U.S. patent application number 14/705382 was filed with the patent office on 2015-08-20 for 3-cycloalkylaminopyrrolidine derivatives as modulators of chemokine receptors.
The applicant listed for this patent is Pfizer Inc.. Invention is credited to Amy Qi Han, Brian W. Metcalf, Darius J. Robinson, Anlai Wang, Chu-Biao Xue, Yingxin Zhang, Changsheng Zheng.
Application Number | 20150231114 14/705382 |
Document ID | / |
Family ID | 34710216 |
Filed Date | 2015-08-20 |
United States Patent
Application |
20150231114 |
Kind Code |
A1 |
Xue; Chu-Biao ; et
al. |
August 20, 2015 |
3-CYCLOALKYLAMINOPYRROLIDINE DERIVATIVES AS MODULATORS OF CHEMOKINE
RECEPTORS
Abstract
The present invention relates to 3-cycloalkylaminopyrrolidine
derivatives of the formula I: ##STR00001## (wherein R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, X, Y and Z
are as defined herein) which are useful as modulators of chemokine
receptor activity. In particular, these compounds, are useful as
modulators of chemokine receptors and more specifically as
modulators of the CCR2 and/or CCR5 receptor. The compounds and
compositions of the invention may bind to chemokine receptors,
e.g., fee CCR2 and/or CCR5 chemokine receptors, and are useful for
treating diseases associated with chemokine, e.g., CCR2 and/or
CCR5, activity, such as atherosclerosis, restenosis, lupus, organ
transplant rejection and rheumatoid arthritis.
Inventors: |
Xue; Chu-Biao; (Hockessin,
DE) ; Metcalf; Brian W.; (Moraga, CA) ; Han;
Amy Qi; (Hockessin, DE) ; Robinson; Darius J.;
(Wilmington, DE) ; Zheng; Changsheng; (Wilmington,
DE) ; Wang; Anlai; (Wilmington, DE) ; Zhang;
Yingxin; (New Castle, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Pfizer Inc. |
New York |
NY |
US |
|
|
Family ID: |
34710216 |
Appl. No.: |
14/705382 |
Filed: |
May 6, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14174300 |
Feb 6, 2014 |
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14705382 |
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12274045 |
Nov 19, 2008 |
8697870 |
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14174300 |
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11014322 |
Dec 16, 2004 |
7576089 |
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12274045 |
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60531270 |
Dec 18, 2003 |
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Current U.S.
Class: |
514/210.18 ;
514/236.8; 514/252.05; 514/255.05; 514/342; 514/343; 514/365;
514/423 |
Current CPC
Class: |
A61P 9/10 20180101; C07D
401/14 20130101; A61K 31/5377 20130101; A61K 31/506 20130101; A61P
17/00 20180101; A61P 19/00 20180101; A61P 43/00 20180101; A61P 3/10
20180101; A61P 9/08 20180101; A61P 25/00 20180101; C07D 401/12
20130101; C07D 403/12 20130101; A61P 19/02 20180101; A61P 31/18
20180101; A61K 31/4439 20130101; A61P 31/12 20180101; C07D 417/14
20130101; C07D 413/14 20130101; A61P 25/04 20180101; C07D 413/12
20130101; A61K 31/427 20130101; A61K 31/497 20130101; C07D 207/14
20130101; A61K 31/4025 20130101; A61P 37/00 20180101; C07D 417/04
20130101; A61P 15/00 20180101; A61K 31/40 20130101; A61K 31/454
20130101; A61P 3/04 20180101; A61K 31/501 20130101; A61P 29/00
20180101; A61P 37/06 20180101; A61P 37/02 20180101; C07D 417/12
20130101; A61P 35/00 20180101; C07D 277/24 20130101 |
International
Class: |
A61K 31/40 20060101
A61K031/40; A61K 31/427 20060101 A61K031/427; A61K 31/506 20060101
A61K031/506; A61K 31/501 20060101 A61K031/501; A61K 31/497 20060101
A61K031/497; A61K 31/4439 20060101 A61K031/4439; A61K 31/5377
20060101 A61K031/5377 |
Claims
1-45. (canceled)
46. A method of treating a disease associated with expression or
activity of a chemokine receptor in a patient comprising
administering to said patient a therapeutically effective amount of
a compound of the formula I: ##STR00212## or pharmaceutically
acceptable salt thereof, wherein: X and Y together are selected
from the group consisting of: ##STR00213## Z is a phenyl, pyridyl
or pyrimidinyl group that is substituted with a CF.sub.3 group;
R.sup.1 is phenyl, pyridyh thiazole, pyrimidine or pyrazine,
wherein said R.sup.1 is substituted with 0-3 R.sup.1a, wherein
R.sup.1a is independently selected from the group consisting of:
CH.sub.2 OMe, alkyl amides, pyrollidinyl amides, morpholinyl,
alkyl, alkoxy, CH.sub.2--NMe.sub.2, pyridyl, carboxylate, phenyl
bearing a CH.sub.2--OH group, pyrimidine, thiazole, oxazole and
pyrazine; R.sup.2 is OH; R.sup.3 and R.sup.4 are H; R.sup.5 is
independently selected from the group consisting of hydrogen,
alkyl, and formyl; R.sup.6 and R.sup.7 are each independently
selected from the group consisting of H, C.sub.1-C.sub.10 alkyl,
OH, CH.sub.2--OMe and hydroxyalkyl; and r=1.
47. The method of claim 46 wherein said chemokine receptor is CCR2
or CCR5.
48. The method of claim 46 wherein said disease is an inflammatory
disease.
49. The method of claim 46 wherein said disease is an immune
disorder.
50. The method of claim 46 wherein said disease is rheumatoid
arthritis, atherosclerosis, lupus, multiple sclerosis, neuropathic
pain, transplant rejection, diabetes, or obesity.
51. The method of claim 46 wherein said disease is cancer.
52. The method of claim 51 wherein said cancer is characterized by
tumor associated macrophages.
53. The method of claim 51 wherein said cancer is breast cancer,
ovarian cancer or multiple myeloma.
54. The method of claim 46 wherein said disease or condition is a
viral infection.
55. The method of claim 54 wherein said viral infection is HIV
infection.
56-57. (canceled)
58. A method of treating cancer in a patient comprising
administering to said patient a therapeutically effective amount of
a compound of the formula I: ##STR00214## or pharmaceutically
acceptable salt thereof, wherein: X and Y together are selected
from the group consisting of: ##STR00215## Z is a phenyl, pyridyl
or pyrimidinyl group that is substituted with a CF.sub.3 group; R1
is phenyl, pyridyl, thiazole, pyrimidine or pyrazine, wherein said
R.sup.1 is substituted with 0-3 R.sup.1a, wherein Rla is
independently selected from the group consisting of: CH.sub.2--OMe,
alkyl amides, pyrollidinyl amides, morpholinyl, alkyl, alkoxy,
CH.sub.2--NMe.sub.2, pyridyl, carboxylate, phenyl bearing a
CH.sub.2--OH group, pyrimidine, thiazole, oxazole and pyrazine;
R.sup.2 is OH; R.sup.3 and R.sup.4 are H; R.sup.5 is independently
selected from the group consisting of hydrogen, alkyl, and formyl;
R.sup.6 and R.sup.7 are each independently selected from the group
consisting of H, C.sub.1-C.sub.10 alkyl, OH, CH.sub.2--OMe and
hydroxyalkyl; and r=1.
59. The method of claim 58, wherein the cancer is characterized by
infiltration of macrophages into tumors or diseased tissue.
60. The method of claim 39, wherein said compound is administered
with a pharmaceutically acceptable carrier.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application Ser. No. 60/531,270, filed Dec. 18, 2003, the
disclosure of which is incorporated herein by reference in its
entirety.
FIELD OF THE INVENTION
[0002] The instant invention is directed to chemokine receptor
modulators, e.g., antagonists, and their use as medicinal agents.
The present invention further relates to novel compounds and
medical methods of treatment of inflammation, and other disorders
especially those associated with lymphocyte or monocyte
accumulation such as rheumatoid arthritis, lupus, graft versus host
diseases and/or transplant rejection. More particularly, the
present invention relates to 3-cycloalkylaminopyrrolidine
derivatives and their use as modulators of chemokine receptors.
[0003] More specifically, the instant invention relates to new
anti-inflammatory and immunomodulatory bioactive compounds and
pharmaceutical compositions thereof that act via antagonism of the
CCR2 receptor, (also known as the MCF-1 receptor), and therefore
leading to the. inhibition of Monocyte Chemtialtractarit Protein-1
(MCP-1). The new compounds are 3-cycloalkylaminopyrrolidine
derivatives. The invention further relates to novel compounds for
use in the compositions, to processes for their preparation, to
intermediates useful in their preparation and to their use as
therapeutic agents.
[0004] The chemokine receptor modulators/antagonists of the
invention may be effective as therapeutic agents and/or preventive
agents for diseases such as atherosclerosis, asthma, pulmonary
fibrosis, myocarditis, ulcerative colitis, psoriasis, asthma,
ulcerative colitis, nephritis (nephropathy), multiple sclerosis,
lupus, systemic lupus erythematosus, hepatitis, pancreatitis,
sarcoidosis, organ transplantation, Crohn's disease, endometriosis,
congestive heart failure, viral meningitis, cerebral infarction,
neuropathy, Kawasaki disease, and sepsis in which tissue
infiltration of blood leukocytes, such as monocytes and
lymphocytes, play a major role in the initiation, progression or
maintenance of the disease.
[0005] The present invention also provides immunomodulatory
bioactive compounds and pharmaceutical compositions thereof that
act via antagonism of the CCR5 receptor.
BACKGROUND OF THE INVENTION
[0006] The migration and transport of leukocytes from blood vessels
into diseased tissues appears to be a critical component to the
initiation of normal disease-lighting. inHammatory responses. The
process, also known as leukocyte recruitment, is also related to
the onset and progression of life-threatening inflammatory, as well
as debilitating autoimmune diseases. The resulting pathology of
these diseases derives from the attack of the body's immune system
defenses on normal tissues. Accordingly, preventing and blocking
leukocyte recruitment to target tissues in inflammatory and
autoimmune disease would be a highly effective approach to
therapeutic intervention.
[0007] The different classes of leukocyte cells that are involved
in cellular immune responses include monocytes, lymphocytes,
neutrophils, eosinophils and basophils. In most cases, lymphocytes
are the leukocyte class that initiates, coordinates, and maintains
chronic inflammatory responses, and thns are generally the most
important class of cells to block, from entering inflammatory
sites. Lymphocytes attract monocytes to the tissue sites, which,
collectively with lymphocytes, are responsible for most of the
actual tissue damage that occurs in inflammatory disease.
Infiltration, of the lymphocytes and/or monocytes is known to lead
to a wide range of chronic, autoimmune diseases, and also organ
transplant rejection. These diseases include, but are not limited
to, rheumatoid arthritic chronic contact dermatitis, inflammatory
bowel disease, lupus, systemic lupus erythematosus, multiple
sclerosis, atherosclerosis, psoriasis, sarcoidosis, idiopathic
pulmonary fibrosis, dermatomyositis, skin pemphigoid and related
diseases, (e.g., pemphigus vulgaris, p. foliacious, p.
erythematosis), glomerulonephritides, vasculitides, hepatitis,
diabetes, allograft rejection, and graft-versus-host disease.
[0008] The process, by which leukocytes leave the bloodstream and
accumulate at inflammatory sites, and start a disease, has at least
three steps which have been, described as (1) rolling, (2)
activation/firm adhesion and (3) transendomelial migration
[Springer, T. A., Nature 346:425-433 (1990); Lawrence and Springer,
Cell 65:859-873 (1991): Butcher, E. C, Cell 67:1033-1036 (1991)].
The second step is mediated at the molecular level by
chemoattractant receptors, Chemoattractant receptors on the
surface, of leukocytes then bind chemoattractant cytokines which
are secreted by cells at the site of damage or infection. Receptor
binding activates leukocytes, increases the adhesiveness of the
adhesion molecules that mediate transendothelial migration, and
promotes directed migration of the cells toward the source of the
chemoattractant cytokine.
[0009] Chemotactic cytokines (leukocyte chemoattracant/activating
(actors) also known as chemokines, also known as intercrines and
SIS cytokines are a group of inflammatory/immunomodulatory
polypeptide factors, of molecular weight 6-15 kDa, that are
released by a wide variety of cells such as macrophages, monocytes,
eosinophils, neutrophils, fibroblasts, vascular endotherial cells,
smooth muscle cells, and mast cells, at inflammatory sites
(reviewed in Luster, New Eng. J Med., 338, 436-445 (1998) and
Rollins, Blood, 90, 909-928 (1997)). Also, chemokines has been
described in Oppenheirn, J. J. et al., Annu. Rev. Immunol.,
9:617-648 (1991); Schall and Bacon, Curr. Opin. Immunol., 6:865-873
(1994); Baggloiini, M., et al., and Adv. Immunol., 55:97-179
(1994). Chemokines have the ability to stimulate directed cell
migration, a process known as chemotaxis. Each chemokine contains
four cysteine residues (C) and two internal disulfide bonds.
Chemokines can be grouped into two subfamilies, based on whether
the two amino terminal cysteine residues are immediately adjacent
(CC family) or separated by one amino acid (CXC family). These
differences correlate with the organization of the two subfamilies
into separate gene clusters. Within each gene cluster, the
chemokines typically show sequence similarities between 25 to 60%.
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,
lymphotacin-2 (both C chemokines), and fractalkine (a CXXXC
chemokine) that do not fall into either of the major chemokine
subfamilies.
[0010] MCP-1 (also known as MCAF (abbreviation for macrophage
chemotactic and activating factor) or JE) is a CC chemokine
produced by monocytes/macrophages, smooth muscle cells,
fibroblasts, and vascular endothelial cells and causes cell
migration and cell adhesion of monocytes (see for example Valente,
A. J., et al. Biochemistry, 1988, 27, 4162; Matsushima, K., et ah,
J. Exp. Med., 1989, 169, 1485; Yoshimura, T., et al., J, Immunol.,
1989, 142, 1956; Rollins, B. J., et al., Proc. Natl. Acad. Sol.
USA, 1988, 85, 3738; Rollins, B. J., et al., Blood, 1991, 78 1112;
Jiang, Y., et al., J. Immunol, 1992, 148, 2423; Yaddi, K., et al.,
J. Immunol., 1994, 153,4721), memory T lymphocytes (see for example
Carr, M. W., et al., Free. Natl. Acad, Set,. USA, 1994, 91, 3652),
T lymphocytes (see for example Loetscher, P., et al., FASEB J.,
1994, 8, 1055) and natural killer cells (see for example Loetscher,
P., et al., J. Immunol, 1996, 156, 322; Allavena, P. et al., Eur.
J. Immunol., 1994, 24, 3233), as well as mediating histamine
release by basophils (see for example Alam, R., et al., J. Clin,
Invest., 1992, 89, 723; Bischoff, S. C., et al. J. Exp. Med., 1992,
175, 1271; Kuna, P., et al. J. Exp. Med., 1992, 175, 489). In
addition, high expression of MCP-1 has been reported in diseases
where accumulation of monocyte/macrophage and/or T cells is thought
to be important in the initiation or progression of diseases, such
as atherosclerosis (see for example Hayes, I. M., et al.,
Arterioseler. Thromb. Vasc. Biol., 1998, 18, 39: Takeya, M. et.
al., Hum. Pathol., 1993, 24, 534; Yla-Herttuala, S., et ah, Proc,
Natl Acad, Scl. USA, 1991, 88, 52; Nelken, M A., J. Clin, invest.,
1991, 88,1121), rheumatoid arthritis (see for example Koch, A. E et
al., J. Clin. Invest., 1992, 90, 772; Akahoshi, Y., et al.
Arthritis Rheum., 1993, 36, 762; Robinson, E., et al., Clin. Exp.
Immunol, 101, 398), nephritis (see for example Noris, M., et al.,
Lab. invest., 1995, 73, 804: Wada, T., at al., Kidney Int., 1996,
49, 761; Gesualdo, L., et al., Kidney Int., 1997, 51, 155),
nephropathy (see for example Saitoh, A., et al, J. Clin; Lab. Anal,
1998, 12, 1: Yoloyams, H., et al., J. Leukoc. Biol, 1998, 63, 493),
pulmonary fibrosis, pulmonary sarcoidosis (see for example
Sugiyania. Y., et al., Internal Medicine, 1997, 36, 856), asthma
(see for example Karma, M., et al., J. Invest. Allergol. Clin.
Immunol., 1997, 7, 254; Stephene T. H., Am. J, Respir. Crit. Care
Med., 1997, 156, 1377; Sousa, A. R. et al. Am. J. Respir. Cell Mol.
Biol, 1994, 10,142), multiple sclerosis (see for example McManus,
C., et al., J. Neuroimmunol., 1998, 86, 20), psoriasis (see lor
example Gillitzer, R., et al., J. Invest. Dermatol, 1993, 101,
127), inflammatory bowel disease (see for example Grimm, M. C, et
al., J. Leukoc. Biol, 1996, 59, 804; Reinecker, H. C., et al.,
Gastroenterology, 1995, 106, 40), myocarditis (see for example
Seino, Y., et al., Cytokine, 1995, 7, 301), endometriosis (see for
example Jolicoeur, C., et al., Am. J. Pathol, 1998. 152, 125),
intraperitoneal adhesion (see for example Zeyneloglu, H. B., et
al., Human Reproduction, 1998, 13, 1194), congestive heart failure
(see for example Aurust, P., et al., Circulation, 1998, 97, 1136),
chronic liver disease (see for example Marra, F., et al., Am. J.
Pathol, 1998, 152, 423), viral meningitis (see for example Lahrtz,
F., et al., Eur. J. Immunol, 1997, 27, 2484), Kawasaki disease (see
for example Wong, M.; et al., J. Rheumatol, 1997, 24, 1179) and
sepsis (see for example Salkowski, C. A.; et al, Infect, immun.,
1998, 66, 3569). Furthermore, anti-MCP-1 antibody has been reported
to show an inhibitory effect or a therapeutic effect in animal
models of rheumatoid arthritis (see for example Schimmer, R. C, et
al., J. Immunol, 1998, 160, 1466; Schrier, D, J., J. Leukoc. Biol.,
1998, 63, 359; Ogata, H., et al., J. Pathol, 1997, 182, 106),
multiple sclerosis (see for example Karpus, W. J., et al., J.
Leukoc. Biol., 1997, 62, 681), nephritis (see for example Lloyd, C.
M, et al., J. Exp. Med., 1997, 185, 1371; Wada, T. et al., FASEB
J., 1996, 10, 1418), Asthma (see for example Gonzalo, J.-A., et
al., J. Exp. Med., 1998, 188, 157; Lukacs, N. W., J. Immunol.,
1997, 158, 4398), atherosclerosis (see for example Guzman, L. A.,
et al., Circulation, 1993, 88 (suppl.), 1-371), delayed type
hypersensitivity (see for example Rand, M. L., et al., Am. J.
Pathol., 1996, 148, 855), pulmonary hypertension (see for example
Kimura, H,, et al., Lab. Invest., 1998, 78, 571), and
intraperitoneal adhesion (see for example Zeyneloglu, H. B., et
al., Am. J. Obstet. Gynecol., 1998, 179, 438). A peptide antagonist
of MCP, MCP-1 (9-76), has been also reported to inhibit arthritis
in the mouse model (see Gong, J.-H. J. Exp., 4ed., 1997, 186, 131),
as well as studies in MCP-1-deficient mice have shown that MCP-1 is
essential for monocyte recruitment in vivo (see Lu, B, et al., J.
Exp. Med., 1998, 187, 601; Gu, L., et al., Moll Cell, 1998, 2,
275),
[0011] The published literature indicate that chemokines such as
MCP-1 and MIP-1.alpha. attract monocytes and lymphocytes to disease
sites and mediate their activation and thus are thought to be
intimately involved in the initiation, progression and maintenance
of diseases deeply involving monocytes and lymphocytes, such as
atherosclerosis, restenosis, rheumatoid arthritis, psoriasis,
asthma, ulcerative colitis, nephritis (nephropathy), multiple
sclerosis, pulmonary fibrosis, myocarditis, hepatitis,
pancreatitis, sarcoidosis, Crohn's disease, endometriosis,
congestive heart failure, viral meningitis, cerebral infarction,
neuropathy, Kawasaki disease, and sepsis (see for example Rovin, B.
H., et al., Am. J. Kidney, Dis., 1998, 31, 1065; Lloyd, C., et al.,
Curr. Opin. Nephrol. Hypertens., 1998, 7, 281; Conti, P.,et al.,
Allergy and Asthma Proc, 1998, 19, 121; Ransohoff, M., et al.,
Trends Neurosci., 1998, 21, 154; MacDermott, R. P., et al.,
Inflammatory Bowel Diseases, 1998, 4, 54).
[0012] 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., 15, 159-165 (1994)) which are termed "chemokine
receptors." On binding their cognate ligands, chemokine receptors
transduce an intracellular signal through 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, degranuiation,
and promotion of cell migration.
[0013] Genes encoding receptors of specific chemokines have been
cloned, and it is now known that these receptors are G
protein-coupled seven-transmembrane receptors present on various
leukocyte populations. So far, at least five CXC chemokme receptors
(CXCR1-CXCR5) and eight CC chemokine receptors (CCR1-CCR8) have
been identified. For example IL-8 is a iigand for CXCR1 and CXCR2,
MIP-1.alpha. is that for CCR1 and CCR5, and MCP-1 is that for CCR2A
and CCR2B (for reference, see for example, Holmes, W. E., et ah,
Science 1991, 253, 1278-1280; Murphy P. M., et al., Science, 253,
1280-1283; Neote, K. et al, Cell, 1993, 72, 415-425; Charo, I. F.,
et al., Proc, Natl. Acad. Sci. USA, 1994, 91, 2752-2756; Yamagami,
S., et al., Biochem. Biophys, Res. Commun., 1994, 202, 1156-1162;
Combadier, C., et. al., The Journal of Biological Chemistry, 1995,
270, 16491-16494, Power, C. A., et al., J. Biol. Chem., 1995, 270,
19495-19500; Samson, M, et al., Biochemistry, 1996, 35, 3362-3367;
Murphy, P. M., Annual Review of Immunology, 1994, 12, 592-633). It
has been reported that lung inflammation and granuroma formation
are suppressed in CCR1-deficient mice (see Gao, J.-L., et al., J.
Exp. Med., 1997, 185, 1959, Gerard, C., et al., J. Clin. Invest.,
1997, 100, 2022), and that recruitment of macrophages and formation
of atherosclerotic lesion decreased in CCR2-deficient mice (see
Boring, L., et al., Nature, 1998, 394, 894; Kuziel, W. A., et al.,
Proc. Natl. Acad. Sci., USA, 1997, 94, 12053; Kurihara, T., et al.,
J. Exp, Med., 1997, 186, 1757; Boring, L., et al., J. Clin.
Invest., 1997, 100, 2552).
[0014] Accordingly, drugs which inhibit the binding of chemokines
such as MCP-1 and/or MIP-1.alpha. to these receptors, e.g.,
chemokine receptor antagonists, may be useful as pharmaceutical
agents which inhibit the action of chemokines such as MCP-1 and/or
MIP-1.alpha. on the target cells, but the prior art is silent
regarding 3-cycloakylaminopyrrolidine derivatives having such
pharmacological effects. The identification of compounds that
modulate the function of CCR2 and/or CCR5 represents an excellent
drug design approach to the development of pharmacological agents
for the treatment of inflammatory conditions and diseases
associated with CCR2 and/or CCR5 activation, such as rheumatoid
arthritis, lupus and other inflammatory diseases. The present
invention provides solutions to a long felt need in the field of
chemokine receptor modulators and antagonists.
OBJECTS OF THE INVENTION
[0015] With the foregoing in mind, it is an object of the present
invention to provide chemokine receptor antagonists and chemokine
receptor modulators for treating rheumatoid arthritits.
[0016] Another main object of the invention is to provide chemokine
receptor antagonists and their use as medicinal agents.
[0017] An additional object of the invention is to provide
chemokine receptor modulators and their use as medicinal
agents.
[0018] A further object of the present invention is to provide
3-cycloalkylaminopyrrolidine derivatives.
[0019] Another object of the invention relates to novel compounds
and medical methods of treatment of inflammation.
[0020] A still further object of the invention provides new
anti-inflammatory and immunomodulatory hioactive compounds and
pharmaceutical compositions thereof that act via antagonism of the
CCR2 receptor.
[0021] An additional object of the invention provides
3-cycloalkylaminopyrrolidine derivatives and their use as
modulators of chemokine receptors.
[0022] A still additional object of the invention provides
3-cycloalkylaminopyrrolidine derivatives and their use in treating
and preventing atherosclerosis and restenosis.
[0023] A further object of the invention provides
3-cycloalkylaminopyrrolidine derivatives and their use as
modulators of the CCR5 receptor.
[0024] Another main object of the invention provides
3-cycloalkylaminopyrrolidine bioactive. compounds and
pharmaceutical compositions thereof that act via antagonism of the
CCR5 receptor.
[0025] Other objects and embodiments of the present invention will
be discussed below. However, it is important to note that many
additional embodiments of the present invention not described in
this specification may nevertheless fall within the spirit and
scope of the present Invention and/or the claims.
SUMMARY OF THE INVENTION
[0026] The present invention is directed to compounds of formulas I
and II:
##STR00002##
[0027] or enantiomers, diastereomers, enantiomerically enriched
mixtures, racemic mixtures thereof, prodrugs, crystalline forms,
non-crystalline forms, amorphous forms thereof, solvates thereof,
metabolites thereof, and pharmaceatically acceptable salts thereof,
wherein constituent variables are provided herein.
[0028] The instant invention also relates to pharmaceutical
compositions which comprise anti-inflammatory and/or
immunomodulatory compounds of formula I and II as shown above, that
act via antagonism of the CCR2 receptor, (also known as the MCP-1
receptor), therefore inhibiting the Monocyte Chemoattractant
Protein-1 (MCP-1),
[0029] The instant invention is also directed to pharmaceutical
compositions which comprise anti-inflammatory and/or
immunomodulatory compounds of formula I and II as shown above, that
act via antagonism of the CCR5 receptor (also known as the MCP-1
receptor), therefore inhibiting the Monocyte Chemoattractant
Protein-1 (MCP-1).
[0030] The present invention is also directed to compounds of
formula I and II which are modulators of CCR2 chemokine. receptor
function and are useful in the prevention or treatment of
inflammatory conditions and diseases such as rheumatoid arthritis,
allergic diseases, psoriasis, atopic dermatitis, lupus and
asthma.
[0031] The present invention also describes compounds of formula I
and II which are modulators of CCR5 chemokine receptor function and
are useful in the prevention or treatment of inflammatory
conditions and diseases such as rheumatoid arthritis, allergic
diseases, psoriasis, atopic dermatitis, lupus and asthma.
[0032] The invention is also provides pharmaceutical compositions
comprising compounds selected from the group of formula I and II,
and the use of these compounds and compositions in the prevention
or treatment of diseases in which CCR2 chemokine receptors are
involved.
[0033] The invention further provides pharmaceutical compositions
comprising compounds selected from the group of formula I and II,
and the use of these compounds and compositions in the prevention
or treatment of diseases in which CCR5 chemokine receptors are
involved.
[0034] The invention additionally provides a method for foe
treatment of inflammation, rheumatoid arthritis, lupus, systemic
lupus erythematosus, atherosclerosis, restenosis, immune disorders,
and transplant rejection in a mammal in need thereof comprising
administering to such mammal a therapeutically effective amount of
a pharmaceutical composition containing a compound according to
formula I and II in admixture with a pharmaceutically acceptable
excipient, diluent, or carrier.
[0035] The present invention further provides compositions
comprising a compound of the invention and a pharmaceutically
acceptable carrier.
[0036] The present invention further provides methods of modulating
activity of a chemokine receptor comprising contacting said
chemokine receptor with a compound of the invention.
[0037] The present invention further provides methods of treating a
disease associated with expression or activity of a chemokine
receptor in a patient comprising administering to the patient a
therapeutically effective amount of a compound of the
invention.
DETAILED DESCRIPTION
[0038] The instant invention is directed to a compound of the
formula I:
##STR00003##
including its enantiomers, diastereomers, enantiomerically enriched
mixtures, racemic mixtures thereof, prodrugs, crystalline forms,
non-crystalline forms, amorphous forms thereof, solvates thereof,
metabolites thereof, and pharmaceutically acceptable salts,
wherein:
[0039] X is selected from the group consisting of a bond, aryl,
mono or poly substituted aryl, heterocycle, mono or poly
substituted heterocycle, heteroaryl, mono or poly substituted
heteroaryl, carbocycle, mono or poly substituted carbocycle and
(CR.sup.8R.sup.9).sub.n, wherein n=0-5;
[0040] Y is a bond, or is selected from the group consisting of
oxygen, sulfur, nitrogen, amide bond, thioamide bond, sulfonamide,
ketone, --CHOH--, --CHO-alkyl-, -alkyl-O-alkyl, oxime, and a
urea;
[0041] Z is selected from the group consisting of carbocycle, aryl,
heterocycle and heteroaryl, each having 0-3 R.sup.10 substituents,
wherein R.sup.10 is independently selected from the group
consisting of: halogen, alkyl, alkenyl, alkynyl, alkoxy, cyclic
alkoxy, heterocyclic alkoxy, alkoxyalkyl, cyclic alkoxyalkyl,
heterocyclic alkoxyalkyl, alkylthioalkyl, cyclic alkylthioalkyl,
heterocyclic alkylthioalkyl, thioalkyl, mono-, di- or
tri-haloalkyl, mono-, di- or tri-haioalkoxy, nitro, amino, mono- or
di-substituted amino, mono- or di-substituted aminoalkyl, carboxyl,
esterified carboxyl, carboxamido, mono- or di-substituted
carboxamido, carbamate, mono- or di-substituted carbamate,
sulfonamide, mono-or di-substituted sulfonamide, alkylsulfonyl,
cyclic alkylsulfonyl, heterocyclic alkylsulfonyl, arylsulfonyl,
heteroarylsulfonyl, alkylcarbonyl, chyclic alkylcarbonyl,
heterocyclic alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl,
thiocarboxamido, cyano, R.sup.10a-carbocycle,
R.sup.10a-heterocycle, R.sup.10a-aryl and R.sup.10a-heteroaryl,
wherein R.sup.10a is H, halogen, OH, amino, mono- or di-substituted
amino, mono-, di- or tri-haloalkyl, alkoxy, mono-, di- or
tri-haloalkoxy, carboxamide, sulfonamide, carbamate, urea or
cyano;
[0042] R.sup.1 is independently selected from the group consisting
of: carbocycle, heterocycle, aryl, heteroaryl, arylalkyl,
heteroarylalkyl, arylalkenyl, heteroarylalkenyl, arylalkynyl,
heteroarylalkynyl, arylaminocarbonyl, heteroarylaminocarbonyl,
arylcarboxamido, heteroarylcarboxamido, arylureido,
heteroarylureido, aryloxy, heteroaryloxy, arylalkoxy,
heteroarylalkoxy, arylamino and heteroarylamino, wherein said
carbocycle, heterocycle, aryl or heteroaryl is substituted with 0-3
R.sup.1a, wherein R.sup.1a is independently selected from the group
consisting of: halogen, alkyl, alkenyl, alkynyl, alkoxy, cyclic
alkoxy, heterocyclic alkoxy, alkoxyalkyl, cyclic alkoxyalkyl,
heterocyclic alkoxyalkyl, alkylthioalkyl, cyclic alkylthioalkyl,
heterocyclic alkylthioalkyl, hydroxyalkyl, mono-, di- or
tri-haloalkyl mono-, di- or tri-haloalkoxy, nitro, amino, mono- or
di-substituted amino, mono- or di-substituted aminoalkyl,
aminocarbonyl, mono- or di-substituted animocarbonyl, cyclic
aminocarbonyl, aminosulfonyl, mono- or di-substituted
aminosulfonyl, alkylcarbonyl, cyclic alkylcarbonyl, heterocyclic
alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl, formyl,
alkylsulfonyl, cyclic alkylsulfonyl, heterocyclic alkylsulfonyl,
aryisulfonyl, heteroarylsulfonyl, carboxylic acid, esterifed
carboxylic acid, alkylcarbonylamino, cyclic alkylcarbonylamino,
heterocyclic alkylcarbonylamino, arylcarbonylamino,
heteroarylcarbonylamino, cyano, arylalkyl, heteroarylalkyl,
aryloxyalkyl, heteroaryloxyalkyl, arylthioalkyl,
heteroarylthioalkyl, carbamate, mono- or di-substituted carbamate,
R.sup.1b-carbocycle, R.sup.1b-heterocycle, R.sup.1b-aryl and
R.sup.1b-heteroaryl, wherein R.sup.1b is H, halogen, OH, amino,
mono- or di-substituted amino, mono-, di- or tri-haloalkyl, alkoxy,
mono-, di- or tri-haloalkoxy, hydroxyalkyl, alkoxyalkyl,
aminoalkyl, mono- or di-substituted aminoalkyl, carboxamide,
sulfonamide, carbamate, urea or cyano;
[0043] R.sup.2 is independently selected from the group consisting
of: H, amino, mono- or di-substituted amino, OH, carboxyl,
esterified carboxyl, carboxamide, N-monosusbstituted carboxamide,
and N,N-disubstituted carboxamide, cyano, alkyl, alkenyl, alkynyl,
cycloalkyl, cycloalkenyl, alkoxy, thioalkyl, mono-, di- or
tri-haloalkyl, halogen, aryl and heteroaryl;
[0044] optionally R.sup.1 and R.sup.2 can be bonded to each other
to form a spirocycle;
[0045] R.sup.3 and R.sup.4, are independently selected form the
group consisting of: H, amino, OH, alkyl, haloalkyl, dihaloalkyl,
trihaloalkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl,
heteroarylalkyl, alkoxy and thioalkyl;
[0046] optionally R.sup.3 and R.sup.4 can occupy multiple positions
in the cycloalkyl ring;
[0047] optionally R.sup.1 and R.sup.3 can be cyclized to form a
carbocycle or heterocycle having 0-3 R.sup.a substituents, wherein
R.sup.a is selected from the group consisting of halogen, alkyl,
alkoxy, thioalkyl, mono-, di- or tri-haloalkyl, mono-, di- or
tri-haloalkoxy, nitro, amino, carboxyl, esterified carboxyl,
carboxamido, thiocarboxamido, cyano, mono-, di-, or
poly-susbstituted aryl or mono-, di-, or poly-susbstituted
heterocycle optionally, wherein said substituted aryl and
substituted heterocycle are substituted with 0-3 R.sup.b, wherein
R.sup.b is selected from the group consisting of halogen, alkyl,
alkoxy, thioalkyl, mono-, di- or trihaloalkyl, mono-, di- or
trihaloalkoxy, nitro, amino, carboxyl, esterified carboxyl,
carboxamido, thiocarboxamido and cyano;
[0048] optionally R.sup.3 and R.sup.4 can be cyclized to form a
bridged bicyclic system having a methylene group or an ethylene
group or a heteroatom selected form the group consisting of N, O
and S; [0049] optionally R.sup.3 and R.sup.4 can be cyclized to
form a spirocycle;
[0050] R.sup.5 is independently selected from the group consisting
of hydrogen, alkyl and formyl; and when R.sup.5 is alkyl the
nitrogen may optionally be in the N-oxide form;
[0051] R.sup.6 and R.sup.7 are each independently selected from the
group consisting of H; C.sub.1-C.sub.10 alkyl, wherein said
C.sub.1-C.sub.10 alkyl can be optionally interrupted by oxygen (O),
nitrogen (NH), or sulfur (S); carbocycle; heterocycle; alkoxy;
cycloalkoxy; heterocyclcalkoxy; mono-, di- or tri-haloalkyl; mono-,
di- or tri-haloalkoxy; aryloxy; heteroaryloxy; arylalkoxy;
heteroarylalkoxy; aryloxyalkyl; heteroaryloxyalkyl;
arylalkoxyalkyl; heteroarylalkoxyalkyl; aryl; heteroaryl,
arylalkyl; heteroarylalkyl; hydroxyalkyl; alkoxyalkyl;
cycloalkyloxyalkyl; heterocycloalkyloxyalkyl; aminoalkyl; mono- or
di-substituted aminoalkyl; arylaminoalkyl; heteroarylaminoalkyl;
alkylthioalkyl; cycloalkylthioalkyl, heterocycloalkylthioalkyl;
arylthioalkyl; heteroarylthioalkyl; alkylsulfonylalkyl;
cycloalkylsulfonylalkyl; heterocycloalkylsulfonylalkyl;
arylsulfonylalkyl; heteroarylsulfonylalkyl; aminocarbonyl; mono- or
di-substituted aminocarbonyl; aminocarbonylalkyl; mono- or
di-substituted aminocarbonylalkyl; alkylcarbonylalkyl;
cycloalkylcarbonylalkyl; heterocycloalkylcarbonylalkyl;
alkylcarbonylaminoalkyl; cycloalkylcarbonylaminoalkyl;
heterocycloalkylcarbonylaminoalkyl; arylcarbonylaminoalkyl;
heteroarylcarbonylaminoalkyl; arylsulfonylaminoalkyl; and
heteroarylsulfonylaminoalkyl;
[0052] optionally, R.sup.6 and R.sup.7 can be cyclized to form a
carbocycle or heterocycle, or a spirocycle or spiroheterocycle;
[0053] R.sup.8 and R.sup.9 are independently selected from the
group consisting of H, OH, amino, alkyl, arylalkyl,
heteroarylalkyl, aryl, heteroaryl, alkoxy, alkenyl, alkynyl,
alkoxyalkyl, mono- or di-substituted amino, a carbocycle and a
heterocycle;
[0054] optionally R.sup.8 and R.sup.9 can be cyclized to form a 3-7
membered carbocycle or heterocycle; and
r=0-3.
[0055] In a further embodiment, the invention relates to a compound
of the formula II:
##STR00004##
including its enantiomers, diasteromers, enantiomerically enriched
mixtures, racemic mixtures thereof, prodrugs, crystalline forms,
non-crystalline forms, amorphous forms thereof, solvates thereof,
metabolites thereof, and pharmaceutically acceptable salts, wherein
constituent variables are provided hereinabove.
[0056] In some embodiments, X can be selected from aryl, mono or
poly substituted aryl, heterocycle, heteroaryl, mono or poly
substituted heteroaryl, carbocycle, mono or poly substituted
carbocycle, and (CR.sup.8R.sup.9).sub.n wherein n=0-5 (e.g., n is
0, 1, 2, 3, 4, or 5).
[0057] In some embodiments, X is a bond, heterocycle, mono or poly
substituted heterocycle heteroaryl, mono or poly substituted
heteroaryl, or (CR.sup.8R.sup.9).sub.n wherein n=0-3.
[0058] In some embodiments, X is a heterocycle, mono or poly
substituted heterocycle. heteroaryl, or mono or poly substituted
heteroaryl.
[0059] In some embodiments, X is (CR.sup.8R.sup.9).sub.n wherein
n=0-3.
[0060] In some embodiments, X is CH.sub.2.
[0061] In some embodiments, Y is a bond or -alkyl-O-alkyl-.
[0062] In some embodiments, --X--Y-- is
--(CR.sup.8R.sup.9).sub.n--HNH--CO--, -alkyl-O-alkyl-, heterocycle,
or heteroaryl.
[0063] In some embodiments, --X--Y-- is --CH.sub.2--NH--CO--,
--CH.sub.2--O--CH.sub.2--, azetidine, pyrrolidine, piperidine,
imidazole, or 4,5-dihydroisoxazole.
[0064] In some embodiments,--X--Y-- is --CH.sub.2--NH--CO--.
[0065] In some embodiments, Z is aryl or heteroaryl, each
substituted with 0-3 R.sup.10 substituents.
[0066] In some embodiments, Z is 6-memebered aryl or 6-membered
heteroaryl, each substituted with 0-3 R.sup.10 substituents.
[0067] In some embodiments, Z is phenyl, pyridyl or pyrimidinyl,
each substituted with 0-3 R.sup.10 substituents.
[0068] In some embodiments, Z is phenyl, pyridyl or pyrimidinyl,
each substituted with at least one mono-, di- or tri-haloalkyl.
[0069] In some embodiments, Z is:
##STR00005##
[0070] In some embodiments, Z is:
##STR00006##
[0071] In some embodiments, the carbocycle substituent of R.sup.1
is intended to include, for example, cycloalkyl of 3-10 carbon
atoms, and bicyclic and multicyclic bridged systems such as
norbornanyl, adamantyl and bicyclo[2.2.2]octyl. The carbocycle of
R.sup.1 may also be further substituted with a heterocycle or
heteroaryl ring such as pyridyl, pyrrolidinyl, and all those
defined under X above.
[0072] Specific examples of R.sup.1 substituents include phenyl,
pyridin-2-yl, 4-methylphenyl, 3-methyl-phenyl, 2-methylphenyl,
4-bromophenyl, 3-bromophenyl, 4-chlorophenyl, 3chlorophenyl,
4-trifluoromethylphenyl, 3-trifluoromethylphenyl,
2-trifluoromethylphenyl, 2-methoxyphenyl, 3-pyridyl, 4-pyridyl,
2-methoxy-5-pyridyl, 2-ethoxy-5-pyridyl, 3,4-methylenedioxyphenyl,
4-fluorophenyl, 3-trifluoromethyl-1H-pyrazol-1-yl, 3-fluorophenyl,
4-methoxyphenyl, 3-methoxyphenyl, pyridin-4-yl, pyridin-3-yl,
5-methylpyridin-2-yl, 6-methylpyridin-2-yl, quinolin-4-yl,
3-methyl-1H-pyrazol-1-yl, 3,5-dimethyl-1H-pyrazol-1-yl,
4-trifluoromethylphenyl, 3-trifluoromethylphenyl,
3,4-methylene-dioxyphenyl, 4-cyanophenyl, cyanophenyl,
4-(methylaminocarbonyl)phenyl, 1-oxidopyridin-4-yl, pyridin-2-yl,
pyridin-3-yl, pyridin-4-yl, 4-methylpyridin-2-yl,
5-methyl-pyridin-2-yl, 6-methylpyridin-2-yl, 6-methoxypyridin-2-yl,
6-methoxypyridin-3-yl, 6-methylpyridin-3-yl, 6-ethylpyridin-3-yl,
6-isopropylpyridin-3-yl, 6-cyclopropylpyridin-3-yl,
1-oxidopyridin-3-yl, 1-oxidopyridin-2-yl, 3-cyanophenyl,
3-(methylaminocarbonyl)-phenyl, 4-(morpholin-4-ylcarbonyl)-phenyl,
5-(morpholin-4-ylcarbonyl)pyridin-2-yl,
6-(morpholin-4-ylcarbonyl)pyridin-3-yl,
4-(4-methylpiperazin-1-yl-carbonyl)phenyl,
6-(azetin-1-yl)pyridin-3-yl, 5-cyanopyridin-2-yl,
6-cyanopyridin-3-yl, 5-(methoxy-methyl)pyridin-2-yl,
5-(1-hydroxy-1-methylethyl)pyridin-2-yl, 5-dimethylaminomethyl,
4-ethylaminocarbonylphenyl, 4-isopropylaminocarbonylphenyl,
4-tert-butylamino-carbonylphenyl, 4-dimethylaminocarbonyl-phenyl,
4-(azetidin-1-yl)carbonylphenyl, 4-(pyrrolidin-1-yl)carbonylphenyl,
4-(morpholin-4-yl)carbonylphenyl,
4-(dimethyl-aminocarbonyl)-2-methylphenyl,
2-methyl-4-(methylamino-carbonyl)phenyl,
3-methyl-4-(methylaminocarbonyl)phenyl,
4-(dimethylaminocarbonyl)-3-methylphenyl,
3-methyl-4-(pyrrolidin-1-ylcarbonyl)phenyl,
4-(dimethylaminocarbonyl)-3-fluorophenyl,
4-[(2,2,2-trifluoroethyl)aminocarbonyl]phenyl,
3-fluoro-4-methylaminocarbonyl-phenyl,
4-ethyl-aminocarbonyl-3-fluorophenyl, 3-methylaminocarbonylphenyl,
3-dimethyl-aminocarbonylphenyl,
5-dimethylaminocarbonyl-2-methoxyphenyl,
2-methoxy-5-methyl-aminocarbonylphenyl,
3-(methylaminocarbonylamino)phenyl,
6-(morpholin-4-yl)-pyridin-3-yl, 6-dimethylaminopyridin-3-yl,
6-isopropylaminopyrid-3-yl, 6-(pyrrolidin-1-yl)pyridin-3-yl,
6-cyclopropylaminopyridin-3-yl, 6-ethoxypyridin-3-yl,
6-(2-fluoroethoxy)pyridin-3-yl, 6-(2,2-difluoroethoxy)pyridin-3-yl,
6-(2,2,2-trifluoroethoxy)-pyridin-3-yl, 4-iodophenyl,
5-(pyrrolidin-1-ylcarbonyl)-2-pyridyl,
5-(morpholin-4-yl-carbonyl)-2-pyridyl,
5-dimethylaminocarbonyl-2-pyridyl,
4-methylaminocarbonyl-aminophenyl,
6-(1-hydroxy-1-methylethyl)pyridin-3-yl,
4-(1-hydroxy-1-methylethyl)-phenyl, 4-(methoxymethyl)phenyl,
3-fluoro-4-(methoxymethyl)phenyl, 4-(dimethyl-amino)phenyl,
4-(dimethylamino)-3-fluorophenyl, 1H-indazol-5-yl,
1-methyl-1H-indazol-5-yl, 2-methyl-1H-indazol-5-yl,
1,3-thiazol-2-yl, 5-ethyl-1,3-thiazol-2-yl,
5-(methyl-aminocarbonyl)-1,3-thiazol-2-yl, 1,3-thiazole-5-yl,
2-(methoxycarbonylamino)-1,3-thiazol-5-yl,
2-isopropyl-1,3-thiazol-5-yl, 5-(pyridin-3-yl)-1,3-thiazol-2-yl,
5-(morpholin-4-ylcarbonyl)-1,3-thiazol-2-yl,
5-aminocarbonyl-1,3-thiazol-2-yl,
5-dimethylaminocarbonyl-1,3-thiazol-2-yl,
5-(pyrrolidin-1-ylcarbonyl)-1,3-thiazol-2-yl,
5-allyl-1,3-thiazol-2-yl, 5-propyl-1,3-thiazol-2-yl,
5-ethylaminocarbonyl-1,3-thiazol-2-yl, 5-phenyl-1,3-thiazol-2-yl,
5-methyl-1,3-thiazol-2-yl, 5-hydroxymethyl-1,3-thiazol-2-yl,
5-(1-hydroxy-1-methylethyl)-1,3-thiazol-2-yl,
5-methoxy-methyl-1,3-thiazol-2-yl, 5-(2-pyridyl)-1,3-thiazol-2-yl,
2-(pyrrolidin-1-yl)-1,3-thiazol-4-yl,
2-(morpholin-4-yl)-1,3-thiazol-4-yl, 2-methyl-1,3-thiazol-5-yl,
2-(1-hydroxy-1methylethyl)-1,3-thiazol-5-yl,
2-(pyrrolidin-1-yl)-1,3-thiazol-5-yl, 2-ethoxy-1,3-thiazol-5-yl,
2-ethyl-1,3-thiazol-5-yl,
2-(pyrrolidin-1-ylmethyl)-1,3-thiazol-5-yl,
2-(morpholin-4-yl)-1,3-thiazol-5-yl,
2-methoxy-methyl-1,3-thiazol-5-yl, 2-isobutyl-1,3-thiazol-5-yl,
2ethylaminocarbonyl-1,3-thiazol-5-yl,
2-(pyrrolidin-1-ylcarbonyl)-1,3-thiazol-5-yl,
2-(morpholin-4ylcarbonyl-1,3-thiazol-5-yl,
2-(3-pyridyl)-1,3-thiazol-5-yl, 2-(2-pyridyl)-1,3-thiazol-5-yl,
4-methyl-1,3-thiazol-2-yl, 1,3-benzo-thiazol-2-yl, pyrimidin-5-yl,
pyrimidin-2-yl, pyridazin-4-yl, pyridazin-3-yl, pyrazin-2-yl, 2-yl,
2-methoxypyrimidin-5-yl, 2-ethoxypyrimidin-5-yl,
2-(2-fluoroethoxy)pyrimidin-5-yl, 2-methylpyrimidin-5-yl,
2-ethylpyrimidin-5-yl, 2-isopropylpyrimidin-5-yl,
2-cyclopropylpyrimidin-5-yl, pyrimidin-4-yl,
4-(pyrimidin-5-yl)phenyl, 4-(1,3-oxazol-2-yl)phenyl,
4-(1H-imidazol-1-yl)phenyl, 4-(morpholin-4-yl)phenyl,
5-(pyrazin-2-yl)pyridin-2-yl, 4-(1-methyl-1H-imidazol-5-yl)phenyl,
4-(4,6-dimethylpyrimidin-5-yl)phenyl, 6-bromopyridin-3-yl,
5-bromopyridin-2-yl, 4'-(methylsulfonyl)biphenyl-4-yl,
3'-(methylsulfonyl)biphenyl-4-yl,
3'-(methoxy-carbonyl)-biphenyl-4-yl,
4-(2,3-dihydro-1,4-benzodioxin-6-yl)phenyl,
4'-(dimethyl-amino)-biphenyl-4-yl, 4-(pyridin-3-yl)phenyl,
4-(1H-pyrazol-4-yl)phenyl, 4-(3,3'-bipyridin-6-yl,
4-(3,4'-bipyridin-6-yl, 5-(3-acetylphenyl)pyridin-2-yl,
5-[3-(dimethyl-amino)phenyl]pyridin-2-yl,
5-[3-(trifluoromethyl)phenyl]pyridin-2-yl,
5-[4-(methyl-sulfonyl)phenyl]pyridin-2-yl,
5-(4-methoxy-phenyl)pyridin-2-yl,
5-(3-methoxy-phenyl)-pyridin-2-yl,
5-[3-(aminocarbonyl)-phenyl]pyridin-2-yl,
5-(4-fluoro-phenyl)pyridin-2-yl,
5-(3,4-difluorophenyl)pyridin-2-yl,
5-(3,5-dimethylisoxazol-4-yl)pyridin-2-yl,
5-(1-methyl-1H-pyrazol-4-yl)pyridin-2-yl,
5-(1H-pyrazol-4-yl)pyridin-2-yl, 5-(1-benzofuran-2-yl)pyridin-2-yl,
5-(1,3-benzodioxol-5-yl)pyridin-2-yl,
5-(2-formyl-phenyl)pyridin-2-yl, 4-(2'-formylbiphenyl-4-yl,
5-(1,3-oxazol-2-yl)pyridin-2-yl, 6-(1,3-oxazol-2-yl)pyridin-3-yl,
4-(1,3-thizol-2-yl)phenyl, 5-(1,3-thiazol-2-yl)pyridin-2-yl,
6-(1,3-thiazol-2-yl)pyridin-3-yl,
6-(1H-imidazol-1-yl)pyridin-3-yl],
5-(1H-imidazol-1-yl)pyridin-2-yl, 6-phenylpyridin-3-yl,
5-(pyrimidin-5-yl)pyridin-2-yl, 5-(pyrimidin-2-yl)pyridin-2-yl,
5-(3-aminocarbonylphenyl)pyridin-2-yl,
4-(1-methyl-1H-imidazol-4-yl)phenyl, 4-(1H-imidazol-4-yl)phenyl],
5-[2-(hydroxymethyl)phenyl]pyridin-2-yl,
2'-(hydroxymethyl)biphenyl-4-yl,
5-{2-[(dimethylamino)methyl]phenyl}pyridin-2-yl,
2'-[(dimethylamino)methyl]biphenyl-4-yl,
5-fluoromethylpyrazin-2-yl, 5-difluoro-methyl-pyrazin-2-yl,
5-methylpyrazin-2-yl, 2-methyl-pyrimidin-5-yl,
2-fluoromethyl-pyrimidin-5-yl, 2-difluoromethylpyrimidin-5-yl,
2-trifluoro-methylpyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl,
isothiazol-5-yl, 3-methylisothiazol-5-yl,
3-fluoromethyl-isothiazol-5-yl, 4-(dimethylamino-carbonyl)phenyl,
4-(methylaminocarbonyl)-phenyl, 4-(morpholin-4-ylcarbonyl)phenyl,
4-(piperidin-1-ylcarbonyl)phenyl,
3-fluoro-4-(pyrrolidin-1-ylcarbonyl)phenyl,
5-(pyrrolidin-1-yl-carbonyl)pyridin-2-yl,
5-dimethyl-aminocarbonyl)pyridin-2-yl,
5-(morpholin-4-yl-carbonyl)-pyridin-2-yl, quinolin-4-yl,
6-methoxypyridin-3-yl, 6-(morpholin-4-yl)pyridin-3-yl,
4-(dimethyl-aminomethyl)phenyl,
5-(dimethylaminomethyl)pyridin-2-yl,
5-(dimethyl-aminocarbonyl)-pyridin-2-yl,
4-[hydroxyl-(pyridin-3-yl)methyl]phenyl,
6-[(hydroxy-(pyridin-3-yl)methyl]pyridin-3-yl,
6-(dimethyl-aminocarbonyl)pyridin-3-yl,
4-(4-hydroxypiperidin-1-ylcarbonyl)phenyl,
4-(4-methoxy-piperidin-1-ylcarbonyl)phenyl,
5-(4-methoxypiperidin-1-ylcarbonyl)-pyridin-2-yl,
6-(4-methoxy-piperidin-1-ylcarbonyl)pyridin-3-yl, phenoxy,
benzyloxy, 2-thienyl, 5-(methoxy-methyl)-1,3-thiazol-2-yl,
5-(morpholin-4-ylcarbonyl)-1,3-thiazol-2-yl,
2-isopropyl-1,3-thiazol-5-yl, 2-(methoxymethyl)-1,3-thiazol-5-yl,
5-(methoxymethyl)-1,3-thiazol-2-yl, 4-(pyrimidin-2-yl)phenyl,
4-(pyrimidin-4-yl)phenyl, and 5-(methoxymethyl)pyridin-2-yl.
[0073] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted with 0-3 R.sup.1a.
[0074] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted, with 0-3 R.sup.1a.
[0075] In some embodiments, R.sup.1 is phenyl, pyridyl,
pyrimidinyl, pyridazinyl, or thiazolyl, each substituted with 0-2
R.sup.1a.
[0076] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted with 0-3 R.sup.1a alkyl, alkoxy, alkoxyalkyl,
hydroxyalkyl, mono- or di-substituted aminoalkyl, aminocarbonyl,
mono- or di-substituted aminocarbonyl, cyclic aminocarbonyl,
alkylcarbonyl, formyl, carboxylic acid, carbamate, mono- or
di-substituted carbamate, R.sup.1b-aryl or R.sup.1b-heteroaryl.
[0077] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted with 0-1 R.sup.1baryl or R.sup.1b-heteroaryl.
[0078] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted with phenyl, pyridyl, pyrimidinyl, oxazolyl, thiazolyl,
or imidazolyl.
[0079] In some embodiments, R.sup.1 is heteroaryl substituted with
phenyl, pyridyl, pyrimidinyl, oxazolyl, thiazolyl, or
imidazolyl.
[0080] In some embodiments, the R.sup.2 group can be selected fro
H, amino, mono- or di-substituted amino, OH, carboxyl, esterified
carboxyl, carboxamide, N(C.sub.1-C.sub.5)-monosusbstituted
carboxamide, and N(C.sub.1-C.sub.5),
N(C.sub.1-C.sub.5)-disubstituted carboxamide, cyano,
(C.sub.1-C.sub.8)alkyl, (C.sub.2-C.sub.8)-alkenyl,
(C.sub.2-C.sub.8)alkynyl, (C.sub.5-C.sub.7)-cycloalkyl,
(C.sub.5-C.sub.7)-cycloalkenyl, alkoxy, alkoxyalkyl, thioalkyl,
mono-, di- or trihaloalkyl, halogen, aryl or heteroaryl.
[0081] In some embodiments, R.sup.2 is H or OH.
[0082] In some embodiments, R.sup.2 is OH.
[0083] In some embodiments, R.sup.1 is aryl or heteroaryl, each
substituted with 0-1 R.sup.1b-aryl or R.sup.1b-heteroaryl; and
R.sup.2 is OH.
[0084] In some embodiments, the R.sup.3 and R.sup.4 group
susbtituents can be independently selected form the group
consisting of: H, amino, OH, (C.sub.1-C.sub.8)alkyl,
halo(C.sub.1-C.sub.5)alkyl, dihalo(C.sub.1-C.sub.5)alkyl,
trihalo(C.sub.1-C.sub.5)alkyl, (C.sub.2-C.sub.8)alkenyl,
(C.sub.2-C.sub.8)alkynyl, aryl, heteroaryl, arylalkyl,
heteroarylalkyl, (C.sub.1-C.sub.5)alkoxy and
thio(C.sub.1-C.sub.5)alkyl.
[0085] In some embodiments, R.sup.3 and R.sup.4 are both H.
[0086] In some embodiments, the R.sup.5 substituent can be
independently selected from hydrogen, (C.sub.1-C.sub.8)alkyl,
formyl; and when R.sup.5 is alkyl, the nitrogen may optionally be
in the N-oxide form.
[0087] In some embodiments, R.sup.5 is H.
[0088] In some embodiments, the R.sup.6 and R.sup.7 substituents
are each independently selected from the group consisting of H,
C.sub.1-C.sub.10 alkyl, optionally C.sub.1-C.sub.10 alkyl can be
interrupted by oxygen, nitrogen or sulfur, carbocycle, heterocycle,
alkoxy, mono-, di- or tri-haloalkyl, mono-, di- or tri-haloalkoxy,
cycloalkoxy, heterocycloalkoxy, aryloxy, heteroaryloxy, arylalkoxy,
heteroarylalkoxy, aryloxyalkyl, heteroaryloxyalkyl, arylalkoxyalkyl
or heteroarylalkoxyalkyl; aryl, heteroaryl, arylalkyl,
heteroarylalkyl, hydroxyalkyl, alkoxyalkyl, cycloalkyloxyalkyl,
heterocycloalkyloxyalkyl, aminoalkyl, mono- or di-substituted
aminoalkyl, arylaminoalkyl, heteroarylaminoalkyl, alkylthioalkyl,
cycloalkylthioalkyl, heterocycloalkylthioalkyl, arylthioalkyl,
heteroarylthioalkyl, alkylsulfonylalkyl, cycloalkylsulfonylalkyl,
heterocycloalkylsulfonylalkyl, arylsulfonylalkyl,
heteroarylsulfonylalkyl, aminocarbonyl, mono- or di-substituted
aminocarbonyl, aminocarbonylalkyl, mono- or di-substituted
aminocarbonylalkyl, alkylcarbonylalkyl, cycloalkylcarbonylalkyl,
heterocycloalk6ylcarbonylalkyl, alkylcarbonylaminoalkyl,
arylcarbonylaminoalkyl, heteroarylcarbonylaminoalkyl,
arylsulfonylaminoalkyl, and heteroarylsulfonylaminoalkyl. Specific
examples of R.sup.6 and R.sup.7 substituents are the same as those
defined for R.sup.1 above.
[0089] In some embodiments, R.sup.6 and R.sup.7 are independently
selected from H, C.sub.1-C.sub.10 alkyl, hydroxyalkyl, and
alkoxyalkyl.
[0090] In some embodiments, one of R.sup.6 and R.sup.7 is H and the
other is H, C.sub.1-C.sub.10 alkyl, hydroxyalkyl, or
alkoxyalkyl.
[0091] In some embodiments, R.sup.6 and R.sup.7 are both H.
[0092] In some embodiments, the R.sup.8 and R.sup.9 substituents
are independently selected from the group consisting of H, OH,
amino, (C.sub.1-C.sub.8)-alkyl, arylalkyl, heteroarylalkyl, aryl,
heteroaryl, (C.sub.1-C.sub.8)-alkoxy, (C.sub.2-C.sub.8)-alkenyl,
(C.sub.2-C.sub.8)-alkynyl, (C.sub.1-C.sub.8)alkoxyalkyl,
mono(C.sub.1-C.sub.8)- or di(C.sub.1-C.sub.8)-substituted amino, a
carbocycle, and a heterocycle. When R.sup.8 and R.sup.9 are
cyclized to form a 3-7 membered carbocycle or heterocycle, such
groups can be, for example, cyclopropyl. cyclobutyl, cyclopentyl,
cyclohexyl, cyclopentyl, isoxazolyl thiazolyl, dihydrooxazolyl,
pyridyl, pyrimidyl, or imidazolyl.
[0093] In some embodiments, R.sup.8 and R.sup.9 are both H.
[0094] In some embodiments, r is 0, 1, 2, or 3. In further
embodiments, r is 1.
[0095] In some embodiments:
[0096] X is a bond, heterocycle, mono or poly substituted
heterocycle, heteroaryl, mono or poly substituted heteroaryl, or
(CR.sup.8R.sup.9).sub.n, wherein n=0-3;
[0097] Y is a bond or -alkyl-O-alkyl-;
[0098] Z is aryl or heteroaryl, each substituted with 0-3 R.sup.10
substituents;
[0099] R.sup.1 is aryl or heteroaryl, each substituted with 0-3
R.sup.18;
[0100] R.sup.2 is H or OH;
[0101] R.sup.3 and R.sup.4 are both H;
[0102] R.sup.5 is hydrogen, alkyl, or formyl;
[0103] R.sup.6 and R.sup.7 is H, C.sub.1-C.sub.10 alkyl,
hydroxyalkyl, or alkoxyalkyl;
[0104] R.sup.8 and R.sup.9 both H; and
[0105] r is 1.
[0106] In some embodiments:
[0107] --X--Y-- is --(CR.sup.8R.sup.9).sub.n--NH--CO--,
-alkyl-O-alkyl-, heterocycle, or heteroaryl;
[0108] Z is aryl or heteroaryl, each substituted with 0-3 R.sup.10
substituents;
[0109] R.sup.1 is aryl or heteroaryl, each substituted with 0-3
R.sup.18;
[0110] R.sup.2 is H or OH;
[0111] R.sup.3 and R.sup.4 are both H;
[0112] R.sup.5 is hydrogen;
[0113] R.sup.6 and R.sup.7 are both H;
[0114] R.sup.8 and R.sup.9 both H; and
[0115] r is 1.
[0116] In some embodiments:
[0117] --X--Y-- is --CH.sub.2--NH--CO--;
[0118] Z is phenyl, pyridyl or pyrimidinyl, each substituted with
at least one mono-, di- or tri-haloalkyl;
[0119] R.sup.1 is aryl or heteroaryl, each substituted with phenyl,
pyridyl, pyrimidinyl, oxazolyl, thiazolyl, or imidazolyl;
[0120] R.sup.2 is OH;
[0121] R.sup.3 and R.sup.4 are both H;
[0122] R.sup.5 is hydrogen;
[0123] R.sup.6 and R.sup.7 are both H
[0124] R.sup.8 and R.sup.9 both H; and
[0125] r is 1.
[0126] In some embodiments;
[0127] --X--Y-- is --CH.sub.2--NH--CO--;
[0128] Z is phenyl substituted with at least one mono-, di- or
tri-haloalkyl;
[0129] R.sup.1 is heteroaryl substituted with pyridyl, pyrimidinyl,
oxazolyl, thiazolyl, or imidazolyl;
[0130] R.sup.2 is OH;
[0131] R.sup.3 and R.sup.4 are both H;
[0132] R.sup.5 is hydrogen;
[0133] R.sup.6 and R.sup.7 are both H; R.sup.8 and R.sup.9 both H;
and
[0134] r is 1.
[0135] At various places is the present specification, substituents
of compounds of the invention are disclosed in groups or in ranges.
It is specifically intended that the invention include each and
every individual subcombination of the members of such groups and
ranges. For example, the term "C.sub.1-6 alkyl" is specifically
intended to individually disclose methyl, ethyl, C.sub.3 alkyl,
C.sub.4 alkyl, C.sub.5 alkyl, and C.sub.6 alkyl.
[0136] For compounds of the invention in which a variable appears
more than once, each variable can be a different moiety selected
from the Markush group defining the variable. For example, where a
structure is described having two R groups that are simultaneously
present on the same compound; the two R groups can represent
different moieties selected from the Markush group defined for
R.
[0137] It is further appreciated that certain features of the
invention, which are, for clarity, described in the context of
separate embodiments, can also be provided in combination in a
single embodiment. Conversely, various features of the invention
which are, for brevity, described in the context of a single
embodiment, can also be provided separately or in any suitable
subcombination.
[0138] The term aryl groups is intended to include aromatic
carbocylic groups such as phenyl, biphenylyl, indenyl, naphthyl as
well as aromatic carbocycles fued to a heterocycle such as
benzothienyl, benzofuranyl, indolyl, quinolinyl, benzothiazole,
benzooxazole, benzimidazole, isoquinolinyl, isoindolyl,
benzotriazole, indazole, and acridinyl.
[0139] The term heteroaryl is intended to include mono- and
poly-cyclic aromatic rings containing from 3 to 20, preferably from
4 to 10 ring atoms, at least one of which is a heteroatom such as
oxygen, sulphur, phosphorus or nitrogen. Examples of such groups
include furyl, thienyl, pyrrolyl, imidazolyl, triazolyl, thiazolyl,
tetrazolyl, oxazolyl, isoxazolyl, pyrazolyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridazinyl, triazinyl, quinolinyl, iosquinolinyl,
quinoxalinyl, benzthiazolyl, benzoxazolyl, benzothienyl or
benzofuryl.
[0140] The terms "cyclic alkyl," "cycloalkyl," and "carbocycle" are
used interchangably herein to refer to non-aromatic, cyclized
hydrocarbons (mono and polycyclic) such as cyclized alkyl, alkenyl,
or alkynyl groups. In some embodiments, the cycloalkyl group is
C.sub.3-14, C.sub.3-10, C.sub.3-8, C.sub.3-7, C.sub.3-6, or
C.sub.3-5. In some embodiments, cycloalkyl moieties each have from
3 to 14, from 3 to 10, or from 3 to 7 ring-forming carbon atoms. In
some embodiments, the cycloalkyl group has 0, 1, or 2 double or
triple bonds. Examples of cycloalkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl,
etc. In the present application, cycloalkyl is also intended to
include bridged cyclic hydrocarbons such as adamantyl groups and
the like.
[0141] Heterocycles are non-aromatic carbocyclic rings (mono or
polycyclic) which include one or more heteroatoms such as nitrogen,
oxygen or sulfur in the ring. In some embodiments, the ring can be
three, four, five, six, seven or eight-membered. In some
embodiments, the heterocycle contains 1, 2, or 3 heteroatoms.
Heterocycles can be saturated or unsaturated. In some embodiments,
heterocycles contain 0, 1 or 2 double bonds or triple bonds.
Ring-forming carbon atoms and heteroatoms can also bear oxo or
sulfide substituents (e.g., CO, CS, SO, SO.sub.2, NO, etc. ).
Examples of heterocycles include tetrahydrofuranyl,
tetrahydrothiophenyl, morpholino, thiomorpholino, azetidiny,
pyrrolidinyl, piperazinyl, piperidinyl, pyrane, dioxane, and
thiazolidinyl.
[0142] Additionally, when the heteroaryl or heterocyclic groups are
nitrogen containing heterocycles, the nitrogen may be modified to
exist in the form of the N.fwdarw.O (N oxides) and such oxides are
intended to be included within the scope of the instant invention.
In the cases of sulfur containing heterocycles, the sulfur oxides
are also intended to be included within the scope of the present
invention.
[0143] Monosubstituted aryl refers to an aryl group having one
substituent. Poly substituted aryl refers to aryl having 2 or more
substitutents (such as 2-4 substituents). Monosubstituted
heteroaryl refers to a heteroaryl group having one substituent.
Polysubstituted heteroaryl refers to heteroaryl having 2 or more
substitutents (such as 2-4 substituents). Monosubstituted
cycloalkyl (or carbocycle) refers to a cycloalkyl group having one
substituent. Polysubstituted cycloalkyl (or carbocycle) refers to
cycloalkyl having 2 or more substitutents (such as 2-4
substituents). Monosubstituted heterocycle refers to a heterocycle
having one substituent. Polysubstituted heterocycle refers to
heterocycle having 2 or more substitutents (such as 2-4
substituents).
[0144] The substituents on the aryl groups, arylalkyl groups,
heteroaryl groups, heteroarylalkyl groups, carbocycle (cycloalkyl)
groups and heterocyclic groups of the invention can be selected
from the group consisting of halogen, alkyl alkoxy, monohaloalkoxy,
dihaloalkoxy, trihaloalkoxy, thioalkyl and monohaloalkyl,
dihaloalkyl, trihaloalkyl, nitro, amino, carboxyl, esterified
carboxyl, carboxamide, thiocarboxamido and cyano. More in
particular, the substituents can also be selected from the group
consisting of trifluoromethyl, C.sub.1-4 alkyl, halo,
trifluoromethoxy, fluoromethoxy, difluoromethoxy, C.sub.1-5 alkoxy,
C.sub.1-5 alkanoyl, C.sub.1-5 alkanoyloxy, C.sub.1-5 alkylamino,
di(C.sub.1-5 alkyl)-amino, C.sub.1-5 alkanoylamino, nitro, carboxy,
carbamoyl, C.sub.1-5 alkoxycarbonyl, thiol, C.sub.1-5,
sulphon-amido, carbamoyl C.sub.1-5 alkyl, N--(C.sub.1-5
alkyl)carbamoyl C.sub.1-5 alkyl, N--(C.sub.1-5 alkyl).sub.2
carbamoyl- C.sub.1-5 alkyl, hydroxy C.sub.1-5 alkyl, and C.sub.1-5
alkoxy C.sub.1-4 alkyl.
[0145] The terms halo or halogen, by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine. Similarly, terms such as haloalkyl,
are meant to include monohaloalkyl and polyhaloalkyl. For example,
the term haloalkyl, such as halo(C.sub.1-C.sub.4)alkyl, is meant to
include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,
3-bromopropyl, and the like.
[0146] The term alkyl when used either alone or as a suffix
includes straight chain and branched structures such as primary
alkyl groups, secondary alkyl groups and tertiary alkyl groups.
These groups may contain up to 15, preferably up to 8 and more
preferably up to 4 carbon atoms. In some embodiments, the alkyl
group is C.sub.1-10, C.sub.1-8, C.sub.1-6, C.sub.1-5, C.sub.1-4, or
C.sub.1-3. Examples of alkyl radicals include groups such as
methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, and
sec-butyl. Similarly the terms alkenyl and alkynyl refer to
unsaturated straight or branched structures containing for example
from 2 to 12, preferably from 2 to 6 carbon atoms. In some
embodiments, the alkenyl or alkynyl group is C.sub.2-10, C.sub.2-8,
C.sub.2-6, C.sub.2-5, C.sub.2-4, or C.sub.2-3. Examples of alkenyl
and alkynyl groups include vinyl, 2-propenyl, crotyl,
2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,
3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the
higher homologs and isomers.
[0147] Aralkyl or arylalkyl is meant to refer to an alkyl group
substituted by an aryl group. An example arylalkyl group is benzyl.
Arylalkenyl refers to an alkenyl group substituted by aryl.
Arylalkynyl refers to an alkynyl group substituted by an aryl
group. Heteroarylalkyl is meant to refer to an alkyl group
substituted by heteroaryl. Heteroarylalkenyl refers to an akenyl
group substituted by a heteroaryl. Heteroarylalkynyl refers to an
alkynyl group substituted by heteroaryl. Heterocycloalkyl or
heterocyclicalkyl is meant to refer to an alkyl group substituted
by a heterocycle. Cycloalkylalkyl or cyclic alkyl alkyl is meant to
refer to an alkyl group substituted by a cycloalkyl group. Examples
of cycloalkylalkyl groups include (cyclohexyl)methyl,
cyclopropylmethyl, and the like.
[0148] The terms alkoxy, alkylamino and alkylthio (or thioalkoxy)
are used in their conventional sense, and refer to those alkyl
groups attached to the remainder of the molecule via an oxygen
atom, an amino group, or a sulfur atom, respectively. Therefore,
terms such as alkoxy and thioalkyl comprise alkyl moieties as
defined above, attached to the appropriate functionality.
[0149] other suitable substituents which can be used in the many
carbon rings of the present invention such as cycloaliphatic,
aromatic, non-aromatic heterocyclic ring or benzyl group include,
for example, --OH, halogen (--Br, --Cl, --I and --F) -O(aliphatic,
substituted aliphatic, benzyl, substituted benzyl, phenyl,
substituted phenyl, aromatic or substituted aromatic group), --CN,
--NO.sub.2, --COOH, --NH.sub.2, --NH(aliphatic group, substituted
aliphatic, benzyl, substituted benzyl, phenyl, substituted phenyl,
aromatic or substituted aromatic group), --N(aliphatic group,
substituted aliphatic, benzyl, substituted benzyl, phenyl,
substituted phenyl, aromatic or substituted aromatic group).sub.2,
--COO(aliphatic group, substituted aliphatic, benzyl, substituted
benzyl, phenyl, substituted phenyl, aromatic or substituted
aromatic group), --CONH.sub.2, --CONH(aliphatic, substituted
aliphatic group, benxyl, substituted benzyl, phenyl, substituted
phenyl, aromatic or substituted aromatic group)), --SH,
--S(aliphatic, substituted aliphatic, benzyl, substituted benzyl,
phenyl, substituted phenyl, aromatic or substituted aromatic group)
and --NH--C.dbd.NH)--NH.sub.2. A substituted non-aromatic
heterocyclic ring, benzylic group or aromatic group can also have
an aliphatic or substituted aliphatic group as a substituent. A
substituted alkyl or aliphatic group can also have a non-aromatic
heterocyclic ring, benzyl, substituted benzyl, aromatic or
substituted aromatic group as a substituent. A substituted
non-aromatic heterocyclic ring can also have .dbd.O, .dbd.S,
.dbd.NH or .dbd.N(aliphatic, aromatic, substituted non-aromatic
heterocyclic ring or substituted benzyl group can have more than
one substituent.
[0150] For bivalent moieties such as X and Y, the term "amide bond"
refers to --NHCO--; the term "thiamide bond" refers to --NHCS--;
the term "sulfonamide" refers to --NHSO.sub.2--; the term "ketone"
refers to --OC--; the term "oxime" refers to --C(.dbd.N--OH)--; and
the term "urea" refers to --NHCONH--.
[0151] "Cyclic alkoxy" refers to --O-(cycloalkyl). "Heterocyclic
alkoxy" refers to --O-(heterocycle). "Alkoxyalkyl" refers to alkyl
substituted by alkoxy. "Cyclicalkoxyalkyl" refers to alkyl
substituted by --O-(cycloalkyl). "Heterocyclic alkoxy alkyl" refers
to alkyl substituted by --O-(heterocycle. "Alkylthioalkyl" refers
to alkyl substituted by thioalkyl. "Cyclic alkyl thioalkyl" refers
to alkyl substituted by --S-(cycloalkyl). "Heterocyclic alkyl
thioalkyl" refers to alkyl substituted by --S-(heterocycle). "Mono-
or di-substituted amino" refers to --NH.sub.2 wherein either one
(e.g., mono) or both (e.g., di) hydrogens are replaced with a
substituent such as C.sub.1-8 alkyl, OH, CO--(C.sub.1-4 alkyl),
etc. "Mono- or di-substituted aminoalkyl" refers to alkyl
substituted by mono or di-substituted amino. "Esterified carboxyl"
refers to COOH where the hydrogen atom is replaced by a substituent
such as C.sub.1-8 alkyl, carbocycle, heterocycle, aryl or
heteroaryl. "Carboxamido" refers to --CONH.sub.2. "Mono or
di-substituted carboxamide" refers to --CONH.sub.2 werein either
one (e.g., mono) or both (e.g., di) hydrogens are replaced with a
substituent such as C.sub.1-8 alkyl, OH, CO--(C.sub.1-4 alkyl),
etc. "Carbamate" refers to --OCONH.sub.2 and "mono or
di-substituted carbamate " refers to --OCONH.sub.2 where either one
(e.g., mono) or both (e.g., di) hydrogens are replaced with a
substituent such as C.sub.1-8 alkyl, OH, CO--(C.sub.1-4 alkyl),
etc. "Sulfonamide" refers to --SO.sub.2NH.sub.2 and "mono or
di-substituted sulfonamide" refers to --SO.sub.2NH.sub.2 wherein
either one (e.g., mono) or both (e.g., di) hydrogens are replaced
with a substituent such as C.sub.1-8 alkyl, OH, CO--(C.sub.1-4
alkyl), etc. "Alkylsulfonyl" refers to --SO.sub.2-(alkyl). "Cyclic
alkylsulfonyl" refers to --SO.sub.2-(carbocycle). "Hetercyclic
sulfonyl" refers to --SO.sub.2-(heterocycle). "Aryl sulfonyl"
refers to --SO.sub.2-(aryl). "Heteroaryl sulfonyl" refers to
--SO.sub.2-(heteroaryl). "Alkylcarbonyl" refers to --CO-(alkyl).
"Cyclic alkylcarbonyl" refers to --CO-(cycloalkyl). "Heterocyclic
alkylcarbonyl" refers to --CO-(heterocycle). "Arylcarbonyl" refers
to --CO-(aryl). "Heteroarylcarbonyl" refers to --CO-(heteroaryl).
"Thiocarboxamido" refers to --CSNH.sub.2. "Arylaminocarbonyl"
refers to --CO--NH-(aryl). "Heteroarylaminocarbonyl" refers to
--CO--NH-(heteroaryl). "Arylcarboxamido" refers to --CO--NH-(aryl).
"Heteroarylcarboxamido" refers to --CO--NH-(heteroaryl).
"Arylureido" referst to ureido substituted by aryl.
"Heteroarylureido" refers to ureido substituted by heteroaryl.
"Aryloxy" refers to --O-(aryl). "Heteroaryloxy" refers to
--O-(heteroaryl). "Arylalkoxy" refers to alkoxy substituted by
aryl. "Heteroarylalkoxy" refers to alkoxy substituted by
heteroaryl. "Arylamino" refers to --NH-(aryl). "Heteroarylamino"
refers to --NH-(heteroaryl). "Hydroxylalkyl" refers to alkyl
substituted by hydroxyl (OH). "Aminocarbonylalkyl" refers to alkyl
substituted by aminocarbonyl. "Mono- or di-substituted
aminocarbonlyalkyl" refers to alkyl substituted by mono- or
di-substituted aminocarbonyl. "Alkylcarbonlyalkyl" refers to alkyl
substituted by alkylcarbonyl. "Cycloalkylcarbonylalkyl" refers to
alkyl substituted by --CO-(cycloalkyl).
"Heterocycloalkylcarbonylalkyl" refers to alkyl substituted by
--CO-(heterocyle). "Alkylcarbonylaminoalkyl" refers to alkyl
substituted by --NH--CO-(alkyl). "Cycloalkylcarbonylaminoalkyl"
refers to alkyl substituted by --NH--CO-(cycloalkyl).
"Heterocycloalkylcarbonylaminoalkyl" refers to alkyl substituted by
--NH--CO-(heterocycle). "Arylcarbonylaminoalkyl" refers to alkyl
substituted by --NH--CO-(aryl). "Heteroarylcarbonylaminoalkyl"
refers to alkyl substituted by --NH--CO-(heteroaryl).
"Arylsulfonylaminoalkyl" refers to alkyl substituted by
--NH--SO.sub.2-(aryl). "Heteroaylsulfonylaminoalkyl" refers to
alkyl substituted by --NH--SO.sub.2-(heteroaryl).
[0152] "Spirocycle" refers to a cycloalkyl group sharing one of its
ring-forming atoms with another cycloalkyl or heterocyclyl group.
"Spiroheterocycle" refers to a heterocycle group sharing one of its
ring-forming atoms with another cycloalkyl or heterocyclyl
group.
[0153] The phrase "optionally R.sup.3 and R.sup.4 can be cyclized
to form a bridged bicyclic system having a methylene group or an
ethylene group or a heteroatom selected form the group consisting
of N, O and S" refers to when R.sup.3 and R.sup.4, residing on
different atoms, together form a divalent bridging moiety such as,
for example, methylene, ethylene, NH, O, S, methylene-O,
methyiene-S, or methylene-NH.
[0154] Unless otherwise indicated, the compounds provided in the
above formula are meant to include pharmaceutically acceptable
salts, prodrugs thereof, enantiomers, diastereomers, racemic
mixtures thereof, crystalline forms, non-crystalline forms,
amorphous forms thereof and solvates thereof.
[0155] The term "pharmaceutically acceptable salts" is meant to
include salts of the active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituents found on the compounds described herein. When
compounds of the present invention contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired base, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable base addition salts include sodium,
potassium, calcium, ammonium, organic amino, or magnesium salt, or
a similar salt. When compounds of the present invention contain
relatively basic functionalities, acid addition salts can be
obtained by contacting the neutral form of such compounds with a
sufficient amount of the desired acid, either neat or in a suitable
inert solvent. Examples of pharmaceutically acceptable acid
addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, phosphoric, partially
neutralized phosphoric acids, sulfuric, partially neutralized
sulfuric, hydroiodic, or phosphorous acids and the like, as well as
the salts derived from relatively nontoxic organic acids like
acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,
suberic, fumaric, mandelic, phthalic, benzenesulfonic,
p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
Also included are salts of amino acids such as arginate and the
like, and salts of organic acids like glucuronic or galactunoric
acids and the like. Certain specific compounds of the present
invention may contain both basic and acidic functionalities that
allow the compounds to be converted into either base or acid
addition salts. Lists of suitable salts are found in Remington's
Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton,
Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2
(1977), each of which is incorporated herein by reference in its
entirety.
[0156] The neutral forms of the compounds of the present invention
may be regenerated by contacting the salt with a base or acid and
isolating the parent compound in the conventional manner. The
parent form of the compound differs from the various salt forms in
certain physical properties, such as solubility in polar solvents,
but otherwise the salts are equivalent to the parent form of the
compound for the purposes of the present invention.
[0157] As noted above, some of the compounds of the present
invention possess chiral or asymmetric carbon atoms (optical
centers) or double bonds; the racemates, diastereomers, geometric
isomers and individual optical isomers are all intended to be
encompassed within the scope of the present invention.
[0158] Some of the compounds of formula I or II can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are substantially equivalent for the uses
contemplated by the present invention and are intended to be within
the scope of the present invention.
[0159] In addition to salt forms, the present invention provides
compounds that may be in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an ex-vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent. Prodrugs can be prepared by modifying functional
groups present in the compounds in such a way that the
modifications are cleaved, either in routine manipulation or in
vivo, to the parent compounds. Prodrugs include compounds wherein
hydroxyl, amino, sulfhydryl, or carboxyl groups are bonded to any
group that, when administered to a mammalian subject, cleaves to
form a free hydroxyl, amino, sulfhydryl, or carboxyl group
respectively. Examples of prodrugs include, but are not limited to,
acetate, formate and benzoate derivatives of alcohol and amine
functional groups in the compounds of the invention. Preparation
and use of prodrugs is discussed in T. Higuchi and V. Stella,
"Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S.
Symposium Series, and in Bioreversible Carriers in Drug Design, ed.
Edward B. Roche, American Pharmaceutical Association and Pergamon
Press, 1987, both of which are hereby incorporated by reference in
their entirety.
[0160] Compounds of the invention, including salts, hydrates, and
solvates thereof, can be prepared using known organic synthesis
techniques and can be synthesized according to any of numerous
possible synthetic routes.
[0161] The reactions for preparing compounds of the invention can
be carried out in suitable solvents which can be readily selected
by one of skill in the art of organic synthesis. Suitable solvents
can be substantially nonreactive with the starting materials
(reactants), the intermediates, or products at the temperatures at
which the reactions are carried out, e.g., temperatures which can
range from the solvent's freezing temperature to the solvent's
boiling temperature. A given reaction can be carried out in one
solvent or a mixture of more than one solvent. Depending on the
particular reaction step, suitable solvents for a particular
reaction step can be selected.
[0162] Preparation of compounds of the invention can involve the
protection and deprotection of various chemical groups. The need
for protection and deprotection, and the selection of appropriate
protecting groups can be readily determined by one skilled in the
art. The chemistry of protecting groups can be found, for example,
in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic
Synthesis, 3rd. Ed., Wiley & Sons, Inc., New York (1999), which
is incorporated herein by reference in its entirety.
[0163] Reactions can be monitored according to any suitable method
known in the art. For example, product formation can be monitored
by spectroscopic means, such as nuclear magnetic resonance
spectroscopy (e.g., .sup.1H or .sup.13C) infrared spectroscopy,
spectrophotometry (e.g., UV-visible), or mass spectrometry, or by
chromatography such as high performance liquid chromatography
(HPLC) or thin layer chromatography.
[0164] A variety of 4,4-disubstituted cyclohexanone derivatives can
be synthesized using the protocols described in Schemes 1.
Compounds of formula 1-2 can be prepared by addition of arylMgX or
ArX/BuLi to 1,4-cyclohexanedione 1-1. Alternatively, compounds of
formula 1-2 can be prepared by treatment of 1,4-cyclohexanedione
mono-ethylene ketal 1-3 with arylMgX, ArX/BuLi or
heteroarylH/lithium tetramethylpiperidine followed by converting
the ketal in 1-4 to a ketone using an acid such as HCl in aqueous
solution.
##STR00007##
[0165] 4-Arylcyclohexanone derivatives of formula 2-3 can be
synthesized following the procedures shown in Scheme 2. The
intermediate 1-4 is subjected to a treatment with a dehydrating
agent such as thionyl chloride/pyridine followed by reduction of
the resulting olefin by hydrogenation using a catalyst such as
Pd--C or PtO.sub.2. Conversion of the ketal in 2-2 by treatment
with an acid provides the ketones of formula 2-3.
##STR00008##
[0166] Alternatively, compounds of formula 2-3 can be synthesized
according to Scheme 3. Reduction of ketone 1-3 using a reducing
agent such as sodium borhydride produces the alcohol 3-1 which is
converted to a mesylate 3-2 by treating with methanesulfonyl
chloride. Displacement of the mesylate 3-2 with a heterocycle such
as pyrazole, imidazole, triazole or tetrazole provides the
intermediate 2-2 which is converted to compounds of formula 2-3 by
treatment with an acid such as HCl.
##STR00009##
[0167] Ar=substituted pyrazole, imidazole, triazole or
tetrazole
[0168] Introduction of a substituent on the aromatic ring in
ketones of formula 1-2 or 2-3 can be accomplished starting from the
ketal intermediate 1-4 or 2-2 using the methods described in
Schemes 4-8. When the aromatic ring in 1-4 or 2-2 bears a cyano
group, the ketal 4-1 is subjected to a hydrolysis using a base such
as sodium or potassium hydroxide to give the carboxylic acid 4-2.
Coupling of 4-2 with an amine using a coupling agent such as BOP
provides the amide 4-3. Treatment of 4-3 with an acid such as HCl
affords the ketones of formula 4-4.
##STR00010##
[0169] When the aromatic ring in the ketal intermediate 1-4 or 2-2
bears a halide such as bromo or iodo, the halide can be transformed
to a substitutent using the procedures described in Scheme 5.
Treatment of 5-1 with butyl lithium followed by quenching with an
electrophile such as alkyl halide, aldehyde, ketone, chloroformate,
or carbonate provides the R-substituted ketal 5-2. Suzuki coupling
of 5-1 with a boronic acid ArB(OH).sub.2 (Ar=aryl or heteroaryl) or
coupling of 5-1 with ArZnCl which can be generated in situ by
treating ArX (X.dbd.Br, I) with butyl lithium followed by
quenching, with zinc chloride or treating 5-1 with iPrMgCl followed
by coupling with ArX (X--Br, I) in the presence of a catalyst such
as
Ni(CH.sub.3COCH(OH)CH.sub.3).sub.2-1,2bis(diphenylphosphino)ethane
provides the Ar-substituted ketal intermediate 5-4. Treatment of
5-2 and 5-4 with an acid affords their corresponding ketones 5-3
and 5-5.
##STR00011##
[0170] Alternatively, ketones of formula 5-5 can be obtained using
the protocol depicted in Scheme 6. Following conversion of 5-1 to a
boronic acid ester, the resulting boronic acid ester 6-1 is coupled
with ArX (X.dbd.Br, I) using a palladium catalyst such as
Pd(PPh.sub.3).sub.4 to give the Ar-substituted ketal 5-4 from which
ketones of formula 5-5 are obtained by treatment with an acid such
as HCl.
##STR00012##
[0171] When the Ar group in ketones of formula 1-2 or 2-3 is a
2-thiazole residue, introduction of a substituent at the 5-position
on the thiazole 7-1 with butyl lithium followed by quenching with
1,4-cyclohexanedione mono-ethylene ketal 1-3 gives rise to the
teritiary alcohol 7-2. Treatment of 7-2 with butyl lithium followed
by quenching the anion 7-3 with an electrophile such as alkyl
halide, aldehyde, ketone, chloroformate or carbonate produces the
ketal 7-4 with an R substituent at the 5-position on thiazole.
Alternatively, the anion 7-3 can be quenched with zinc chloride and
the resulting intermediate is coupled with ArX (X.dbd.Br, I) using
a palladium catalyst such as PdCl.sub.2(PPh.sub.3).sub.2 to give
the ketal 7-6 with an Ar residue at the 5-position on thiazole.
Ketals 7-4 and 7-6 are then converted to their corresponding
ketones of formula 7-5 and 7-7 by treatment with an acid such as
HCl.
##STR00013##
[0172] When the Ar group in ketones of formula 1-2 or 2-3 is a
5-thiazole residue, introduction of a substituent at the
2-trimethylsilyl protected thiazole 8-1 followed by quenching with
1-3 gives rise to the intermediate 8-2. Following removal of the
trimethylsilyl group using TBAF, lithiation of 8-3 followed by
quenching with an electrophile such as alkylhalide, aldehyde,
ketone, isocyanate, chloroformate or carbonate provides the
5-R-substituted thiazole derivative 8-4. Treatment of 8-4 with an
acid such as HCl affords the ketones of formula 8-5.
##STR00014##
[0173] A variety of 3-aminopyrrolidine intermediates can be
prepared as shown in Schemes 6-17. Coupling of a carboxylic acid of
formula 9-1 with a commercially available pyrrolidine derivative of
formula 9-2 using a coupling agent such as BOP gives rise to the
amide 9-3. Removal of the protecting group P (P=Boc, benzyl or Cbz)
using an acid such as TFA or HCl or by hydrogenation using a
palladium catalyst provides the pyrrolidine intermediates of
formula 9-4.
##STR00015##
[0174] 4-Amino-2-methylpyrrolidine derivatives of formula 10-8 can
be prepared using the sequence described in Scheme 10. Following
Boc protection at the amine and TBS protection at the hydroxyl of
trans-4-hydroxy-L-proline methyl ester 10-1, the ester in 10-2 is
reduced to an alcohol and the resulting alcohol is converted to a
tosylate. Detosylation in 10-3 can be achieved by reduction using
lithium triethylborohydride (LiEt.sub.3BH). The resulting
intermediate 10-4 is subjected to a deprotection using an acid such
as HCL to remove the Boc and the TBS groups. Following coupling of
the resulting amine 10-5 with a carboxylic acid of formula 9-1
using a coupling agent such as EDC, conversion of the hydroxyl to a
mesylate is followed by displacement with sodium axide. The
resulting azido group is then reduced to an amine by hydrogenation
to give the pyrrolidine intermediates of formula 10-8.
##STR00016##
[0175] 4-Aminopyrrolidine derivatives of formula 11-6 can be
prepared according to Scheme 11. Alkylation of the intermediate
10-2 with an alkyl halide (RX) using LHMDS provides the
R-substituted intermediate 11-1. Following reduction of the ester
to an alcohol using diisobutylaluminun hydride (DIBAL), the alcohol
is converted to a tosylate and the resulting tosylate is reduced
using LiIt.sub.3BH to give 11-2 is then converted to compounds of
formula 11-6 in a manner similar to that described in Scheme
10.
##STR00017##
[0176] 4-Aminopyrrolidine derivatives of formula 12-5 can be
synthesized using the method shown in Scheme 12. The intermediate
10-2 is reduced to an alchol using a reducing agent such as DIBAL
and the resulting alcohol is alkylated with an alkyl halide (RX)
using sodium hydride to give intermediate 12-1. Using procedures
similar to those described in Scheme 10, compounds of formula 12-5
are obtained from the intermediate 12-1.
##STR00018##
[0177] 4-Aminopyrrolidine derivatives of formula 13-7 can be
generated according to Scheme 13. The intermediate 10-2 is reduced
to an alcohol using a reducing agent such as DIBAL and the
resulting alcohol is oxidized to an aldehyde using a oxidizing
agent such as Swern oxidation. Addition of a Grignard reagent RMgX
to the aldehyde 13-1 is followed by alkylation of the resulting
alcohol with an alkyl halide (RX) using sodium hydride. After
removal of the Boc and TBS protecting groups in 13-2 or 13-3 using
an acid such as HCl, the resulting amine 13-4 is condensed with a
carboxylic acid of formula 9-1. Mesylation at the 4-hydroxy on the
pyrroldine followed by displacement of the resulting mesylate with
sodium azide and reduction of the azido by hydrogenation provides
compounds of formula 13-7.
##STR00019##
[0178] 4-Aminopyrrolidine derivatives of formula 14-6 can be
synthesized using a protocol depicted in Scheme 14. After double
addition of a Grignard reagent RMgX to the intermediate 10-2, the
resulting tertiary alcohol 14-1 is subjected to an alkylation with
an alkyl halide (R'X) to give 14-2. Intermediates 14-1 and 14-2 are
then converted to compounds of formula 14-6 in a manner similar to
that described in Scheme 13.
##STR00020##
[0179] The synthesis of 4-ammopyrrolidine derivative's of formula
15-5 is given in Scheme 15. After dehydration of the intermediate
14-1 followed by reduction of the olefin by hydrogenatiort, the
resulting intermediate 15-1 is converted to compounds of formula
15-5 in a fashion similar to that described in Scheme 10.
##STR00021##
[0180] Compounds of formula I can be obtained by assembling the
aminopyrrolidine derivatives of formula 16-1 with a ketone of
formula 16-2 by reductive animation using a reducing agent such as
sodium triacetoxyborohydride or through hydrogenation followed by
treating the resulting secondary amine 16-3 via reductive amination
with an aldehyde or by alkylation with an alkyl halide (RX).
##STR00022##
[0181] Alternatively, compounds of formula I can be prepared using
a sequence outlined in Scheme 17. Reductive animation of the
aminopyrrolidine derivatives of formula 17-1 with a ketone of
formula 16-2 gives rise to the secondary amine 17-2. After removal
of the protecting group P (P=Boc, benzyl or Cbz) using an acid or
through hydrogenation using a catalyst such as Pd--C, the resulting
amine 17-3 is condensed with a carhoxylic acid of formula 9-1 to
provide compounds of formula 17-4.
##STR00023##
[0182] Alternatively, compounds of formula I can be prepared using
s sequence outlined In Scheme 18. Reduction of the cyclohexanone
1-2 with a reducing agent such as lithium aluminum hydride produces
the cis diol 18-1. After converting the secondary alcohol to a
mesylate, the resulting mesylate 18-2 is displaced with an
aminopyrrolidine derivative of formula 17-1 to give the tram
4-amino-1-cyclohexanol derivative of formula 18-3. Removal of the
protecting group using an acid or through hydrogenation followed by
coupling of the resulting amine with a carboxyiie acid of formula
9-1 affords compounds of formula 18-5.
##STR00024##
[0183] Alternatively, compounds of formula I can be synthesized
according to Scheme 19. Displacement of the mesylate 18-2 with
sodium azide gives rise to the azido Intermediate 19-1 which is
reduced to an amine by hydrogenation using a catalyst such as
Pd--C. Displacement of the mesylate of formula 19-3 with the
resulting amine 19-2 or reductive animation of 19-2 with a ketone
of formula 19-4 affords compounds of formula 19-5.
##STR00025##
[0184] The compounds of the present invention are MCP-1 receptor
modulators, e.g., antagonists, and are capable of inhibiting the
binding of MCP-1 to its receptor. Surprisingly, the compounds block
T cell migration in vitro, and have dramatic effects on the
recruitment of inflammatory cells in multiple models of
inflammatory diseases. Therefore, the compounds of formula I are
useful as agents for the treatment of inflammatory disease,
especially those associated with lymphocyte and/or monocyte
accumulation, such as arthritis, rheumatoid arthritis, multiple
sclerosis, neuropathic pain, atherosclerosis and transplant
rejection. In addition, these compounds can be used in the
treatment of allergic hypersensitivity disorders such as asthma and
allergic rhinitis characterized by basophil activation and
eosinophil recruitment, as well as for the treatment of restenosis
and chronic or acute immune disorders.
[0185] Modulation of chemokine receptor activity, as used in the
context of the present invention, is intended to encompass
antagonism, agonism, partial antagonism and/or partial agonism of
the activity associated with a particular chemokine receptor,
preferably the CCR2 receptor. The term composition as used herein
is intended to include a product comprising the specified
ingredients in the specified amounts, as well as any product which
results, directly or indirectly, from combination of the specified
ingredients in the specified amounts. By pharmaceutically
acceptable it is meant the carrier, diluent or excipient must be
compatible with the other ingredients of the formulation and not
deleterious to the recipient thereof.
[0186] The compounds of formula I of the present invention, and
compositions thereof are useful in the modulation of chemokine
receptor activity, particularly CCR2. Accordingly, the compounds of
the present invention are those which inhibit at least one function
or characteristic of a mammalian CCR2 protein, for example, a human
CCR2 protein. The ability of a compound to inhibit such a function
can be demonstrated, in a binding assay (e.g., ligand binding or
promoter binding), a signalling assay (e.g., activation of a
mammalian G protein, induction of rapid and transient increase in
the concentration of cytosolic free calcium), and/or cellular
response function (e.g., stimulation of chemotaxis, exocytosis or
inflammatory mediator release by leukocytes).
[0187] The invention is illustrated by the following examples,
which are not intended to be limiting in any way.
EXAMPLES
[0188] Reagents and solvents used below can be obtained from
commercial sources such as Aidrich Chemical Co. (Milwaukee, Wis.,
USA), Mass spectrometry results are reported as the ratio of mass,
over charge, followed by the relative abundance of each ion (in
parentheses), in tables, a single m/e value is reported far the M+H
(or, as noted, M-H) ion containing the most common atomic isotopes.
Isotope patterns correspond to the expected formula in all
cases.
Example 1
Step A
##STR00026##
[0190] (3-Trifluoromethyl-benzoylamino)acetic acid. To a rapid
stirring solution of glycine (15.014 g, 0.20 mol) in MeCN (400 mL)
and 2 M NaOH (250 ml) at 0.degree. C. was slowly added a solution
of 3-(trifluoromethyl)-benzoyl chloride (41.714 g, 0.20 mol) in 75
mL of MeCN over 30 min. The cloudy yellow solution was stirred at
0.degree. C. for 30 min. The reaction mixture was acidified with 3
M HCl to pH=3, followed by removal of MeCN on rotary evaporator.
The resulting mixture was then extracted with EtOAc (400
mL.times.3). The combined organic layers were dried, filtered and
concentrated to give a light yellow solid (48.53 g), which was
triturated with toluene (500 mL). After filtration, the solid
product was washed with cold toluene until the filtrate was
colorless. After dried under high vacuum Over the weekend, a white
powder product 44.60 g (90%) was afforded. MS (M+H.sup.+)=248.1.
.sup.1H NMR (DMSO-d.sub.6) .delta. 12.70 (br s, 1 H), 9.17 (m, 1H),
8.20 (dd, 2H), 7.94 (dd, 1H), 7.78 (m, 1H), 3.97 (d, 2H),
Step B
##STR00027##
[0192] tert-Butyl
[(3S)-1-({[3-(Trifluoromethyl)benzoyl]amino}acetyl)
pyrrolidin-3-yl]carbamate. To a solution of the carbosyiic add (2.7
g, 11 mmol) from step A and tert-butyl
(3S)-pyrrolidin-3-ylcarbamate (2.0 g, 11 mmol) in DMF (30 ml,)
cooled in an ice bath was added BOP (5 g, 11 mmol) followed by
triethylamine (3 mL, 22 mmol). The mixture was allowed to warm to
temperature and stirred overnight. Ethyl acetate (150 mL) was
added. The resulting solution was washed with NaHCO.sub.3 and brine
each three times, dried over MgSO.sub.4 and concentrated.
Chromatography on silica gel eluting with EtOAc provided 4.4 g
(96%) of the desired product. MS (M-Boc+H).sup.+ 316.
Step C
##STR00028##
[0194]
N-{2-[(3S)-3-Aminopyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)
benzamide. The above product (4.2 g) was dissolved in 4 N
HCl/dioxane (30 mL). After being stirred for 1 hour at room
temperature, the solution was concentrated to provide 4.0 g of the
title compound. MS (M+H).sup.+ 316.
Step D
##STR00029##
[0196] 8-Phenyl-1,4-dioxaspiro[4.5]decan-8-ol. To a solution of
1,4-cyclohexanone mono-ethylene ketal (8.1 g, 50 mmol) in THF (20
mL) at 10.degree. C. was added a 1 M solution of phenyl magnesium
bromide in THF (70 mL, 70 mmol). The resulting mixture was stirred
at room temperature for 2 hours before quenching with saturated
NHaCl solution. The solution was extracted with EtOAc 3 times. The
combined organic phase was washed with brine, dried over MgSO.sub.4
and concentrated. Chromatography on silica gel elating with 40%
EtOAc/hexanes provided 9.5 g (81%) of the desired product MS
(M+H).sup.+ 234.
Step E
##STR00030##
[0198] 4-Hydroxy-4-phenylcyclohexanone. The above product was
dissolved in THF (50 ml). To it was added 10% HCl/H.sub.2O (50 mL).
The solution was stirred at room temperature overnight and
extracted with EtOAc three times. The combined extracts were washed
with brine, dried over MgSO.sub.4 and concentrated to give the
title compound as a white solid. MS (M+H).sup.+ 191.
Step F
##STR00031##
[0200]
N-(2-{(3S)-3-[(4-Hydroxy-4-phenylcyclohexyl)amino]pyrrolidin-1-yl}--
2-oxoethyl)-3-(trifluoromethyl)benzamide. To a solution of the
pyrrolidine intermediate from step C (0.3 g, 0.85 mmol) and the
ketone from step E (0.16 g, 0.85 mmol) in THF (5 ml) was added
Na(OAc).sub.3BH (0.35 g, 2.5 mmol) followed by triethylamine (0.2
ml, 1.5 mmol). The reaction was continued at room temperature
overnight and quenched by addition of a saturated NaHCO.sub.3
solution. The resulting solution was extracted with EtOAc arid the
EtOAc layer was dried over MgSO.sub.4 and concentrated. Separation
on silica gel eluting with 10% to 30% MeOH/EtOAc provided the cis
(fast moving spot) and trans (slow moving spot) isomers of the
title compound. MS (M+H).sup.+ 490.0.
Example 2
Step A
##STR00032##
[0202] 8-Pyridin-2-yl-1,4-dioxaspiro[4.5]decan-8-ol. To a solution
of 2-bromopyridine (14 g, 88.6 mmol) in anhydrous ether (300 mL)
cooled at --78.degree. C. was slowly added a solution of 2.5 M
n-butyl lithium (36 mL). After the addition, stirring was continued
at -78.degree. C. for 1 hour. To it was slowly added a solution, of
1,4 -eyciohexanedione mono-ethylene ketal (15 g, 96 mmol) in
anhydrous ether (300 ml). When the addition was complete, the
mixture was allowed to warm to 0.degree. C. and stirring was
continued for I hour. The reaction was quenched by the addition of
an aqueous solution (100 mL) of ammonium chloride (4.5 g). The
organic phase was separated and. the aqueous phase was extracted
with methylene chloride 4 times. The combined organic phases were
dried over MgSO.sub.4 and concentrated. Crystallization from EtOAc
provided 7 gofthe desired product. The mother liquid was purified
on silica gel elating with 10% MeOH/EtOAe to give 3 g of the
desired product. MS (M+H).sup.+ 236.0.
Step B
##STR00033##
[0204] 4-Hydroxy-4-(pyridin-2-yl)cyclohexanone. The above product
was dissolved in THF (30 mL) and a 3 N solution of HCl in water (30
mL). The mixture was stirred at 50.degree. C. for 3 hours. After
cooling to room, temperature, NaHCO.sub.3 was added to the solution
with stirring until no bubbling occurred. The organic phase was
separated and the aqueous layer was extracted with EtOAc three
times. The combined organic phase was dried over MgSO.sub.4 and
concentrated. The residue was triturated with EtOAc to give 5.5 g
of the title compound. MS (M+H).sup.+0 192.
Step C
##STR00034##
[0206]
N-(2-{(3S)-3-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]pyrrolidin--
1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The title compound
was prepared by reductive amination of the ketone obtained above
with the pyrrolidine derivative obtained from step C in Example 1
using a procedure analogous to that described in step F, Example 1.
MS (M+H).sup.+ 491.
Example 3
##STR00035##
[0208]
N-(2-{(3S)-3-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)(methyl)amino]pyr-
rolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. To a
solution of
N-(2-{(3S)-3-[(4-hydroxy-4-pyridin-2-ylcyclohexyl)amino]pyrrolidin-1-yl}--
2-oxoethyl)-3-(trifluoromethyl)benzamide (49 mg, 0.1 mmol) and
formaldehyde (0.3 mL, 37% water solution) in THE (2 mL) was added
Na(OAc).sub.3BH (64 mg, 0.3 mmol). After being stirred at room
temperature overnight, the reaction was quenched by addition of a
saturated NaHCO.sub.3 solution. The resulting solution was
extracted with EtOAc and the EtOAc layer was dried (MgSO.sub.4) and
concentrated. Purification by prep HPLC provided the title compound
as a TFA salt. MS (M+H).sup.+ 505.
Example 4
Step A
##STR00036##
[0210] 2-Bromo-5-bromomethylpyridine. 2-Bromo-5-methylpyridine
(5.00 g, 29.1 mmoles) and N-bromosuccinimide (5.22 g, 29.3 mmoles)
were dissolved in carbon tetrachloride (40 mL) under nitrogen.
Benzoyl peroxide. (0.35 g, 1.4 mmoles) was added and the mixture
heated at reflux for four hours. The mixture was cooled to room
temperature, filtered, and washed with NaHCO.sub.3/H.sub.2O. The
mixture was adsorbed onto silica gel and then chromatographed,
eluting with a gradient of hexane to 10% ethyl acetate/hexane. Pure
fractions were combined and concentrated to provide the desired
mono-brominated product as a pale yellow solid, 3.60 g (49%). LC/MS
(M+H).sup.+ m/z=249.8, 251.8, 253.8.
Step B
##STR00037##
[0212] 2-Bromo-5-(methoxymethyl)pyridine.
2-Bromo-5-bromomethyl-pyridine, 4 (3.58 g, 14.3 mmoles) was
dissolved in methanol (20 mL) under nitrogen. Sodium methoxide
(0.89 g, 15.7 mmoles, 95%) was added and the mixture stirred at
room temperature. After 3 hours, the methanol was rotovapped off
and the residue dissolved in dichloromethane and washed with water.
The organic extract was adsorbed onto silica gel and
chromatographed. The column was eluted with a gradient of hexane to
20% ethyl acetate/hexane. Pure fractions were combined and
concentrated to provide the title compound as a colorless oil, 2.62
g (90%). LC/MS (M+H).sup.+m/z=202.0.
Step C
##STR00038##
[0214] 4-Hydroxy-4-[5-(methoxymethyl)pyridin-2-yl]cyclohexanone. A
solution of 2-bromo-5-(methoxymethyl)pyridine (2.61 g, 12.9 mmoles)
was dissolved in dry THF (40 mL) under nitrogen and cooled to
-78.degree. C. n-Butyllithium (6.20 mL, 15.5 mmoles, 2.5 M in
hexane) was added dropwise over 10 minutes to form a black
solution. After 15 minutes, a solution of
1,4-dioxa-spiro[4.5]decan-8-one (2.21 g, 14.1 mmoles) in THF was
added dropwise over 2 minutes and the mixture was gradually warmed
to room temperature over 3 hours. TLC (50% ethyl asetate/hexane)
and LC/MS indicated complete conversion. Aqueous HCl (14 ml, 6.0 M)
was added and the mixture was stirred for 3 hours at room
temperature and then neutralized with NaHCO.sub.3/H.sub.2O. The
mixture was extracted 3 times with ethyl acetate and the combined
extracts were adsorbed onto silica gel and chromatographed. The
column was eluted with a gradient of hexane to 40% ethyl
acetate/hexane. Pure fractions were combined and concentrated to
provide the title compound as a pale yellow solid, 1.00 g (33%).
LC/MS (M+H).sup.+m/z=236.1.
Step D
##STR00039##
[0216]
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(methoxymethyl)pyridin-2-yl]cyclohexy-
l}amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
The title compound was prepared from the ketone of step C using a
procedure analogous to that described for Example 1. MS (M+H).sup.+
535.
Example 5
Step A
##STR00040##
[0218] 6-Bromo-pyridine-3-carbaldehyde. 2,5-Dibromopyridine 9.48 g
(40 mmol) was dissolved in 60 mL of THF and 150 mL of anhydrous
ether. After the solution was cooled to -78.degree. C., 16 mL of
n-butyllithium (2.5 M, 40 mmol) was slowly dropped through a
syringe in 30 mm. After being stirred at -78.degree. C. for 30
minutes, N,N-dimethylformamide (3.5 g, 48 mmol) was added. The
reaction mixture was warmed up to room temperature during two hours
and then quenched by addition of 10 ml water. The mixture was
extracted twice using EtOAc. The combined extracts were dried and
concentrated. After flash column using 30-40% EtOAc in hexane, 2.80
g white solid was obtained (28% yield), MS; (M+H).sup.+ 86.0,
188.0.
Step B
##STR00041##
[0220] 1-(6-Bromopyridin-3-yl)-N,N-dimethylmethanamine. To a
solution of titanium tetraisopropoxide (6.4 g, 22 mmol) and 2.0 M
of dimethylamine in methanol (22 mL, 44 mmol),
6-bomo-pyridine-3-carbaldehyde (2.10 g, 11 mmol) in 20 mL of
methanol was added. After being stirred at r. t. for 5 hrs, sodium
borohydride (0.43 g, 11 mmol) was added and the mixture was stirred
overnight. The reaction was quenched by addition of 10 mL of water
and extracted twice using EtOAc. The combined extracts were dried
and concentrated. After flash column using 20-40% methanol in EtOAc
and 0.5% NH.sub.4OH, 1.15 g oil was obtained (47% yield), MS:
(M+H).sup.+ 214.0, 216.0.
Step C
##STR00042##
[0222]
8-{5-[(Dimethylamino)methyl]pyridin-2-yl}-1,4-dioxaspiro[4,5]decan--
8-ol. 1-(6-Bromopyridin-3-yl)-N,N dimethylmethanamine (1.15 g, 5.4
mmol) was dissolved in 30 mL of THF and 80 mL of anhydrous ether.
After the solution was cooled to -78.degree. C. 2.60 mL of
n-butyllithium (2.5 M, 6.40 mmol) was slowly dropped through a
syringe in 10 min. After being stirred at -78.degree. C. for 30
minutes, 1,4-cyclohexanedione mono-ethylene ketal (1.01 g, 6.4
mmol) was added. The reaction mixture was allowed to warm up to
room temperature during two hours and then quenched by addition of
10 mL of water. The mixture was extracted twice using EtOAc. The
combined extracts were dried and concentrated. After flash column
using 20-40% methanol in EtOAc and 0.5% NH.sub.4OH, 0.85 g oil was
obtained (54% yield), MS: (M+H).sup.+ 293.2.0.
Step D
##STR00043##
[0224]
4-{5-[(Dimethylamino)methyl]pyridin-2-yl}-4-hydroxycyclohexanone.
8-{5-[(Dimethylamino)methyl]pyridin-2-yl}-1,4-dioxaspiro[4,5]decan-8-ol
(0.85 g, 2.9 mmol) was dissolved in 10 mL of THF and 10 mL of 2 N
HCl solution was added. After being stirred for two hours, the
reaction mixture was neutralized to pH-8-9 by addition of a
saturated NaHCO.sub.3 aqueous solution and extracted twice using
EtOAc. The combined extracts were dried and concentrated to obtain
0.37 g white solid (51% yield), MS: (M+H).sup.+ 249.2.
Step E
##STR00044##
[0226]
N-(2-{(3S)-3-[(4-{5-[(Dimethylamino)methyl]pyridin-2-yl}-4-hydroxyc-
yclohexyl)amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
[0227] The title compound was prepared from the above ketone
following the procedure described for Example 1. MS (M+H).sup.+
548.
[0228] The following Examples 6-13 were prepared in a fashion
similar to the previous 5 examples.
Example 6
##STR00045##
[0230]
N-[2-((3S)-3-{[4-Hydroxy-4-(4-methylphenyl)cyclohexyl]amino}pyrroli-
din-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS (M+H).sup.+
504.
Example 7
##STR00046##
[0232]
N-(2-{(3S)-3-[(4-Hydroxy-4-pyridin-3ylcyclohexyl)amino]pyrrolidin-1-
-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. MS (M+H).sup.+
491.
Example 8
##STR00047##
[0234]
N-(2-{(3S)-3-[(4-Hydroxy-4-pyridin-4ylcyclohexyl)amino]pyrrolidin-1-
-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. MS (M+H).sup.+
491.
Example 9
##STR00048##
[0236]
N-[2-((3S)-3-{[(4-Hydroxy-4-(5-methlpyridin-2-yl)cyclohexyl])amino}-
pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 505.
Example 10
##STR00049##
[0238]
N-[2-((3S)-3-{[4-Hydroxy-4-(4-methylpyridin-2-yl)cyclohexyl)]amino}-
pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 505.
Example 11
##STR00050##
[0240]
N-[2-((3S)-3-{[4-Hydroxy-4-(6-methylpyridin-2-yl)cyclohexyl]amino}p-
yrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 505.
Example 12
##STR00051##
[0242]
N-[2-((3S)-3-{[4-Hydroxy-4-(6-methoxypyridin-2-yl)cyclohexyl]amino}-
pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 521.
Example 13
##STR00052##
[0244]
N-[2-((3S)-3-{[4-Hydroxy-4-(6-methoxypyridin-3-yl)cyclohexyl]amino}-
pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 521.
Example 14
Step A
##STR00053##
[0246] 8-(1,3-Thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol. A
solution of n-butyllithium (8.1 mL of 1.6 M solution in hexane,
12.92 mmol) was added to thiazole (1.0 g, 11.75 mmol) in THF (10
mL) at -78.degree. C. with stirring under N.sub.2. After being
stirred at -78.degree. C. for 1 h, a solution of
1,4-cyclohexanedione mono-ethylene ketal (1.84 g, 11.75 mmol) in
THF (10 mL) was added to the lithiated compound solution via
syringe and stirred for 3 h at -78.degree. C. Water (5mL) was
added, and the reaction mixture was warmed to room temperature and
extracted using EtOAc (3.times.). The combined organic layers were
dried (MgSO.sub.4), filtered, concentrated in vacuo and
chromatographed to yield 2.531 g of
8-(1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol in 89% yield.
MS(EI) (M+H).sup.+=242.2.
Step B
##STR00054##
[0248] 8-(5-Methyl-1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol.
A solution of n-butyllithium (5.70 mL of 1.6 M solution in hexane,
9.12 mmol) was added to
8-(1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol (1.00 g, 4.14
mmol) in THF (10 mL) at -78.degree. C. with stirring under N2.
After being stirred at -78.degree. C. for 1 h, methyl iodide (0.71
mL, 9.12 mmol) was added to the lithiated compound solution via
syringe at -78.degree. C. The reaction mixture was allowed to warm
to room temperature slowly and stirred overnight. Water and EtOAc
were added. The aqueous layer was extracted with EtOAc (3.times.).
The combined organic layers were washed with saturated NaCl, dried
(MgSQ.sub.4), concentrated and flash chromatographed using 20%
EtOAc/hexane to give 0.77 g of the title compound in 71% yield. MS
(EI) (M+H).sup.+=256.1.
Step C
##STR00055##
[0250] 4-Hydroxy-4-(5-methyl-1,3-thiazol-2-yl)cyclohexanone. A
solution of
8-(5-Methyl-1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol (1.0 g,
4.14 mmol) in 20 mL of THF/3 N HCl (1:1) was stirred for 1 h at
50.degree. C. After cooling to room temperature, the mixture was
treated with Na.sub.2CO.sub.3 to pH 8 and extracted with EtOAc
(3.times.). The combined organic layers were washed with saturated
NaCl solution, dried (MgSO.sub.4), and concentrated to give 0.82 g
of 4-hydroxy-4-(5-methyl-1,3-thiazol-2-yl)cyclohexanone in 99%
yield. MS (EI) (M+H).sup.+=212.2
Step D
##STR00056##
[0252]
3-(Trifluoromethyl)-N-[2-((3S)-3-{[4-hydroxy-4-(5-methyl-1,3-thiazo-
l-2-yl)cyclohexyl]amino}pyrrolidin-1-yl)-2-oxoethyl]benzamid. The
title compound was prepared from the ketone of step C using
aprocedure similar the that described for Example 1. MS
(EI):(M.sub.+H).sup.+ 511.1.
[0253] The followning Examples 15-16 were prepared in a fashion
similar to Example 14.
##STR00057##
3-(Trifluoromethyl)-N-{2-[(3S)-3-({4-hydroxy-4-[5-(1-hydroxy-1-methylethy-
l)-1,3-thiazol-2-yl]cyclohexyl}amino)pyrrolidin-1-yl]-2-oxoethyl}benzamide-
. MS (EI): (M+H)+ 555.2.
Example 16
##STR00058##
[0254]
3-(Trifluoromethyl)-N-{2-[(3S)-3-({4-hydroxy-4-[5-(methoxymethyl)-1-
,3-thiazol-2yl]cyclohexyl}amino)pyrrolidin-1-yl]-2-oxoethyl}benzamide.
MS (EI): (M+H.sup.+ 541.1.
Example 17
Step A
##STR00059##
[0256]
2-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-1,3-thiazole-4-carboxylic
acid. A solution of n-butyllithium (17.1 mL of 1.6 M solution in
hexane, 27.35 mmol) was added to
8-(1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol (3.00 g, 12.43
mmol) in THF (50 mL) at -78.degree. C. with stirring under N.sub.2.
After being stirred at -78.degree. C. for 1 h, dry ice (10 g, 227
mmol) was added to the lithiated compound solution and stirred for
2 h at -78.degree. C. Water was added and the solution was warmed
to room temperature. The mixture was then treated with 1N HCl to pH
3 to 4 and extracted with EtOAc (3.times.). The combined organic
layers were washed with saturated NaCl solution, dried
(MgSO.sub.4), and concentrated and chromatographed (EtOAc to 1%
AcOH/EtOAc) to give 3.23 g of
2-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-1,3-thiazole-4-carboxylic
acid. MS (EI) (M+H).sup.+=286.0.
Step B
##STR00060##
[0258]
2-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-N-methyl-1,3-thiazole-4-c-
arboxamide. To a stirred solution of
2-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-1,3-thiazole-4-carboxylic
acid (0.30 g, 1.05 mmol) and methylamine (2M in THF, 2 mL, 4 mmol)
in CH.sub.2Cl.sub.2 (10 mL) (0.193 g, 1.26 mmol). The mixture was
stirred at room temperature overnight. Then the reaction mixture
was diluted with EtOAc and washed with saturated Na.sub.2CO.sub.3
and brine. The organic layer was dried (MgSO.sub.4), concentrated
and flash chromatographed (50% EtOAc/hexanes) to give 0.16 g of the
title compound in 50% yield. MS (EI) (M+H).sup.+=299.0.
Step C
##STR00061##
[0260]
2-(1-Hydroxy-4-oxocyclohexyl)-N-methyl-1,3-thiazole-4-carboxamide.
The title compound was prepared by conversion of the ketal of step
B to a ketone using a procedure similar to that described in step C
of Example 14. MS (EI) (M+H).sup.+=255.0.
Step D
##STR00062##
[0262]
2-(1-Hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino}acetyl)-
pyrrolidin-3-yl]amino}cyclohexyl)-N-methyl-1,3-thiazole-5-carboxamide.
The title compound was prepared from the ketone of step C using the
method described for Example 1. MS (EI): (M+H)+553.
[0263] The following Examples 18-19 were prepared in fashion
similar to Example 17.
Example 18
##STR00063##
[0264]
N-Ethyl-2-(1-hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino-
}acetyl)pyrrolidin-3-yl]amino}cyclohexyl)-1,3-thiazole-5carboxamide.
MS (EI): (M+H).sup.+ 567.1.
Example 19
##STR00064##
[0265]
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(pyrrolidin-1-ylcarbonyl)-1,3-thiazol-
-2-yl]cyclohexyl}-amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)be-
nzamide. MS(EI): (M+H).sup.+ 594.1.
Example 20
Step A
##STR00065##
[0267] 8-(1,3-Thiazol-5-yl)-1,4-dioxaspiro[4,5]decan-8-ol.
2-TMS-thiazole (2.5 g, 15.89 mmol) was added to a solution of
n-butyllithium (11.9 mL of 1.6 M solution in hexane, 19.07 mmol) In
THF (20 mL) at -78.degree. C. with stirring under N.sub.2. After
being stirred at -78.degree. C. for 0.5 h, a solution of
1,4-cyclohexanedione mono-ethyiene ketal (2.48 g, 15.89 mmol) in
THF (20 mL) was added to the iithlated compound solution via
syringe and stirred for 1 h at -78.degree. C. Water (5 mL) and
EtOAc were added, and the reaction mixture was warmed to room
temperature and extracted using EtOAc (3.times.). The combined
organic layers were dried (MgSO.sub.4), filtered, and crystallised
from EtOAc to yield 3.4 g of
8-(1,3-thiazol-5-yl)-1,4-dioxaspiro[4,5]decan-8-ol in 90% yield, MS
(EI) (M+H).sup.+=242.1.
Step B
##STR00066##
[0269]
4-Hydroxy-4-[2-(morpholin-4-ylcarbonyl)-1,3-thiazol-5-yl]cyclohexan-
one. A solution of n-butyllithium (2.90 mL of 1.6 M in hexane, 4.64
mmol) was added to 8-(1,3-thiazol-5yl)-1,4-dioxaspiro[4,5]
decan-8-ol (1.00 g, 4.10 mmol) in THF (20 ml) at -78.degree. C.
under N.sub.2. After being stirred at -78.degree. C. for 1 h,
4-morpholinecarbonyl chloride (0.93 g, 6.15 mmol) was added to the
lithiated compound solution via syringe and stirred for 2 h at
-78.degree. C. Water (5 mL) was added, and the reaction mixture was
warmed to room temperature. The reaction mixture was diluted with
water and EtOAc. The aqueous layer was extracted with EtOAc
(3.times.). The combined organic layers were washed with brine,
dried (Na.sub.2SO.sub.4), and concentrated to give the ketal
Intermediate. Then this intermediate was treated with 20 ml, of
THF/1N HCl (1:1) overnight at room temperature. The reaction
solution was justified, to pH 10 with Na.sub.2CO.sub.3 and
extracted with EtOAc (3.times.). The combined organic layers were
washed with brine, dried (Na.sub.2SO.sub.4), concentrated and flash
chromatographed using 20% EtOAc/hexanes to yield 309 mg of the
title compound, MS (EI) (M+H).sup.+=311.0.
Step C
##STR00067##
[0270]
3-(Trifluoromethyl)-N-{2-[(3S)-3-({4-hydroxy-4-[2-(methoxymethyl)-1-
,3-thiazol-5-yl]cyclohexyl}amino)pyrrolidin-1-yl]-2-oxoethyl}benzamide.
The title compound was prepared from the ketone of step B using
procedures similar to that for Example 14. MS (EI): (M+H).sup.+
541.1.
[0271] The following Examples 21-23 were prepared in fashion
similar to Example 20
Example 21
##STR00068##
[0272]
3-(Trifluoromethyl)-N-[2-((3S)-3-{[4-hydroxy-4-(2-methyl-1,3-thiazo-
l-5-yl)cyclohexyl]-amino}pyrrolidin-1-yl)-2-oxoethyl]benzamide. MS
(EI): (M+H).sup.+ 511.1.
Example 22
##STR00069##
[0273]
3-(Trifluoromethyl)-N-[2-((3S)-3-{[4-(2-ethyl-1,3-thiazol-5-yl)-4-h-
ydroxycyclohexyl]-amino}pyrrolidin-1-yl)-2-oxoethyl]benzamide. MS
(EI): (M+H).sup.+ 525.2.
Example 23
##STR00070##
[0274]
N-[2-((3S)-3-{[4-Hydroxy-4-(2-isopropyl-1,3-thiazol-5-yl)cyclohexyl-
]amino}pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide.
MS (EI): (M+H).sup.+ 539.2.
Example 24
A
##STR00071##
[0276]
8-(5-Pyridin-3-yl-1,3-thiazol-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol.
A solution of n-butyllithium (7.8 mL of 1.6 M solution in hexane,
12.45 mmol) was added to
8-(1,3-thiazol-5-yl)-1,4-dioxaspiro[4,5]decan-8-ol (1.0 g, 4.15
mmol) in THF (20 mL) at -78.degree. C. with stirring under N.sub.2.
After being stirred at -78.degree. C. for 0.5 h, 12.5 mL of 0.5 M
solution of ZnCl.sub.2 (6.23 mmol) in THF was added. The resulting
mixture was stirred at room temperature for 0.5 h and a mixture of
3-bromopyridine (0.40 mL, 4.15 mmol) and
PdCl.sub.2(PPh.sub.3).sub.2 (0.11 g, 0.16 mmol) in 5 mL of THF was
added via syringe. After refluxing overnight the reaction was
quenched with 10 mL of saturated NH.sub.4Cl solution. The aqueous
layer was extracted using EtOAc (3.times.). The combined organic
layers were dried (MgSO.sub.4), filtered, concentrated in vacuo and
chromatographed to yield 0.68 g of the title compound in 52% yield.
MS (EI) calcd: (M+H).sup.+=319.1; found: 319.1.
Step B
##STR00072##
[0277]
N-[2-(3S)-(3-{[4-Hydroxy-4-(5-pyridin-3-yl-1,3-thiazol-2-yl)cyclohe-
xyl]methyl}-pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide.
The title compound was prepared from the ketal of step A following
the procedures described for Example 14. MS (EI): (M+H).sup.+
574.2.
Example 25
##STR00073##
[0279]
N-[2-({(3S)-1-[4-Hydroxy-4-(5-pyridin-2-yl-1,3-thiazol-2-yl)cyclohe-
xyl]pyrrolidin-3yl}amino)-2-oxoethyl]-3-(trifluoromethyl)benzamide.
The title compound was prepared following the procedures described
for Example 24. MS (EI): (M+H).sup.+ 574.2.
Example 26
Step A
##STR00074##
[0281] 8-Pyridazin-3-yl-1,4-dioxaspiro[4.5]decan-8-ol. To a
solution of pyridazine (17.7 mmol, 1.28 mL) in THF (60 mL) was
added lithium 2,2,6,6-tetramethylpiperidine (71 mmol, 10 g) at
-78.degree. C. The reaction was then stirred for 5 h at -78.degree.
C. at which point the reaction was quenched using a solution of
ethanol, hydrochloric acid and THF (30 mL, 1:1:1). The resulting
solution was extracted using EtOAc. The organic layers were
combined, dried over MgSO.sub.4 and concentrated. The residue was
purified using flash chromatography to afford the desired alcohol
(44%, 1.84 g). MS (M+H).sup.+ 237.1.
Step B
##STR00075##
[0283] 4-Hydroxy-4-pyridazin-3-ylcyclohexanone. To the product from
step A (7.79 mmol, 1.84 g) in THF (15 mL) was added HCl (45 mmol,
15 mL). The reaction was stirred overnight and subsequently
quenched using Na.sub.2CO.sub.3. The solution was then extracted
using EtOAc (3.times.100 mL). The organic layers were combined,
dried and concentrated in vacuo to afford the desired ketone (780
mg, 52%). MS (M+H).sup.+ 193.1
Step C
##STR00076##
[0285]
N-(2-{3S)-3-[(4-Hydroxy-4-pyridazin-3-ylcyclohexyl)amino]pyrrolidin-
-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The title compound
was prepared from the ketone of step B using a procedure similar to
that described for Example 1. MS (M+H).sup.+ 492.2.
Example 27
##STR00077##
[0287]
N-(2-{(3S)-3-[(4-hydroxy-4-pyrazin-2-ylcyclohexyl)amino]pyrrolidin--
1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The title compound
was prepared in a manner similar to that for Example 26. MS
(M+H).sup.+ 492.2.
Example 28
Step A
##STR00078##
[0289] 8-Pyrimidin-2-yl-1,4-dioxa-spiro[4. 5]decan-8-ol (1a). To a
solution of 2-bromopyrimidine (0.20 g, 1.258 mmol) in dry methylene
chloride (3.0 mL) was dropwise added 1.6 M of n-butyllithium in
hexane (0.86 mL) at -78.degree. C. The reaction mixture was stirred
for 29 min at -78.degree. C. and 1,4-dioxa-spiro[4.5]decan-8-one
(0.196 g, 1.26 mmol) in CH.sub.2Cl.sub.2 (3 mL) was added dropwise.
The reaction was stirred at -78.degree. C. for 50 min and quenched
with an aqueous solution of NH.sub.4Cl. After being woarmed to room
temperature, the mixture was extracted with CH.sub.2Cl.sub.2 three
times. The combined extracts were dried over MgSO.sub.4, filtered
and concentrated in vacuo to provide 0.50 g of crude product.
Purification by column chromatography on silica gel eluting with
0->50% EtOAc in hexanes provide 0.159 g (54%) of desired product
as a light brown-yellow solid. MS (M+H).sup.+237.2.
Step B
##STR00079##
[0291] 4-Hydroxy-4-pyrimidin-2ylcyclohexanone. To the product from
step A (190 mmol, 44 g) in THF (200 mL) was added HCl solution (300
mmol, 100 mL). The reaction was stirred over 2 days after which the
reaction was washed using diethyl ether. The aqueous layer was then
quenched using NaOH (50%) to obtain a pH of 11. The aqueous layer
was extracted using EtOAc (6.times.300 mL). The organic layers were
combined and dried over MgSO.sub.4 and concentrated in vacuo. The
reaction was purified via flash chromatography to afford the
desired ketone (18 g, 49%). MS (M+H).sup.+ 193.1.
Step C
##STR00080##
[0293]
N-(2-{(3S)-3-[(4-Hydroxy-4-pyrimidin-2-ylcyclohexyl)amino]pyrrolidi-
n-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The title
compound was prepared from the ketone of step B using a procedure
similar to that for Example 1. MS (M+H).sup.+ 492.2.
Example 29
Step A
##STR00081##
[0295] 6-Bromonicotinonitrile. 6-Chloronicotinonitrile (13.8 g, 100
mmol) was heated at 145.degree. C. in phosphorus tribromide (150
mL) for 32 h. After cooling, the mixture was concentrated in vacuo.
To the residue was added phosphorus tribromide (150 mL), and the
mixture was heated at 145.degree. C. for another 32 h. After
cooling, the mixture was concentrated in vacuo, and an ice-water
mixture (500 mL) was added. Sodium bicarbonate was added to
neutralize the mixture, and the product was extracted with ethyl
acetate (3.times.250 mL). The combined organic extracts were washed
with brine and dried over magnesium sulfate. The solvent was
removed in vacuo, and the residue was chromatographed
(hexanes-ethyl acetate) to give 14.9 g (81%) of
6-bromonieotlnonitrile as a white solid: 5H NMR (400 MHz.
CDCl.sub.3) .delta. 7.66 (d, J=11.0 Hz, 1H), 7.80 (dd, J=3.1, 11.0
Hz, 1 H), 8.67 (d, J=3.1 Hz, 1 H); MS (M+H).sup.+ m/z=183.0,
185.0.
Step B
##STR00082##
[0297] 6-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)nicotinonitrile. A
solution of 6-bromonicotinonitrile (2 g, 11 mmol) in 50 mL of dry
THF and 15 mL of dry hexane under argon was cooled to -100.degree.
C. in a liquid nitrogen-Et.sub.2O bath. n-Butyllithium (7.5 mL, 11
mmol, 1.6 M solution in hexane) was added dropwise so that the
internal temperature did not exceed -95.degree. C. The orange
solution was stirred for an additional 10 min at -100.degree. C. to
-95.degree. C. and then treated dropwise over 10 min with a
solution of 1,4-cyciohexanedione monoethylene ketai (1.8 g, 11
mmol; in 55 mL of dry THF, again carefully maintaining the
temperature below -95.degree. C. The reaction mixture was stirred
for 10 min at -100.degree. C. to -95.degree. C., allowed to warm to
20.degree. C. and poured into ice water (400 mL). The organic layer
was separated, and the aqueous layer was extracted twice with
Et.sub.2O (200 mL). The combined organic extracts were dried over
MgSO.sub.4 and evaporated to give 2.8 g of white crystalline solid.
Trituration with Et.sub.2O afforded 1.9 g (67% yield) of white
crystals: MS: (M+H).sup.+ 261.
Step C
##STR00083##
[0299] 6-(8-hydroxy-1,4dioxaspiro[4.5]dec-8-yl)nicotinic acid. A
mixture of 6-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)nicotinonitrile
(1.9 g, 7.3 mmol) in 50 mL of 2-methoxyethanol and 50 mL of 2.5 N
NaOH was heated on a steam bath for 15 h. The solution was cooled
in an ice hath, adjusted to pH 7-8 with concentrated HCl, and
evaporated to driness. Water (375 mL) was added, and the pH was
adjusted to 2 with HCl. The tan solid was filtered off and washed
with water to give 1.92 g (6. 9 mmol, 94% yield) of
6-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)nicotinic acid: MS:
(M+H).sup.+ 280.
Step D
##STR00084##
[0301]
6-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-N-methylnicotinamide.
6-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)nicotinic acid (560 mg, 2
mm0l), methylamine (1.2 mL, 2.0 M THF solution), BOP reagent (1.07
g, 2.4 mmol) and 0.8 mL (6 mmol) of triethylamine were dissolved in
15 mL of DMG at room temperature. The reaction mixture was stirred
at room temperature overnight. Direct chromatography on silical gel
(flash chromatography grade) with 50% ethyl acetate- hexane gave
410 mg (70%) of the desired product,
6-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-N-methylnicotinamide: MS:
(M+H).sup.+ 293.
Step E
##STR00085##
[0303] 6-(1-Hydroxy-4-oxocyclohexyl)-N-methylnicotinamide.
6-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-N-methylnicotinamide (410
mg, 1.4 mmol) was dissolved in the mixture solvent of 7 mL of THF
and 7 mL of 1 N HCL aqueous solution at room temperature. The
reaction mixture was then stirred at 60.degree. C. for 1 h. The
solution was cooled down to room temperature, adjusted to pH 7-8
with saturated NaHCO.sub.3 aqueous solution. The organic layer was
separated, arid the aqueous layer was extracted twice with EA (20
ml.times.2). The combined organic extracts were dried over
MgSO.sub.4 and evaporated to give an oil residue. Chromatography on
silica gel (flash chromatography grade) with 40% ethyl
acetate-hexane gave 410 mg (90%) of the desired product,
6-(1-hydroxy-4-oxocyclohexyl)-N-methylnicotinamide: MS: (M+H).sup.+
249.
Step F
##STR00086##
[0305]
6-(1-Hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino}acetyl)-
pyrrolidin-3-yl]amino}cyclohexyl)-N-methylnicotinamide.
6-(1-Hydroxy-4-oxocyclohexyl)-N-methylnicotinamide (100 mg, 0.4
mmol) and 126 mg (0.4 mmol) of
N-{2-[(3S)-3-aminopyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzami-
de were dissolved in 10.0 mL of methylene chloride. To the solution
was added 170 mg (0.8 mmol) of sodium, triacetoxyborohydride. The
reaction mixture was stirred at room temperature for 2 h. Direct
chromatography on silica gel gave 48 mg (23%) of the final desired
product (top spot on TLC and first peak on HPLC). MS: (M+H).sup.+
547.
[0306] The following Examples 30-31 were prepared in fashion
similar to Example 29.
Example 30
##STR00087##
[0308]
6-(1-Hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino}acetyl)-
pyrrolidin-3-yl]amino}cyclohexyl)-N,N-dimethylnicotinamide. MS
(M+H).sup.+ 562.
Example 31
##STR00088##
[0309]
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(pyrrolidin-1-ylcarbonyl)pyridin-2-yl-
]cyclohexyl}-amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzami-
de. MS (M+H).sup.+ 588.
Example 32
Step A
##STR00089##
[0311] 8-(5-Bromopyridin-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol. To a
solution of 2,5-dibromopyridine (4.10 g, 17 mmol) in anhydrous
toluene (250 mL) at -78.degree. C. was dropwise added n-BuLi (1.6
M, 12 mL). After stirred at -78.degree. C. for 2.5 hours, a
solution of 1,4-dioxaspiro[4.5]decan-8-one (2.73 g, 17 mmol) in
methylene chloride (25 mL) was added into the reaction mixture, and
the resulting mixture was stirred for additional one hour and
allowed to warm up to room temperature slowly. The reaction mixture
was poured into aqueous NaHCO.sub.3 (200 mL) and then extracted
with EtOAc (2.times.50 mL). The organic extracts were combined,
washed with saline solution (2.times.50 mL), dried over MgSO.sub.4,
and concentrated in vacuo. The resulting solid was triturated with
ether and the solid was collected by filtration. The ether solution
was concentrated and the solid was chromatographed on silica gel,
eluting with hexane/ethyl acetate (2 to 1) to give a pale yellow
solid. Weight of combined solids: 4.26 g. LCMS: 316.10/314.10
(M+H.sup.+, 100%). .sup.1HNMR: .delta. 8.6 (s, 1 H), 7.82 (d, 1 H),
7.38 (d, 1 H), 4.6 (s, 1 H), 4.0 (m, 4 H), 2.2 (m, 4 H), 1.7 (m, 4
H).
Step B
##STR00090##
[0313] 4-(5-Bromopyridin-2-yl)-4-hydroxycyclohexanone. The title
compound was prepared by treating the ketal of step A with HCl in
water following the procedure described in step B of Example 2. MS
(M+H).sup.+ 271.
Step C
##STR00091##
[0315]
N-[2-((3S)-3-[4-(5-bromopyridin-2-yl)-4-hydroxycyclohexyl]aminopyrr-
olidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. To a 1-neck
round-bottom flask charged with isopropanol (6mL) was added
4-(5-bromopyridin-2-yl)-4-hydroxycyclohexanone (497.6 mg, 1.85
mmol),
N-2-[(3S)-3-aminopyrrolidin-1-yl]-2-oxoethyl-3-(trifluoromethyl)-benzamid-
e hydrochloride (651 mg. 1.85 mol), and triethylamine (0.851 mL,
6.11 mol). The resulting mixture was stirred for 30 minutes at
25.degree. C. Then to it was added sodium triaceloxyhorohydride
(619 mg. 2.78 mmol) and the mixture was stirred at room temperature
overnight. The reaction mixture was concentrated, and the residue
was chromatographed on SiO.sub.2, eluting with acetone/methanol
(100% to 90%/10%) to give two fractions, F1 (404 mg) and F2 (368
mg) in a total of 73% yield. LCMS: (M+H).sup.+ 571.1/569.1 for both
isomers. Isomer 1 .sup.1H NMR (CD.sub.3OD) .delta. 8.65 (t, 1H),
8.21 (s, 1H), 8.14 (d, 1H), 8.03 (dt, 1H), 7.88 (d, 1H), 7.69 (m,
2H), 4.23 (dd, 1H), 4.16 (s, 1H), 4.10 (m, 2H), 3.90 (m, 2H), 3.70
(m, 2H), 3.60 (dd, 1H), 3.52 (m, 2H), 2.55 (m, 1H), 2.42 (m, 2H),
2.22 (m, 3H), 1.80 (m, 4H).
Example 33
##STR00092##
[0317]
N-{2-[(3S)-3-({4-[5-(2-formylphenyl)pyridin-2-yl]-4-hydroxycyclohex-
yl}-amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
A solution of
N-[2-((3S)-3-[4-(5-bromopyridin-2-yl)-4-hydroxycyclohexyl]aminopyrrolidin-
-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide (30.0 mg, 0.0527
mmol) and (2-formylphenyl)boronic acid (8.6 mg, 0.052 mmol) in DMf
(0.60 mL) and aqueous sodium carbonate (2-formylphenyl)boronic acid
8.6 mg, 0.052 mmol) in DMF (0.60 mL) and aqueous sodium carbonate
(2M, 0.198 mL) was degassed with N.sub.2 for 5 minutes. The
[1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II),
complex with dichloromethane (1:1) (2.2 mg, 0.0026 mmol) was added
in under N.sub.2 flush. The reaction mixture was degassed with
N.sub.2 for another 5 minutes and then the tube was sealed. The
reaction mixture was heated under microwave at 130.degree. C. for 5
minutes. After cooling down, the reaction mixture was filtered
through a short pad of silica gel and washed with CH.sub.3CN. The
resulting solution was acidified with TFA to pH 1.about.2, then was
subjected to purification on Prep-HPLC. The appropriate fractions
were lypholized to give the product (23 mg, 53%) as a white powder.
MS: (M+H).sup.+ 595.
Example 34
##STR00093##
[0319]
N-(2-(3S)-3-[(4-Hydroxy-4-5-[2-(hydroxymethyl)phenyl]pyridin-2-ylcy-
clohexyl)amino]pyrrolidin-1-yl-2-oxoethyl)-3-(trifluoromethyl)benzamide
bis(trifluoroacetate). To a solution of
N-2-[(3S)-3-(4-[5-(2-formylphenyl)pyridin-2-yl]-4-hydroxycyclohexylamino)-
pyrrolidin-1-yl]-2-oxoethyl-3-(trifluoromethyl)benzamide
bis(trifluoroacetate) (salt) (3.3 mg, 0.004 mmol) in methanol (0.50
mL) at 0.degree. C. was added sodium borohydrkie (0.455 mg, 0.0120
mmol). The reaction mixture was allowed to warm up to room
temperature and stirred at: room temperature for 60 minutes and
then at 60.degree. C. for 60 minutes. The mixture was purified by
prep-HPLC to afford the product as a TEA salt (1.1 mg, 33%). LCMS:
(M+H).sup.+ 597.2.
Example 35
Step A
##STR00094##
[0321] 8-(4-Iodo-phenyl)-1,4-dioxa-spiro[4.5]decan-8-ol. To a
solution of 1,4-diiodobenzene (16.5 g, 50 mmol) in THE (350 mL) at
-78.degree. C. was added n-BuLi (2.5 M, 24 mL) over 1 hour. After
being stirred additional 30 minutes, a solution of
1,4-dioxaspiro[4.5]decan-8-one (7.8 g, 50 mmol) in THF (30 mL) was
added in and the resulting mixture was stirred for 3 hours. To the
mixture was added TMSCl (5.4 g, 50 mmol) and the resulting mixture
was allowed to warm to room temperature and stirred at room
temperature for 18 hours. The reaction mixture was neutralized to
pH 6.0, and extracted with ethyl acetate (3.times.50 mL). The
organic extracts were combined, washed with saline solution
(2.times.50 mL), dried over sodium sulfate, and concentrated in
vacuo. The residue was chromatographed on silica gel, eluting with
hexane/ethyl acetate (95/5 to 100/0). The appropriate fractions
were combined to give
8-(4-Iodo-phenyl)-1,4-dioxa-spiro[4.5]decan-8-ol (12 g, 66.6%) with
LCMS: 361.2 (M+H.sup.+, 100%) and
{[8-(4-iodophenyl)-1,4-dioxaspiro[4.5]dec-8-yl]oxy}(trimethyl)silane
(6 g, 27%) with LCMS: 433.1 (M+H).sup.+, 100%).
Step B
##STR00095##
[0323] 8-(4-pyrimidin-2-ylphenyl)-1,4-dioxaspiro[4.5]decan-8-ol. To
a solution of 8-(4-iodo-phenyl)-1,4-dioxa-spiro[4.5]decan-8-ol
(450.0 mg, 1.249 mmol) in THF (1.0 mL) at room temperature was
added dropwise isopropyimagnesiuro chloride (2.0 M in THF, 1.37 mL)
and the reaction mixture was stirred at room temperature for 30
mins. To another flask charged with nickel acetylacetonate (20 mg,
0.06 mmol) and 1,3-bis(diphenylphosphino)-propane propane (26 mg,
0.062 mmol) suspended in THF (3 mL) under N.sub.2 was added
2-bromopyrimidine (199 mg, 1.25 mmol). The resulting mixture was
stirred at room temperature until it is clear. This mixture was
transferred into the degassed Grignard solution prepared above. The
resulting mixture was stirred at room temperature overnight. The
reaction mixture was diluted with EtOAc, quenched with water,
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated.
The residue was columned on silica gel, eluting with hexane/EtOAc
(2/1), to gave the desired compound (270 mg, 69%) as a white solid,
LCMS: 313.1, (M+H, 100%). .sup.1H NMR (CDCl.sub.3): .delta. 8.86
(d, 2H), 8.46 (dd, 2H), 7.71 (dd, 2H), 7.24 (t, 1H), 4.05 (d, 4H),
2.30 (dt, 2H), 2.18 (dt, 2H), 1.90 (m, 2H), 1.78 (m, 2H).
Step C
##STR00096##
[0325] 4-Hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexanone. The title
compound was prepared by treating the ketal of step B with HCl in
water following the procedure described in step B of Example 2, MS
(M+H).sup.+ 269.
[0326] Step D
##STR00097##
N-[2-((3S)-3-[4-hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexyl]aminopyrroli-
din-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide
bis(trifluoroacetate) (salt). To a 1-neck round-bottom flask
charged with methylene chloride (1 mL) was added
4-hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexanone (50.0 mg, 0.186
mmol),
N-2-[(3S)-3-aminopyrrolidin-1-yl]-2-oxoethyl-3-(trifluoromethyl)benzamide
hydrochloride (65.5 mg, 0.186 mmol), and triethylamine (85.7 uL,
0.615 mmol). The resulting mixture was stirred at 25.degree. C. for
30 minutes, and to it was added sodium triacetoxyborohydride (62.4
mg, 0.28 mmol) in portion. The reaction mixture was stirred at room
temperature overnight and concentrated. The residue, was
chromatographed on SiO.sub.2, eluting with acetone/methanol (100%
to 90%/10%) to give two fractions, which were further purified on
prep-LCMS separately to afford F1 (24.2 mg ) and F2 (25.9 mg) as
white powder in a total of 34% yield. LCMS: 568.2 (M+H, 100%) for
both isomers.
[0327] The following Examples 36-37 were prepared in a similar
manner.
Example 36
##STR00098##
[0329]
N-[2-((3S)-3-{[4-Hydroxy-4-(5-phenylpyridin-2-yl)cyclohexyl]amino}p-
yrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 567.
Example 37
##STR00099##
[0331]
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(1,3-thiazol-2-yl)pyridine-2-yl]cyclo-
hexyl}amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 573.
Example 38
Step A
##STR00100##
[0332]
8-(5-Pyrimidin-2-ylpyridin-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol.
[0333] A solution of
8-(5-bromopyridin-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol (168.5 g,
0.5363 mol) in THF (2000 mL) was degassed with nitrogen for 30
minutes. A 2.0 M solution of isopropylmagnesiurn chloride in THF
(563 mL) was added dfopwise over 70 mins at room, temperature to
the above solution. The reaction mixture (light brownish color) was
stirred for 180 minutes at 25.degree. C.
Into another flask was charged with THF (500 mL) that was degassed
with nitrogen for 10 min. To it were added Nickel acetylacetonate
(6.9 g, 0.027 mol) and 1.2-bis(diphehylphosphino)-ethane (11 g,
0.027 mol) under nitrogen flush, and 10 minutes later
2-iodopyrimidine (113 g, 0.536 mol). After being stirred for 30
minutes at 25.degree. C., the resulting light green suspension was
transfered to the above solution. The reaction mixture was stirred
at room temperature overnight and the reaction was found to be
complete by HPLC. LC-MS: found (M+H) 314.20 for desired product.
The reaction mixture was directly used for next reaction.
[0334] Step B
##STR00101##
4-Hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexanone.
[0335] About half of the THF in the reaction mixture from step A
was removed by evaporation under reduced pressure. To the remaining
reaction mixture was added a 4.00 M solution, of HCl in water (900
mL). After being stirred for 1 hour, the mixture was diluted with
1000 mL water and neutralized with solid Na.sub.2CO.sub.3 to pH
8.about.9. Large ammount of yellow solid precipitated out. The
solid was filtered off and washed with ethyl aceate containing 1%.
aqueous NH.sub.4OH (about 2000 mL) until no desired product was
detected by TLC. The filtrate was partitioned and the aqueous layer
was extracted with ethyl acetate (1200 mL.times.3). The combined
organic layers were washed with brine, dried, over magnesium
sulfate and concentrated to half of the volume. The solid
precipitating out was filtered and dissolved in dichloromethane
(600 mL). The resulting solution was heated to reflux for 30
minutes and filtered. The filtrate was cooled in an ice bath. The
solid precipitating out was collected by filtration to give 30 g of
pure product. The mother liquids from the two crystallizations were
combined and evaporated. The residue was taken into acetonitrik
(500 mL). The resulting solution was heated to reflux until all
solid was dissolved. Once Insolubles were filtered off, the
filtrate was allowed to stand at room temperature and solid was
precipitated out. The solid was filtered and suspended in
dichloromethane (700 mL). After being heated to reflux, the
solution was filtered, evaporated to half of the volume, and cooled
in an ice bath. The light brownish solid precipitating out was
collected by filtration to give the second batch of solid (58 g).
MS (M+H) 270.2.
[0336] Step C
##STR00102##
N-[2-((3S)-3-{[4-Hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexyl]amino-
}pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide.
[0337] To a solution of
N-{2-[(3S)-3-aminopyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzami-
de hydrochloride (22.10 g, 47.1 mmol) and
4-hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexanone (12.7 g.
47.1 mmol) in isobutyl alcohol (80.0 mL) was added triethyiamlne
(19.7 mL, 141 mmol). The reaction mixture was cooled in an ice bath
and stirred for 30 minutes. To it was added sodium
triaeetoxyborohydride (11.0 g, 51.8 mmol) in portion. After being
stirred at room temperature for 4 hours, the solvent was removed by
evaporation under reduced pressure. Saturated aqueous NaHCO.sub.3
solution, was added and the solution was extracted with ethyl
acetate (150.times.3). The combined extracts were washed with
brine, dried (Na.sub.2SO.sub.4), filtered, and concentrated. The
residue was columned on silica get, eluting with ethyl acetate (1%
NH40H aqueous solution)/ methanol (95/5 to 80/20). The appropriate
fractions were combined and concentrated to give the title,
compound as a white powder (17.77 g), MS (M+H) 569.
[0338] The following examples were prepared in a similar
manner.
Example 38
##STR00103##
[0339]
N-(2-{(3S)-3-[(4-[5-[3-(Aminocarbonyl)phenyl]pyridin-2-yl]-4-hydrox-
ycyclohexyl)-amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzami-
de. MS (M+H).sup.+ 610.
Example 40
##STR00104##
[0341]
N-(2-{(3S)-3-[(4-{5-[2-(Aminocarbonyl)phenyl]pyridin-2-yl}-4-hydrox-
ycyclohexyl)amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamid-
e. MS (M+H).sup.+ 610.
Example 41
##STR00105##
[0343]
N-{2-[(3S)-3-({4-[5-(3-Acetylphenyl)pyridin-2-yl]-4-hydroxycyclohex-
yl}amino)-pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 609.
Example 42
##STR00106##
[0345]
3-[6-(1-Hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino}acet-
yl)-pyrrolidin-3-yl]amino}cyclohexyl)pyridin-3-yl]benzoic acid. MS
(M+H).sup.+ 611.
Example 43
##STR00107##
[0347]
N-(2-{(3S)-3-[(4-Hydroxy-4-{5-[3-(hydroxymethyl)phenyl]pyridin-2-yl-
}cyclohexyl)amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamid-
e. MS (M+H).sup.+ 597.
Example 44
##STR00108##
[0349]
N-[2-((3S)-3-{[4-Hydroxy-4-(5-pyrimidin-5-ylpyridin-2-yl)cyclohexyl-
]amino}-pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 569.
Example 45
##STR00109##
[0351]
N-[2-((3S)-3-{[4-(3,3'-Bipyridin-6-yl)-4-hydroxycyclohexyl]amino}py-
rrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 568.
Example 46
##STR00110##
[0353]
N-[2-((3S)-3-{[4-(3,4'-Bipyridin-6-yl)-4-hydroxycyclohexyl]amino}py-
rrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 568.
Example 47
##STR00111##
[0354]
N-[2-((3S)-3-{[4-Hydroxy-4-(5-pyrazin-2-ylpyridin-2-yl)cyclohexyl]a-
mino}pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 569.
Example 48
##STR00112##
[0356]
N-[2-((3S)-3-{[4-Hydroxy-4-(4isoxazol-4-ylphenyl)cyclohexyl]amino}p-
yrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 557.
Example 49
##STR00113##
[0357]
N-{2-[(3S)-3-({4-Hydroxy-4-[4-1H-imidazol-1-yl)phenyl]cyclohexyl}am-
ino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 556.
Example 50
##STR00114##
[0359]
4'-(1-Hydroxy-4-{[(3S)-1-({[3-(trifluoromethyl)benzoyl]amino})acety-
l)pyrrolidin-3-yl]amino}cyclohexyl)biphenyl-2-carboxamide. MS
(M+H).sup.+ 609.
Example 51
##STR00115##
[0361]
N-[2-((3S)-3-{[4-(2'-Formylbiphenyl-4-yl)-4-hydroxycyclohexyl]amino-
}-pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide. MS
(M+H).sup.+ 594.
Example 52
##STR00116##
[0362]
N-{2-[(3S)-3-({4-Hydroxy-4-[2'-(hydroxymethyl)biphenyl-4-yl]cyclohe-
xyl}amino)-pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 596.
Example 53
##STR00117##
[0363]
N-{2-[(3S)-3-({4-[5-(3,5-Dimethylisoxazol-4-yl)pyridin-2-yl]-4-hydr-
oxycyclohexyl}-amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benza-
mide. MS (M+h).sup.+586.
Example 54
##STR00118##
[0365]
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(1,3-oxazol-2-yl)pyridin-2-yl]cyclohe-
xyl}amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 574.
Example 55
Step A
##STR00119##
[0367] 3-(Trifluoromethyl)benzaldehyde oxime. To a flask containing
3-trifluorobenzaldehyde (1.74 g, 10 mmol) and hydroxyiamine
hydrochloride (0.76 g, 11 mmol) in methanol (25 mL) was added TEA
(0.65 g, 11 mmol). The reaction mixture was heated to reflux for 3
h, neutralized to pH 6.0, and extracted with ethyl acetate
(3.times.20 mL), The organic extracts were combined, washed with
saline solution (20 mL), dried over sodium sulfate, concentrated in
vacuo to give the oxime (1.9 g) as a colorless oil. LCMS:
(M+H).sup.+ 190.2.
[0368] Step B
##STR00120##
[0369] 3-(Trifluoromethyl)benzaldehyde oxime. To a dried flask
containing 3-(trifluoromethyl)benzaldehyde oxime (1.89 g, 10 mmol)
in methylene chloride (100 mL) was added N-chlorosuccinimide (1.40
g, 10.5 mmol) slowly at 0.degree. C. The reaction mixture was
warmed to 45.degree. C. for 2 h, poured over ice, diluted with H2O
(20 mL), and extracted, with EtOAc (100 mL). The organic phase was
washed with H2O (2.times.25 mL) and saline solution (25 mL), dried
over sodium sulfate, concentrated in vacuo to give the oxime (2 g,
90%). LCMS: (M+H).sup.+0 224.4.
Step C
##STR00121##
[0371] Methyl
3-[3-(Trifluoromethyl)phenyl]-4,5-dihydroisoxazole-5-carboxylate.
To a flask containing
N-hydroxy-3-(trifluoromethyl)benzenecarboximidoyl chloride (2.0 g,
8.9 mmol) and methyl acrylate (0.7 g, 8 mmol) in methylene chloride
(100 mL) at 0.degree. C. under an inert atmosphere was added TEA
(0.90 g, 8.8 mmol). The reaction mixture was slowly wanned to
ambient temperature, stirred for 20 h, quenched with water (30 mL),
and extracted with methylene chloride (2.times.50 mL). The organic
extracts were combined, washed with saline solution (50 mL), dried
over sodium sulfate, concentrated in vacuo, and chromatographed on
silica gel, eiuting with methylene chioride/methanoi (100/1 to
95/5). The appropriate fractions were combined and concentrated in
vacuo to give the title compound (2.3 g; 100%); LCMS:
(M+H).sup.+274.2. .sup.1H NMR: (CDCl.sub.3) .delta. 8.03 (s, 1H),
7.92 (d, 1H), 7.71 (d, 1H), 7.59 (dd, 1H), 5.28 (dd, 1H), 3.86 (s,
3H), 3.71 (dd, 2H).
Step D
##STR00122##
[0373]
3-[3-(Trifluoromethyl)phenyl]-4,5-dihydroisoxazole-5-carboxylic
Acid. To a solution of methyl
3-[3-(trifluoromethyl)phenyl]-4,5-dihydroisoxazole-5-carboxylate
(2.3 g, 8.4 mmol) in THF (10 mL) was added a 2 M solution of sodium
hydroxide in water (10 mL) at 0.degree. C. The reaction mixture was
slowly warmed to ambient temperature, stirred for 2 h, neutralised
with 2 N HCl to pH 7, and extracted with, ethyl acetate (2.times.50
mL). The organic extracts were combined, washed with saline
solution (50 mL), dried over sodium sulfate, and concentrated in
vacuo. The residue was chromatographed on silica gel, eluting with
methylene chloricle/methanol (95/5 to 80/20). The appropriate
tractions were combined and concentrated in vacuo to give the title
compound (2.18 g, 100%) as a white crystalline solid. LCMS;
(M-H).sup.+ 258.2.
Step E
##STR00123##
[0375] tert-Butyl
[(3S)-1-(3-[3-(Trifluoromethyl)phenyl]-4,5-dihydroisoxazol-5-ylcarbonyl)p-
yrrolidin-3-yl]carbamate. To a solution of
3-[3-(trifluoromethyl)phenyl]-4,5-dihydroisoxazole-5-carboxylic
acid (259 mg, 1 mmol) and tert-butyl (3S)-pyrrolidin-3-ylcarbamate
( (186 mg, 1 mmol) in DMF (0.5 mL) and methylene chloride (5 mL) at
0.degree. C. was added triethylamine (120 mg, 1.2 mmol) and
benzotriazol-1-yloxytris(dimethylamino)-phosphonium
hexafluorophosphate (442 mg, 1 mmol). The mixture was allowed to
warm to room temperature over 1 h and stirred at room temperature
for 1 h. The mixture was concentrated in vacuo, and the residue was
chromatographed on silica gel, eluting with 1% NH4OH in ethyl
acetate to give the desired coupling intermediate (410 mg) as a
white solid. LCMS: (M+H).sup.+ 428.4.
Step F
##STR00124##
[0377]
3S)-1-(3-[3-(Trifluoromethyl)phenyl]-4,5-dihydroisoxazol-5-ylcarbon-
yl)-pyrrolidin-3-amine hydrochloride To a solution of the
intermediate of step E in methylene chloride (5 mL) was added 4 M
HCl in dioxane (5 mL). After stirred at room temperature for 2 h,
the resulting solution wa. concentrated in vacuo to give the HCl
salt (350 mg) of the amine, as a white solid. LCMS: (M+H).sup.+
364.4.
Step G
##STR00125##
[0379]
1-Pyridin-2-yl-4-[(3S)-1-(3-[3-(trifluoromethyl)phenyl]-4,5-dihydro-
isoxazol-5-ylcarbonyl)pyrrolidin-3-yl]aminocyclohexanol. to a
solution of
(3S)-1-(3-[3-(trifluoromethyl)phenyl]-4,5-dihydroisoxazol-5-ylcarbonyl)py-
rrolidin-3-amine hydrochloride (178 mg, 0.489 mmol) and and
4-hydroxy-4-pyridin-2-yl-cyclohexanone (95.1 mg, 0.498 mmol) in
methylene chloride (6 mL) was added triethylamine (50.3 mg, 0.498
mmol) and then NaBH(OAc).sub.3 (120 mg, 0.54 mmol). After being
stirred at room temperature for 2 h, the reaction mixture was
neutralized with 1 N NaOH to pH 7, and extracted with, ethyl
acetate (2.times.25 mL). The organic extracts were combined, washed
with saline solution (20 mL), dried over sodium sulfate,
concentrated in vacuo, and chromatographed on silica gel, eluting
with 1% NH.sub.4OH in ethyl acetate/methanol (95/5 to 80/20). The
appropriate fractions were combined and concentrated in vacuo to
give two fractions of the desired compounds: peak 1 (100 mg) and
peak 2 (85 mg). Both fractions were further purified by HPLC on a
C18 column, during with 1% NH4OH in water/acetonitrile, to give
peak 1 (68 mg) and peak 2 (65 mg) as white solids. Both compounds
have LCMS: (M+H).sup.+ 503.3. Peak 1 shows two peaks in a 1 to 1
ratio in a chiral analytical -column. Peak 2 shows two peaks in a 1
to 10 ratio in a chiral analytical column.
[0380] The following Examples 56-58 were prepared in a fashion
similar to Example 55.
Example 56
##STR00126##
[0382]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({3-[3-(trifluoromethyl-
)phenyl]-4,5-dihydroisoxazol-5-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexa-
nol. MS (M+H).sup.+ 581.
Example 57
##STR00127##
[0384]
1-{5-[(Dimethylamino)methyl]pyridin-2-yl}-4-{[(3S)-1-({3-[3-(triflu-
oromethyl)phenyl]-4,5-dihydroisoxazol-5-yl}carbonyl)pyrrolidin-3-yl]amino}-
cyclohexanol. MS (M+H).sup.+ 560.
Example 58
##STR00128##
[0385]
1-[5-(1,3-Oxazol-2-yl)pyridin-2-yl]-4-{[(3S)-1-({3-[3-(trifluoromet-
hyl)phenyl]-4,5-dihydroisoxazol-5-yl}carbonyl)pyrrolidin-3-yl]amino}cycloh-
exanol. MS (M+H) 570.3.
Example 59
Step A
##STR00129##
[0387] Methyl (2S,4R
)-N-tert-Butoxycarbonyl-4-hydroxy-2-pyrrolidinecarboxylate.
L-trans-4-Hydroxyproline methyl ester hydrochloride (25.00 g, 138.0
mmol) was dissolved dichloromethane (300 mL) and triethyiamine
(58.0 mL. 413 mmol). The solution was cooled to 0.degree. C. and
then di-tert-butyldicarbonate: (33.00 g, 151.0 mmol) was added in
small portions. After stirring at room temperature overnight, the
mixture was concentrated to a thick white sludge. The residue was
dissolved in ethyl acetate and the organic layer was washed
successively with NH.sub.4Cl/H.sub.2O, NaHCO.sub.3/H.sub.2O and
brine. The organic extracts were dried over MgSO.sub.4, filtered,
and concentrated to give 33.0 g (99%) of desired product as a
colorless oil. LC/MS (M+Na).sup.+ m/z=267,9. .sup.1H
NMR(CDCl.sub.3) .delta. 4.50 (m, 1H), 4.40 (m, 1H), 3.75 (s, 3H),
3.43-3.68 (ms 2H), 2.30 (in, 1H), 1.95-2.15 (m, 2H), 1.42 and 1.45
(s, 9H).
Step B
##STR00130##
[0389] 1-tert-Butyl 2-Methyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}pyrolidine-1,2-dicarboxylate.
Methyl
(2S,4R)-N-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinecarboxylate
(22.1 g, 82.6 mmol) was dissolved in dry DMF (100 mL) under
nitrogen. Imidazole (16.8 g, 248 mmol) was added and the mixture
cooled to 0.degree. C. tert-Butyldimethylsilyl chloride (13.1 g,
86.7 mmol) was added in small portions and then the mixture was
allowed to warm to room temperature. After stirring, overnight, the
mixture was
[0390] diluted with 300 ml, ethyl acetate and washed with water
three times (500 mL, 200 ml, 200 mL). The organic extracts were
washed one final time with brine and then dried over MgSO.sub.4,
filtered and concentrated to give 29.5 g (99%) of desired product
as a colorless oil. LC/MS (M-Boc+H).sup.+m/z=260.2. .sup.1H NMR
(CDCl.sub.3) .delta. 4.30-4.47 (m, 2H), 3.73 and 3.75 .(s, 3H),
3.60 (m, 1H), 3.28-3.45 (m, 1H), 2.18 (m, 1H), 2.03 (m, 1H), 1.42
and 1.47 (s, 9H), 0.87 (s, 9H), 0.06 (s, 6H).
Step C
##STR00131##
[0392] tert-Butyl
(2S,4R)-4-{[tert-Butyl(dimethyl)silyl]oxy}-2-(hydroxymethyl)-pyrrolidine--
1-carboxylate. 1-tert-Butyl 2-methyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]-oxy}pyrrolidine-1,2-dicarboxylate
(5.00 g, 13.91 mmol) was dissolved in dry THF (50 mL) under
nitrogen and cooled to -78.degree. C. Diisobutylaluminum hydride
solution (31.0 mL, 31.0 mmol, 1.0 M in toluene) was added dropwise
over 30 minutes. After stirring for ten minutes, the mixture was
slowly warmed to room temperature at which point TLC indicated
complete conversion. The mixture was diluted with ethyl acetate
(200 mL) and saturated aqueous sodium potassium tartrate. (200 mL).
The mixture was stirred vigorously for 30 minutes until two phases
were apparent. The aqueous layer was then extracted twice with
ethyl acetate and washed with brine. The organic layer was dried
over MgSO.sub.4, filtered and concentrated to give 4.91 g of the
crude alcohol as a pale yellow oil, LC/MS (M-Boc+H).sup.+
m/z=232.2. .sup.1H NMR (CDCl) .delta. 4.88 (d, 1H), 4.27 (bs, 1H),
4.14 (m, 1H), 3.69 (t. 1H), 3.54 (m, 1H), 3.42 (d, 1H), 3.34 (dd,
1H), 1.96 (m, 1H), 1.58 (m, 1H), 1.47 (s, 9H), 0.87 (s, 9H), 0.06
(s, 6H).
Step D
##STR00132##
[0394] tert-Butyl
(2S,4R)-4-{[tert-Butyl(dimethyl)silyl]oxy}-2-({[(4-methylphenyl)-sulfonyl-
]-oxy}methyl)pyrrolidine-1-carboxylate. tert-butyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-(hydroxymethyl)pyrrolidine-1-
-carboxylate (4.91 g, 14.8 mmol) was dissolved in dichloromethane
(70 mL) under nitrogen. Triethylamine (5.8 mL, 41.7 mmol) was added
followed by p-toluenesulfonyl chloride (3.18 g, 16.7 mmol) and the
mixture was stirred at room temperature overnight. TLC revealed
about half conversion, pyridine (3.4 mL, 41 mmol) was added to the
mixture which turned dark orange after 20 minutes. After two more
days, the mixture was diluted with ethyl acetate and the organic
layer was washed successively with NaHCO.sub.3/H.sub.2O,
NH.sub.4Cl/H.sub.2O, water, and brine. The organic extract was
dried over MgSO.sub.4, filtered and concentrated to a red oil which
was chromatographed on silica gel (10% to 20% ethyl
acetate/hexane). Pure fractions were combined to give the tosylate
as a yellow oil 6.32 g (93%, 2 steps). .sup.1H NMR (CDCl.sub.3)
.delta. 7.77 (d, 2H), 7.34 (L 2H), 4.30 (m, 2H), 4.10 (ms 2H), 3.30
(m, 2H), 2.45 (s, 3H), 1.07 (ms 2H), 1.41 and 1.37 (s, 9H), 0.85
(s, 9H), 0.06 (s, 6H).
Step E
##STR00133##
[0396] tert-Butyl
(2R,4R)-4-{[tert-Butyl(dimethyl)silyl]oxy}-2-methylpyrrolidine-1-carboxyl-
ate. tert-Butyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-({[(4-methylphenyl)-sulfonyl-
]-oxy}methyl)pyrrolidine-1-carboxylate (6.32 g, 13.01 mmol) was
dissolved in THF (50 mL) under nitrogen and cooled to 0.degree. C.
Lithium triethylborohydride solution (Super Hydride, 14.3 mL, 1.0 M
in THF) was added dropwise and the mixture was then slowly warmed
to room temperature. After 2 hours, TLC revealed half conversion.
More lithium triethylborohydride solution (12.0 mL) was added and
the solution stirred at room temperature overnight. Diluted with
NaHCO.sub.3/H.sub.2O and extracted twice with ethyl acetate. Washed
organic layer with NH.sub.4Cl/H.sub.2O and brine. Dried organic
extracts over MgSO.sub.4, filtered and concentrated to give a
colorless oil. Chromatographed on silica gel eluting with 10%
ethyl, acetate/hexane. Pure fractions were combined to give the
desired product as a colorless oil, 3.74 g (91%). LC/MS
(M+Na).sup.+ m/z=338.2. .sup.1H NMR (CDCl.sub.3) .delta.4.34 (m,
1H), 3.95 (m, 1H), 3.35 (m, 2H), 1.98 (m, 1H), 1.65 (m, 1H), 1.47
(s, 9H). 1.20 (bs, 3H), 0.87 (s, 9H), 0.06 (s, 6H).
Step F
##STR00134##
[0398] (3,5R)-5-Methylpyrrolidin-3-ol hydrochloride. tert-Butyl
(2R,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-methylpyrrolidine-1-carboxyl-
ate (3.74 g, 11.85 mmol) was dissolved in dry THF (20 mL) under
nitrogen. Hydrogen chloride solution (40 mL, 4.0 M solution in
1,4-dioxane) was added and the mixture was stirred at room
temperature for four hours. The solution was concentrated on the
rotovap to an oil which was azeotroped with toluene and pumped
under vacuum to provide the hydrochloride salt as an off white
solid, 1 80 g (100%) which was used for the next step without
further purification. .sup.1H HMR (CD.sub.3OD) .delta. 4.54 (m,
1H)33.95 (m, 1H), 3.44 (dds 1H), 3.18 (d, 1H), 2.19 (dd, 1H), 1.76
(m, 1 H), 1.44 (d; 3H).
Step G
##STR00135##
[0400]
N-{2-[(2R,4R)-4-Hydroxy-2-methylpyrrolidin-1-yl]-2-oxoethyl}-3-(tri-
fluoromethyl)benzamide. (3R,5R)-5-Methylpyrrolidin-3-ol
hydrochloride (1.80 g, 13 mmol) was dissolved in dichloromethane
(50 mL) and diisopropylethylamine (2.1 mL, 12.0 mmol) under
nitrogen. (3-Trifluoromethyl-benzoylamino)-acetic acid (2.93 g,
11.85 mmol) was added followed by EDC (3.41 g, 17.8 mmol) and the
mixture was stirred at room temperature for four hours. The mixture
was diluted with NH.sub.4Cl/H.sub.2O and extracted twice with ethyl
acetate. The combined extracts were washed with
NaHCO.sub.3/H.sub.2O and brine, dried over MgSO.sub.4, filtered and
concentrated to give a dark orange oil. Chromatography on silica
gel eluting with ethyl acetate to 5% methanol/ethyl acetate gave
the coupled product as a pale orange solid, 3.19 g (81%, 2 steps),
LC/MS (M+H).sup.+ m/z=331.1. .sup.1H NMR (CDCl.sub.3, major
rotamer) .delta. 8.12 (s, 1H), 8.01 (d, 1H), 7.76 (d, 1H), 7.57 (t,
1H), 7.50 (m, 1H), 4.56 (m, 1H), 4.34 (m, 1H), 4.23 (m, 1H), 4.11
(m, 1H), 3.61 (dd, 1H), 3.51 (d, 1H), 2.71 (d, 1H), 2.17 (m, 1H),
1.81 (m, 1H), 1.32 (d, 3H).
Step H
##STR00136##
[0402]
(3R,5R)-5-Methyl-1-({[3-(trifluoromethyl)benzoyl]amino}acetyl)pyrro-
lidin-3-yl methanesulfonate. To a solution of
N-{2-[(2R,4R)-4hydroxy-2-methylpyrrolidin-1-yl]-2-oxoethyl}-3-(trifluorom-
ethyl)benzamide (1.50 g, 4.54 mmol) in dichloromethane (30 mL) and
pyridine (1.83 mL, 22.7 mmol) under nitrogen at 0.degree. C. was
added methanesulfonyl chloride (0.42 mL, 5.45 mmol) dropwise. After
being stirred at 0.degree. C. for two hours, the reaction was
allowed to slowly warm to room temperature and stirred overnight.
The mixture was diluted with NaHCO.sub.3/H.sub.2O and brine, dried
over MgSO.sub.4, filtered and concentrated to give the mesylate as
a brown oil, 1.87 g (100%). LC/MS (M+H).sup.+ m/z=409.0. .sup.1H
NMR (CDCl.sub.3, major rotamer) .delta.8.12 (s, 1H), 8.01 (d, 1H),
7.78 (d, 1H), 7.59 (t, 1H), 7.29 (bs, 1H), 5.33 (m, 1H), 4.37 (m,
1H), 4.18 (m, 2H), 3.86 (d, 1H), 3.76. (dd, 1H), 3.08 (s, 3H), 2.51
(m, 1H), 1.94 (m, 1H), 1.38 (d, 3H).
Step I
##STR00137##
[0404]
N-{2-[(2R,4S)-4-Azido-2-methylpyrrolidin-1-yl]-2-oxoethyl}-3-(trifl-
uoromethyl)benzamide. To a solution of the crude mesylate (1.87 g)
in dry DMF (20 mL) was added sodium azide (1.50 g, 22.7 mmol). The
mixture was stirred at 60-65.degree. C. for five hours, then
50.degree. C. for twenty hours. Ethyl acetate was added. The
organic layer was separated, washed twice with water and then with
brine, dried over MgSO.sub.4, filtered and concentrated to an
orange oil. Chromatography on silica gel emting with 80% ethyl
acetate/hexane gave the aside as a yellow oil, 1.33 g (82%). LC/MS
(M+H).sup.+ m/z=356.1. .sup.1H NMR (CDCl.sub.3, major rotamer)
.delta. 8.12 (s, 1H), 8.00 (t, 1H), 7.77 (d, 1H), 7.58 (t, 1H),
7.37 (bs, 1H), 4.35 (m, 2H), 4.17 (m, 2H), 3.73 (dd, 1H), 3.50 (d,
1H), 2.39 (m, 1H), 1.87 (d, 1H), 1.43 (d, 3H).
Step J
##STR00138##
[0406]
N-{2-[(2R,4S)-4-Amino-2-methylpyrrolidin-1-yl]-2-oxoethyl}-3-(trifl-
uoromethyl)benzamide.
N-{2-[(2R,4S)-4-Azido-2-methylpyrrolidin-1-yl]-2oxoethyl}-3-(trifluoromet-
hyl)benzamide (1.33 g, 3.74 mmol) was dissolved in ethanol (50 mL)
and then 10% Pd--C (130 mg) was added to the solution. The flask
was purged With hydrogen and then stirred under an atmosphere of
hydrogen using a balloon for four hours at which point, TLC
indicated complete consumption of starting material. The reaction
was then freshed with nitrogen and filtered through Celite on a
glass hit and washed with methanol. The filtrate was concentrated
to give the desired amine as a dark brown oil, 1.21 g (98%). LC/MS
(M+H).sup.+ m/z=330.1. .sup.1H NMR (CDCl.sub.3) .delta. 8.12 (s,
1H), 8.02 (d, 1H), 7.77 (d, 1H), 7.58 (t, 1H), 7.37 (bs, 1H), 4.16
(m, 3H), 3.72 (m, 1H), 3.61 (m, 1H): 3.1 5 (m, 1H), 2.44 (m, 1H),
1.70-1.20 (m, 3H), 1.43 (d, 3H); .sup.19F NMR (CDCl.sub.3) .delta.
-63.12 (s).
Step K
##STR00139##
[0408]
N-(2-{(2R,4S)-4-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-methy-
l-pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
N-{2-[(2R,4S)-4-Amino-2-methylpyrrolidin-1-yl]-2-oxoethyl}-3-(trifluorome-
thyl)benzamide (200 mg, 0.607 mmol) and 4-hydroxy-4-pyridin-2-yl
-cyclohexanone (116 mg, 0.607 mmol) were dissolved in 2-propanol
(10 mL). After stirring for 30 minutes, sodium
triacetoxyborohydride (257 mg, 1.21 mmol) was added and the mixture
was stirred at room temperature overnight. TLC indicated complete
conversion to desired products in about a 1:1 ratio of two isomers.
The reaction mixture was chromatographed on silica gel eiuting with
dichloromethane to 10% methanol/dichloromethane/0.5% ammonium
hydroxide to give 229 mg (75%) as a mixture of isomers. .sup.1H NMR
(CDCl.sub.3, mixture of isomers) .delta. 8.53 (m, 1H), 8.13 (bs,
1H), 8.02 (d, 1H), 7.75 (m, 2H), 7.58 (t, 1H), 7.40 (m, 2H), 7.22
(ms 1H), 4.05-438 (m, 3H), 3.80 (m, 1H), 3.56 (m, 1H), 3.42 (m,
1H), 3.19 (m, 1H), 3.04 (m, 1H), 2.65 (m, 1H), 2.47 (m, 1H), 2.16
(m, 2H), 1.40-2.00 (m, 7H), 1.43 (d, 3H). LCMS (M+H).sup.+: Higher
Rf isomer m/z=505.2; Lower Rf isomer m/z=505.2.
Example 60
Step A
##STR00140##
[0410] tert-Butyl
(2S,4R)-4-{[tert-Butyl(dimethyl)silyl]oxy}-2-(methoxymethyl)-pyrrolidine--
1-carboxylate. Iodomethane (0.85 mL, 13.6 mmol) was added to a
solution of tert-butyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-(hydroxymethyl)pyrrolidine-1-
-carboxylate (1.50 g, 4.52 mmol) in dry DMF (15 mL) under nitrogen.
Sodium hydride (0.22 g, 5.42 mmol, 60% dispersion in mineral oil)
was added in portions and the mixture was stirred overnight at room
temperature-. The mixture was diluted with ethyl acetate. The
organic layer was separated, washed twice with water and then
brine, dried over MgSO.sub.4, filtered and concentrated to give
1.51 g. (96%) of methyl ether as a yellow oil. LC/MS
(M-Boc+H).sup.30 m/z=246.2. .sup.1H NMR (CDCl.sub.3) .delta. 4.38
(m, 1), 4.05 (m, 1H), 3050 (m, 2H), 3.25-3.45 (m, 2H), 3.34 (s,
3H), 1.87-2.06 (m, 2H), 1.47 (s, 9H), 0.87 (s, 9H), 0.06 (s,
6H).
B
##STR00141##
[0412]
N-{2-[(2S,4S)-4-[(4-hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-(meth-
oxymethyl)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
The title compound was prepared from the intermediate of step A
following the procedures described for Example 59. Higher Rf
isomer; LCMS m/z=532.2 (M+H); .sup.1H NMR (CDCl.sub.3) .delta. 8.53
(d, 1H, 8.12 (s, 1H), 8.03 (d, 1H), 7.77 (m, 1H), 7.72 (m, 1H),
7.58 (t, 1H), 7.47 (m, 1H), 7.34 (m, 1H), 7.21 (M, 1H), 4.90 (m,
1H), 4.12-4.47 (m, 4H), 3.89 (dd, 1H), 3.79 (dd, 1H), 3.54 (m, 2H),
3.38 (s, 3H), 3.03 (m, 1H), 2.40 (m, 1H), 2.18 (m, 3h), 1.90 (m,
1H), 1.75 (m, 1H), 1.60 (m, 2H), 1.50 (m, 2H); .sup.19F NMR
(CDCl.sub.3) .delta. -63.11 (s). Lower Rf isomer: LCMS (M+H).sup.+
m/z=535.2; .sup.1H NMR (CDCl.sub.3) .delta. 8.53 (d, 1H), 8.12 (s,
1H), 8.02 (d, 1H), 7.78 (m, 1H), 7.72 (m, 1H), 7.58 (t, 1H), 7.42
(m, 1H), 7.34 (m, 1H), 7.21 (m, 1H), 4.12-4.48 (m, 4H), 3.83 (m,
2H), 3.68 (m, 1H), 3.56 (m, 1H), 3.38 (s, 3H), 2.72 (m, 1H), 1H),
2.38 (m, 1H), 1.60-2.20 (m, 10H; .sup.19F NMR (CDCl.sub.3) .delta.
-63.12 (s).
Example 61
##STR00142##
[0414]
N-(2-{(2S,4S)-2-(Ethoxymethyl)-4-[(4-hydroxy-4-pyridin-2-ylcyclohex-
yl)-amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
The title compound was prepared following the procedures described
for Example 60. Higher Rf isomer: LCMS (M+H).sup.+ m/z=549.1;
.sup.1H NMR (CDCl.sub.3) .delta. 8.51 (m, 1H), 8.10 (m, 1H), 7.999
(m, 1H), 7.70 (m, 2H), 7.32-7.60 (m, 3H), 7.18 (m, 1H), 4.03-4.47
(m, 3H), 3.22-3.91 (m, 5H), 3.04 (m, 1H), 1.70-2.47 (m, 7H), 1.51
(m, 4H), 1.21 (m, 4H).
Lower Rf isomer: LCMS (M+H).sup.+ m/z=549.1; .sup.1H NMR
(CDCl.sub.3) .delta. 8.52 (m, 1H), 8.11 (m, 1H), 8.00 (m, 1H), 7.73
(m, 2H), 7.55 (m, 1H), 7.39 (m, 2H), 7.20 (m, 1H), 4.11-4.48 (m,
3H), 3.46-3.88 (m, 5H), 3.21 (m, 1H), 2.63 (m, 1H), 2.38 (m, 1H),
1.55-1.98 (m, 10H), 1.20 (m, 3H).
Example
Step A
##STR00143##
[0416] tert-Butyl
(2S,4R)-4-{[tert-Butyl(dimethyl)silyl]oxy}-2-(1-hydroxy-1-methylethyl)pyr-
rolidine-1-carboxylate. To a solution of 1-tert-butyl 2-methyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}pyrrolidine-1,2-dicarboxylate
(1.00 g, 2.78 mmol) in dry THF (20 mL) at 0.degree. C. was dropwise
added methylmagnesium bromide solution (2.0 mL, 6.0 mmol, 3.0 M in
ether) over 5 minutes. After stirring for four hours, the mixture
was wanned to room temperature and quenched with
NH.sub.4Cl/H.sub.2O and extacted twice with ethyl acetate. The
organic extracts were dried over MgSO.sub.4filtered and
concentrated to give 1.00 g (100%) of the title compound as a white
solid. .sup.1 NMR (CDCl.sub.3) .delta. 5.85 (s, 1H), 4.25 (s, 1H),
4.08 (t, 1H), 3.67 (d, 1H), 3.18 (d, 1H), 1.94 (m, 1H), 1.60 (m,
1H), 1.45 (s, 9H), 1.15 (s, 3H), 1.05 (s, 3H), 0.87 (s, 9H), 0.06
(s, 6H).
Step B
##STR00144##
[0418]
N-(2-{(2S,4S)-2-(1-Hydroxy-1-methylethyl)-4-[(trans-4-hydroxy-4-pyr-
idin-2-ylcyclohexyl)amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)-
benzamide. The title compound was prepared from the alcohol of step
A following the procedures described for Example 59. Higher Rf
isomer: LCMS (M+H).sup.+ m/z=549.3; .sup.1H NMR (CDCl.sub.3)
.delta. 8.53 (m, 1H), 8.13 (s, 1H), 8.01 (d, 1H), 7.78 (d, 1H),
7.74 (t, 1H), 7.59 (t, 1H), 7.48 (d, 1H), 7.32 (m, 1H), 7.22 (m,
1H), 4.19-4.40 (m, 3H), 3.98 (dd, 1H), 3.49 (m, 2H), 3.29 (m, 1H),
3.08 (m, 1H), 2.10-2.45 (m, 8H), 1.71 (m, 2H), 1.24 (s, 3H), 1.21
(s, 3H); .sup.19F NMR (CDCl.sub.3) .delta. -63.12 (s). Lower Rf
isomer: LCMS (M+H).sup.+ m/z=549.3: .sup.1H NMR (CDCl.sub.3)
.delta. 8.52 (d, 1H), 8.12 (s, 1H), 8.01 (d, 1H), 7.77 (d, 1H),
7.73 (m, 1H), 7.59 (t, 1H), 7.40 (d, 1H), 7.37 (m, 1H), 7.22 (m,
1H), 5.14 (bs, 1H), 4.39 (m, 1H), 4.33 (m, 1H), 4.20 (m, 1H), 3.97
(m, 1H), 3.72 (m, 1H), 3.40 (m, 1H), 2.74 (m, 1H), 1.70-2.35 (m,
12H), 1.24 (s, 3H), 1.21 (s, 3H); .sup.19F NMR (CDCl.sub.3) .delta.
-63.12 (s).
Example 63
##STR00145##
[0420]
N-(2-{(2S,4S)-2-[1-Hydroxyethyl]-4-[(4-hydroxy-4-pyridin-2-ylcycloh-
exyl)-amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
The title compound was prepared in a manner similar to that for
Example 62. MS (M+H).sup.+ 535.
Example 64
##STR00146##
[0422]
N-{2-[(2S,4S)-4-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-(1-me-
thoxy-1-methylethyl)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzam-
ide. The title compound was prepared starting from (tert-butyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-(1-hydroxy-1-methylethyl)pyr-
rolidine-1-carboxylate following the procedures described for
Example 60. Higher Rf isomer: LC/MS (M+H).sup.+ m/z=563.3; .sup.1H
NMR (CDCl.sub.3) .delta.8.55 (m, 1H), 8.14 (m, 1H), 8.04 (m, 1H),
7.74 (m, 2H), 7.58-7.63 (m, 3H), 7.22 (m, 1H). 5.42-5.80 (bs, 1H),
4.84 (bs, 1H), 4.15-4.43 (m, 3H), 3.96 (m, 1H), 3.42 (m, 1H), 3.22
(m, 4H), 3.02 (m, 1H), 1.89-2.34 (m, 6H), 1.46-1.67 (m, 4H), 1.22
(m, 6H). Lower Rf isomer: LC/MS (M+H).sup.+ m/z=563.3; .sup.1H NMR
(CDCl.sub.3) .delta. 8.53 (m, 1H), 8.15 (m, 1H), 8.03 (m, 1H), 7.74
(m, 2H), 7.35-7.61 (m, 3H), 7.22 (m, 1H), 3.87-4.43 (m, 4H), 3.50
(m, 1H), 3.21 (m, 4H), 2.64 (m, 1H), 2.27 (m, 1H), 1.67-1.98 (m,
9H), 1.22 (m, 6H).
Example 65
##STR00147##
[0424]
N-(2-{(2S,4S)-4[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-[(1S)--
1-methoxyethyl]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
The title compound was prepared in a fashion similar to that for
Example 64. MS (M+H).sup.+ 549.
Example 66
Part A
##STR00148##
[0425] 1-tert-Butyl 2-Methyl
(4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-methylpyrrolidine-1,2-dicarboxy-
late. To a solution of 1-tert-butyl 2-methyl
(2S,4R)-4-{[tert-butyl(dimethyl)silyl]oxy}pyrrolidine-1,2-dicarboxylate
(5.11 g, 14.2 mmol) in dry THF (60 mL) at -78.degree. C. was
dropwise added lithium bistrimethylsilylamide (17.0 mL, 17.0 mmol,
1.0 M in THF), After being stirred for 30 minutes, lodomethane
(1.77 mL, 28.4 mmol) was then added, The mixture was stirred at
-78.degree. C. for one hour, warmed to 0.degree. C. for one hour
and finally quenched with NaHCO.sub.3/H.sub.2O. The resulting
mixture was extracted twice with ethyl acetate. The combined
extracts were dried over MgSO.sub.4, filtered and concentrated. The
residue was chromatographed on silica gel eiuting with hexane to 5%
ethyl acetate/hexane to provide 2.66 g (50%) of a mixture of
product isomers as a colorless oil. LC/MS (M-Boc+H).sup.+
m/z=274.1. .sup.1H NMR (CDCl.sub.3) .delta. 4.38 (m, 1H), 3.71 (m,
4H), 3.36 (m, 1H), 1.84-2.35 (m, 2H), 1.61 (m, 3H), 1.44 (m, 9H),
0.88 (m, 9H), 0.07 (m, 6H).
[0426] Step B
##STR00149##
[0427]
N-(2-{(4S)-4-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2,2-dimeth-
yl-pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The
title compound was prepared from 1-tert-butyl 2-methyl
(4R)-4-{[tert-butyl(dimethyl)silyl]oxy}-2-methylpyrrolidine-1,2-dicarboxy-
late following the procedures described for Example 59, Higher Rf
homer: LC/MS (M+H).sup.+ m/z=519.2; .sup.1H NMR (CD.sub.3OD,
bis-trifluoroacetate salt) .delta. 8.51 (m, 1H), 8.18 (m, 2H),
7.63-7.90 (m, 4H), 7.27 (m, 1H), 4.15 (dd, 2H), 3.98 (m, 1H), 3.55
(m, 1H), 3.28 (m, 2H), 2.92 (m, 1H), 2.38 (m, 2H), 1.96-2.20 (m,
3H), 1.50-1.79 (m, 7H), 1.42 (s, 311). Lower Rf isomer: LC/MS
(M+H).sup.+ m/z=519.2; .sup.1H NMR (CD.sub.3OD,
bis-trifluoroacetate salt) .delta. 8.49 (m, 1H), 8.21 (m, 1H), 8.14
(m, 1H), 7.65-7.90 (at, 4H), 7.25 (m, 1H), 4.10 (m, 3H), 3.72 (m,
1H), 3.28 (m, 2H), 2.73 (m, 1H), 2.10 (m, 3H), 1.82 (m, 2H), 1,73
(m, 4H), 1.58 (s, 3H), 1.45 (s, 3H).
Example 67
Step A
##STR00150##
[0429] 1-Benzyl 2-Methyl
(2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate.
L-trans-4-Hydroxyproline methyl ester hydrochloride (9.70 g, 54.0
mmol) was dissolved in dry THF (180 mL) and triethylamine (7.53 mL,
54.0 mmol). N-(Benzyloxycarbonyloxy)succinimide (13.5 g, 54.0 mmol)
dissolved is THF (70 mL) was slowly added to the solution. After
stirring at room temperature overnight, the mixture was diluted
with ethyl acetate and the organic layer was washed successively
with water and brine. The organic extracts were dried over
Na.sub.2SO.sub.4, filtered, and concentrated. The residue was
chromatographed on silica gel (30% to 70% ethyl acetate/hexane) to
provide 12.8 g (85%) of desired product as a colorless oil. LC/MS
(M+H).sup.+ m/z=280.0; .sup.1H NMR (CDCl.sub.3) .delta. 7.33 (m,
5H), 5.00-5.25 (m, 2H), 4.52 (m, 2H), 3.69 (m, 2H), 3.56 and 3.78
(s, 3H), 2.05-2.40 (m, 2H).
Step B
##STR00151##
[0431] 1Benzyl 2-Methyl
(2S,4R)-4-(benzyloxy)pyrrolidine-1,2-dicarboxylate. 1-Benzyl
2-methyl (2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (6.60 g,
23.6 mmol) was dissolved in dry THF (100 mL) and cooled to
0.degree. C. under nitrogen. Sodium hydride (1.04 g, 26.0 mmol, 60%
dispersion in mineral oil) was added in portions and the mixture
was stirred for 15 minutes. Tetra-n-butylammonium iodide (0.40 g,
1.0 mmol) and benzyl bromide (3.15 mL, 26.0 mmol) were added and
the mixture stirred for one hour at 0.degree. C. and then one hour
at room temperature. The mixture was diluted with ethyl acetate.
The organic layer was washed with water and then brine, dried over
MgSO4, filtered, and concentrated. The residue was chromatographed
on silica gel (20% to 50% ethyl acetate/hexane) to give 4.21 g
(4-8%) of benzyl ether, LC/MS (M+H).sup.+ m/z=370.2; .sup.1H NMR
(CDCl.sub.3) .delta. 7.34 (m, 10H), 5.13 (m, 2H), 4.51 (m, 3H),
4.20 (m, 1H), 3.68 (m, 2H), 3.54 and 3.78 (s, 3H), 2.45 (m, 1H),
2.11 (m, 1H).
Step C
##STR00152##
[0433] Benzyl
(2S,4R)-4-(Benzyloxy)-2-(1-hydroxy-1-methylethyl)pyrrolidine-1-carboxylat-
e. 1-Benzyl 2-methyl
(2S,4R)-4-(benzyloxy)pyrrolidine-1,2-dicarboxylate (4.21 g, 11.4
mmol) was dissolved in dry THF (20 mL) under nitrogen and cooled to
0.degree. C. Methylmagnesium bromide solution (8.4 mL, 25 mmol, 3.0
M in ether) was added dropwise. After stirring for twelve hours at
0.degree. C., the mixture was Warmed to room, temperature and
quenched with NH.sub.4Cl/H.sub.2O and extracted twice with ethyl
acetate. The organic extracts were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
chromatographed on silica gel (20% to 30% ethyl acetate/hexane) to
give 2.47 g (59%) of the alcohol as a viscous oil. LC/MS
(M+H).sup.+ m/z=370.1; .sup.1H NMR (CDCl.sub.3) .delta. 7.33 (m,
10H), 5.55 (bs, 1H), 5.20 (s, 2H), 4.50 (s, 2H), 4.19 (m, 1H), 4.05
(m, 2H), 3.31 (m, 1H), 2.27 (m, 1H), 1.73 (m, 1H), 1.21 (s, 3H),
1.13 (s, 3H).
Step D
##STR00153##
[0435] Benzyl
(2S,4R)-4-(Benzyloxy)-2-isopropenylpyrrolidine-1carboxylate. Benzyl
(2S,4R)-4-(benzyloxy)-2-(1-hydroxy-1-methylethyl)pyrrolidine-1-car-
boxylate (2.22 g, 6.01 mmol) was dissolved in toluene (40 ml) and
triethylamine (10.0 mL, 72 mmol) under nitrogen. The mixture was
cooled to -50.degree. C. and thionyl chloride (0.44 mL, 6.0 mmol)
was added dropwise. Alter stirring tor three hoots at. -30.degree.
C. the mixture was quenched by addition of water. The resulting
mixture was extracted twice with ethyl acetate and the organic
extracts were washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was ehromatographed on
silica gel (10% to 20% ethyl acetate/hexane) to give 1.10 g (52%)
of the olefin as a pale yellow oil. LC/MS M+H).sup.+ m/z=352.2;
.sup.1H NMR (CDCl.sub.3) .delta. 7.35 (m, 10H), 5.16 (m, 2H), 4.84
(m, 2H), 4.52 (m, 3H), 4.16 (m, 1H), 3.87 (m, 1H), 3.58 (m, 1H),
2.29 (m, 1H), 1.94 (m 1H), 1.69 (m, 3H).
[0436] Step E
##STR00154##
[0437] (2S,4R)-4-(Benzyloxy)-2-isopropylpyrrolidine. Benzyl
(2S,4R)-4-(benzyloxy)-2-isopropenylpyrrolidine-1-carboxylate (100
g, 2.84 mmol) was dissolved in ethanol (40 mL) and then 5% Pd--C
(100 mg) was added to the solution. The flask was purged with
hydrogen and then shaken on a Parr under 53 psi atmosphere of
hydrogen for 17 hours. The reaction was then flushed with nitrogen
and filtered through Cellte on a glass frit and washed with
methanol. The filtrate was concentrated arid ehromatographed on
silica gel (1% rriethylamine/10% methanol/89% ethyl acetate) to
famish the amine as a pale yellow oil, 0.53 g (85%). LC/MS
(M+H).sup.+ m/z=220.2; .sup.1H NMR (CDCl.sub.3) .delta. 7.33 (m,
5H), 4.49 (m, 2H)< 4.12 (m, 1H), 3.19 (dd, 1H), 3.00 (m, 2H),
2.05 (m, 1H), 1.96 (bs, 1H)S L49 (m, 2H), 1.00 (d, 3H), 0.91 (d,
3H).
[0438] Step F
##STR00155##
[0439]
N-{2-[(2S,4R)-4-Benzyloxy-2-isopropylpyrrolidin-1-yl]-2-oxoethyl}-3-
-(trifluoromethyl)benzamide.
(2S,4R)-4-(Benzyloxy)-2-isopropylpyrrolidine (0.410 g, 1.90 mmol)
was dissolved in dichloromethane (30 mL) under nitrogen.
(3-Trifluoromethyl-benzoylamino)-acetic acid (0.462 g, 1.90 mmol)
was added followed by EDC (0.394 g, 2.06 mmol) and the mixture was
stirred at room temperature overnight. LC/MS revealed the reaction
was not yet complete. More (3-Trifluoromethyl-benzoylamino)-acetic
acid (0.12 g, 0.48 mmoles) and more EDC (0.30 g, 1.6 mmoles) were
added and stirring continued for 3 hours at room temperature, then
at reflux for 1.5 hours. The mixture was chromatographed cm silica
gel eluting with 30% ethyl acetate/hexane to provide 0.66 g (79%)
of the coupled product as a colorless oil. LC/MS (M+H).sup.+
m/z=449.2; .sup.1H NMR (CDCl.sub.3) .delta. 8.03 (m, 1H), 7.76 (m,
1H), 7.58 (m, 2H), 7.34 (m, 5H), 4.52 (m, 2H), 4.03-4,34 (m, 4H)s
3.65 (m, 1H), 3.48 (m, 1H), 2.54 (m, 1H), 2.12 (m, 1H), 1.92 (m,
1H), 0.92 (d, 3H), 0.77 (d, 3H).
Step G
##STR00156##
[0441]
N-{2-[(2S,4R)-4-Hydroxy-2-isopropylpyrrolidin-1-yl]-2-oxoethyl}-3-(-
trifluoromethyl)benzamide.
N-{2-[(2S,4R)-4-Benzyloxy-2-isopropylpyrrolidin-1-yl]-2-oxoethyl}-3-(trif-
luoromethyl)benzamide (0.630 g, 1.40 mmol) was dissolved In
methanol (60 mL) and then palladium hydroxide (90 mg) was added to
the solution. The flask was purged with hydrogen, and then stirred
under an atmosphere of hydrogen using a balloon. After three hours,
TLC indicated complete consumption of starting material. The
reaction was then flushed with nitrogen and filtered through Ceiite
on a glass hit and washed with methanol. The filtrate was
concentrated to give the desired alcohol as a white solid, 0.52 g
(100%). LC/MS (M+H).sup.+ m-z=359.2; .sup.1H NMR (CDCl.sub.3)
.delta. 8.11 (m, 2H), 7.53-7.82 (m, 3H), 4.04-4.52 (m, 4H), 3.63
(m, 1H), 3.43 (m, 1H), 2.50 (ms 1H), 1.86-2.25 (m, 2H), 0.89 (d,
3H), 0.78 (d, 3H).
Step H
##STR00157##
[0443]
N-(2-{(2S,4S)-4-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-isopr-
opyl-pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide. The
title compound was prepared from the above intermediate following
the procedures described for Example 59. Higher Rf Isomer: LC/MS
(M+H).sup.+ m/z=533.3; .sup.1H NMR (CD.sub.3OD,
bis-trifluoroacetate salt) .delta. 8.66 (m, 1H), 8.20 (m, 3H), 7.94
(m, 2H), 7.74 (m, 1H), 7.59 (m, 1H), 4.36 (m, 2H), 4.06-4.27 (m,
2H), 4.00 (m, 1H), 3.63 (m, 1H), 3.46 (m, 1H), 2.63 (m, 1H), 2.50
(m, 1H), 2.34 (m, 4H), 1.76-2.05 (m, 5H), 0.96 (d, 3H), 0.93 (d,
3H); Lower Rf isomer: LC/MS (M+H).sup.+ m/z=533.2; .sup.1H NMR
(CD.sub.3OD, bis-trifluoroacetate salt) .delta. 8.66 (m, 1H), 8.24
(m, 3H), 7.96 (m, 2H), 7.72 (m, 2H), 4.00-4.42 (m, 5H), 3.45 (m,
2H), 2.65 (m, 1H), 2.49 (m, 1H), 2.22 (m, 4H), 1.95 (m, 5H), 0.96
(d, 3H), 0.91 (d, 3H).
[0444] The following Examples 68-71 were prepared in a manner
similar to Example 67.
Example 68
##STR00158##
[0446]
N-{2-[(2S,4S)-4-({4-Hydroxy-4-[5-(methoxymethyl)pyridin-2-yl]cycloh-
exyl}-amino)-2-(methoxymethyl)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromet-
hyl)benzamide. MS (M+H).sup.+ 579.
Example 69
##STR00159##
[0448]
N-{2-[(2S,4S)-4-[(4-{5-[(Dimethylamino)methyl]pyridin-2-yl}-4-hydro-
xy-cyclohexyl)amino]-2-(methoxymethyl)pyridin-1-yl]-2-oxoethyl}-3-(trifluo-
romethyl)benzamide. MS (M+H).sup.+ 592.
Example 70
##STR00160##
[0450]
N-{2-[(2S,4S)-4-[(4-Hydroxy-4-pyridin-2-ylcyclohexyl)amino]-2-(isop-
ropoxy-methyl)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide.
MS (M+H).sup.+ 563.
Example 71
##STR00161##
[0451]
N-{2-[(2S,4S)-4-{[4-Hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohe-
xyl]amino}-2-(methoxymethyl)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethy-
l)benzamide. MS (M+H) 613.3.
Example 72
##STR00162##
[0452]
N-(2-{(3S)-3-[[4-Hydroxy-4-(5-pyrazin-2-ylpyridin-2-yl)cyclohexyl](-
methyl)amino-9
pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamide.
N-[2-((3S)-3-{[4-Hydroxy-4-(5-pyrazin-2-ylpyridin-2-yl)cyclohexyl]amino}p-
yrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide (29.0 mg,
0.051 mmol) and 37% aqueous formaldehyde (21 uL, 0.26 mmol) were
dissolved in THF (1.0 mL). The mixture was evaporated to dryness.
Then the residue was taken up in THF (1 mL) and sodium,
triacetoxyborohydride (24 mg, 0.11 mmol) was added. After being
stirred at room temperature overnight, the mixture was purified by
HPLC to provide the title compound (5.9 mg). MS (M+H) 583.3.
Example 73
##STR00163##
[0453]
N-(2-{(3S)-3-[{4-Hydroxy-4-[5-(1,3-oxazol-2-yl)pyridin-2-yl]cyclohe-
xyl}(methyl)amino]pyrrolidin-1-yl}-2-oxoethyl)-3-(trifluoromethyl)benzamid-
e.
N-{2-[(3S)-3-({4-Hydroxy-4-[5-(1,3-oxazol-2-yl)pyridin-2-yl]cyclohexyl}-
amino)pyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)benzamide (45
mg, 0.081 mmol) and 37% aqueous formaldehyde (30 mg, 1.0 mmol) were
dissolved in methylene chloride (5.6 mL). lite mixture was
evaporated to dryness. Then the residue was taken up in THF (1 mL)
and sodium triacetoxyborohydride (38 mg, 0.18 mmol) was added.
After being stirred at room temperature overnight, the mixture was
purified by HPLC to provide the title compound (27 mg). MS (M+H)
572.3.
Example 74
Step A
##STR00164##
[0454] Methyl
1-[3-(Trifluoromethyl)phenyl]piperidine-4-carboxylate. Methyl
piperidine-4-carboxylate (2.0 g, 14 mmol),
1-bromo-3-(trifluoromethyl)benzene (1.5 g, 6.8 mmol), and potassium
tert-butoxide (0.76 g, 6.8 mmol) m a mixed solvent of toluene (20
mL) and DMF (4 ml) was added
[1,1'-bis(diplhenylphosphino)ferrocene]dichloropalladium(II),
complex with dichloromethane (1:1) (0.3 g, 0.4 mmol) under
nitrogen. The mixture was heated at 130.degree. C. in an oil bath
overnight. After cooling to room temperature, the mixture was
filtered through celite and diluted with EtOAc. The resulting
solution was washed with saturated NaHCO.sub.3. The aqueous layer
was extracted with EtOAc twice. The combined organic layers were
dried (MgSO.sub.4), concentrated and flash chromatographed with
EtOAc/hexanes (20% to 40%) to give 0.90 g of product. MS (M+H)
288.2.
Step B
##STR00165##
[0456] 1-[3-(Trifluoromethyl)phenyl]piperidine-4-carboxylic Acid.
Methyl 1-[3-(trifluoromethyl)Phenyl]piperidine-4-carboxylate (0.9
g, 3 mmol). was treated with the mixture of 2 M of sodium,
hydroxide in water (10 mL), THF (10 mL) and methanol (10 mL) at
50.degree. C. for 1 h. After being neutralized with concentrated
HCl (pH=3), the solution was concentrated. The resulting residue
was azeotropically treated with benzene for 3 times to give the
title compound which was used for the next reaction without
purification. MS (M+H) 274.1.
Step C
##STR00166##
[0457] tert-Butyl
[(3S)-1-({1-[3-(Trifluoromethyl)phenyl]piperidin-4-yl}carbonyl)pyrrolidin-
-3-yl]carbamate. tert-Butyl (3S)-pyrrolidin-3-ylcarbamate (0.65 g,
3.5 mmol), 1-[3-(trifluoromethyl)phenyl]piperidine-4-carboxylic
acid (0.80 g, 2.9 mmol), triethylamine (0.82 mL, 5.8 mmol) and
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (1.4 g, 3.2 mmol) were mixed in dry methylene
chloride (10 mL). After being stirred overnight, the reaction
mixture was diluted with EtOAc and washed with saturated
NaHCO.sub.3. The aqueous layer was extracted with EtOAc three
times. The combined organic layers were dried (MgSO.sub.4),
concentrated and flash chromatographed (20% EtOAc/hexanes to 40%
EtOAc/hexanes) to give 0.975 g of the desired product MS (M+H)
442.1.
Step D
##STR00167##
[0458]
(3S)-1-({1-[3-(Trifluoromethyl)phenyl]piperidin-4-yl}carbonyl)pyrro-
lidin-3-amine bis(trifluoroacetate. tert-Butyl
[(3S)-1-({1-[3-(trifluoromethyl)phenyl]piperidin-4-yl}carbonyl)pyrrolidin-
-3-yl]carbamate (0.975 g, 2.21 mmol) was treated with
trifluoroacetic acid (5 mL) and methylene chloride (5 mL) for 1 h
at room temperature. The solution was concentrated to give 1.75 g
of product which was used for the next step without
purification.
Step E
##STR00168##
[0459]
1-Pyridin-2-yl-4-{[(3S)-1-({1-[3-(trifluoromethyl)phenyl]piperidin--
4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
(3S)-1-({1-[3-(Trifluoromethyl)phenyl]piperidin-4-yl}carbonyl)pyrrolidin--
3-amine bis(trifluoroacetate) (110 mg, 0.20 mmol),
4-hydroxy-4-pyridin-2-ylcyclohexanone (45 mg, 0.24 mmol),
triethylamine (0.082 mL, 0.59 mmol), and sodium
triacetoxybornhydride (83 mg, 0.39 mmol) were mixed in methylene
chloride (6 mL). After being stirred overnight, the reaction
mixture was diluted with EtOAc and washed with saturated
Na.sub.2CO.sub.3. The aqueous layer was extracted with EtOAc three
times. The combined organic layers were dried (MgSO.sub.4),
concentrated and purified by silica gel column (EtOAc to 1%
Et.sub.3N/EtOAc to 5% Et.sub.3N/EtOAc) to provide the title
compound. LCMS (M+H)=517.2.
Example 75
##STR00169##
[0460]
1-(5-pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[3-(trifluoromethyl-
)phenyl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
The title compound was prepared in a manner analogous to that
described for Example 74. MS (M+H) 595.2.
Example 76
##STR00170##
[0461]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[3-(trifluoromethyl-
)phenyl]pyrrolidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
The title compound was prepared in a manner analogous to that
described for Example 74. MS (M+H) 581.2.
Example 77
##STR00171##
[0463]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[3-(trifluoromethyl-
)phenyl]azetidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
The title compound was prepared in a manner analogous to that
described for Example 74. MS (M+H) 567.2.
[0464] Example 78
Step A
##STR00172##
[0466] 1-[(Benzyloxy)carbonyl]piperidine-4-carboxylic acid.
Triethylamine (8.1 ml, 58 mmol) was added to a solution of
piperidine-4-carboxylic acid (5 g, 40 mmol) and benzyl
chloroformate (7.9 g, 46 mmol) in dichloromethane (100 mL) in an
ice-water batb. After being stirred overnight, the solution was
washed-with concentrated HQ arid brioe, dried over Na.sub.2SO.sub.4
and concentrated. Chromatography on silica gel gave the title
compound (10 g) as an oil. MS (M+H) 264.2.
Step B
##STR00173##
[0467] Benzyl
4-({(3S)-3-[(tert-Butoxycarbonyl)amino]pyrrolidin-1-yl}carbonyl)piperidin-
e-1-carboxylate. A mixture of
1-[(benzyloxy))carbonyl]piperidine-4-carboxylic acid (5 g, 20
mmol), tert-butyl (3S)-pyrrolidin-3-ylcarbamate (3.9 g, 21 mmol),
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (9.2 g, 21 mmol), and triethylamine (3.8 g, 38
mmol) in dichloromethane (100 mL) was stirred at room temperature
overnight. The reaction solution was washed with water, dried over
Na.sub.2SO.sub.4, and concentrated. The residue was chromatographed
on silica gel to give 7.5 g of product. MS (M+H) 432.2.
Step C
##STR00174##
[0468] tert-Butyl
[(3S)-1-Piperidin-4-ylcarbonyl)pyrrolidin-3-yl]carbamate. A mixture
of benzyl
4-({(3S)-3-[(tert-butoxycarbonyl)amino]pyrrolidin-1-yl}carbonyl)pi-
peridine-1-carboxylate (7.5 g, 17 mmol) and palladium on carbon
(800 mg, 8 mmol) in methanol (100 mL) was shaken under hydrogen at
50 psi overnight. The mixture was filtered through celite and the
filtrate was concentrated to give 5.1 g of product as a white
solid. MS (M+H) 298.2.
Step D
##STR00175##
[0469] tert-Butyl
[(3S)-1-({1-[6-(Trifluoromethyl)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrr-
olidin-3-yl]carbamate. A solution of
2-chloro-6-(trifluoromethyl)pyridine (1.8 g, 9.9 mmol), tert-butyl
[(3S)-1-(piperidin-4-ylcarbonyl)pyrrolidin-3-yl]carbamate (2.97 g,
10. 0 mmol) and triethylamine (4.1 mL, 30 mmol) in DMF (50 mL) was
heated at 100.degree. C. for 4 hrs. After cooling down, ethyl
acetate was added. The resulting solution was washed with brine
several times, dried over Na.sub.2SO.sub.4 and concentrated. The
residue was chromatographed on silical gel to give the title
compound (1.3 g) as a yellow solid. MS (M+H) 443.2.
Step E
##STR00176##
[0470]
(3S)-1-({1-[6-(Trifluoromethyl)pyridin-2-yl]piperidin-4-yl}carbonyl-
)pyrrolidin-3-amine. tert-Butyl
[(3S)-1-({1-[6-(trifluoromethyl)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrr-
olidin-3-yl]carbamate (1.3 g, 2.9 mmol) was dissolved in a 4 M
solution of HCl in 1,4-dioxane (10 mL). After being stirred at room
temperature for 1 hr, the solution was concentrated to give the
desired product as HCl salt (0.6 g). MS (M+H) 343.1.
Step F
##STR00177##
[0471]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[6-(trifluoromethyl-
)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
A solution of
(3S)-1-({1-[6-(trifluoromethyl)pyridin-P-2-yl]piperidin-4-yl}carbonyl)pyr-
rolidin-3-amine (40 mg, 0.1 mmol),
4-hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexanone (47 mg,
0.18 mmol), sodium triacetoxyborohydride (50 mg, 0.23 mmol), and
triethylamine (35 mg, 0.35 mmol) in dichloromethane (10 mL) was
stirred at room temperature overnight. The reaction mixture was
passed through a silica gel pad. The filtrate was concentrated and
purified by HPLC to give the cis- and trans-isomers. MS (M+H) 596.2
for both isomers. The following examples were prepared in a manner
analogous to that for Example 78.
Example 79
##STR00178##
[0472]
1-Pyridin-2-yl-4-{[(3S)-1-({1-[6-(trifluoromethyl)pyridin-2-yl]pipe-
ridin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS (M+H)
518.2.
Example 80
##STR00179##
[0473]
1-(6-Pyrimidin-2-ylpyridin-3-yl)-4-{[(3S)-1-({1-[6-(trifluoromethyl-
)pyrimidin-4-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol-
. MS (M+H) 597.3.
Example 81
##STR00180##
[0474]
1-Pyridin-2-yl-4-{[(3S)-1-({1-[6-(trifluoromethyl)pyrimidin-4-yl]pi-
peridin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS (M+H)
51.9.2.
Example 82
##STR00181##
[0475]
1-Pyridin-2-yl-4-{[(3S)-1-({1-[4-(trifluoromethyl)pyridin-2-yl]pipe-
ridin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS (M+H)
518.2.
Example 83
##STR00182##
[0476]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl-
)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 596.2.
Example 84
##STR00183##
[0477]
1-[5-(1,3-Oxazol-2-yl)pyridin-2-yl]-4-{[(3S)-1-({1-[4-(trifluoromet-
hyl)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexano-
l. MS (M+H) 584.2.
Example 85
##STR00184##
[0478]
1-(5-Pyrazin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl)p-
yridin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 596.2.
Example 86
##STR00185##
[0479]
1-(5Methylpyridin-2-yl)-4-{[(3S)-1-({1-[4-)trifluoromethyl)pyridin--
2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS
(M+H) 532.2.
Example 87
##STR00186##
[0480]
1-(3,3'-Bipyridin-6-yl)-4-{[(3S)-1-({1-[4-(trifluoromethylpyridin-2-
-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS
(M+H) 595.3.
Example 88
##STR00187##
[0481]
1-(3,4'-Bipyridin-6-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl)pyridin--
2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol. MS
(M+H) 595.3.
Example 89
##STR00188##
[0482]
1-(5-Methoxypyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl)pyridi-
n-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 548.2.
Example 90
##STR00189##
[0483]
1-[5-(Methoxymethyl)pyridin-2-yl]-4-{[(3S)-1-({1-[4-(trifluoromethy-
l)pyridin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 562.2.
Example 91
##STR00190##
[0484]
6-(1-Hydroxy-4-{[(3S)-1-({1-[4-(trifluoromethyl)pyridin-2-yl]piperi-
din-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexyl)nicotinamide. MS
(M+H) 561.3.
Example 92
##STR00191##
[0485]
6-(1-Hydroxy-4-{[(3S)-1-({1-[4-(trifluoromethyl)pyridin-2-yl]piperi-
din-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexyl)-N-methylnicotinamide.
MS (M+H) 575.3.
Example 93
##STR00192##
[0486]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl-
)pyrimidin-2-yl]piperidin-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol-
. MS (M+H) 597.4.
Example 94
##STR00193##
[0487]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[6-(trifluoromethyl-
)pyridin-2-yl]pyrrolidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 582.2
Example 95
##STR00194##
[0488]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[5-(trifluoromethyl-
)pyridin-2-yl]pyrrolidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 582.3.
Example 96
##STR00195##
[0489]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl-
)pyrimidin-2-yl]pyrrolidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexano-
l. MS (M+H) 583.3.
Example 97
##STR00196##
[0490]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({(3R)-1-[4-(trifluorom-
ethyl)pyridin-2-yl]piperidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexa-
nol. MS (M+H) 596.4.
Example 98
##STR00197##
[0491]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({(3S)-1-[4-(trifluorom-
ethyl)pyridin-2-yl]piperidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexa-
nol. MS (M+H) 596.4.
Example 99
##STR00198##
[0492]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl-
)pyridin-2-yl]azetidin-3-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexanol.
MS (M+H) 568.1.
Example 100
[0493] Step A
##STR00199##
Ethyl
1-[4-(Trifluoromethyl)pyridin-2-yl]-1H-imidazole-4-carboxylate. To
a solution of methyl 1H-imidazole-4-carboxylate (417 mg, 3.3 mmol)
in DMF (10 mL) was added sodium hydride (130 mg, 3.3 mmol). After
being stir for 1 h at room temperature,
2-chloro-4-(trifluoromethyl)pyridine (500 mg, 2.8 mmol) was added.
The mixture was stirred at 80.degree. C. overnight. After being
cooled to room temperature, ethyl acetate was added. The solution
was washed with brine several times, dried (MgSO.sub.4) and
concentrated. Chromatography on silica gel eluting with
EtOAc/hexanes (1:1) afforded the title compound (120 mg). MS (M+H)
272.1.
Step B
##STR00200##
[0494]
1-[4-(Trifluoromethyl)pyridin-2-yl]-1H-imidazole-4-carboxylic Acid.
To a solution of methyl
1-[4-(trifluoromethyl)pyridin-2-yl]-1H-imidazole-4-carboxylate (120
mg, 0.44 mmol) in methanol (2.5 mL) was added a 5 M solution of
sodium hydroxide in water (2.5 mL) and the mixture was stirred at
room temperature for 1 h. After removal of methanol under vacuum,
the resulting solution was acidified with concentrated HCl (pH=5)
and concentrated. The residue was taken up in acetone and
insolubles were filtered off. The filtrate was evaporated to give
the title compound (120 mg). MS (M+H)258.2.
Step C
##STR00201##
[0495] tert-Butyl
[(3S)-1-({1-[4-(Trifluoromethyl)pyridin-2-yl]-1H-imidazol-4-yl}carbonyl)p-
yrrolidin-3-yl]carbamate. To a solution of
1-[4-(trifluoromethyl)pyridin-2-yl]-1H-imidazole-4-carboxylic acid
(120 mg, 0.47 mmol) in DMF (3 mL) was added
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (210 mg, 0.47 mmol) followed by triethylamine
(0.20 mL, 1.4 mmol). The reaction was stirred at room temperature
overnight and purified by HPLC to give the title compound. MS (M+H)
426.3.
Step D
##STR00202##
[0496]
1-({(3S)-1-[4-(trifluoromethyl)pyridin-2-yl]-1H-imidazol-4-yl}carbo-
ny)pyrrolidin-3-amine. To a soluton of tert-butyl
[(3S)-1-({1-[4-(trifluoromethyl)pyridin-2-yl]-1H-imidazol-4-yl}carbonyl)p-
yrrolidin-3-yl]carbamate (120 mg, 0.28 mmol) in methanol (2 mL) was
added a 4.0 M soluton of HCl in 1,4-dioxane (3.0 mL). After being
stirred for 0.5 h, the solution was concentrated under vacuum to
give the title compound. MS (M+H) 326.2.
Step E
##STR00203##
[0497]
1-(6-Pyrimidin-2-ylpyridin-3-yl)-4-{[(3S)-1-({1-[4-(trifluoromethyl-
)pyridin-2-yl]-1H-imidazol-4-yl}carbonyl)pyrrolidin-3-yl]amino}cyclohexano-
l. To a soluton of
(3S)-1-({1-[4-(trifluoromethyl)pyridin-2-yl]-1H-imidazol-4-yl}carbonyl)py-
rrolidin-3-amine (50 my, 0.15 mmol) and
4-hydroxy-4-(6-pyrimidin-2-ylpyridin-3-yl)cyclohexanone (41 mg,
0.15 mmol) in methylene chloride (3 mL) was added sodium
triacetoxyborohydride (36 mg, 0.17 mmol) followed by triethylamine
(0.086 mL, 0.61 mmol). After being stirred at room temperature for
2 h, EtOAc (50 mL) was added. The solution was washed with
NaHCO.sub.3 soluton and water, dried (MgSO.sub.4) and concentrated.
Purification by HPLC provided two isomers. MS (M+H) 579.3 for both
isomers.
Example 101
Step A
##STR00204##
[0498] 2-Methyl-4-trifluoromethyl)pyridine 1-Oxide. to a soluton of
2-methyl-4-(trifluoromethyl)pyridine (3.9 g, 24 mm0l) in methylene
chloride (50 mL) was added m-chloroperbenzoic acid (7.0 g, 31
mmol). After being stirred at room temperature over night, the
solution was washed with 50 mL of 1 N NaOH. the water phase was
back-extracted with methylene chloride. The combined organic phases
were dried over Na.sub.2SO.sub.4 and concentrated under vacuum to
give the title compound. MS (M+H) 178.1.
Step B
##STR00205##
[0499] [4-(Trifluoromethyl)pyridin-2-yl]methyl Acetate.
2-Methyl-4-(trifluoromethyl)pyridine 1-oxide (4.0 g, 22 mmol) was
added to acetic anhydride (12 mL) at 120.degree. C. The mixture was
reflued for 1 h. To it was carefully added 10 mL of ethanol. Reflux
was continued for 10 min. the mixture was poured into ice,
neutralized with NaHCO.sub.3, and extracted with Et.sub.2O. The
organic layer was dried (MgSO.sub.4) and concentrated.
Chromatography on silica gel (5:2 hexanes/EtOAc) provided the
product (3.4 g) as a brown oil. MS (M+H) 220.1.
Step C
##STR00206##
[0500] [4-(Trifluoromethyl)pyridin-2-yl]methanol. To a solution of
[4-(trifluoromethyl)pyridin-2-yl]methyl acetate (1.0 g, 3.2 mmol)
in methanol (10 mL) was added a 1.0 M soluton of sodium hydroxide
in water (10 mL). After being stirred at room temperature
overnight, the solution was diluted with 20 mL of water and
extracted with EtOAc twice. The combined organic layers were dried
(MgSO.sub.4) and concentrated under vacuum. Chromatography on
silica gel eluting with hexanes/EtOAc (1.1) afforded the title
compound (0.34 g) as a clear oil. MS (M+H) 178.1.
Step D
##STR00207##
[0501] {[4-(Trifluoromethyl)pyridin-2-yl]methoxy}acetic Acid. to a
solution of [4-(trifluoromethyl)pyridin-2-yl]methanol (340 mg, 1.9
mmol) in DMF (10 mL) was added sodium hydride (150 mg, 3.8 mmol).
After being stirred at room temperature for 5 min.
1,1-dimethylethyl bromoacetate (0.28 mL, 1.9 mmol) was added.
Stirring was continued at room temperature for 1 h. Water (20 mL)
was added and the resulting solution was extracted with EtOAc. The
water layer was neutralized to pH=5 with HCl and extracted with
EtOAc twice. The combined organic layers were dried (MgSO.sub.4)
and concentrated under vacuum to give the title compound which was
used for the next reaction without purification. MS (M+H)
292.2.
Step E
##STR00208##
[0502] tert-Butyl
[(3S)-1-({[4-(Trifluoromethyl)pyridin-2-yl]methoxy}acetyl)pyrrolidin-3-yl-
]carbamate. To a solution of
{[4-(trifluoromethyl)pyridin-2-yl]methoxy}acetic acid (450 mg, 1.9
mmol) and tert-butyl (3S)-pyrrolidin-3-ylcarbamate (360 mg, 1.9
mmol) in DMF (10 mL) was added
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (880 mg, 2.0 mmol) followed by triethylamine
(0.80 mL, 5.7 mmol). After being stirred at room temperature
overnight, ethyl acetate was added. The solution was washed with 1
N NaOH and water. Purification on silica gel column eluting with
EtOAc provided the title compound (300 mg) as a clear oil. MS (M+H)
404.3.
Step F
##STR00209##
[0503]
1-({[4-(Trifluoromethyl)pyridin-2-yl]meth9oxy}acetyl)pyrrolidin-3-a-
mine. To a solution of tert-butyl
[(3S)-1-({[4-(trifluoromethyl)pyridin-2-yl]methoxy}acetyl)pyrrolidin-3-yl-
]carbamate (300 mg, 0.74 mmol) in methanol (3 mL) was added a 4.0 M
soluton of HCl in 1,4-dioxane (6 mL). After being stirred for 0.5 h
at room temperature, the solution was concentrated under vacuum to
give the title compound. MS (M+H) 304.2.
Step G
##STR00210##
[0504]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({[4-(trifluoromethyl)p-
yridin-2-yl]methoxy}acetyl)pyrrolidin-3-yl]amino}cyclohexanol. to a
solution of
(3S)-1-({[4-(trifluoromethyl)pyridin-2-yl]methoxy}acetyl)pyrrolidine-3-am-
ine (47 mg, 0.15 mmol) and
4-hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexanone (41 mg,
0.15 mmol) in methanol (2 mL) and isopropanol (2 mL) was added
sodium triacetoxyborohydride (36 mg, 0.17 mmol). After being
stirred at room temperature overnight, EtOAc was added. The
solution was washed with NaHCO.sub.3 solution and water, dried
(MgSO.sub.4) and concentrated. Purification by HPLC provided two
isomers of the title compoind. MS (M+H) 557.2 for both isomers.
Example 102
##STR00211##
[0505]
1-(5-Pyrimidin-2-ylpyridin-2-yl)-4-{[(3S)-1-({[4-(trifluoromethyl)2-
-phenyl]methoxy}acetyl)pyrrolidin-3-yl]amino}cyclohexanol. The
title compound was prepared in a manner analogous to that for
Example 101. MS (M+H) 556.3.
Pharmaceutical Applications of the Compounds of the Invention
[0506] The capacity of the novel compounds of the invention to
antagonize CCR2 function can be determined using a suitable screen
(e.g., high through-put assay). For example, an agent can be tested
in an extracellular acidification assay, calcium flux assay, ligand
binding assay or chemotaxis assay (see, for example, Hesselgesser
et al., J Biol. Chem. 273(25): 15687-15692 (1998); WO 00/05265 and
WO 98/02151).
[0507] In a practical assay, a CCR2 protein which can be isolated
or recombinantly derived is used which has at least one property,
activity or functional charateristic of a mammalian CCR2 protein.
The specific property can be a binding property (to, for example, a
ligand or inhibitor), a signalling activity (e.g., activation of a
mammalian G protein, induction of rapid and transient increase in
the concentration of cytosolic free calcium [Ca.sup.++]i, cellular
response function (e.g., stimulation of chemotaxis or inflammatory
mediator release by leukocytes), and the like.
[0508] In one embodiment, a composition containing a CCR2 protein
or variant thereof is maintained under conditions suitable for
binding. The CCR2 receptor is contacted, with a compound to be
tested, arid binding is detected, or measured.
[0509] In alternate embodiments, the assay is a cell-based assay
and cells are used which are stably or transiently transfected with
a vector or expression cassette having a nucleic acid sequence
which encodes the CCR2 receptor. The cells are maintained under
conditions appropriate for expression of the receptor and are
contacted with an agent under conditions appropriate for binding to
occur. Binding can be detected using standard techniques. For
example, the extent of binding can be determined relative to a
suitable control. Also, a cellular fraction, such as a membrane
fraction, containing the receptor can be used in lieu of whole
cells.
[0510] Detection of binding or complex formation can be detected
directly or indirectly. For example, the agent can be labeled with
a suitable label (e.g., fluorescent label, label, isotope label,
enzyme label, and the like) and binding can be determined by
detection of the label. Specific and/or competitive binding can be
assessed by competition or displacement studies, using unlabeled
agent or a ligand as a competitor.
[0511] The CCR2 antagonist activity of test agents (e.g., the
3-cycloakylaminopyrrolidine compounds of formula I or II of the
invention) can be reported as the inhibitor concentration required
for 50% inhibition (IC.sub.50 values) of specific binding in
receptor binding assays using .sup.125I-labeled MCP-1, as ligand,
and Peripheral Blood Mononuclear Cells (PBMCs) prepared from normal
human whole blood via density gradient centrifugation. Specific
binding is preferably defined as the total binding (e.g., total cpm
on filters) minus the non-specific binding. Non-specific binding is
defined as the amount of cpm still detected in the presence of
excess unlabeled competitor (e.g., MCP-1).
[0512] The human PBMCs described above can be used in a suitable
binding assay. For example, 200,000 to 500,000 cells can be
incubated with 0.1 to 0.2 nM .sup.125 I-labeled MCP-1, with or
without unlabeled competitor (10 nM MCP-1) or various
concentrations of compounds to be tested, .sup.125I-labeled MCP-1,
can be prepared by suitable methods or purchased from commercial
vendors (Perkin Elmer, Boston Mass.), The binding reactions can be
performed in 50 to 250 .mu.l of a binding buffer consisting of 1M
HEPES pH 7.2, and 0.1% BSA (bovine serum albumin), for 30 min at
room temperature. The blading reactions can be terminated by
harvesting the membranes by rapid filtration through glass fiber
filters (Perkin Elmer) which can be presoaked in 0.3%
polyethyleneimine or Phosphate Buffered Saline (PBS). The filters
can be rinsed with approximately 600 .mu.l of binding buffer
containing 0.5 M NaCl or PBS, then dried, and the amount of bound
radioactivity can be determined by counting on a Gamma Counter
(Perkin Elmer).
[0513] The capacity of compounds to antagonize CCR2 function can
also be determined in a leukocyte chemotaxis assay using suitable
cells. Suitable cells include, Err example, cell lines, recombinant
cells or isolated cells which express CCR2 and undergo CCR2
ligand-induced (e.g., MCP-1) chemotaxis. The assay in use, utilizes
human, peripheral blood mononuclear cells, in a modified Boyden
Chamber (Neuro Probe), 500,000 cells in serum free DMEM media (in
Vitrogen.) are Incubated with or without the inhibitors and warmed
to 37.degree. C. The chemotaxis chamber (Neuro Probe) is also
prewarmed. 400 ul of warmed 10 nM MCP-1 is added to the bottom
chamber in all wells expect the negative control which has DMEM
added. An 8 micron membrane filter (Neuro Probe) is place on top
and the chamber lid is closed. Cells are then added to the holes in
the chamber lid which are associated with the chamber wells below
the filter membrane. The whole chamber is incubated at 37.degree.
C., 5% CO2 for 30 minutes. The cells are then aspirated off the
chanber lid opened, and the filter gently removed. The top of the
filter is washed 3 times with PBS and the bottom is left,
untouched. The filter is air dried and stained with Wright Geirasa
stain (Sigma). Filters are counted by microscopy. The negative
control wells serve as background and are subtracted from all
values. Antagonist potency can be determined by comparing the
number of cell that migrate to the bottom chamber in wells which
contain antagonist, to the number of cells which migrate to the
bottom chamber in MCP-1 control wells.
[0514] When the binding assay protocol is used, the compounds of
the present invention have IC50 in the range of about 0.01 to about
500 (nM). In chemotaxis assays the compounds of the invention have
IC50's in the range of about 1 to about 3000 (nM).
[0515] A method of modulating activity of a chemokine receptor
comprising, contacting said chemokine receptor with a compound of
claim. Chemokine receptors to which the present compounds bind
and/or modulate include any chemokine receptor. In some
embodiments, the chemokine receptor belongs to the CC family of
chemokine receptors including, for example, CCR1, CCR2, CCR3, CCR4,
CCRS, CCR6, CCR7, and CCR8. In some embodiments, the chemokine
receptor is CCR2. In some embodiments, the chemokine receptor is
CCR5. In some embodiments, the chemokine receptor binds and/or
modulates both CCR2 and CCR5.
[0516] As used herein, the term "contacting" refers to the bringing
together of indicated moieties in an in vitro system or an in vivo
system. For example, "contacting" the chemokine receptor with a
compound of the invention includes the administration of a compound
of the present invention to an individual or patient, such as a
human, having a chemokine receptor, as well as, for example,
introducing a compound of the invention into a sample containing a
cellular or purified preparation containing the chemokine
receptor.
[0517] The compounds of the invention can be selective. By
"selective" is meant that a compound binds to or inhibits a
chemokine receptor with greater affinity or potency, respectively,
compared to at least one other chemokine receptor, or preferably
compared to all other chemokine receptors of the same class (e.g.,
all othe CC-type receptors). In some embodiments, the compounds of
the invention have binding or inhibition selectivity for CCR2 or
CCR5 over any other chemokine receptor. Selectivity can be at least
about 10-fold. at least about 20-fold, at least about 50-fold, at
least about 100-fold, at least about 200-fold, at least about
500-fold or at, least about 1000-fold. Binding affinity and
inhibitor potency can be measured according to routine methods its
die art, such as according to the assays provided herein.
[0518] The present invention further provides methods of treating a
chemokine receptor-associated disease or disorder in an individual
(e.g., patient) by administering to the individual in need of such
treatment a therapeutically effective amount or dose of a compound
of the present invention or a pharmaceutical composition thereof. A
chemokine receptor-associated disease can include any disease,
disorder or condition that is directly or indirectly linked to
expression or activity of the chemokine receptor. A chemokine
receptor-associated disease can also include arty disease, disorder
or condition that can be prevented, ameliorated, or cured by
modulating chemokine receptor activity. A chemokine
receptor-associated disease can farther include any disease,
disorder or condition that is characterized by binding of an
infectious agent such as a virus or viral protein with a chemokine
receptor. In some embodiments, the chemokme receptor-associated
disease is a CCR5-associated disease such as HIV infection.
[0519] As used herein, the term "individual" or "patient," used
interchangeably, refers to any animal, including mammals,
preferably mice, rats, other rodents, rabbits, dogs, cats, swine,
cattle, sheep, horses, or primates, and most preferably humans. The
compounds of the invention can be administered to a mammal, such,
as a human, but can also be other mammals such as an animal in need
of veterinary treatment, e.g., domestic animals (e.g., dogs, cats,
and the like), farm animals (e.g., cows, sheep, pigs, horses, and
the like) and laboratory animals (e.g., rats, mice, guinea pigs,
and the like). The mammal treated in the methods of the invention
is a mammal, male or female, in whom modulation of chemokine
receptor activity is desired. The term modulation is intended to
encompass antagonism (e.g., inhibition), agonism, partial
antagonism, and/or partial agonism. In some embodiments, compounds
of the present invention are antagonists (e.g., inhibitors) of
chemokine receptors.
[0520] In the present specification, the term "therapeutically
effective amount" means the amount of the subject compound that
will elicit the biological or medical response of a tissue, system,
animal or human that is being sought by the researcher,
veterinarian, medical doctor or other clinician.
[0521] The compounds of the invention are administered in
therapeutically effective amounts to treat a disease for example
such as rheumatoid arthritis. A therapeutically effective amount of
a compound is that amount which results in the inhibition of one or
more of the processes mediated by the binding of a chemokine to a
receptor such as CCR2 in a subject with a disease associated with
aberrant leukocyte recruitment and/or activation. Typical examples
of such processes include leukocyte migration, integrin activation,
transient increases in the concentration of intracellular free
calcium [Ca.sup.2+]i and granule release of proinflammatory
mediators. Alternatively, a therapeutically effective amount of a
compound is the quantity required to achieve a desired therapeutic
and/or prophylactic effect, such as an amount which results in the
prevention of or a decrease in the symptoms associated with a
disease associated with aberrant leukocyte recruitment and/or
activation.
[0522] Additional diseases or conditions of human or other species
which can be treated with the inhibitors or modulators of chemokine
receptor function of the invention, 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, Sjogren'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 grafs-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, repertusion injury, atherosclerosis,
restenosis, certain hematologic malignancies, cytokine-induced
toxicity (e.g., septic shock, endotoxic shock), polymyositis,
dermatomyositis.
[0523] In some embodiments, the chemokine receptor-associated
diseases, disorders and conditions include inflammation and
inflammatory diseases, immune disorders, cancer, and viral
infections. Example inflammatory diseases include diseases having
an inflammatory component such us asthma, allergic rhinitis,
restenosis, atherosclerosis, multiple sclerosis, Crohn's disease,
ulcerative colitis, hypersensitivity lung diseases, neuropathic
pain, hypersensitivity pneumonitis, eosinophilic pneumonias,
delayed-type hypersensitivity, asthma, interstitial lung disease
(ILD) (e.g., idiopathic pulmonary fibrosis, or ILD associated with
rheumatoid arthritis, systemic lupus erythematosus, ankylosing
spondylitis, systemic sclerosis, Sjogren's syndrome, polymyositis
or dermatomyositis), eye disorders (e.g., retinal
neurodegeneration, choroidal neovascularization, etc.), obesity,
and the like. Example immune disorders include rheumatoid
arthritis, psoriatic arthritis, systemic lupus erythematosus,
myastenia gravis, diabetes (e.g., juvenile onset diabetes), insulin
resistance; glomerulonephritis, autoimmune throiditis, organ
transplant rejection including allograft rejection and
graft-versus-host disease. Example cancers include cancers such as
breast cancer, ovarian cancer, multiple myeloma and the like that
are characterised by infiltration of macrophages (e.g., tumor
associated macrophages, TAMs) into tumors or diseased tissues.
Example viral infections include HIV infection.
[0524] One or more additional pharmaceutical agents such as, for
example, antibodies, anti-inflammatory agents, immunosuppressants,
chemotherapeutics can be used in combination with the compounds of
the present invention for treatment of chemokine
receptor-associated diseases, disorders or conditions. The agents
can be combined with the present compounds in a single dosage form,
or the agents can be administered simultaneously or sequentially as
separate dosage forms.
[0525] One or more additional pharmaceutical agents such as, for
example, anti-viral agents, antibodies, anti-inflammatory agents,
and/or immunosuppressants can be used in combination with the
compounds of the present invention for treatment of chemokine
receptor-associated diseases, disorders or conditions. Tire agents
can be combined with the present compounds in a single dosage form,
or the agents can be administered simultaneously or sequentially as
separate dosage forms.
[0526] Suitable antiviral agents contemplated for use in
combination with the compounds of the present invention can
comprise nucleoside and nucleotide reverse transcriptase inhibitors
(NRTIs) non-nucleoside reverse transcriptase inhibitors (NNRTIs),
protease inhibitors and other antiviral drugs.
[0527] Example suitable NRTIs include zidovudine (AZT); didanosine
(ddl); zalcitabine (ddC); stavudine (d4T); lamivudine (3TC);
abacavir (1592U89); adefovir dipivoxil [bis(POM)-PMEA]; lobucavir
(BMS-180194); BCH-10652, emitricitabine [(-)-FTC]; beta-L-FD4 (also
called beta-L-D4C and named bets-L-2 ',
3'-dicleoxy-5-fluoro-cytidene); DAPD,
((-)-beta-D-2,6,-diamino-purine dioxolane); and lodenosine
(EddA).
[0528] Typical suitable NNRTIs include nevirapine (BI-RG-587);
delaviradine (BHAP, U-90152); efavirenz (DMP-266); PNU-142721;
AG-1549; MKC-442
(1-(ethoxy-methyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(1H,3H--
pyrimidi nedione); and (+)-calanolide A (NSC-675451) and B.
[0529] Typical suitable protease inhibitors include saquinavir (Ro
31-8959); ritonavir (ABT-538); indinavir (MK-639); nelfnavir
(AG-1343); amprenavir (141W94); lasinavir (BMS-234475); DMP-450;
BMS-2322623, ABT-378; and AG-1 549.
[0530] Other antiviral agents include hydroxyurea, ribavirin, IL-2,
IL-12, pentafuside and Yissum Project No. 11607.
[0531] In some embodiments, anti-inflammatory or analgesic agents
contemplated for use in combination with the compounds of the
present invention can comprise, for example, an opiate agonist, a
lipoxygenase inhibitor such as an inhibitor of 5-lipoxygenase, a
cyclooxygenase inhibitor such as a cyclooxygenase-2 inhibitor, an
interleukin inhibitor such as an interleukin-1 inhibitor, an NNMA
antagonist, an inhibitor of nitric oxide or an inhibitor of the
synthesis of nitric oxide, a non-steroidal antiinflammatory agent,
or a cyctokine-suppressing antiinflammatory agent, for example,
such as acetaminophen, asprin, codiene, fentanyl, ibuprofen,
indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam,
a steroidal analgesic, sufentanyl, sunlindac, tenidap, and the
like. Similarly, the instant compounds can 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,
propylhexedfine, or levo-desoxyephedrine; an antitussive such as
codeine, hydrocodone, caramiphen, carbetapentane, or
dextramethorphan; a diuretic; and a sedating or non-sedating
antihistamine.
[0532] In some embodiments, pharmaceutical agents contemplated for
use in combination with the compounds of the present invention can
comprise, (a) VLA-4 antagonists such as those described in U.S.
Pat. No. 5,510,332, W095/15973, W096/01644, W096/06108, W096/20216,
W096/229661, W096/31206, W096/4078, W097/O30941, W097/022897 WO
98/426507 W098/53814, W098/53817, W098/538185, W098/54207, and
W098/58902; (b) steroids such as beclornethasone,
methylpi-ednisoione, betamethasone, prednisone, dexamethasone, and
hydrocortisone; (e) immunosuppressants such as cyclosporin ,
tacrolimus, rapamycin and other FK506 type immunosuppressants; (d)
antihistamines (H1-histamine antagonists) such as bromopheniramine,
chlorpheniramine, dexchlorpheniramine, triproiidine, clemastine,
diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine,
methdilazine, promethazine, trimeprazine, azatadine,
cyproheptadine, antazoline, pheniramine pyrilarnine, asternizole,
terfenadine, loratadine, cetirizine, fexofenadine,
desearboethoxyloratadine, and the like; (e) non-steroidal
anti-asthmatics such as terbutaline, metaproterenol, fenoterol,
isoethaiine, albuterol, bitolterol, pirbuterol, theophylline,
cromolyn sodium, atropine, ipratropium bromide, leukotriene
antagonists (e.g., zafirlnkast, montelukast, pranlukast, iralukast
pobilukast, SKB-106,203), leukotriene biosynthesis inhibitors
(e.g., zileuton, BAY-1005); (f) nonsteroidal antiinflammatory
agents (NSAIDs) such as propionic acid derivatives (e.g.,
aiminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen,
fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen,
ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen,
pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen), acetic
acid derivatives (e.g., indomethacin, acernetacin, alclofenac,
clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac,
furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac,
tolmetin, zidometacin, and zomepirac), fenamic acid derivatives
(flufenarnic acid, meclofenamic acid, mefenamic acid, niflumic acid
and tolfenarnic acid), biphenylcarboxylic acid derivatives
(diflunisal and flufenisal), Oxicarns (isoxicarn, 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, especially CXCR-4, CCR1, CCR2, CCR3 and CCR5;
(j) cholesterol lowering agents such as HMG-CoA reductase
inhibitors (lovastatin, sirrivastatin and pravastatin, fluvastatin,
atorvastatin, and other statins), sequestrants (cholestyramine and
colestipol), nicotinic acid, fenofibric acid derivatives
(gemfibrozil, clofibrat, fenofibrate and benzafibrate), and
probucol; (k) anti-diabetic agents such as insulin, sulfonylureas,
biguanides (metformin), U-glucosidase inhibitors (acarbose) and
orlitazones (troglitazone and pioglitazone); (1) preparations of
interferon beta (interferon beta-lo, interferon-beta-1 P); (m)
other compounds such as aminosalicylic acids, antimetabolites such,
as azathioprine and 6-mercaptopurine, and cytotoxic cancer
chemotherapeutic agents. The weight ratio of the compound of the
compound of the present invention to the second active ingredient
may be varied and will depend upon the effective dose of each
ingredient.
[0533] Rheumatoid arthritis (RA) patients, treated aggressively
with disease modifying agents (methotrexate, antimalarials, gold,
penicillamine, sulfasalazine, dapsone, leflunamide, or
bloiogicals), can achieve varying degrees of disease control,
including complete remissions. These clinical responses are
associated with improvement in standardized scores of disease
activity, specifically the ACR criteria which includes: pain,
function, number of tender joints, number of swollen joints,
patient global assessment, physician global assessment, laboratory
measures of inflammation (CRP and ESR), and radiologic assessment
of joint structural damage. Current disease-modifying drugs
(DMARDs) require continued administration to maintain optimal
benefit. Chronic dosing of these agents is associated with
significant toxicity and host defense compromise. Additionally,
patients often become refractory to a particular therapy and
require an alternative regimen. For these reasons, a novel,
effective therapy which allows withdrawal of standard DMARDs would
be a clinically important advance.
[0534] Patients with significant response to anti-TNF therapies
(infliximab, etanercept, adalimumab), anti-IL-1 therapy (kinaret)
or other disease modifying anti-rheumatic drugs (DMARDs) including
but not limited to methotrexate, cyclosporine, gold salts,
antimalarials, penicillamine or leflunamide, who have achieved
clinical remission of disease can be treated with a substance that
inhibits expression and/or activity of CCR2 including, for example,
nucleic acids (e.g., antisense or siRNA molecules), proteins (e.g.,
anti-CCR2 antibodies), small molecule inhibitors (e.g., the
compounds disclosed herein and other chemokine receptor inhibitors
known in the art).
[0535] In some embodiments, the substance that inhibits expression
and/or activity of CCR2 is a small molecule CCR2 inhibitor (or
antagonist). The CCR2 antagonist can be dosed orally q.d. or b.i.d
at a dose not to exceed about 500 mgs a day. The patients can be
withdrawn from or have a decrease In the dosage of their current
therapy and would be maintained on treatment with she CCR2
antagonist. Treating patients with a combination of CCR2 antagonist
and their current therapy can be carried out for, for example,
about one to about two days, before discontinuing or dose reducing
the DMARD and continuing on CCR2 antagonist.
[0536] Advantages of substituting traditional. DMARDS with CCR2
antagonists are numerous. Traditional DMARDs have serious
cumulative dose-limiting side effects, the most common being damage
to the liver, as well as immunosuppressive actions. CCR2 antagonism
is expected to have an improved long-term safety profile and will
not have similar immunosuppressive liabilities associated with
traditional DMARDs. Additionally, the half-life of the biologicals
is typically days or weeks, which is an issue when dealing with
adverse reactions. The half-life of an orally bioavailable CCR2
antagonist is expected to be on the order of hours so the risk of
continued exposure to the drug after an adverse event is very
minimal as compared to biological agents. Also, the current
biologic agents (infliximab, etanercept, adalimumab, kinaret) are
typically given either i.v. or s.c, requiring doctor's
administration or patient self-injection. This leads to the
possibility of infusion reaction or injection site reactions. These
are avoidable using an orally administered CCR2 antagonist.
[0537] The compounds of the 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, ah 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.
[0538] 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 metabolic stability, rate
of excretion, drug combination, and length of action of that
compound 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
specific route of administration, the renal and hepatic function of
the patient, and the desired effect. A physician or veterinarian
can determine and prescribe the effective amount of the drug
required to prevent, counter, or arrest, the progress of the
specific disorder for which treatment is necessary.
[0539] Generally, the daily oral dosage of each active ingredient,
when used for the indicated effects, will range between about
0.0001 to 1000 mg/kg of body weight, preferably between about 0.001
to 100 mg/kg of body weight per day, and most preferably between
about 0.1 to 20 mg/kg/day. For intravenous use, the most preferred
doses will range from about 0.1 to about 10. mg/kg/minute during a
constant rate infusion. For oral administration, the compositions
are preferably provided in the form of tablets containing 1.0 to
1000 milligrams of the active ingredient, particularly 1.0, 5.0,
10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0,
300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 000.0, and 1000.0
milligrams of the active ingredient, for the symptomatic adjustment
of the dosage to the patient to be treated. The compounds may be
administered on a regimen of 1 to 4 times per day, preferably once
or twice per day.
[0540] The compounds of the instant invention can also be
administered in intranasal form via topical use of suitable
intranasal vehicles, or via transdermal routes, e.g., by using
transdermal skin patches. When administered in the form of a
transdermal delivery system, the dosage administration can be
continuous rather than intermittent throughout the dosage
regimen.
[0541] The compounds of the invention are typically administered in
admixture with suitable pharmaceutical diluents, excipients, or
carriers (collectively referred, to herein as pharmaceutical
carriers) suitably selected with respect to tire intended form of
administration, that Is, oral tablets, capsules, elixirs, syrups
and die like, and consistent with conventional pharmaceutical
practices.
[0542] 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, pharniaceuticaLly acceptable. Inert carrier
such as lactose, starch, sucrose, glucose, methyl cellulose,
magnesium stearate, dicalium 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. Additionally, when desired or
necessary, suitable hinders, lubricants, disintegrating agents, and
coloring agents can also be incorporated into the mixture. Suitable
hinders 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 benaoate, sodium acetate, sodium chloride, and the
like. Disintegrators include, without limitation, starch, methyl
cellulose, agar, bentonite, xanthan gum, and the like.
[0543] The compounds of the present invention can also be provided
to a patient in the form of liposome delivery systems, such as
small unilameliar vesicles, large unilamellar vesicles, and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as cholesterol, stearylamine, or
phosphatidylcholines.
[0544] The 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 poly-ethyleneoxide-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, polylactlc acid, polyglycolic acid, copolymers of
polylactic and polyglycolic acid, polyepsilon caprolactone,
polyhydroxy butyric acid, polyorthoesters, polyacetals,
polydihydropyrans, and crosslinked or amphipathic block copolymers
of hydrogels.
[0545] Dosage forms for the compounds of the invention suitable for
administration may contain from about 0.1 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.
[0546] Gelatin capsules can also be used as dosage forms and 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.
[0547] When using liquid dosage forms for oral administration they
can contain coloring and flavoring to increase patient
acceptance.
[0548] Generally, 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
preferably contain a water soluble salt of the active ingredient,
suitable stabilizing agents, and if necessary, buffer substances.
Aniloxidizing agents such as sodium bisulfite, 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. Suitable pharmaceutical carriers are described in
Remington's Pharmaceutical Sciences, Mack Publishing Company, a
standard reference text in the field of pharmacology. The
pharmaceutical compositions of the invention may also be in the
form of oil-in-water emulsions. The oily phase may be a vegetable
oil, for example olive oil or arachis oil, or a mineral oil, for
example liquid paraffin or mixtures of these. Suitable emulsifying
agents may be naturally-occurring gums, for example gum acacia or
gum tragacanth, naturally-occurring phosphatides, for example soy
bean, lecithin, and esters or partial esters derived from fatty
acids and hexitol anhydrides, for example sorbitan monooleate, and
condensation products of the said partial esters with ethylene
oxide, for example polyoxy ethylene sorbitan monooleate. The
emulsions may also contain sweetening and flavoring agents.
[0549] The compounds of the present invention may also be
administered in the form of suppositories for rectal administration
of the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient which is solid at ordinary
temperatures bar liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols.
[0550] For topical use, creams, ointments, jellies, solutions or
suspensions, etc., containing the compounds of the present
invention are employed. As used herein, topical application is also
meant to include the use of mouth washes and gargles.
[0551] The pharmaceutical compositions and methods of the present
invention may further comprise other therapeutically active
compounds which are usually applied in the treatment of the above
mentioned pathological conditions.
[0552] Representative useful pharmaceutical dosage-forms for
administration of the compounds of this invention can be
illustrated as follows:
Capsules
[0553] A large number of unit capsules can be prepared by filling
standard two-piece hard gelatin capstdes each with 50 milligrams of
powdered active ingredient, 100 milligrams of lactose, 25
milligrams of cellulose, and 3 milligrams magnesium stearate.
Soft Gelatin Capsules
[0554] 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
farm soil, gelatin capsules containing 75 milligrams of the active
ingredient. The capsules should be washed and dried.
Tablets
[0555] Tablets may be prepared by conventional procedures so that
the dosage unit is 75 milligrams of active ingredient, 0.15
milligrams of colloidal silicon dioxide, 4 milligrams of magnesium
stearate, 250 milligrams of microcrystalhnc cellulose, 9 milligrams
of starch and 75 milligrams of lactose. Appropriate coatings well
known to one skilled in the art may be applied to increase
palatabiiity or delay absorption.
Injectable
[0556] A parenteral composition suitable for administration by
injection may be prepared by stirring 1.0% by weight of active
ingredient in 8% by volume propylene glycol and water. The solution
should be made isotonic with sodium chloride and sterilized.
Suspension
[0557] An aqueous suspension can be prepared for oral
administration so that each 5 mL contain 75 mg of finely divided
active ingredient, 150 mg of sodium carboxymethyl cellulose, 3.75
mg of sodium benzoate, 0.75 g of sorbitol solution, U.S.P., and
0.015 mL, of vanillin.
Example A
[0558] This example describes a procedure to evaluate the efficacy
of CCR2 antagonists for treatment of rheumatoid arthritis.
[0559] An animal model of rheumatoid arthritis can be induced in
rodents by injecting them with type II collagen in selected
adjuvants. Three series of rodent groups consisting 15
genetically-susceptible mice or rats per group are injected
sub-cutaneously or intra-dermally with type II collagen emulsified
in Complete Freund's Adjuvant at days 0 and 21. One series of
rodents additionally receives phosphate buffered saline (PBS) and
Tween 0.5% i.p. at the initial sensitization, and at different
dosing schedules thereafter. A second series consists of groups of
rodents receiving different doses of the CCR2 antagonists) given
either intra-peritoneally, intravenously, sub-cutaneously,
intra-muscularly, orally, or via any other mode of administration
at the initial sensitization, and at different dosing schedules
thereafter. A third series of rodents, serving as positive control,
consists of groups treated with either mouse IL-10 i.p., or
anti-TNF antibodies i.p. at the initial sensitization, and at
different dosing schedules thereafter.
[0560] Animals are monitored from weeks 3 til 8 for the development
of swollen joints or paws, and graded on a standard disease
severity scale. Disease severity is confirmed by histological
analysis of joints.
[0561] Another aspect of the present invention relates to
radio-labeled compounds of the invention that would be useful not
only in radio-imaging but also in assays, both, in vitro and in
vivo, for localising and quantitating the chemokine receptor in
tissue samples, including human, and for identifying chemokine
receptor ligands by inhibition binding of a radio-labeled compound.
Accordingly, the present invention includes, chemokine receptor
assays that contain such radio-labeled compounds.
[0562] The present invention further includes isotopically-isbeled
compounds of the invention. An "isotopically" or "radio-labeled"
compound is a compound of the invention where one or more atoms are
replaced or substituted by an atom having an atomic mass or mass
number different from tire atomic mass or mass number typically
found in nature (i.e., naturally occurring). Suitable radionuclides
that may be incorporated in compounds of the present invention
include but are not limited to .sup.2H (also written as D for
deuterium), .sup.3H (also written as T for tritium), .sup.11C,
.sup.13C, .sup.14C, .sup.--N, .sup.15N, .sup.15O, .sup.17O,
.sup.18O, .sup.18F, .sup.35S, .sup.36Cl, .sup.82Br,.sup.75Br,
.sup.76Br, .sup.77Br, .sup.123I, .sup.124I, .sup.125I and
.sup.131I. The radionuclide that is incorporated in the instant
radio-labeled compounds will depend on the specific application, of
that radio-labeled compound. For example, for in vitro chemokine
receptor labeling and competition assays, compounds that
incorporate .sup.3H, .sup.14C, .sup.82Br, .sup.125I, .sup.131I,
.sup.35S or will generally be most useful. For radio-imaging
applications .sup.11C, .sup.18F, .sup.125I, .sup.123, .sup.124I,
.sup.131I, .sup.75Br, .sup.76Br or .sup.77Br will generally be most
useful.
[0563] It is understood that a "radio-labeled" or "labeled
compound" is a compound that has incorporated at least one
radionuclide. In some embodiments the radionuclide is selected from
the group consisting of .sup.3H, .sup.14C, .sup.125I, .sup.35S and
.sup.82Br.
[0564] Synthetic methods for incorporating radio-isotopes into
organic compounds are applicable to compounds of the invention and
are well known in the art.
[0565] A radio-labeled compound of the Invention can be used in a
screening assay to identify/evaluate compounds, in general terms, a
newly synthesized or identified compound (i.e., test compotmd) can
be evaluated for its ability to reduce binding of fee radio-labeled
compound of the invention to the chemokine receptor. Accordingly,
the ability of a test compound to compete with the radio-labeled
compound for binding to the chemokine receptor directly correlates
to its binding affinity.
[0566] The present invention also includes pharmaceutical kits
useful- for example, in. the treatment or prevention of
chernoklne-associated diseases which include one or more containers
containing a pharmaceutical composition comprising a
therapeutically effective amount of a compound of Formula I. Such
kits can further include, if desired, one or more of various
conventional pharmaceutical kit components, such, as, for example,
containers with one or more pharmaceutically acceptable carriers,
additional containers, etc., as will be readily apparent to those
skilled in the art. Instructions, either as inserts or as labels,
indicating quantities of the components to be administered,
guidelines for administration, and/or guidelines for mixing the
components. Can also be included in the kit.
[0567] All publications, patents, and patent applications including
all cited art and bibliographic references cited herein are hereby
incorporated by reference in their entirety for all purposes.
[0568] While the many forms of the invention herein disclosed
constitute presently preferred embodiments, many others are
possible and further details of die preferred embodiments and other
possible embodiments are not to be construed as limitations. It is
understood that the terms used herein are merely descriptive rather
than limiting and thai various changes many equivalents may be made
without departing from the spirit or scope of the claimed
Invention.
* * * * *