was diluted with diethyl ether, and stirred
vigorously. The mixture was then filtered through a pad of silica
gel and the collected solids were rinsed with additional diethyl
ether. The combined filtrates were concentrated under vacuum to
provide 4-chloro-3-methoxybenzaldehyde as a pale tan-yellow solid
(630 mg, 87%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.98
(s, 3H) 7.41 (dd, J=7.9, 1.8 Hz, 1H) 7.45 (d, J=2.0 Hz, 1H) 7.55
(d, J=7.6 Hz, 1H) 9.95 (s, 1H).
[0269] Step 3.
[0270] Following the procedure described in Step 1, Example 5-1,
4-chloro-3-methoxybenzaldehyde (132 mg, 772 .mu.mol) was converted
to racemic methyl
2-(3-methoxy-4-chlorophenyl)-4-oxochroman-3-carboxylate. In
contrast to the procedure described in Step 1, Example 5-1, the
crude product was not a solid, and rather was purified by column
chromatography (silica gel, 5-20% ethyl acetate-hexane) to provide
a colorless gum (172 mg, 64%). By NMR, this material was a mixture
of the enol and keto forms (ratio about 7:3).
[0271] Step 4.
[0272] Following the procedures described in Steps 2 and 3 of
Example 5-1, racemic methyl
2-(3-methoxy-4-chlorophenyl)-4-oxochroman-3-carboxylate (169 mg,
487 .mu.mol) was converted to Example 7-1 as a white solid (72 mg,
47%). .sup.1H NMR (400 MHz, CDCl.sup.3) .delta. ppm 3.80 (s, 3H)
6.22 (s, 1H) 6.79-6.86 (m, 2H) 6.95 (t, J=7.4 Hz, 1H) 7.19 (d,
J=1.5 Hz, 1H) 7.25 (td, J=7.6, 1.5 Hz, 1H) 7.35 (d, J=8.1 Hz, 1H)
7.40 (dd, J=7.4, 1.3 Hz, 1H) 7.74 (s, 1H) 12.98 (s, 1H). High
resolution mass spectrum m/z 315.0430 (M-H).sup.-, calc.
315.0424.
Examples 7-2 to 7-5
[0273] Examples 7-2 to 7-5 set forth in Table 3 below were prepared
using the methods described in Example 7-1:
TABLE-US-00003 TABLE 3 Example Compound name Mass spectrum 7-2
racemic 2-(6-chloropyridin-3-yl)-2H- 288.2 (M + H).sup.+
chromene-3-carboxylic acid 7-3 racemic
2-(5,6-dichloropyridin-3-yl)-2H- 322.0 (M + H).sup.+
chromene-3-carboxylic acid 7-4 racemic
2-(3-bromo-4-chlorophenyl)-2H- 362.9(M - H).sup.-
chromene-3-carboxylic acid 7-5 racemic
2-(3-chloro-4-trifluoromethyl- 353.0(M - H).sup.-
phenyl)-2H-chromene-3-carboxylic acid
Example 8
Preparation of racemic
2-(3,4-dichlorophenyl)-6-methoxy-2H-chromene-3-carboxylic acid
[0274] Step 1.
[0275] A solution of lithium bis(trimethylsilyl)amide (1.0 M in
tetrahydrofuran, 61.5 mL, 61.5 mmol) was stirred at -78.degree. C.
and treated over a 5 min period with a solution of tert-butyl
acetate (5.2 mL, 38.4 mmol) in tetrahydrofuran (13 mL). The
resulting mixture was stirred at -78.degree. C. for 55 min, and
then treated dropwise with a solution of methyl
2-hydroxy-5-methoxybenzoate (1.63 mL, 10.98 mmol) in
tetrahydrofuran (13 mL) over a 5 min period. The resulting solution
was warmed slowly to rt where it stirred for 3 days. At the
conclusion of this period, the solution was treated with ice-cold
saturated aqueous ammonium chloride, and then extracted three times
with ethyl acetate. The combined organic phases were dried over
sodium sulfate and concentrated under vacuum to provide an oil. The
oil was purified by column chromatography (silica gel, 20-30% ethyl
acetate-hexane), and the effluent was concentrated to yield a
residue. The residue was recrystallized from hexane to provide
tert-butyl 3-(2-hydroxy-5-methoxyphenyl)-3-oxopropanoate as yellow
crystals (2.40 g, 82%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
ppm 1.46 (s, 9H) 3.79 (s, 3H) 3.89 (s, 2H) 6.95 (d, J=9.2 Hz, 1H)
7.11 (d, J=3.1 Hz, 1H) 7.14 (dd, J=8.6, 2.5 Hz, 1H) 11.54 (s, 1H).
Mass spectrum m/z 289.19 (M+Na).sup.+.
[0276] Step 2.
[0277] A solution of tert-butyl
3-(2-hydroxy-5-methoxyphenyl)-3-oxopropanoate (500 mg, 1.88 mmol)
in isopropanol (5.0 mL) was treated with 3,4-dichlorobenzaldehyde
(329 mg, 1.88 mmol), piperidine (20 .mu.L, 188 .mu.mol) and acetic
acid (10 .mu.L, 188 .mu.mol). The reaction mixture was heated at
reflux overnight, and then treated with water (1.0 mL). The
resulting mixture was cooled to rt and then concentrated under
vacuum to yield a residue. The residue was purified by column
chromatography (silica gel, 2-25% ethyl acetate-hexane) to provide
racemic tert-butyl
2-(3,4-dichlorophenyl)-6-methoxy-4-oxochroman-3-carboxylate as a
yellow oil (380 mg, 48%). This material was used without further
purification in Step 3 below.
[0278] Step 3.
[0279] A solution of racemic tert-butyl
2-(3,4-dichlorophenyl)-6-methoxy-4-oxochroman-3-carboxylate (100
mg, 236 .mu.mol) in tetrahydrofuran (1.0 mL) was diluted with
methanol (1.0 mL) and stirred at 0.degree. C. Sodium borohydride
(12 mg, 473 .mu.mol) was added and the resulting mixture was
stirred for 30 min. After this time, water (2.0 mL) was added and
the mixture was stirred at rt. The resulting white solid was
collected by filtration and dried under vacuum, and then purified
by column chromatography (silica gel, 5-55% ethyl acetate-hexane)
to provide a diastereomeric mixture of tert-butyl
2-(3,4-dichlorophenyl)-4-hydroxy-6-methoxychroman-3-carboxylate (21
mg, 21%). This material was used without further purification in
Step 4 below.
[0280] Step 4.
[0281] A mixture of tert-butyl
2-(3,4-dichlorophenyl)-4-hydroxy-6-methoxychroman-3-carboxylate (21
mg, 49 .mu.mol) and p-toluenesulfonic acid hydrate (5 mg, 25
.mu.mol) in toluene (5 mL) was heated at reflux for 30 min. At the
conclusion of this period, the solution was cooled to rt and then
concentrated under vacuum to yield a residue. The residue was
treated with water and the resulting solid was collected by
filtration and then dried under vacuum. The resulting material was
purified by column chromatography (silica gel, 0-50% ethyl
acetate-hexane) and then again by rotary thin-layer chromatography
(silica gel, hexane-ethyl acetate) to provide Example 8 (5 mg,
28%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 3.77 (s, 3H)
6.16 (s, 1H) 6.74-6.78 (m, 2H) 6.83 (dd, J=9.2, 3.1 Hz, 1H) 7.21
(dd, J=8.6, 2.0 Hz, 1H) 7.35 (d, J=8.1 Hz, 1H) 7.44 (d, J=2.0 Hz,
1H) 7.76 (s, 1H). Mass spectrum m/z 349.2 (M-H).sup.-.
Example 9-1
Preparation of racemic
2-(3,4-dichlorophenyl)-7-fluoro-2H-chromene-3-carboxylic acid
[0282] Step 1.
[0283] A 1.0 M tetrahydrofuran solution of lithium
bis(trimethylsilyl)amide (39.0 mL, 39.0 mmol) was stirred at
-78.degree. C. and treated over a 25 min period with a solution of
1-(4-fluoro-2-hydroxyphenyl)ethanone (2.00 g, 13.0 mmol) in 50 mL
of tetrahydrofuran. The resulting solution was stirred at
-78.degree. C. for 1 h, and then at -15.degree. C. for 2 h. After
being cooled again to -78.degree. C., the solution was treated with
a solution of dimethyl carbonate (1.20 mL, 14.3 mmol) in
tetrahydrofuran (5 mL). The resulting mixture was allowed to warm
to rt where it stirred overnight. After this time, the mixture was
poured over ice and then concentrated hydrochloric acid (10 mL) was
added. The resulting mixture was extracted three times with
dichloromethane. The combined organic phases were washed twice with
water, dried over sodium sulfate and then concentrated under vacuum
to yield a residue. The residue was purified by radial thin-layer
chromatography (silica gel, 2-40% ethyl acetate-hexane) to provide
methyl 3-(4-fluoro-2-hydroxyphenyl)-3-oxopropanoate as a white
solid (1.76 g, 64%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm
3.78 (s, 3H) 3.98 (s, 2H) 6.62-6.71 (m, 2H) 7.69 (dd, J=8.9, 6.4
Hz, 1H) 12.15 (d, J=1.5 Hz, 1H). Mass spectrum m/z 213.14
(M+H).sup.+.
[0284] Step 2.
[0285] Following the procedure described in Step 1, Example 5-1,
methyl 3-(4-fluoro-2-hydroxyphenyl)-3-oxopropanoate (500 mg, 2.36
mmol) was converted to racemic methyl
2-(3,4-dichlorophenyl)-7-fluoro-4-oxochroman-3-carboxylate. In
contrast to the procedure described in Step 1, Example 5-1, the
crude product was purified by column chromatography (silica gel,
ethyl acetate-hexane) to provide a white solid (630 mg, 72%). By
NMR this material was a mixture of the enol and keto forms (ratio
about 7:3).
[0286] Step 3.
[0287] Following the procedures described in Steps 2 and 3, Example
5-1, racemic methyl methyl
2-(3,4-dichlorophenyl)-7-fluoro-4-oxochroman-3-carboxylate (100 mg,
271 .mu.mol) was converted to Example 9-1 as a pale yellow solid
(37 mg, 40%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. ppm 6.20
(s, 1H) 6.56 (dd, J=9.7, 2.0 Hz, 1H) 6.67 (td, J=8.4, 2.5 Hz, 1H)
7.19-7.23 (m, 2H) 7.38 (d, J=8.1 Hz, 1H) 7.44 (d, J=2.0 Hz, 1H)
7.77 (s, 1H). Mass spectrum m/z 337.2 (M-H).sup.-.
Examples 9-2 to 9-6
[0288] Examples 9-2 to 9-6 set forth in Table 4 below were prepared
using the methods described in Example 9-1:
TABLE-US-00004 TABLE 4 Example Compound name Mass spectrum 9-2
racemic 2-(3,4-dichlorophenyl)-6-chloro-2H- 353.0 (M - H).sup.-
chromene-3-carboxylic acid 9-3 racemic 2-(3,4-dichlorophenyl)-2H-
369.1 (M - H).sup.- benzo[h]chromene-3-carboxylic acid 9-4 racemic
2-(3,4-dichlorophenyl)-6-fluoro-2H- 337.1 (M - H).sup.-
chromene-3-carboxylic acid 9-5 racemic
2-(3,4-dichlorophenyl)-7-bromo-2H- 396.9 (M - H).sup.-
chromene-3-carboxylic acid 9-6 racemic
2-(3,4-dichlorophenyl)-7-methoxy- 349.1 (M - H).sup.-
2H-chromene-3-carboxylic acid
Utility
[0289] In general, compounds of the present invention, such as
particular compounds disclosed in the preceding examples, have been
shown to be modulators of chemokine receptor activity (for example,
by displaying Ki values <10,000 nM in a binding assay such as
those set forth below). By displaying activity as modulators of
chemokine receptor activity, compounds of the present invention are
expected to be useful in the treatment of human diseases associated
with chemokines and their cognate receptors.
Antagonism of MCP-1 Binding to Human PBMC
[0290] (Yoshimura et al., J. Immunol. 1990, 145, 292)
[0291] Millipore filter plates (#MABVN1250) are treated with 100
.mu.l of binding buffer (0.5% bovine serum albumin, 20 mM HEPES
buffer and 5 mM magnesium chloride in RPMI 1640 media) for thirty
minutes at room temperature. To measure binding, 50 .mu.l of
binding buffer, with or without a known concentration compound, is
combined with 50 .mu.l of .sup.125-I labeled human MCP-1 (to give a
final concentration of 150 pM radioligand) and 50 .mu.l of binding
buffer containing 5.times.10.sup.5 cells. Cells used for such
binding assays can include human peripheral blood mononuclear cells
isolated by Ficoll-Hypaque gradient centrifugation, human monocytes
(Weiner et al., J. Immunol. Methods. 1980, 36, 89), or the THP-1
cell line which expresses the endogenous receptor. The mixture of
compound, cells and radioligand are incubated at room temperature
for thirty minutes. Plates are placed onto a vacuum manifold,
vacuum applied, and the plates washed three times with binding
buffer containing 0.5M NaCl. The plastic skirt is removed from the
plate, the plate allowed to air dry, the wells punched out and
counted. The percent inhibition of binding is calculated using the
total counts obtained in the absence of any competing compound and
the background binding determined by addition of 100 nM MCP-1 in
place of the test compound.
Antagonism of MCP-1-Induced Calcium Influx
[0292] (Sullivan et al., Methods Mol. Biol. 1999, 114, 125-133)
[0293] Calcium mobilization is measured using the fluorescent
Ca.sup.2+ indicator dye, Fluo-3. Cells are incubated at
8.times.10.sup.5 cells/ml in phosphate-buffered saline containing
0.1% bovine serum albumin, 20 mM HEPES buffer, 5 mM glucose, 1%
fetal bovine serum, 4 .mu.M Fluo-3 AM and 2.5 mM probenecid for 60
minutes at 37.degree. C. Cells used for such calcium assays can
include human monocytes isolated as described by Weiner et al., J.
Immunol. Methods 1990, 36, 89-97 or cell lines which expresses the
endogenous CCR2 receptor such as THP-1 and MonoMac-6. The cells are
then washed three times in phosphate-buffered saline containing
0.1% bovine serum albumin, 20 mM HEPES, 5 mM glucose and 2.5 mM
probenecid. The cells are resuspended in phosphate-buffered saline
containing 0.5% bovine serum albumin, 20 mM HEPES and 2.5 mM
probenecid at a final concentration of 2-4.times.10.sup.6 cells/ml.
Cells are plated into 96-well, black-wall microplates (100
.mu.l/well) and the plates centrifuged at 200.times.g for 5
minutes. Various concentrations of compound are added to the wells
(50 .mu.l/well) and after 5 minutes, 50 .mu.l/well of MCP-1 is
added to give a final concentration of 10 nM. Calcium mobilization
is detected by using a fluorescent-imaging plate reader. The cell
monolayer is excited with an argon laser (488 nM) and
cell-associated fluorescence measured for 3 minutes, (every second
for the first 90 seconds and every 10 seconds for the next 90
seconds). Data are generated as arbitrary fluorescence units and
the change in fluorescence for each well determined as the
maximum-minimum differential. Compound-dependent inhibition is
calculated relative to the response of MCP-1 alone.
THP-1 Monocytic Cell Binding: Whole Cell-Based Assay
[0294] The human CCR2 binding assay was also established with the
THP-1 human monocytic leukemic cell line, which expresses
endogenous CCR2, using .sup.125I-human MCP-1 as the tracer ligand.
Radioligand competition binding assays were used for assessment of
binding affinity of test compounds to the CCR2 receptor. For
radioligand competition studies, 100 .mu.l containing
2.5.times.10.sup.5 THP-1 cells/well (in assay buffer containing 50
mM HEPES, pH7.4, 5 mM MgCl.sub.2, 1 mM CaCl.sub.2 and 0.5% BSA)
were added to 96-well assay plates containing the test compounds in
3-fold serial dilution, with final concentrations ranging from 5
.mu.M to 100 pM. Subsequently, 50 .mu.l .sup.125I-MCP-1 radioligand
at a final concentration of 0.2 nM in assay buffer were added to
the reaction. After a 90 minute incubation period at room
temperature, the binding reaction was terminated by harvesting on
GF/B filter plates (PerkinElmer Cat. No. 6005177) followed by
washing with ice-cold wash buffer (50 mM HEPES, pH 7.4, 0.1% BSA,
0.5 M NaCl) to remove unbound ligand. After washing, the plate was
dried for 45 minutes at 60.degree. C. followed by addition of 40
.mu.l MicroScint 20 scintillation fluid, sealed and analyzed by the
Packard TopCount reader. Non-specific binding was determined in the
presence of 10 .mu.M (a molar excess of 5000 fold) of an in-house
CCR2 small molecule antagonist (CCR2IC50=2 nM). Specific binding of
.sup.125I-MCP-1 was calculated as the difference between total and
non-specific binding. The competition data was plotted as a
percentage inhibition of radioligand specific bound in the absence
of test compound (percent of total signal). After correcting for
non-specific binding, IC50 values were determined. The IC50 is
defined as the concentration of test compound required to reduce
.sup.125I-MCP-1 specific binding by 50% and was calculated using
the four parameter logistic equation to fit the normalized
data.
Antagonism of MCP-1-Induced Human PBMC Chemotaxis
[0295] (Bacon et al., Brit. J. Pharmacol. 1988, 95, 966)
[0296] Neuroprobe MBA96-96-well chemotaxis chamber, Polyfiltronics
MPC 96 well plate, and Neuroprobe polyvinylpyrrolidone-free
polycarbonate PFDS 8-micron filters are warmed in a 37.degree. C.
incubator. Human Peripheral Blood Mononuclear Cells (PBMCs) (Boyum
et al., Scand. J. Clin. Lab Invest. Suppl. 1968, 97, 31), freshly
isolated via the standard ficoll density separation method, are
suspended in DMEM at 1.times.10 .sup.7 c/ml and warmed at
37.degree. C. A 60 nM solution of human MCP-1 is also warmed at
37.degree. C. Dilutions of test compounds are made up at 2.times.
the concentration needed in DMEM. The PBMC suspension and the 60 nm
MCP-1 solution are mixed 1:1 in polypropylene tubes with prewarmed
DMEM with or without a dilution of the test compounds. These
mixtures are warmed in a 37.degree. C. tube warmer. To start the
assay, add the MCP-1/compound mixture into the wells of the
Polyfiltronics MPC 96 well plate that has been placed into the
bottom part of the Neuroprobe chemotaxis chamber. The approximate
volume is 400 .mu.l to each well and there should be a positive
meniscus after dispensing. The 8 micron filter is placed gently on
top of the 96 well plate, a rubber gasket is attached to the bottom
of the upper chamber, and the chamber is assembled. A 200 .mu.l
volume of the cell suspension/compound mixture is added to the
appropriate wells of the upper chamber. The upper chamber is
covered with a plate sealer, and the assembled unit is placed in a
37.degree. C. incubator for 45 minutes. After incubation, the plate
sealer is removed and all the remaining cell suspension is
aspirated off. The chamber is disassembled and the filter gently
removed. While holding the filter at a 90 degree angle, unmigrated
cells are washed away using a gentle stream of phosphate buffered
saline and the top of the filter wiped with the tip of a rubber
squeegee. Repeat this wash twice more. The filter is air dried and
then immersed completely in Wright Geimsa stain for 45 seconds. The
filter is then washed by soaking in distilled water for 7 minutes,
and then a 15 second additional wash in fresh distilled water. The
filter is again air dried. Migrated cells on the filter are
quantified by visual microscopy.
[0297] Mammalian chemokine receptors provide a target for
interfering with or promoting immune cell function in a mammal,
such as a human. Compounds that inhibit or promote chemokine
receptor function are particularly useful for modulating immune
cell function for therapeutic purposes. Accordingly, the present
invention is directed to compounds which are useful in the
prevention and/or treatment of a wide variety of inflammatory,
infectious, and immunoregulatory disorders and diseases, including
asthma and allergic diseases, infection by pathogenic microbes
(which, by definition, includes viruses), as well as autoimmune
pathologies such as the rheumatoid arthritis and
atherosclerosis.
[0298] For example, an instant compound which inhibits one or more
functions of a mammalian chemokine receptor (e.g., a human
chemokine receptor) may be administered to inhibit (i.e., reduce or
prevent) inflammation or infectious disease. As a result, one or
more inflammatory process, such as leukocyte emigration, adhesion,
chemotaxis, exocytosis (e.g., of enzymes, histamine) or
inflammatory mediator release, is inhibited.
[0299] Similarly, an instant compound which promotes one or more
functions of the mammalian chemokine receptor (e.g., a human
chemokine) as administered to stimulate (induce or enhance) an
immune or inflammatory response, such as leukocyte emigration,
adhesion, chemotaxis, exocytosis (e.g., of enzymes, histamine) or
inflammatory mediator release, resulting in the beneficial
stimulation of inflammatory processes. For example, eosinophils can
be recruited to combat parasitic infections. In addition, treatment
of the aforementioned inflammatory, allergic and autoimmune
diseases can also be contemplated for an instant compound which
promotes one or more functions of the mammalian chemokine receptor
if one contemplates the delivery of sufficient compound to cause
the loss of receptor expression on cells through the induction of
chemokine receptor internalization or the delivery of compound in a
manner that results in the misdirection of the migration of
cells.
[0300] In addition to primates, such as humans, a variety of other
mammals can be treated according to the method of the present
invention. For instance, mammals, including but not limited to,
cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other
bovine, ovine, equine, canine, feline, rodent or murine species can
be treated. However, the method can also be practiced in other
species, such as avian species. The subject treated in the methods
above is a mammal, male or female, in whom modulation of chemokine
receptor activity is desired. "Modulation" as used herein is
intended to encompass antagonism, agonism, partial antagonism
and/or partial agonism.
CCR5Binding and Functional Assays
Cell Derivation and Cell Culture
[0301] A pool of HT1080 cells stably expressing endogenous CC
chemokine receptor 5 (CCR5) were developed using the methods
outlined by Harrington, Sherf, and Rundlett (see U.S. Pat. Nos.
6,361,972 and 6,410,266). The highest-expressing clones were
isolated using repetitive flow cytometry, followed by sub-cloning.
These cells were then cultured in 6-well dishes at 3.times.10.sup.5
cells/well and transfected with a DNA vector containing the
chimeric HA-tagged G protein Gqi5 (Molecular Devices; 5 micrograms
of linearized vector DNA in 15 microL of Ex-Gen from Fermentes was
used for the transfection). Two days after transfection, the wells
were combined and plated into P100 plates. Seven days after
plating, colonies were picked, expanded, and analyzed for Gqi5
content by Western blot. A clone (designated as 3559.1.6) having
high expression of Gqi5 (from transfection) and of CCR5
(endogenous) was selected and used for the experiments described
below. The HT1080 cells (clone 3559.1.6) were cultured with
alpha-MEM supplemented with 10% dialyzed fetal bovine serum, 2%
penicillin/streptomycin/glutamine, and 500 microgram/mL hygromycin
B (final concentration) at 37.degree. C. with 5% CO.sub.2 in a
humidified atmosphere.
Membrane Preparation
[0302] A cell pellet containing 1.times.10.sup.8 HT1080 cells
(clone 3559.1.6) was resuspended in 5 mL of ice-cold Membrane Prep
Buffer (50 mM HEPES, 5 mM MgCl.sub.2, 1 mM CaCl.sub.2) and
homogenized at high-speed on a Polytron homogenizer for 20 sec on
ice. The homogenate was diluted with another 25 mL of Membrane Prep
Buffer and centrifuged for 12 min (48,000.times.g at 4.degree. C.).
The cell pellet was resuspended in 5 mL of Membrane Prep Buffer
before being rehomogenized as described previously. The homogenate
was diluted with 5 mL of Membrane Prep Buffer and assayed for CCR5
protein concentration.
Binding Assay
[0303] The freshly-prepared homogenate from the Membrane
Preparation described above was diluted in Binding buffer (50 mM
HEPES, 5 mM MgCl.sub.2, 1 mM CaCl.sub.2, 0.1% BSA; one complete
protease inhibitor tablet was added before assay) to achieve a
final protein concentration of 10 micrograms/well (solid white
96-well plates from Corning, Inc.). This membrane preparation was
mixed with WGA-SPA beads (Amerhsam; pre-soaked in Binding buffer)
to give a concentration of 200 micrograms/well. The membrane/SPA
bead mix (100 microliters/well) was then added to a plate that had
been pre-dotted with 2 microliters DMSO containing various
concentrations of test articles (pure DMSO for negative control;
various concentrations of examples of this invention for test
articles; 500 nM MIP-1 beta as a positive control). The binding
assay was initiated through the addition of 50 microliters of
[.sup.125I]-MIP-1 beta (Perkin Elmer; material was diluted in
Binding buffer such that the addition of 50 microliters/well gives
a final concentration of 0.1 nM [.sup.125I]-MIP-1 beta). The plate
was sealed and allowed to stand at room temperature for 4-6 h
before being counted on a Packard TopCount. The percentage bound
for the test article was calculated, using negative and positive
controls to define the window for each experiment.
Fluorometric Imaging Plate Reader (FLIPR)-Based Functional
Assay
[0304] HT1080 cells (clone 3559.1.6) were plated at 10,000
cells/well (30 microliters) in 384-well plates (black/clear bottom
Biocoat PDL, Beckton Dickinson) and charged with 30
microliters/well of Fluoro-4 AM fluorescent dye (prepared by
dissolving 1 mg Fluoro-4 AM in 440 microliters DMSO and diluting
with 100 microliters of pluronic solution before diluting further
with 10 mL of Hanks buffer). The cells were incubated at 37.degree.
C. with 5% CO.sub.2 for 30 min before being washed three times and
suspended in Assay Buffer (20 mM HEPES, 1.2 mM CaCl.sub.2, 5 mM
MgCl.sub.2, 2.5 mM Probenecid, 0.5% BSA, 1.times. Hanks). The test
article was serially diluted in DMSO and then diluted 1:10 with
Assay Buffer before being added to the cells (10 microliters/well).
Using FLIPR, the plates were read (10-70 sec) for induction of flux
(i.e., agonist activity). The cells were then further charged with
Agonist Solution (30 microliters/well; prepared by diluting 30
microliters of 100 microMolar MIP-1 beta in 100 mL of Assay Buffer;
this protocol delivers a final concentration of 5 nM MIP-1 beta in
the assay) and the plates were read using FLIPR for one minute.
Antagonist activity of the test article was determined relative to
0.4% DMSO/Buffer negative control.
[0305] At least one compound of the disclosure is an inhibitor of
both CCR2 and CCR5 and may be used to treat diseases associated
with either chemokine. These compounds of the present invention are
considered dual antagonists.
[0306] Compounds of the present invention were tested in one of the
assays described above and the results shown in Table 5 below were
obtained.
TABLE-US-00005 TABLE 5 CCR2 Binding Ki, nM Example (n = 1 unless
otherwise noted) 1 2000 2-1 75 2-2 950 2-3 5300 2-4 4500 3 1100 4-1
220 4-2 1200 5-1 140 5-2 1900 5-3 1800 5-4 1800 5-5 1700 5-6 5700
5-7 290 5-8 560 5-9 1600 5-10 920 5-11 8600 5-12 2800 5-13 230 5-14
160 5-15 430 5-16 1700 5-17 85 5-18 720 5-19 4500 5-20 4200 5-21
1400 5-22 2100 5-23 4300 5-24 1100 5-25 8300 5-26 2600 6a 80 6b
1700 7-1 2900 7-2 4500 7-3 570 7-4 170 7-5 390 8 360 9-1 350 9-2
300 9-3 1200 9-4 340 9-5 720 9-6 300
[0307] Mammalian chemokine receptors provide a target for
interfering with or promoting immune cell function in a mammal,
such as a human. Compounds that inhibit or promote chemokine
receptor function are particularly useful for modulating immune
cell function for therapeutic purposes.
[0308] Accordingly, the present invention is directed to compounds
which are useful in the prevention and/or treatment of a wide
variety of inflammatory, infectious, and immunoregulatory disorders
and diseases, including asthma and allergic diseases, infection by
pathogenic microbes (which, by definition, includes viruses), as
well as autoimmune pathologies such as the rheumatoid arthritis and
atherosclerosis.
[0309] For example, an instant compound which inhibits one or more
functions of a mammalian chemokine receptor (e.g., a human
chemokine receptor) may be administered to inhibit (i.e., reduce or
prevent) inflammation or infectious disease. As a result, one or
more inflammatory process, such as leukocyte emigration, adhesion,
chemotaxis, exocytosis (e.g., of enzymes, histamine) or
inflammatory mediator release, is inhibited.
[0310] Similarly, an instant compound which promotes one or more
functions of the mammalian chemokine receptor (e.g., a human
chemokine) as administered to stimulate (induce or enhance) an
immune or inflammatory response, such as leukocyte emigration,
adhesion, chemotaxis, exocytosis (e.g., of enzymes, histamine) or
inflammatory mediator release, resulting in the beneficial
stimulation of inflammatory processes. For example, eosinophils can
be recruited to combat parasitic infections. In addition, treatment
of the aforementioned inflammatory, allergic and autoimmune
diseases can also be contemplated for an instant compound which
promotes one or more functions of the mammalian chemokine receptor
if one contemplates the delivery of sufficient compound to cause
the loss of receptor expression on cells through the induction of
chemokine receptor internalization or the delivery of compound in a
manner that results in the misdirection of the migration of
cells.
[0311] In addition to primates, such as humans, a variety of other
mammals can be treated according to the method of the present
invention. For instance, mammals, including but not limited to,
cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other
bovine, ovine, equine, canine, feline, rodent or murine species can
be treated. However, the method can also be practiced in other
species, such as avian species.
[0312] The subject treated in the methods above is a mammal, male
or female, in whom modulation of chemokine receptor activity is
desired. "Modulation" as used herein is intended to encompass
antagonism, agonism, partial antagonism and/or partial agonism.
[0313] Diseases or conditions of human or other species which can
be treated with inhibitors of chemokine receptor function, include,
but are not limited to: inflammatory or allergic diseases and
conditions, including respiratory allergic diseases such as asthma,
allergic rhinitis, hypersensitivity lung diseases, hypersensitivity
pneumonitis, eosinophilic cellulitis (e.g., Well's syndrome),
eosinophilic pneumonias (e.g., Loeffler's syndrome, chronic
eosinophilic pneumonia), eosinophilic fasciitis (e.g., Shulman's
syndrome), delayed-type hypersensitivity, interstitial lung
diseases (ILD) (e.g., idiopathic pulmonary fibrosis, or ILD
associated with rheumatoid arthritis, systemic lupus erythematosus,
ankylosing spondylitis, systemic sclerosis, Sjorgren's syndrome,
polymyositis or dermatomyositis); systemic anaphylaxis or
hypersensitivity responses, drug allergies (e.g., to penicillin,
cephalosporins), eosinophilia-myalgia syndrome due to the ingestion
of contaminated tryptophan, insect sting allergies; autoimmune
diseases, such as rheumatoid arthritis, psoriatic arthritis,
multiple sclerosis, systemic lupus erythematosus, myasthenia
gravis, juvenile onset diabetes; glomerulonephritis, autoimmune
thyroiditis, Behcet's disease; graft rejection (e.g., in
transplantation), including allograft rejection or
graft-versus-host disease; inflammatory bowel diseases, such as
Crohn's disease and ulcerative colitis; spondyloarthropathies;
scleroderma; psoriasis (including T-cell mediated psoriasis) and
inflammatory dermatoses such as an dermatitis, eczema, atopic
dermatitis, allergic contact dermatitis, urticaria; vasculitis
(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);
eosinophilic myositis, eosinophilic fasciitis; cancers with
leukocyte infiltration of the skin or organs. Other diseases or
conditions in which undesirable inflammatory responses are to be
inhibited can be treated, including, but not limited to,
reperfusion injury, atherosclerosis, certain hematological
malignancies, cytokine-induced toxicity (e.g., septic shock,
endotoxic shock), polymyositis, dermatomyositis. Infectious
diseases or conditions of human or other species which can be
treated with inhibitors of chemokine receptor function, include,
but are not limited to, HIV.
[0314] Diseases or conditions of humans or other species which can
be treated with promoters of chemokine receptor function, include,
but are not limited to: immunosuppression, such as that in
individuals with immunodeficiency syndromes such as AIDS or other
viral infections, individuals undergoing radiation therapy,
chemotherapy, therapy for autoimmune disease or drug therapy (e.g.,
corticosteroid therapy), which causes immunosuppression;
immunosuppression due to congenital deficiency in receptor function
or other causes; and infections diseases, such as parasitic
diseases, including, but not limited to helminth infections, such
as nematodes (round worms); (Trichuriasis, Enterobiasis,
Ascariasis, Hookworm, Strongyloidiasis, Trichinosis, filariasis);
trematodes (flukes) (Schistosomiasis, Clonorchiasis), cestodes
(tape worms) (Echinococcosis, Taeniasis saginata, Cysticercosis);
visceral worms, visceral larva migraines (e.g., Toxocara),
eosinophilic gastroenteritis (e.g., Anisaki sp., Phocanema sp.),
cutaneous larva migraines (Ancylostona braziliense, Ancylostoma
caninum). The compounds of the present invention are accordingly
useful in the prevention and treatment of a wide variety of
inflammatory, infectious and immunoregulatory disorders and
diseases.
[0315] In addition, treatment of the aforementioned inflammatory,
allergic and autoimmune diseases can also be contemplated for
promoters of chemokine receptor function if one contemplates the
delivery of sufficient compound to cause the loss of receptor
expression on cells through the induction of chemokine receptor
internalization or delivery of compound in a manner that results in
the misdirection of the migration of cells.
[0316] In another aspect, the instant invention may be used to
evaluate the putative specific agonists or antagonists of a G
protein coupled receptor. The present invention is directed to the
use of these compounds in the preparation and execution of
screening assays for compounds that modulate the activity of
chemokine receptors. Furthermore, the compounds of this invention
are useful in establishing or determining the binding site of other
compounds to chemokine receptors, e.g., by competitive inhibition
or as a reference in an assay to compare its known activity to a
compound with an unknown activity. When developing new assays or
protocols, compounds according to the present invention could be
used to test their effectiveness. Specifically, such compounds may
be provided in a commercial kit, for example, for use in
pharmaceutical research involving the aforementioned diseases. The
compounds of the instant invention are also useful for the
evaluation of putative specific modulators of the chemokine
receptors. In addition, one could utilize compounds of this
invention to examine the specificity of G protein coupled receptors
that are not thought to be chemokine receptors, either by serving
as examples of compounds which do not bind or as structural
variants of compounds active on these receptors which may help
define specific sites of interaction.
[0317] The compounds of the present invention are used to treat or
prevent disorders selected from rheumatoid arthritis,
osteoarthritis, septic shock, atherosclerosis, aneurysm, fever,
cardiovascular effects, haemodynamic shock, sepsis syndrome, post
ischemic reperfusion injury, malaria, Crohn's disease, inflammatory
bowel diseases, mycobacterial infection, meningitis, psoriasis,
congestive heart failure, fibrotic diseases, cachexia, graft
rejection, autoimmune diseases, skin inflammatory diseases,
multiple sclerosis, radiation damage, hyperoxic alveolar injury,
HIV, HIV dementia, non-insulin dependent diabetes mellitus, asthma,
allergic rhinitis, atopic dermatitis, idiopathic pulmonary
fibrosis, bullous pemphigoid, helminthic parasitic infections,
allergic colitis, eczema, conjunctivitis, transplantation, familial
eosinophilia, eosinophilic cellulitis, eosinophilic pneumonias,
eosinophilic fasciitis, eosinophilic gastroenteritis, drug induced
eosinophilia, cystic fibrosis, Churg-Strauss syndrome, lymphoma,
Hodgkin's disease, colonic carcinoma, Felty's syndrome,
sarcoidosis, uveitis, Alzheimer, Glomerulonephritis, and systemic
lupus erythematosus.
[0318] In another aspect, the compounds are used to treat or
prevent inflammatory disorders selected from rheumatoid arthritis,
osteoarthritis, atherosclerosis, aneurysm, fever, cardiovascular
effects, Crohn's disease, inflammatory bowel diseases, psoriasis,
congestive heart failure, multiple sclerosis, autoimmune diseases,
skin inflammatory diseases.
[0319] In another aspect, the compounds are used to treat or
prevent inflammatory disorders selected from rheumatoid arthritis,
osteoarthritis, atherosclerosis, Crohn's disease, inflammatory
bowel diseases, and multiple sclerosis.
[0320] Combined therapy to prevent and treat inflammatory,
infectious and immunoregulatory disorders and diseases, including
asthma and allergic diseases, as well as autoimmune pathologies
such as rheumatoid arthritis and atherosclerosis, and those
pathologies noted above is illustrated by the combination of the
compounds of this invention and other compounds which are known for
such utilities. For example, in the treatment or prevention of
inflammation, the present compounds may be used in conjunction with
an anti-inflammatory or analgesic agent such as an opiate agonist,
a lipoxygenase inhibitor, a cyclooxygenase-2 inhibitor, an
interleukin inhibitor, such as an interleukin-1 inhibitor, a tumor
necrosis factor inhibitor, an NMDA antagonist, an inhibitor nitric
oxide or an inhibitor of the synthesis of nitric oxide, a
non-steroidal anti-inflammatory agent, a phosphodiesterase
inhibitor, or a cytokine-suppressing anti-inflammatory agent, for
example with a compound such as acetaminophen, aspirin, codeine,
fentaynl, ibuprofen, indomethacin, ketorolac, morphine, naproxen,
phenacetin, piroxicam, a steroidal analgesic, sufentanyl,
sunlindac, interferon alpha and the like. Similarly, the instant
compounds may be administered with a pain reliever; a potentiator
such as caffeine, an H2-antagonist, simethicone, aluminum or
magnesium hydroxide; a decongestant such as phenylephrine,
phenylpropanolamine, pseudophedrine, oxymetazoline, ephinephrine,
naphazoline, xylometazoline, propylhexedrine, or
levodesoxy-ephedrine; and antitussive such as codeine, hydrocodone,
caramiphen, carbetapentane, or dextramethorphan; a diuretic; and a
sedating or non-sedating antihistamine. Likewise, compounds of the
present invention may be used in combination with other drugs that
are used in the treatment/prevention/suppression or amelioration of
the diseases or conditions for which compound of the present
invention are useful. Such other drugs may be administered, by a
route and in an amount commonly used therefore, contemporaneously
or sequentially with a compound of the present invention. When a
compound of the present invention is used contemporaneously with
one or more other drugs, a pharmaceutical composition containing
such other drugs in addition to the compound of the present
invention may be used. Accordingly, the pharmaceutical compositions
of the present invention include those that also contain one or
more other active ingredients, in addition to a compound of the
present invention.
[0321] Examples of other active ingredients that may be combined
with a compound of the present invention, either administered
separately or in the same pharmaceutical compositions, include, but
are not limited to: (a) integrin antagonists such as those for
selectins, ICAMs and VLA-4; (b) steroids such as beclomethasone,
methylprednisolone, betamethasone, prednisone, dexamethasone, and
hydrocortisone; (c) immunosuppressants such as cyclosporin,
tacrolimus, rapamycin and other FK-506 type immunosuppressants; (d)
antihistamines (H1-histamine antagonists) such as bromopheniramine,
chlorpheniramine, dexchlorpheniramine, triprolidine, clemastine,
diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine,
methdilazine, promethazine, trimeprazine, azatadine,
cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,
terfenadine, loratadine, cetirizine, fexofenadine,
descarboethoxyloratadine, and the like; (e) non-steroidal
anti-asthmatics such as b2-agonists (terbutaline, metaproterenol,
fenoterol, isoetharine, albuteral, bitolterol, and pirbuterol),
theophylline, cromolyn sodium, atropine, ipratropium bromide,
leukotriene antagonists (zafirlukast, montelukast, pranlukast,
iralukast, pobilukast, SKB-102,203), leukotriene biosynthesis
inhibitors (zileuton, BAY-1005); (f) non-steroidal
anti-inflammatory agents (NSAIDs) such as propionic acid
derivatives (alminoprofen, benxaprofen, bucloxic acid, carprofen,
fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,
indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen,
pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen), acetic
acid derivatives (indomethacin, acemetacin, alclofenac, clidanac,
diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac,
ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin,
zidometacin, and zomepirac), fenamic acid derivatives (flufenamic
acid, meclofenamic acid, mefenamic acid, niflumic acid and
tolfenamic acid), biphenylcarboxylic acid derivatives (diflunisal
and flufenisal), oxicams (isoxicam, piroxicam, sudoxicam and
tenoxican), salicylates (acetyl salicylic acid, sulfasalazine) and
the pyrazolones (apazone, bezpiperylon, feprazone, mofebutazone,
oxyphenbutazone, phenylbutazone); (g) cyclooxygenase-2 (COX-2)
inhibitors; (h) inhibitors of phosphodiesterase type IV (PDE-IV);
(i) other antagonists of the chemokine receptors; (j) cholesterol
lowering agents such as HMG-COA reductase inhibitors (lovastatin,
simvastatin and pravastatin, fluvastatin, atorvastatin, and other
statins), sequestrants (cholestyramine and colestipol), nicotonic
acid, fenofibric acid derivatives (gemfibrozil, clofibrat,
fenofibrate and benzafibrate), and probucol; (k) anti-diabetic
agents such as insulin, sulfonylureas, biguanides (metformin),
a-glucosidase inhibitors (acarbose) and glitazones (troglitazone
and pioglitazone); (1) preparations of interferons (interferon
alpha-2a, interferon-2B, interferon alpha-N3, interferon beta-1a,
interferon beta-1b, interferon gamma-1b); (m) antiviral compounds
such as efavirenz, nevirapine, indinavir, ganciclovir, lamivudine,
famciclovir, and zalcitabine; (n) other compound such as
5-aminosalicylic acid an prodrugs thereof, anti-metabolites such as
azathioprine and 6-mercaptopurine, and cytotoxic cancer
chemotherapeutic agents. The weight ratio of the compound of the
present invention to the second active ingredient may be varied and
will depend upon the effective doses of each ingredient.
[0322] Generally, an effective dose of each will be used. Thus, for
example, when a compound of the present invention is combined with
an NSAID the weight ratio of the compound of the present invention
to the NSAID will generally range from about 1000:1 to about
1:1000, or alternatively from about 200:1 to about 1:200.
Combinations of a compound of the present invention and other
active ingredients will generally also be within the aforementioned
range, but in each case, an effective dose of each active
ingredient should be used.
[0323] The compounds are administered to a mammal in a
therapeutically effective amount. By "therapeutically effective
amount" it is meant an amount of a compound of Formula I that, when
administered alone or in combination with an additional therapeutic
agent to a mammal, is effective to prevent or ameliorate the
thromboembolic disease condition or the progression of the
disease.
Dosage and Formulation
[0324] The compounds of this invention can be administered in such
oral dosage forms as tablets, capsules (each of which includes
sustained release or timed release formulations), pills, powders,
granules, elixirs, tinctures, suspensions, syrups, and emulsions.
They may also be administered in intravenous (bolus or infusion),
intraperitoneal, subcutaneous, or intramuscular form, all using
dosage forms well known to those of ordinary skill in the
pharmaceutical arts. They can be administered alone, but generally
will be administered with a pharmaceutical carrier selected on the
basis of the chosen route of administration and standard
pharmaceutical practice.
[0325] The dosage regimen for the compounds of the present
invention will, of course, vary depending upon known factors, such
as the pharmacodynamic characteristics of the particular agent and
its mode and route of administration; the species, age, sex,
health, medical condition, and weight of the recipient; the nature
and extent of the symptoms; the kind of concurrent treatment; the
frequency of treatment; the route of administration, the renal and
hepatic function of the patient, and the effect desired. A
physician or veterinarian can determine and prescribe the effective
amount of the drug required to prevent, counter, or arrest the
progress of the thromboembolic disorder.
[0326] By way of general guidance, the daily oral dosage of each
active ingredient, when used for the indicated effects, will range
between about 0.001 to 1000 mg/kg of body weight, or between about
0.01 to 100 mg/kg of body weight per day, or alternatively, between
about 1.0 to 20 mg/kg/day. Intravenously, the doses will range from
about 1 to about 10 mg/kg/minute during a constant rate infusion.
Compounds of this invention may be administered in a single daily
dose, or the total daily dosage may be administered in divided
doses of two, three, or four times daily.
[0327] Compounds of this invention can be administered in
intranasal form via topical use of suitable intranasal vehicles, or
via transdermal routes, using transdermal skin patches. When
administered in the form of a transdermal delivery system, the
dosage administration will, of course, be continuous rather than
intermittent throughout the dosage regimen.
[0328] The compounds are typically administered in admixture with
suitable pharmaceutical diluents, excipients, or carriers
(collectively referred to herein as pharmaceutical carriers)
suitably selected with respect to the intended form of
administration, that is, oral tablets, capsules, elixirs, syrups
and the like, and consistent with conventional pharmaceutical
practices.
[0329] For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with
an oral, non-toxic, pharmaceutically acceptable, inert carrier such
as lactose, starch, sucrose, glucose, methyl cellulose, magnesium
stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol
and the like; for oral administration in liquid form, the oral drug
components can be combined with any oral, non-toxic,
pharmaceutically acceptable inert carrier such as ethanol,
glycerol, water, and the like. Moreover, when desired or necessary,
suitable binders, lubricants, disintegrating agents, and coloring
agents can also be incorporated into the mixture. Suitable binders
include starch, gelatin, natural sugars such as glucose or
beta-lactose, corn sweeteners, natural and synthetic gums such as
acacia, tragacanth, or sodium alginate, carboxymethylcellulose,
polyethylene glycol, waxes, and the like. Lubricants used in these
dosage forms include sodium oleate, sodium stearate, magnesium
stearate, sodium benzoate, sodium acetate, sodium chloride, and the
like. Disintegrators include, without limitation, starch, methyl
cellulose, agar, bentonite, xanthan gum, and the like.
[0330] The compounds of the present invention can also be
administered in the form of liposome delivery systems, such as
small unilamellar vesicles, large unilamellar vesicles, and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as cholesterol, stearylamine, or
phosphatidylcholines.
[0331] Compounds of the present invention may also be coupled with
soluble polymers as targetable drug carriers. Such polymers can
include polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamide-phenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine
substituted with palmitoyl residues. Furthermore, the compounds of
the present invention may be coupled to a class of biodegradable
polymers useful in achieving controlled release of a drug, for
example, polylactic acid, polyglycolic acid, copolymers of
polylactic and polyglycolic acid, polyepsilon caprolactone,
polyhydroxy butyric acid, polyorthoesters, polyacetals,
polydihydropyrans, polycyanoacylates, and crosslinked or
amphipathic block copolymers of hydrogels.
[0332] Dosage forms (pharmaceutical compositions) suitable for
administration may contain from about 1 milligram to about 100
milligrams of active ingredient per dosage unit. In these
pharmaceutical compositions the active ingredient will ordinarily
be present in an amount of about 0.5-95% by weight based on the
total weight of the composition.
[0333] Gelatin capsules may contain the active ingredient and
powdered carriers, such as lactose, starch, cellulose derivatives,
magnesium stearate, stearic acid, and the like. Similar diluents
can be used to make compressed tablets. Both tablets and capsules
can be manufactured as sustained release products to provide for
continuous release of medication over a period of hours. Compressed
tablets can be sugar coated or film coated to mask any unpleasant
taste and protect the tablet from the atmosphere, or enteric coated
for selective disintegration in the gastrointestinal tract.
[0334] Liquid dosage forms for oral administration can contain
coloring and flavoring to increase patient acceptance.
[0335] In general, water, a suitable oil, saline, aqueous dextrose
(glucose), and related sugar solutions and glycols such as
propylene glycol or polyethylene glycols are suitable carriers for
parenteral solutions. Solutions for parenteral administration may
contain a water soluble salt of the active ingredient, suitable
stabilizing agents, and if necessary, buffer substances.
Antioxidizing agents such as sodium bisulfate, sodium sulfite, or
ascorbic acid, either alone or combined, are suitable stabilizing
agents. Also used are citric acid and its salts and sodium EDTA. In
addition, parenteral solutions can contain preservatives, such as
benzalkonium chloride, methyl- or propyl-paraben, and
chlorobutanol.
[0336] Suitable pharmaceutical carriers are described in
Remington's Pharmaceutical Sciences (Mack Publishing Company), a
standard reference text in this field.
[0337] Representative useful pharmaceutical dosage-forms for
administration of the compounds of this invention can be
illustrated as follows:
Capsules
[0338] A large number of unit capsules can be prepared by filling
standard two-piece hard gelatin capsules each with 100 milligrams
of powdered active ingredient, 150 milligrams of lactose, 50
milligrams of cellulose, and 6 milligrams magnesium stearate.
Soft Gelatin Capsules
[0339] A mixture of active ingredient in a digestible oil such as
soybean oil, cottonseed oil or olive oil may be prepared and
injected by means of a positive displacement pump into gelatin to
form soft gelatin capsules containing 100 milligrams of the active
ingredient. The capsules should be washed and dried.
Tablets
[0340] Tablets may be prepared by conventional procedures so that
the dosage unit is 100 milligrams of active ingredient, 0.2
milligrams of colloidal silicon dioxide, 5 milligrams of magnesium
stearate, 275 milligrams of microcrystalline cellulose, 11
milligrams of starch and 98.8 milligrams of lactose. Appropriate
coatings may be applied to increase palatability or delay
absorption.
Injectable
[0341] A parenteral composition suitable for administration by
injection may be prepared by stirring 1.5% by weight of active
ingredient in 10% by volume propylene glycol and water. The
solution should be made isotonic with sodium chloride and
sterilized.
Suspension
[0342] An aqueous suspension can be prepared for oral
administration so that each 5 mL contain 100 mg of finely divided
active ingredient, 200 mg of sodium carboxymethyl cellulose, 5 mg
of sodium benzoate, 1.0 g of sorbitol solution, U.S.P., and 0.025
mL of vanillin.
[0343] Where the compounds of this invention are combined with
other anticoagulant agents, for example, a daily dosage may be
about 0.1 to 100 milligrams of the compound of Formula I and about
1 to 7.5 milligrams of the second anticoagulant, per kilogram of
patient body weight. For a tablet dosage form, the compounds of
this invention generally may be present in an amount of about 5 to
10 milligrams per dosage unit, and the second anti-coagulant in an
amount of about 1 to 5 milligrams per dosage unit.
[0344] Where two or more of the foregoing second therapeutic agents
are administered with the compound of Formula I, generally the
amount of each component in a typical daily dosage and typical
dosage form may be reduced relative to the usual dosage of the
agent when administered alone, in view of the additive or
synergistic effect of the therapeutic agents when administered in
combination. Particularly when provided as a single dosage unit,
the potential exists for a chemical interaction between the
combined active ingredients. For this reason, when the compound of
Formula I and a second therapeutic agent are combined in a single
dosage unit they are formulated such that although the active
ingredients are combined in a single dosage unit, the physical
contact between the active ingredients is minimized (that is,
reduced). For example, one active ingredient may be enteric coated.
By enteric coating one of the active ingredients, it is possible
not only to minimize the contact between the combined active
ingredients, but also, it is possible to control the release of one
of these components in the gastrointestinal tract such that one of
these components is not released in the stomach but rather is
released in the intestines. One of the active ingredients may also
be coated with a material which effects a sustained-release
throughout the gastrointestinal tract and also serves to minimize
physical contact between the combined active ingredients.
Furthermore, the sustained-released component can be additionally
enteric coated such that the release of this component occurs only
in the intestine. Still another approach would involve the
formulation of a combination product in which the one component is
coated with a sustained and/or enteric release polymer, and the
other component is also coated with a polymer such as a
low-viscosity grade of hydroxypropyl methylcellulose (HPMC) or
other appropriate materials as known in the art, in order to
further separate the active components. The polymer coating serves
to form an additional barrier to interaction with the other
component.
[0345] These as well as other ways of minimizing contact between
the components of combination products of the present invention,
whether administered in a single dosage form or administered in
separate forms but at the same time by the same manner, will be
readily apparent to those skilled in the art, once armed with the
present disclosure.
[0346] While the invention has been described in detail and with
reference to specific embodiments thereof, it will be apparent to
one skilled in the art that various changes and modifications can
be made therein without departing from the spirit and scope
thereof.
* * * * *