U.S. patent application number 15/820008 was filed with the patent office on 2018-09-06 for azaindazole compounds and methods of use.
The applicant listed for this patent is CHEMOCENTRYX, INC.. Invention is credited to Wei CHEN, Mark M. GLEASON, Kevin Lloyd GREENMAN, Manmohan R. LELETI, Lianfa LI, Yandong LI, Andrew M.K. PENNELL, John J. Kim WRIGHT, Yuan XU, Yibin ZENG, Penglie ZHANG.
Application Number | 20180250293 15/820008 |
Document ID | / |
Family ID | 40076726 |
Filed Date | 2018-09-06 |
United States Patent
Application |
20180250293 |
Kind Code |
A1 |
ZHANG; Penglie ; et
al. |
September 6, 2018 |
AZAINDAZOLE COMPOUNDS AND METHODS OF USE
Abstract
Compounds are provided that act as potent antagonists of the
CCR1 receptor, and have in vivo anti-inflammatory activity. The
compounds are generally aryl piperazine derivatives and are useful
in pharmaceutical compositions, methods for the treatment of
CCR1-mediated diseases, and as controls in assays for the
identification of competitive CCR1 antagonists.
Inventors: |
ZHANG; Penglie; (Foster
City, CA) ; PENNELL; Andrew M.K.; (San Francisco,
CA) ; WRIGHT; John J. Kim; (Redwood City, CA)
; CHEN; Wei; (Fremont, CA) ; LELETI; Manmohan
R.; (West San Jose, CA) ; LI; Yandong; (San
Diego, CA) ; LI; Lianfa; (Palo Alto, CA) ; XU;
Yuan; (Fremont, CA) ; GLEASON; Mark M.;
(Redwood City, CA) ; ZENG; Yibin; (Foster City,
CA) ; GREENMAN; Kevin Lloyd; (Sunnyvale, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CHEMOCENTRYX, INC. |
Mountain View |
CA |
US |
|
|
Family ID: |
40076726 |
Appl. No.: |
15/820008 |
Filed: |
November 21, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15050691 |
Feb 23, 2016 |
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15820008 |
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13774938 |
Feb 22, 2013 |
9296740 |
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15050691 |
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12829312 |
Jul 1, 2010 |
8383630 |
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13774938 |
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11752201 |
May 22, 2007 |
7777035 |
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12829312 |
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11474132 |
Jun 22, 2006 |
7524845 |
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11752201 |
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60693525 |
Jun 22, 2005 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
A61P 19/02 20180101; A61K 31/5377 20130101; A61P 43/00 20180101;
A61K 31/496 20130101; A61P 25/00 20180101; A61P 1/00 20180101; A61P
29/00 20180101; A61P 25/28 20180101; A61P 17/00 20180101; A61P
25/16 20180101; A61P 37/08 20180101; C07D 487/04 20130101; C07D
471/04 20130101 |
International
Class: |
A61K 31/496 20060101
A61K031/496; A61K 45/06 20060101 A61K045/06; A61K 31/5377 20060101
A61K031/5377; C07D 471/04 20060101 C07D471/04; C07D 487/04 20060101
C07D487/04 |
Claims
1.-75. (canceled)
76. A method of assaying a small organic molecule for CCR1
antagonistic activity, said method comprising (a) contacting the
small organic molecule with cells expressing CCR1 and a radioactive
CCR1 ligand to form a reaction mixture; (b) aspirating the reaction
mixture onto a GF/B glass filter pre-soaked in a 0.3%
polyethyleneimine solution; (c) measuring the amount radioactivity
remaining on the GF/B glass filter, wherein said method comprises
performing steps (a)-(d) with a positive control sample having a
formula selected from the group consisting of ##STR00193## or a
pharmaceutically acceptable salt, hydrate or N-oxide thereof,
wherein the subscript m is an integer of from 0 to 2; each R.sup.1
is independently selected from the group consisting of --CO.sub.2H
and C.sub.1-4 alkyl, optionally substituted with --OH, --OR.sup.m,
--S(O).sub.2R.sup.m, --CO.sub.2H or --CO.sub.2R.sup.m wherein
R.sup.m is an unsubstituted C.sub.1-6 alkyl; R.sup.2a, R.sup.2c and
R.sup.2d are each independently selected from the group consisting
of hydrogen, halogen, cyano, oxazolyl, --NO.sub.2,
--CO.sub.2R.sup.c, --CONR.sup.cR.sup.d, --C(O)R.sup.c,
--S(O).sub.2R.sup.e, --R.sup.e, --C(NOR.sup.c)R.sup.d, --OR.sup.c,
--SR.sup.c, --NR.sup.dC(O)R.sup.c, --X.sup.2OR.sup.c,
--O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--NR.sup.cS(O).sub.2R.sup.e, --OC(O)R.sup.c, and --NR.sup.cR.sup.d;
wherein within each of R.sup.2a, R.sup.2c and R.sup.2d, X.sup.2 is
C.sub.1-4 alkylene and each R.sup.c and R.sup.d is independently
selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, and
C.sub.3-6 cycloalkyl and each R.sup.e is independently selected
from the group consisting of C.sub.1-8 alkyl, C.sub.1-8 haloalkyl,
and C.sub.3-6 cycloalkyl; each of ring vertices a, b, c and d in
formulae Ia and Ib is independently selected from N and CH, and one
of said ring vertices is N; and R.sup.3a is selected from the group
consisting of hydrogen, halogen, --NR.sup.fR.sup.g, --R.sup.h,
--S(O).sub.2R.sup.h, and --Y, wherein Y is selected from the group
consisting of homopiperidinyl, morpholinyl, thiomorpholinyl,
pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl,
piperazinyl, phenyl, thienyl, furanyl, pyridyl, pyrimidinyl,
pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl,
isoxazolyl, isothiazolyl, triazolyl, tetrazolyl and oxadiazolyl,
optionally substituted with from one to three substituents selected
from the group consisting of halogen, --OR.sup.f,
--NR.sup.fR.sup.g, --R.sup.h, --CN, --NO.sub.2, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, and --C(O)R.sup.f, wherein each R.sup.f and
R.sup.g is independently selected from hydrogen, C.sub.1-8 alkyl,
C.sub.1-8 haloalkyl, and C.sub.3-6 cycloalkyl, and each R.sup.h is
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8 haloalkyl, and C.sub.3-6 cycloalkyl.
77. The method of claim 76, wherein said cells expressing CCR1 are
THP-1 cells or isolated human monocytes.
78. The method of claim 76, wherein said radioactive CCR1 ligand is
MIP-1.alpha., MPIF-1, Leukotactin, or a combination thereof.
79. The method of claim 76, wherein said wash buffer comprises 25
mM Hepes, 500 mM NaCl, 1 mM CaCl.sub.2, 5 mM MgCl.sub.2, pH
7.1.
80. The method of claim 76, wherein said measuring comprises adding
scintillation fluid to the aspirated and washed GF/C glass
filter.
81. The method of claim 76, wherein R.sup.3a is a member selected
from the group consisting of homopiperidinyl, morpholinyl,
thiomorpholinyl, pyrrolidinyl, piperidinyl, azetidinyl, pyranyl,
tetrahydrofuranyl and piperazinyl.
82. The method of claim 76, wherein R.sup.3a is a member selected
from the group consisting of oxazolyl, oxadiazolyl, imidazolyl,
pyrazolyl, triazolyl and thiazolyl.
83. The method of claim 76, wherein m is 0.
84. The method of claim 76, wherein m is 1.
85. The method of claim 83, having formula Ia.
86. The method of claim 83, having formula Ib.
87. The method of claim 76, wherein R.sup.3a is selected from the
group consisting of phenyl, pyridyl, pyrimidinyl, and
pyrazinyl.
88. The method of claim 76, wherein m is 0 or 1; and R.sup.2a is
hydrogen.
89. The method of claim 76, wherein R.sup.2a is selected from the
group consisting of hydrogen, F, Cl, Br and I.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 15/050,691, filed Feb. 23, 2016, which is a continuation of
U.S. application Ser. No. 13/774,938, filed Feb. 22, 2013 (now U.S.
Pat. No. 9,296,740), which is a continuation of U.S. application
Ser. No. 12/829,312, filed Jul. 1, 2010 (now U.S. Pat. No.
8,383,630), which is a continuation of U.S. application Ser. No.
11/752,201, filed May 22, 2007 (now U.S. Pat. No. 7,777,035), which
is a continuation-in-part application of U.S. application Ser. No.
11/474,132, filed Jun. 22, 2006 (now U.S. Pat. No. 7,524,845) which
claims priority to U.S. Provisional Application No. 60/693,525
filed on Jun. 22, 2005, the contents of each are incorporated
herein in their entirety for all purposes.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH OR DEVELOPMENT
[0002] NOT APPLICABLE
REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER PROGRAM
LISTING APPENDIX SUBMITTED ON A COMPACT DISK
[0003] NOT APPLICABLE
BACKGROUND OF THE INVENTION
[0004] The present invention provides compounds, pharmaceutical
compositions containing one or more of those compounds or their
pharmaceutically acceptable salts, which are effective in
inhibiting the binding of various chemokines, such as MIP-1.alpha.,
leukotactin, MPIF-1 and RANTES, to the CCR1 receptor. As
antagonists or modulators for the CCR1 receptor, the compounds and
compositions have utility in treating inflammatory and immune
disorder conditions and diseases.
[0005] Human health depends on the body's ability to detect and
destroy foreign pathogens that might otherwise take valuable
resources from the individual and/or induce illness. The immune
system, which comprises leukocytes (white blood cells (WBCs): T and
B lymphocytes, monocytes, macrophages granulocytes, NK cell, mast
cells, dendritic cell, and immune derived cells (for example,
osteoclasts)), lymphoid tissues and lymphoid vessels, is the body's
defense system. To combat infection, white blood cells circulate
throughout the body to detect pathogens. Once a pathogen is
detected, innate immune cells and cytotoxic T cells in particular
are recruited to the infection site to destroy the pathogen.
Chemokines act as molecular beacons for the recruitment and
activation of immune cells, such as lymphocytes, monocytes and
granulocytes, identifying sites where pathogens exist.
[0006] Despite the immune system's regulation of pathogens, certain
inappropriate chemokine signaling can develop and has been
attributed to triggering or sustaining inflammatory disorders, such
as rheumatoid arthritis, multiple sclerosis and others. For
example, in rheumatoid arthritis, unregulated chemokine
accumulation in bone joints attracts and activates infiltrating
macrophages and T-cells. The activities of these cells induce
synovial cell proliferation that leads, at least in part, to
inflammation and eventual bone and cartilage loss (see, DeVries, M.
E., et al., Semin Immunol 11(2):95-104 (1999)). A hallmark of some
demyelinating diseases such as multiple sclerosis is the
chemokine-mediated monocyte/macrophage and T cell recruitment to
the central nervous system (see, Kennedy, et al., J. Clin. Immunol.
19(5):273-279 (1999)). Chemokine recruitment of destructive WBCs to
transplants has been implicated in their subsequent rejection. See,
DeVries, M. E., et al., ibid. Because chemokines play pivotal roles
in inflammation and lymphocyte development, the ability to
specifically manipulate their activity has enormous impact on
ameliorating and halting diseases that currently have no
satisfactory treatment. In addition, transplant rejection may be
minimized without the generalized and complicating effects of
costly immunosuppressive pharmaceuticals.
[0007] Chemokines, a group of greater than 40 small peptides (7-10
kD), ligate receptors expressed primarily on WBCs or immune derived
cells, and signal through G-protein-coupled signaling cascades to
mediate their chemoattractant and chemostimulant functions.
Receptors may bind more than one ligand; for example, the receptor
CCR1 ligates RANTES (regulated on activation normal T cell
expressed), MIP-1.alpha. (macrophage inflammatory protein),
MPIF-1/CK.beta.8, and Leukotactin chemokines (among others with
lesser affinities). To date, 24 chemokine receptors are known. The
sheer number of chemokines, multiple ligand binding receptors, and
different receptor profiles on immune cells allow for tightly
controlled and specific immune responses. See, Rossi, et al., Ann.
Rev. Immunol. 18(1):217-242 (2000). Chemokine activity can be
controlled through the modulation of their corresponding receptors,
treating related inflammatory and immunological diseases and
enabling organ and tissue transplants.
[0008] The receptor CCR1 and its chemokine ligands, including, for
example MIP-1.alpha., MPIF-1/CK.beta.8, leukotactin and RANTES,
represent significant therapeutic targets (see Saeki, et al.,
Current Pharmaceutical Design 9:1201-1208 (2003)) since they have
been implicated in rheumatoid arthritis, transplant rejection (see,
DeVries, M. E., et al., ibid.), and multiple sclerosis (see,
Fischer, et al., J Neuroimmunol. 110(1-2):195-208 (2000); Izikson,
et al., J. Exp. Med. 192(7):1075-1080 (2000); and Rottman, et al.,
Eur. J Immunol. 30(8):2372-2377 (2000). In fact, function-blocking
antibodies, modified chemokine receptor ligands and small organic
compounds have been discovered, some of which have been
successfully demonstrated to prevent or treat some
chemokine-mediated diseases (reviewed in Rossi, et al., ibid.).
Notably, in an experimental model of rheumatoid arthritis, disease
development is diminished when a signaling-blocking,
modified-RANTES ligand is administered (see Plater-Zyberk, et al.,
Immunol Lett. 57(1-3):117-120 (1997)). While function-blocking
antibody and small peptide therapies are promising, they suffer
from the perils of degradation, extremely short half-lives once
administered, and prohibitive expense to develop and manufacture,
characteristic of most proteins. Small organic compounds are
preferable since they often have longer half lives in vivo, require
fewer doses to be effective, can often be administered orally, and
are consequently less expensive. Some organic antagonists of CCR1
have been previously described (see, Hesselgesser, et al., J. Biol.
Chem. 273(25):15687-15692 (1998); Ng, et al., J. Med. Chem.
42(22):4680-4694 (1999); Liang, et al., J. Biol. Chem.
275(25):19000-19008 (2000); and Liang, et al., Eur. J. Pharmacol.
389(1):41-49 (2000)). In view of the effectiveness demonstrated for
treatment of disease in animal models (see, Liang, et al., J. Biol.
Chem. 275(25):19000-19008 (2000)), the search has continued to
identify additional compounds that can be used in the treatment of
diseases mediated by CCR1 signaling.
BRIEF SUMMARY OF THE INVENTION
[0009] The present invention provides compounds having a formula
selected from the group consisting of:
##STR00001##
or a pharmaceutically acceptable salt, hydrate or N-oxide thereof.
In the formulae above, the subscript m is an integer of from 0 to
4.
[0010] The symbol R.sup.1 is a substituent independently selected
from the group consisting of C.sub.1-8 alkyl, C.sub.1-8 haloalkyl,
C.sub.3-6 cycloalkyl, --CO.sub.2R.sup.a, --S(O).sub.3R.sup.a,
--X.sup.1CO.sub.2R.sup.a, --X.sup.1SO.sub.2R.sup.a,
--X.sup.1S(O).sub.3R.sup.a, --X.sup.1OR.sup.a, --COR.sup.a,
--CONR.sup.aR.sup.b, --X.sup.1NR.sup.aR.sup.b,
--X.sup.1NR.sup.aCOR.sup.b, --X.sup.1CONR.sup.aR.sup.b,
X.sup.1S(O).sub.2NR.sup.aR.sup.b, X.sup.1S(O).sub.2R.sup.a,
--OR.sup.a, --NR.sup.aR.sup.b, --NR.sup.aCOR.sup.b,
--CONR.sup.aR.sup.b, --NR.sup.aS(O).sub.2R.sup.b,
--S(O).sub.2NR.sup.aR.sup.b, --S(O).sub.2R.sup.a,
--X.sup.1COR.sup.a, X.sup.1CONR.sup.aR.sup.b, and
--X.sup.1NR.sup.aS(O).sub.2R.sup.b, wherein X.sup.1 is C.sub.1-4
alkylene and each R.sup.a and R.sup.b is independently selected
from the group consisting of hydrogen, C.sub.1-8 alkyl, C.sub.1-8
haloalkyl and C.sub.3-6 cycloalkyl, or optionally R.sup.a and
R.sup.b when attached to the same nitrogen atom are combined to
form a 3- to 7-membered ring having from 0-2 additional heteroatoms
as ring members; and wherein the aliphatic portions of each of said
R.sup.1 substituents is optionally substituted with from one to
three members selected from the group consisting of --OH,
--OR.sup.m, --OC(O)NHR.sup.m, --OC(O)N(R.sup.m).sub.2, --SH,
--SR.sup.m, --S(O)R.sup.m, --S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.m, --S(O).sub.2N(R.sup.m).sub.2,
--NHS(O).sub.2R.sup.m, --NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2,
--C(O)NHR.sup.m, --C(O)N(R.sup.m).sub.2, --C(O)R.sup.m,
--NHC(O)R.sup.m, --NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2,
--NR.sup.mC(O)NH.sub.2, --NR.sup.mC(O)NHR.sup.m,
--NHC(.dbd.NH)NH.sub.2, --NHC(.dbd.NR.sup.m)NH.sub.2,
--NR.sup.mC(.dbd.NR.sup.m)N(R.sup.m).sub.2,
--NR.sup.mC(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(.dbd.NR.sup.m)N(R.sup.m).sub.2,
--NHC(.dbd.NH)N(R.sup.m).sub.2, --NHC(.dbd.NH)NH(R.sup.m),
--C(.dbd.NH)NH.sub.2, --C(.dbd.NR.sup.m)NH.sub.2,
--C(.dbd.NR.sup.m)N(R.sup.m).sub.2, --C(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(O)NHR.sup.m, --NR.sup.mC(O)N(R.sup.m).sub.2,
--NHC(O)N(R.sup.m).sub.2, --CO.sub.2H, --CO.sub.2R.sup.m,
--NHCO.sub.2R.sup.m, --NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2,
--NH.sub.2, --NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2
and --NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl.
[0011] The symbols R.sup.2a, R.sup.2c and R.sup.2d in formulae Ia
and Ib are each substituents independently selected from the group
consisting of hydrogen, halogen, cyano, aryl, heteroaryl,
--NO.sub.2, --CO.sub.2R.sup.c, --CONR.sup.cR.sup.d, --C(O)R.sup.c,
--S(O)R.sup.c, --S(O).sub.2R.sup.c, --S(O).sub.3R.sup.c, --R.sup.c,
--C(NOR.sup.c)R.sup.d, --C(NR.sup.cV).dbd.NV,
--N(V)C(R.sup.c).dbd.NV, --X.sup.2C(NOR.sup.c)R.sup.d,
--X.sup.2C(NR.sup.cV).dbd.NV, --X.sup.2N(V)C(R.sup.c).dbd.NV,
--X.sup.2NR.sup.cR.sup.d, --X.sup.2SR.sup.c, --X.sup.2CN,
--X.sup.2NO.sub.2, --X.sup.2CO.sub.2R.sup.c,
--X.sup.2CONR.sup.cR.sup.d, --X.sup.2C(O)R.sup.c,
--X.sup.2OC(O)NR.sup.cR.sup.d, --X.sup.2NR.sup.dC(O)R.sup.c,
--X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.e).dbd.NH, --X.sup.2S(O)R.sup.e,
--X.sup.2S(O).sub.2R.sup.e, --X.sup.2NR.sup.cS(O).sub.2R.sup.e,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d, --X.sup.2N.sub.3, --OR.sup.c,
--SR.sup.c, --NR.sup.dC(O)R.sup.c, --NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2S(O).sub.3R.sup.c, --S(O).sub.2NR.sup.cR.sup.d,
--X.sup.2OR.sup.c, --O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d, --NR.sup.d--X.sup.2CO.sub.2R.sup.c,
--NR.sup.c--C(O)NR.sup.cR.sup.d, --NH--C(NH.sub.2).dbd.NH,
--NR.sup.eC(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR.sup.e,
--NH--C(NHR.sup.e).dbd.NH, --NR.sup.eC(NHR.sup.e).dbd.NH,
--NR.sup.eC(NH.sub.2).dbd.NR.sup.e,
--NH--C(NHR.sup.e).dbd.NR.sup.e, --NH--C(NR.sup.cR.sup.c).dbd.NH,
NR.sup.cS(O).sub.2R.sup.c, --NR.sup.cC(S)NR.sup.cR.sup.d,
--X.sup.2NR.sup.cC(S)NR.sup.cR.sup.d, --X.sup.2OC(O)R.sup.c,
--O--X.sup.2CONR.sup.cR.sup.d, --OC(O)R.sup.c, --NR.sup.cR.sup.d,
--NR.sup.d--X.sup.2OR.sup.c and
--NR.sup.d--X.sup.2NR.sup.cR.sup.d.
[0012] Within each of R.sup.2a, R.sup.2c and R.sup.2d, X.sup.2 is
C.sub.1-4 alkylene and each R.sup.c and R.sup.d is independently
selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, and
C.sub.3-6 cycloalkyl. Optionally, R.sup.c and R.sup.d when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members. The symbol R.sup.e is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl and heteroaryl, and each of R.sup.c, R.sup.d and
R.sup.e is optionally further substituted with from one to three
members selected from the group consisting of --OH, --OR.sup.n,
--OC(O)NHR.sup.n, --OC(O)N(R.sup.n).sub.2, --SH, --SR.sup.n,
--S(O)R.sup.n, --S(O).sub.2R.sup.n, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.n, --S(O).sub.2N(R.sup.n).sub.2,
--NHS(O).sub.2R.sup.n, --NR.sup.nS(O).sub.2R.sup.n, --C(O)NH.sub.2,
--C(O)NHR.sup.n, --C(O)N(R.sup.n).sub.2, --C(O)R.sup.n,
--NHC(O)R.sup.n, --NR.sup.nC(O)R.sup.n, --NHC(O)NH.sub.2,
--NR.sup.nC(O)NH.sub.2, --NR.sup.nC(O)NHR.sup.n, --NHC(O)NHR.sup.n,
--NR.sup.nC(O)N(R.sup.n).sub.2, --NHC(O)N(R.sup.n).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.n, --NHCO.sub.2R.sup.n,
--NR.sup.nCO.sub.2R.sup.n, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.n, --N(R.sup.n).sub.2, --NR.sup.nS(O)NH.sub.2 and
--NR.sup.nS(O).sub.2NHR.sup.n, wherein each R.sup.n is
independently an unsubstituted C.sub.1-6 alkyl; and wherein V is
independently selected from the group consisting of --R.sup.c,
--CN, --CO.sub.2R.sup.e and --NO.sub.2.
[0013] Each of ring vertices a, b, c and d in formulae Ia and Ib is
independently selected from N and C(R.sup.3a), and from one to two
of said ring vertices is N. The symbol R.sup.3a in formulae Ia and
Ib is independently selected from the group consisting of hydrogen,
halogen, --OR.sup.f, --OC(O)R.sup.f, --NR.sup.fR.sup.g, --SR.sup.f,
--R.sup.h, --CN, --NO.sub.2, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, --C(O)R.sup.f, --OC(O)NR.sup.fR.sup.g,
--NR.sup.gC(O)R.sup.f, --NR.sup.gC(O).sub.2R.sup.h,
--NR.sup.f--C(O)NR.sup.fR.sup.g, --NH--C(NH.sub.2).dbd.NH,
--NR.sup.hC(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR.sup.h,
--NH--C(NHR.sup.h).dbd.NH, --C(.dbd.NR.sup.f)NR.sup.gR.sup.h,
--S(O).sub.3R.sup.f, --S(O)R.sup.h, --S(O).sub.2R.sup.h,
--S(O).sub.3R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --NR.sup.fS(O).sub.2R.sup.h,
--NR.sup.fS(O).sub.2NR.sup.fR.sup.g, --N.sub.3,
--C(C.dbd.NOR.sup.f)NR.sup.fR.sup.g, --X.sub.3SO.sub.3R.sup.f,
--X.sup.3C(.dbd.NR.sup.f)NR.sup.gR.sup.h, --X.sup.3OR.sup.f,
--X.sup.3OC(O)R.sup.f, --X.sup.3NR.sup.fR.sup.g, --X.sup.3SR.sup.f,
--X.sup.3CN, --X.sup.3NO.sub.2, --X.sup.3CO.sub.2R.sup.f,
--X.sup.3CONR.sup.fR.sup.g, --X.sup.3C(O)R.sup.f,
--X.sup.3OC(O)NR.sup.fR.sup.g, --X.sup.3NR.sup.gC(O)R.sup.f,
--X.sup.3NR.sup.gC(O).sub.2R.sup.h,
--X.sup.3NR.sup.f--C(O)NR.sup.fR.sup.g,
--X.sup.3NH--C(NH.sub.2).dbd.NH,
--X.sup.3NR.sup.hC(NH.sub.2).dbd.NH,
--X.sup.3NH--C(NH.sub.2).dbd.NR.sup.h,
--X.sup.3NH--C(NHR.sup.h).dbd.NH, --X.sup.3S(O)R.sup.h,
--X.sup.3S(O).sub.2R.sup.h, --X.sup.3NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, --Y, --X.sup.3Y,
--X.sup.3N.sub.3, --C(O)NR.sup.fS(O)R.sup.h,
--P.dbd.O(OR.sup.f)(OR.sup.g),
--X.sup.3C(O)NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3C(O)NR.sup.fS(O)R.sup.h and
--X.sup.3P.dbd.O(OR.sup.f)(OR.sup.g). The symbol Y is a five to
ten-membered aryl, heteroaryl or heterocycloalkyl ring, optionally
substituted with from one to three substituents selected from the
group consisting of halogen, --OR.sup.f, --NR.sup.fR.sup.g,
--R.sup.h, --SR.sup.f, --CN, --NO.sub.2, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, --C(O)R.sup.f, --NR.sup.gC(O)R.sup.f,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --X.sup.3OR.sup.f,
--X.sup.3NR.sup.fR.sup.g, --X.sup.3NR.sup.fS(O).sub.2R.sup.h and
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, and wherein each X.sup.3 is
independently selected from the group consisting of C.sub.1-4
alkylene, C.sub.2-4 alkenylene and C.sub.2-4 alkynylene; each
R.sup.f and R.sup.g is independently selected from hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
C.sub.2-8 alkenyl, C.sub.2-8 alkynyl, aryl, heteroaryl,
aryl-C.sub.1-4 alkyl, and aryloxy-C.sub.1-4 alkyl, or when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members; and each R.sup.h is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl, heteroaryl, aryl-C.sub.1-4 alkyl, and
aryloxy-C.sub.1-4 alkyl, wherein the aliphatic portions of X.sup.3,
R.sup.f, R.sup.g and R.sup.h are optionally further substituted
with from one to three members selected from the group consisting
of --OH, --OR.sup.o, --OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2,
--SH, --SR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0014] In addition to the compounds provided herein, the present
invention further provides pharmaceutical compositions containing
one or more of these compounds, as well as methods for the use of
these compounds in therapeutic methods, primarily to treat diseases
associated with CCR1 signaling activity.
BRIEF DESCRIPTION OF THE DRAWINGS
[0015] NONE
DETAILED DESCRIPTION OF THE INVENTION
I. ABBREVIATION AND DEFINITIONS
[0016] The term "alkyl", by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain hydrocarbon radical, having the number of carbon atoms
designated (i.e. C.sub.1-8 means one to eight carbons). Examples of
alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl,
t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,
and the like. The term "alkenyl" refers to an unsaturated alkyl
group having one or more double bonds. Similarly, the term
"alkynyl" refers to an unsaturated alkyl group having one or more
triple bonds. Examples of such unsaturated alkyl groups include
vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),
2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,
3-butynyl, and the higher homologs and isomers. The term
"cycloalkyl" refers to hydrocarbon rings having the indicated
number of ring atoms (e.g., C.sub.3-6cycloalkyl) and being fully
saturated or having no more than one double bond between ring
vertices. "Cycloalkyl" is also meant to refer to bicyclic and
polycyclic hydrocarbon rings such as, for example,
bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term
"heterocycloalkyl" refers to a cycloalkyl group that contain from
one to five heteroatoms selected from N, O, and S, wherein the
nitrogen and sulfur atoms are optionally oxidized, and the nitrogen
atom(s) are optionally quaternized. The heterocycloalkyl may be a
monocyclic, a bicyclic or a polycylic ring system. Non limiting
examples of heterocycloalkyl groups include pyrrolidine,
piperidinyl, imidazolidine, pyrazolidine, butyrolactam,
valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide,
piperidine, 1,4-dioxane, morpholine, thiomorpholine,
thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine,
pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran,
tetrahydrothiophene, quinuclidine, and the like. A heterocycloalkyl
group can be attached to the remainder of the molecule through a
ring carbon or a heteroatom.
[0017] The term "alkylene" by itself or as part of another
substituent means a divalent radical derived from an alkane, as
exemplified by --CH.sub.2CH.sub.2CH.sub.2CH.sub.2--. Typically, an
alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with
those groups having 10 or fewer carbon atoms being preferred in the
present invention. A "lower alkyl" or "lower alkylene" is a shorter
chain alkyl or alkylene group, generally having four or fewer
carbon atoms. Similarly, "alkenylene" and "alkynylene" refer to the
unsaturated forms of "alkylene" having double or triple bonds,
respectively.
[0018] 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.
Additionally, for dialkylamino groups, the alkyl portions can be
the same or different and can also be combined to form a 3-7
membered ring with the nitrogen atom to which each is attached.
Accordingly, a group represented as --NR.sup.aR.sup.b is meant to
include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl and the
like.
[0019] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "C.sub.1-4 haloalkyl" is mean to include
trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,
3-bromopropyl, and the like.
[0020] The term "aryl" means, unless otherwise stated, a
polyunsaturated, typically aromatic, hydrocarbon group which can be
a single ring or multiple rings (up to three rings) which are fused
together or linked covalently. The term "heteroaryl" refers to aryl
groups (or rings) that contain from one to five heteroatoms
selected from N, O, and S, wherein the nitrogen and sulfur atoms
are optionally oxidized, and the nitrogen atom(s) are optionally
quaternized. A heteroaryl group can be attached to the remainder of
the molecule through a heteroatom. Non-limiting examples of aryl
groups include phenyl, naphthyl and biphenyl, while non-limiting
examples of heteroaryl groups include pyridyl, pyridazinyl,
pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,
quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl,
benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,
isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl,
thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl,
imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl,
indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl,
pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl,
isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl and
the like. Substituents for each of the above noted aryl and
heteroaryl ring systems are selected from the group of acceptable
substituents described below.
[0021] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both
aryl and heteroaryl rings as defined above. Thus, the term
"arylalkyl" is meant to include those radicals in which an aryl
group is attached to an alkyl group (e.g., benzyl, phenethyl,
pyridylmethyl and the like).
[0022] The above terms (e.g., "alkyl," "aryl" and "heteroaryl"), in
some embodiments, will include both substituted and unsubstituted
forms of the indicated radical. Preferred substituents for each
type of radical are provided below. For brevity, the terms aryl and
heteroaryl will refer to substituted or unsubstituted versions as
provided below, while the term "alkyl" and related aliphatic
radicals is meant to refer to unsubstituted version, unless
indicated to be substituted.
[0023] Substituents for the alkyl radicals (including those groups
often referred to as alkylene, alkenyl, alkynyl and cycloalkyl) can
be a variety of groups selected from: -halogen, --OR', --NR'R'',
--SR', --SiR'R''R''', --OC(O)R', --C(O)R', --CO.sub.2R',
--CONR'R'', --OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''',
--NR''C(O).sub.2R', --NH--C(NH.sub.2).dbd.NH,
--NR'C(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR', --S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R'', --NR'S(O).sub.2R'', --CN and
--NO.sub.2 in a number ranging from zero to (2 m'+1), where m' is
the total number of carbon atoms in such radical. R', R'' and R'''
each independently refer to hydrogen, unsubstituted C.sub.1-8
alkyl, unsubstituted heteroalkyl, unsubstituted aryl, aryl
substituted with 1-3 halogens, unsubstituted C.sub.1-8 alkyl,
C.sub.1-8 alkoxy or C.sub.1-8 thioalkoxy groups, or unsubstituted
aryl-C.sub.1-4 alkyl groups. When R' and R'' are attached to the
same nitrogen atom, they can be combined with the nitrogen atom to
form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, --NR'R'' is
meant to include 1-pyrrolidinyl and 4-morpholinyl.
[0024] Similarly, substituents for the aryl and heteroaryl groups
are varied and are generally selected from: -halogen, --OR',
--OC(O)R', --NR'R'', --SR', --R', --CN, --NO.sub.2, --CO.sub.2R',
--CONR'R'', --C(O)R', --OC(O)NR'R'', --NR''C(O)R',
--NR''C(O).sub.2R', --NR'--C(O)NR''R''', --NH--C(NH.sub.2).dbd.NH,
--NR'C(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR', --S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R'', --NR'S(O).sub.2R'', --N.sub.3,
perfluoro(C.sub.1-C.sub.4)alkoxy, and
perfluoro(C.sub.1-C.sub.4)alkyl, in a number ranging from zero to
the total number of open valences on the aromatic ring system; and
where R', R'' and R''' are independently selected from hydrogen,
C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, unsubstituted aryl and heteroaryl, (unsubstituted
aryl)-C.sub.1-4 alkyl, and unsubstituted aryloxy-C.sub.1-4 alkyl.
Other suitable substituents include each of the above aryl
substituents attached to a ring atom by an alkylene tether of from
1-4 carbon atoms.
[0025] Two of the substituents on adjacent atoms of the aryl or
heteroaryl ring may optionally be replaced with a substituent of
the formula -T-C(O)--(CH.sub.2).sub.q--U--, wherein T and U are
independently --NH--, --O--, --CH.sub.2-- or a single bond, and q
is an integer of from 0 to 2. Alternatively, two of the
substituents on adjacent atoms of the aryl or heteroaryl ring may
optionally be replaced with a substituent of the formula
-A-(CH.sub.2).sub.r--B--, wherein A and B are independently
--CH.sub.2--, --O--, --NH--, --S--, --S(O)--, --S(O).sub.2--,
--S(O).sub.2NR'-- or a single bond, and r is an integer of from 1
to 3. One of the single bonds of the new ring so formed may
optionally be replaced with a double bond. Alternatively, two of
the substituents on adjacent atoms of the aryl or heteroaryl ring
may optionally be replaced with a substituent of the formula
--(CH.sub.2).sub.s--X--(CH.sub.2).sub.t--, where s and t are
independently integers of from 0 to 3, and X is --O--, --NR'--,
--S--, --S(O)--, --S(O).sub.2--, or --S(O).sub.2NR'--. The
substituent R' in --NR'-- and --S(O).sub.2NR'-- is selected from
hydrogen or unsubstituted C.sub.1-6 alkyl.
[0026] As used herein, the term "heteroatom" is meant to include
oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).
[0027] 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 salts derived from pharmaceutically-acceptable inorganic bases
include aluminum, ammonium, calcium, copper, ferric, ferrous,
lithium, magnesium, manganic, manganous, potassium, sodium, zinc
and the like. Salts derived from pharmaceutically-acceptable
organic bases include salts of primary, secondary and tertiary
amines, including substituted amines, cyclic amines,
naturally-occurring amines and the like, such as arginine, betaine,
caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine,
2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,
ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,
glucosamine, histidine, hydrabamine, isopropylamine, lysine,
methylglucamine, morpholine, piperazine, piperadine, polyamine
resins, procaine, purines, theobromine, triethylamine,
trimethylamine, tripropylamine, tromethamine and the like. When
compounds of the present invention contain relatively basic
functionalities, acid addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired acid, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable acid addition salts include those
derived from inorganic acids like hydrochloric, hydrobromic,
nitric, carbonic, monohydrogencarbonic, phosphoric,
monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, malonic, benzoic, succinic,
suberic, fumaric, mandelic, phthalic, benzenesulfonic,
p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
Also included are salts of amino acids such as arginate and the
like, and salts of organic acids like glucuronic or galactunoric
acids and the like (see, for example, Berge, S. M., et al,
"Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977,
66, 1-19). Certain specific compounds of the present invention
contain both basic and acidic functionalities that allow the
compounds to be converted into either base or acid addition
salts.
[0028] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents, but otherwise the
salts are equivalent to the parent form of the compound for the
purposes of the present invention.
[0029] In addition to salt forms, the present invention provides
compounds which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present invention. Additionally, prodrugs can be converted to
the compounds of the present invention by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present invention when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0030] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present invention. Certain compounds of the present invention may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present invention and are intended to be within the scope of the
present invention.
[0031] Certain compounds of the present invention possess
asymmetric carbon atoms (optical centers) or double bonds; the
racemates, diastereomers, geometric isomers, regioisomers and
individual isomers (e.g., separate enantiomers) are all intended to
be encompassed within the scope of the present invention. The
compounds of the present invention may also contain unnatural
proportions of atomic isotopes at one or more of the atoms that
constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example tritium
(.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C). All
isotopic variations of the compounds of the present invention,
whether radioactive or not, are intended to be encompassed within
the scope of the present invention.
II. GENERAL
[0032] The present invention derives from the discovery that
compounds of formula Ia or Ib (as well as the subgeneric formulae
Ia.sup.1-4 and Ib.sup.1-4) act as potent antagonists of the CCR1
receptor. The compounds have in vivo anti-inflammatory activity.
Accordingly, the compounds provided herein are useful in
pharmaceutical compositions, methods for the treatment of
CCR1-mediated diseases, and as controls in assays for the
identification of competitive CCR1 antagonists.
III. COMPOUNDS
[0033] In one aspect, the present invention provides compounds
having a formula selected from the group consisting of:
##STR00002##
or a pharmaceutically acceptable salt, hydrate or N-oxide thereof.
In the formulae above, the subscript m is an integer of from 0 to
4. In certain embodiments, in formulae Ia and Ib the subscript m is
an integer from 0 to 2. In yet another embodiment, the subscript m
in formulae Ia and Ib is an integer of from 0 to 1.
[0034] The symbol R.sup.1 in formulae Ia and Ib is a substituent
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
--CO.sub.2R.sup.a, --S(O).sub.3R.sup.a, --X.sup.1CO.sub.2R.sup.a,
--X.sup.1SO.sub.2R.sup.a, --X.sup.1S(O).sub.3R.sup.a,
--X.sup.1OR.sup.a, --COR.sup.a, --CONR.sup.aR.sup.b,
--X.sup.1NR.sup.aR.sup.b, --X.sup.1NR.sup.a COR.sup.b,
--X.sup.1CONR.sup.aR.sup.b, X.sup.1S(O).sub.2NR.sup.aR.sup.b,
X.sup.1S(O).sub.2R.sup.a, --OR.sup.a, --NR.sup.aR.sup.b,
--NR.sup.aCOR.sup.b, --CONR.sup.aR.sup.b,
--NR.sup.aS(O).sub.2R.sup.b, --S(O).sub.2NR.sup.aR.sup.b,
--S(O).sub.2R.sup.a, --X.sup.1COR.sup.a, X.sup.1CONR.sup.aR.sup.b,
and --X.sup.1NR.sup.aS(O).sub.2R.sup.b. The symbol X.sup.1 is
C.sub.1-4 alkylene and each R.sup.a and R.sup.b substituent is
independently selected from the group consisting of hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl, or
optionally R.sup.a and R.sup.b when attached to the same nitrogen
atom are combined to form a 3- to 7-membered ring having from 0-2
additional heteroatoms as ring members; and wherein the aliphatic
portions of each of said R.sup.1 substituents is optionally
substituted with from one to three members selected from the group
consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(.dbd.NH)NH.sub.2,
--NHC(.dbd.NR.sup.m)NH.sub.2,
--NR.sup.mC(.dbd.NR.sup.m)N(R.sup.m).sub.2,
--NR.sup.mC(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(.dbd.NR.sup.m)N(R.sup.m).sub.2,
--NHC(.dbd.NH)N(R.sup.m).sub.2, --NHC(.dbd.NH)NH(R.sup.m),
--C(.dbd.NH)NH.sub.2, --C(.dbd.NR.sup.m)NH.sub.2,
--C(.dbd.NR.sup.m)N(R.sup.m).sub.2, --C(.dbd.NR.sup.m)NH(R.sup.m),
--NHC(O)NHR.sup.m, --NR.sup.mC(O)N(R.sup.m).sub.2,
--NHC(O)N(R.sup.m).sub.2, --CO.sub.2H, --CO.sub.2R.sup.m,
--NHCO.sub.2R.sup.m, --NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2,
--NH.sub.2, --NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2
and --NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl.
[0035] In another embodiment, R.sup.1 in formulae Ia and Ib is a
substituent independently selected from the group consisting of
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
--CO.sub.2R.sup.a, --X.sup.1CO.sub.2R.sup.a,
--X.sup.1SO.sub.2R.sup.a and --X.sup.1OR.sup.a, wherein the
aliphatic portions of each of said R.sup.1 substituents is
optionally substituted with from one to three members selected from
the group consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(O)NHR.sup.m,
--NR.sup.mC(O)N(R.sup.m).sub.2, --NHC(O)N(R.sup.m).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.m, --NHCO.sub.2R.sup.m,
--NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2 and
--NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl.
[0036] In another embodiment, R.sup.1 in formulae Ia and Ib is a
substituent independently selected from the group consisting of
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl,
wherein the aliphatic portions of each of said R.sup.1 substituents
is optionally substituted with from one to three members selected
from the group consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(O)NHR.sup.m,
--NR.sup.mC(O)N(R.sup.m).sub.2, --NHC(O)N(R.sup.m).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.m, --NHCO.sub.2R.sup.m,
--NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2 and
--NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl.
[0037] In one embodiment of the invention, R.sup.1 in formulae Ia
and Ib, if present, is selected from the group consisting of
--CO.sub.2H or C.sub.1-4 alkyl, optionally substituted with --OH,
--OR.sup.m, --S(O).sub.2R.sup.m, --CO.sub.2H and --CO.sub.2R.sup.m.
In another embodiment of the invention, R.sup.1 is methyl; and m is
0-2.
[0038] The symbols R.sup.2a, R.sup.2c and R.sup.2d in formulae Ia
and Ib are each substituents independently selected from the group
consisting of hydrogen, halogen, cyano, aryl, heteroaryl,
--NO.sub.2, --CO.sub.2R.sup.c, --CONR.sup.cR.sup.d, --C(O)R.sup.c,
--S(O)R.sup.e, --S(O).sub.2R.sup.e, --S(O).sub.3R.sup.c, --R.sup.e,
--C(NOR.sup.c)R.sup.d, --C(NR.sup.cV).dbd.NV,
--N(V)C(R.sup.c).dbd.NV, --X.sup.2C(NOR.sup.c)R.sup.d,
--X.sup.2C(NR.sup.cV).dbd.NV, --X.sup.2N(V)C(R.sup.c).dbd.NV,
--X.sup.2NR.sup.cR.sup.d, --X.sup.2SR.sup.c, --X.sup.2CN,
--X.sup.2NO.sub.2, --X.sup.2CO.sub.2R.sup.c,
--X.sup.2CONR.sup.cR.sup.d, --X.sup.2C(O)R.sup.c,
--X.sup.2OC(O)NR.sup.cR.sup.d, --X.sup.2NR.sup.dC(O)R.sup.c,
--X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.e).dbd.NH, --X.sup.2S(O)R.sup.e,
--X.sup.2S(O).sub.2R.sup.e, --X.sup.2NR.sup.cS(O).sub.2R.sup.e,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d, --X.sup.2N.sub.3, --OR.sup.c,
--SRC, --NR.sup.dC(O)R.sup.c, --NR.sup.dC(O).sub.2R.sup.c,
--X.sup.2S(O).sub.3R.sup.c, --S(O).sub.2NR.sup.cR.sup.d,
--X.sup.2OR.sup.c, --O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d, --NR.sup.d--X.sup.2CO.sub.2R.sup.c,
--NR.sup.c--C(O)NR.sup.cR.sup.d, --NH--C(NH.sub.2).dbd.NH,
--NR.sup.eC(NH.sub.2).dbd.NH, --NH--C(NH.sub.2).dbd.NR.sup.e,
--NH--C(NHR.sup.e).dbd.NH, --NR.sup.eC(NHR.sup.e).dbd.NH,
--NR.sup.eC(NH.sub.2).dbd.NR.sup.e,
--NH--C(NHR.sup.e).dbd.NR.sup.e, --NH--C(NR.sup.eR.sup.e).dbd.NH,
NR.sup.cS(O).sub.2R.sup.e, --NR.sup.cC(S)NR.sup.cR.sup.d,
--X.sup.2NR.sup.cC(S)NR.sup.cR.sup.d, --X.sup.2OC(O)R.sup.c,
--O--X.sup.2CONR.sup.cR.sup.d, --OC(O)R.sup.c, --NR.sup.cR.sup.d,
--NR.sup.d--X.sup.2OR.sup.c and
--NR.sup.d--X.sup.2NR.sub.cR.sup.d.
[0039] In one embodiment, the symbol R.sup.2a in formulae Ia and Ib
is independently selected from the group consisting of hydrogen,
halogen, cyano, heteroaryl, --NO.sub.2, --CO.sub.2R.sup.c,
--CONR.sup.cR.sup.d, --C(O)R.sup.c, --S(O)R.sup.e,
--S(O).sub.2R.sup.e, --R.sup.e, --C(NOR.sup.c)R.sup.d,
--C(NR.sup.cV).dbd.NV, --N(V)C(R.sup.c).dbd.NV,
--X.sup.2C(NOR.sup.c)R.sup.d, --X.sup.2C(NR.sup.cV).dbd.NV,
--X.sup.2N(V)C(R.sup.c).dbd.NV, --X.sup.2NR.sup.cR.sup.d,
--X.sup.2SR.sup.c, --X.sup.2CN, --X.sup.2NO.sub.2,
--X.sup.2CO.sub.2R.sup.c, --X.sup.2CONR.sup.cR.sup.d,
--X.sup.2C(O)R.sup.c, --X.sup.2OC(O)NR.sup.cR.sup.d,
--X.sup.2NR.sup.dC(O)R.sup.c, --X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.e).dbd.NH, --X.sup.2S(O)R.sup.e,
--X.sup.2S(O).sub.2R.sup.e, --X.sup.2NR.sup.cS(O).sub.2R.sup.e,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d and --X.sup.2N.sub.3.
[0040] In another embodiment, the R.sup.2a substitutent in formulae
Ia and Ib is selected from the group consisting of hydrogen, F, Cl,
Br, I, --CO.sub.2R.sup.c, --CONR.sup.cR.sup.d, --CN, a 5- to
6-membered heteroaryl, --X.sub.2NR.sup.cR.sup.d,
--C(NOR.sup.c)R.sup.d. In yet another embodiment, R.sup.2a is
hydrogen. In yet another embodiment, the R.sup.2a substitutent in
formulae Ia and Ib is selected from the group consisting of F, Cl,
Br, I, --CO.sub.2Me, --CONH.sub.2, CN, oxazolyl,
--CH.sub.2NH.sub.2, --CH.sub.2NHMe, --CH.sub.2NMe.sub.2 and
--CH.dbd.N--OH. In yet another embodiment, in compounds having
formulae Ia and Ib, the R.sup.2a substituent is selected from the
group consisting of hydrogen, F, Cl, Br and I.
[0041] In another embodiment, the symbols R.sup.2c and R.sup.2d in
formulae Ia and Ib are each substituents independently selected
from the group consisting of halogen, --OR.sup.c, --SR.sup.c,
--OC(O)R.sup.c, --NR.sup.cR.sup.d, --R.sup.e, --CN, --NO.sub.2,
--CO.sub.2R.sup.c, --C(O)R.sup.c, --NR.sup.dC(O)R.sup.c,
--NR.sup.dC(O).sub.2R.sup.e, --S(O).sub.2R.sup.e,
--S(O).sub.2NR.sup.cR.sup.d, --X.sup.2OR.sup.c,
--O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d and --NR.sup.d--X.sup.2CO.sub.2R.sup.c.
In certain aspects of this embodiment, R.sup.2c and R.sup.2d are
each independently selected from the group consisting of hydrogen,
halogen, F, Cl, Br, I and OR.sup.c.
[0042] Within each of R.sup.2a, R.sup.2c and R.sup.2d, X.sup.2 is
C.sub.1-4 alkylene and each R.sup.c and R.sup.d is independently
selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, and
C.sub.3-6 cycloalkyl. Optionally, R.sup.c and R.sup.d when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members. The symbol R.sup.e is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl and heteroaryl, and each of R.sup.c, R.sup.d and
R.sup.e is optionally further substituted with from one to three
members selected from the group consisting of --OH, --OR.sup.n,
--OC(O)NHR.sup.n, --OC(O)N(R.sup.n).sub.2, --SH, --SR.sup.n,
--S(O)R.sup.n, --S(O).sub.2R.sup.n, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.n, --S(O).sub.2N(R.sup.n).sub.2,
--NHS(O).sub.2R.sup.n, --NR.sup.nS(O).sub.2R.sup.n, --C(O)NH.sub.2,
--C(O)NHR.sup.n, --C(O)N(R.sup.n).sub.2, --C(O)R.sup.n,
--NHC(O)R.sup.n, --NR.sup.nC(O)R.sup.n, --NHC(O)NH.sub.2,
--NR.sup.nC(O)NH.sub.2, --NR.sup.nC(O)NHR.sup.n, --NHC(O)NHR.sup.n,
--NR.sup.nC(O)N(R.sup.n).sub.2, --NHC(O)N(R.sup.n).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.n, --NHCO.sub.2R.sup.n,
--NR.sup.nCO.sub.2R.sup.n, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.n, --N(R.sup.n).sub.2, --NR.sup.nS(O)NH.sub.2 and
--NR.sup.nS(O).sub.2NHR.sup.n, wherein each R.sup.n is
independently an unsubstituted C.sub.1-6 alkyl; and wherein V is
independently selected from the group consisting of --R.sup.c,
--CN, --CO.sub.2R.sup.e and --NO.sub.2.
[0043] In a certain embodiment of a compound having formulae Ia and
Ib, the subscript m is 0 or 1; and the symbol R.sup.2a is hydrogen.
In another embodiment, the subscript m is 0-1; and R.sup.2a is F or
Cl.
[0044] In another embodiment of the invention, R.sup.2c in formulae
Ia and Ib is selected from the group consisting of halogen, --CN,
--NO.sub.2, --CO.sub.2R.sup.c, --COR.sup.c, --S(O)2R.sup.e. In
another embodiments of the invention, the symbol R.sup.2c is
selected from the group consisting of F, Cl, Br, CN, NO.sub.2,
--CO.sub.2CH.sub.3, --C(O)CH.sub.3 and --S(O).sub.2CH.sub.3.
[0045] In yet another embodiment of the invention, the symbol
R.sup.2d in formulae Ia and Ib is selected from the group
consisting of --SR.sup.c, --O--X.sup.2--OR.sup.c,
--X.sup.2--OR.sup.c, --OC(O)R.sup.c, --NR.sup.cR.sup.d, --R.sup.e
and --OR.sup.c. In another embodiment, R.sup.2d is selected from
the group consisting of --SMe, --OCH.sub.2OMe, --CH.sub.2OMe,
--CH.sub.2OEt, methyl, ethyl, methoxy and ethoxy.
[0046] In formulae Ia and Ib, each of the ring vertices a, b, c and
d is independently selected from N and C(R.sup.3a), and from one to
two of said ring vertices is N. In one embodiment of the invention,
the fused six membered ring having vertices a, b, c and d is a
fused pyridine ring or a fused pyrimidine ring. In yet another
embodiment of the invention, the fused six membered ring having
vertices a, b, c and d is a fused pyrazine ring. In yet another
embodiment of the invention, the fused six membered ring having
vertices a, b, c and d is a fused pyridazine ring.
[0047] Turning to the R.sup.3a substituent in formulae Ia and Ib,
at each occurrence, the symbol R.sup.3a is independently selected
from the group consisting of hydrogen, halogen, --OR.sup.f,
--OC(O)R.sup.f, --NR.sup.fR.sup.g, --SR.sup.f, --R.sup.h, --CN,
--NO.sub.2, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--OC(O)NR.sup.fR.sup.g, --NR.sup.gC(O)R.sup.f,
--NR.sup.gC(O).sub.2R.sup.h, --NR.sup.f--C(O)NR.sup.fR.sup.g,
--NH--C(NH.sub.2).dbd.NH, --NR.sup.hC(NH.sub.2).dbd.NH,
--NH--C(NH.sub.2).dbd.NR.sup.h, --NH--C(NHR.sup.h).dbd.NH,
--C(.dbd.NR.sup.f)NR.sup.gR.sup.h, --S(O).sub.3R.sup.f,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --S(O).sub.3R.sup.h,
--NR.sup.fS(O).sub.2R.sup.h, --S(O).sub.2NR.sup.fR.sup.g,
--NR.sup.fS(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2NR.sup.fR.sup.g,
--N.sub.3, --C(C.dbd.NOR.sup.f)NR.sup.fR.sup.g,
--X.sub.3SO.sub.3R.sup.f, --X.sup.3C(.dbd.NR.sup.f)NR.sup.gR.sup.h,
--X.sup.3OR.sup.f, --X.sup.3OC(O)R.sup.f, --X.sup.3NR.sup.fR.sup.g,
--X.sup.3SR.sup.f, --X.sup.3CN, --X.sup.3NO.sub.2,
--X.sup.3CO.sub.2R.sup.f, --X.sup.3CONR.sup.fR.sup.g,
--X.sup.3C(O)R.sup.f, --X.sup.3OC(O)NR.sup.fR.sup.g,
--X.sup.3NR.sup.gC(O)R.sup.f, --X.sup.3NR.sup.gC(O).sub.2R.sup.h,
--X.sup.3NR.sup.f--C(O)NR.sup.fR.sup.g,
--X.sup.3NH--C(NH.sub.2).dbd.NH,
--X.sup.3NR.sup.hC(NH.sub.2).dbd.NH,
--X.sup.3NH--C(NH.sub.2).dbd.NR.sup.h,
--X.sup.3NH--C(NHR.sup.h).dbd.NH, --X.sup.3S(O)R.sup.h,
--X.sup.3S(O).sub.2R.sup.h, --X.sup.3NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, --Y, --X.sup.3Y,
--X.sup.3N.sub.3, --C(O)NR.sup.fS(O)R.sup.h,
--P.dbd.O(OR.sup.f)(OR.sup.g),
--X.sup.3C(O)NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3C(O)NR.sup.fS(O)R.sup.h and
--X.sup.3P.dbd.O(OR.sup.f)(OR.sup.g). The symbol Y is a five to
ten-membered aryl, heteroaryl or heterocycloalkyl ring, optionally
substituted with from one to three substituents selected from the
group consisting of halogen, --OR.sup.f, --NR.sup.fR.sup.g,
--R.sup.h, --SR.sup.f, --CN, --NO.sub.2, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, --C(O)R.sup.f, --NR.sup.gC(O)R.sup.f,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --X.sup.3OR.sup.f,
--X.sup.3NR.sup.fR.sup.g, --X.sup.3NR.sup.fS(O).sub.2R.sup.h and
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, and wherein each X.sup.3 is
independently selected from the group consisting of C.sub.1-4
alkylene, C.sub.2-4 alkenylene and C.sub.2-4 alkynylene; each
R.sup.f and R.sup.g is independently selected from hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
C.sub.2-8 alkenyl, C.sub.2-8 alkynyl, aryl, heteroaryl,
aryl-C.sub.1-4 alkyl, and aryloxy-C.sub.1-4 alkyl, or when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members; and each R.sup.h is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl, heteroaryl, aryl-C.sub.1-4 alkyl, and
aryloxy-C.sub.1-4 alkyl, wherein the aliphatic portions of X.sup.3,
R.sup.f, R.sup.g and R.sup.h are optionally further substituted
with from one to three members selected from the group consisting
of --OH, --OR.sup.o, --OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2,
--SH, --SR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0048] In one embodiment of formulae Ia and Ib, the symbol
R.sup.3a, at each occurrence, is independently selected from the
group consisting of hydrogen, halogen, --OR.sup.f, --OC(O)R.sup.f,
--NR.sup.fR.sup.g, --SR.sup.f, --R.sup.h, --CN, --NO.sub.2,
--CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--OC(O)NR.sup.fR.sup.g, --NR.sup.gC(O)R.sup.f,
--NR.sup.gC(O).sub.2R.sup.h, --NR.sup.f--C(O)NR.sup.fR.sup.g,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --NR.sup.fS(O).sub.2NR.sup.fR.sup.g,
--X.sup.3OR.sup.f, --X.sup.3NR.sup.fR.sup.g, --X.sup.3SR.sup.f,
--X.sup.3CN, --X.sup.3CO.sub.2R.sup.f, --X.sup.3CONR.sup.fR.sup.g,
--X.sup.3C(O)R.sup.f, --X.sup.3NR.sup.gC(O)R.sup.f,
--X.sup.3NR.sup.gC(O).sub.2R.sup.h, --Y , --X.sup.3Y and
--X.sup.3N.sub.3. The symbol Y is a five or six-membered aryl, a
five or six membered heteroaryl, or a three to eight membered
heterocycloalkyl ring, optionally substituted with from one to
three substituents selected from the group consisting of halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --R.sup.h, --SR.sup.f, --CN,
--NO.sub.2, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--NR.sup.gC(O)R.sup.f, --S(O)R.sup.h, --S(O).sub.2R.sup.h,
--NR.sup.fS(O).sub.2R.sup.h and --S(O).sub.2NR.sup.fR.sup.g.
X.sup.3 is independently C.sub.1-4 alkylene. The symbols R.sup.f
and R.sup.g are independently selected from hydrogen, C.sub.1-8
alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl, and each
R.sup.h is independently selected from the group consisting of
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl. The
aliphatic portions of X.sup.3, R.sup.f, R.sup.g and R.sup.h is
optionally further substituted with from one to three members
selected from the group consisting of --OH, --OR.sup.o,
--OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o,
--S(O)R.sup.o, --S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.o, --S(O).sub.2N(R.sup.o).sub.2,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2,
--C(O)NHR.sup.o, --C(O)N(R.sup.o).sub.2, --C(O)R.sup.o,
--NHC(O)R.sup.o, --NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2,
--NR.sup.oC(O)NH.sub.2, --NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein each R.sup.o is
independently an unsubstituted C.sub.1-6 alkyl.
[0049] In another embodiment of the invention, the symbol R.sup.3a
of formulae Ia and Ib is a member independently selected from the
group consisting of hydrogen, halogen, --OR.sup.f,
--NR.sup.fR.sup.g, --R.sup.h, --CN, and --Y, wherein Y is a five to
six-membered aryl ring, a five to six-membered heteroaryl ring, or
a three to eight-membered heterocycloalkyl ring selected from the
group consisting of homopiperidinyl, morpholinyl, thiomorpholinyl,
pyrrolidinyl, piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl,
piperazinyl, phenyl, pyridyl, pyrimidinyl, oxadiazolyl, oxazolyl
and thiazolyl, optionally substituted with from one to three
substituents selected from the group consisting of halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --R.sup.h, --CN, wherein each
R.sup.f and R.sup.g is independently selected from hydrogen,
C.sub.1-6 alkyl, C.sub.1-6 haloalkyl and C.sub.3-6 cycloalkyl, and
each R.sup.h is independently selected from the group consisting of
C.sub.1-6 alkyl, C.sub.1-6 haloalkyl and C.sub.3-6 cycloalkyl,
wherein the aliphatic portions of R.sup.f, R.sup.g and R.sup.h are
optionally further substituted with from one to three members
selected from the group consisting of --OH, --OR.sup.o,
--OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o,
--S(O)R.sup.o, --S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.o, --S(O).sub.2N(R.sup.o).sub.2,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2,
--C(O)NHR.sup.o, --C(O)N(R.sup.o).sub.2, --C(O)R.sup.o,
--NHC(O)R.sup.o, --NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2,
--NR.sup.oC(O)NH.sub.2, --NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0050] In another embodiment of the invention, the R.sup.3a groups
in formulae Ia and Ib is selected from the group consisting of --Y
and --X.sup.3--Y, wherein Y is selected from the group consisting
of homopiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl,
piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl, piperazinyl,
phenyl, thienyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl,
pyrrolyl, pyridizinyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl,
isoxazolyl, isothiazolyl, triazolyl, tetrazolyl and oxadiazolyl,
which is optionally substituted with from one to three substituents
independently selected from the group consisting of halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --COR.sup.f, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, --NO.sub.2, --R.sup.h and --CN, wherein
R.sup.f and R.sup.g are each independently selected from the group
consisting of H, C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl and
C.sub.1-8 haloalkyl, and each R.sup.h is independently selected
from the group consisting of C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl
and C.sub.1-8 haloalkyl. In certain embodiments of the invention,
the symbol Y is selected from the group consisting of imidazolyl,
phenyl, pyridyl, oxazolyl, pyrimidinyl, oxadiazolyl, and thiazolyl,
each of which is optionally substituted with from one to three
substituents independently selected from the group consisting of
halogen, --OR.sup.f, --NR.sup.fR.sup.g, --COR.sup.f,
--CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --NO.sub.2, --R.sup.h and
--CN, wherein R.sup.f and R.sup.g are each independently selected
from the group consisting of H, C.sub.1-8 alkyl, C.sub.3-6
cycloalkyl and C.sub.1-8 haloalkyl, and each R.sup.h is
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.3-6 cycloalkyl and C.sub.1-8 haloalkyl. In certain
other embodiments of the invention, the symbol Y is selected from
the group consisting of phenyl, pyridyl, oxazolyl, pyrimidinyl,
oxadiazolyl, and thiazolyl, each of which is optionally substituted
with from one to three substituents independently selected from the
group consisting of halogen, --OR.sup.f, --NR.sup.fR.sup.g,
--COR.sup.f, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --NO.sub.2,
--R.sup.h and --CN, wherein R.sup.f and R.sup.g are each
independently selected from the group consisting of H, C.sub.1-8
alkyl, C.sub.3-6 cycloalkyl and C.sub.1-8 haloalkyl, and each
R.sup.h is independently selected from the group consisting of
C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl and C.sub.1-8 haloalkyl.
Within this embodiment, in certain aspects of the invention, m is
an integer from 0-2. In other aspect, m is an integer from 0-1.
[0051] In yet another embodiment of the invention, the R.sup.3a
substituent in formulae Ia and Ib is selected from the group
consisting of hydrogen, halogen, C.sub.1-4 alkyl and C.sub.1-4
haloalkyl, wherein the aliphatic portions are optionally
substituted with from one to three members selected from the group
consisting of --OH, --OR.sup.o, --OC(O)NHR.sup.o,
--OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o, --S(O)R.sup.o,
--S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein each R.sup.o is
independently an unsubstituted C.sub.1-6 alkyl. In certain
instances of this embodiment, m is 0 or 1; R.sup.2a is preferably
hydrogen; and additionally in other instances, R.sup.2c is
preferably selected from the group consisting of F, Cl, Br, CN,
NO.sub.2, --CO.sub.2CH.sub.3, --C(O)CH.sub.3 and
--S(O).sub.2CH.sub.3.
[0052] In yet another embodiment, the R.sup.3a substituent in
formulae Ia and Ib is halogen, C.sub.1-4 alkyl or C.sub.1-4
haloalkyl.
[0053] In yet another embodiment, the R.sup.3a moiety on the
pyrazole ring in formulae Ia and Ib is hydrogen, halogen, chloro,
fluoro, bromo, oxazolyl, pyridyl, pyrimidinyl, imidazolyl,
oxadiazolyl thiazolyl, C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl or
C.sub.1-8 haloalkyl or cyano.
[0054] In yet another embodiment, the R.sup.3a moiety on the
pyrazole ring in formulae Ia and Ib is hydrogen, halogen, chloro,
fluoro, bromo, oxazolyl, pyridyl, pyrimidinyl, oxadiazolyl,
thiazolyl, C.sub.1-8 alkyl, C.sub.3-6 cycloalkyl or C.sub.1-8
haloalkyl or cyano.
[0055] In a certain embodiment of the invention, in the compounds
having formulae Ia and Ib, R.sup.3a is a member independently
selected from the group consisting of hydrogen, halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --C(O)R.sup.f, --C(O)OR.sup.f,
--S(O)R.sup.f, --S(O).sub.2R.sup.f, --S(O).sub.3R.sup.f,
--S(O).sub.3R.sup.h, --X.sup.3C(O).sub.2R.sup.f,
X.sup.3S(O).sub.3R.sup.f, --S(O).sub.2NR.sup.fR.sup.g,
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, --R.sup.h, --CN,
X.sup.3NR.sup.fR.sup.g, NR.sup.gC(O)R.sup.f, X.sup.3N.sub.3 and Y.
The symbol Y is a five to six-membered aryl, a five or six-membered
heteroaryl ring or a three to eight-membered heterocycloalkyl ring
selected from the group consisting of homopiperidinyl, morpholinyl,
thiomorpholinyl, pyrrolidinyl, piperidinyl, azetidinyl, pyranyl,
tetrahydrofuranyl, piperazinzyl, phenyl, pyridyl, oxazolyl,
pyrimidinyl, oxadiazolyl, imidazolyl, pyrazolyl, triazolyl and
thiazolyl, optionally substituted with from one to three
substituents selected from the group consisting of halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --R.sup.h, --CN. Each R.sup.f and
R.sup.g is independently selected from hydrogen, C.sub.1-6 alkyl,
C.sub.1-6 haloalkyl and C.sub.3-6 cycloalkyl, and each R.sup.h is
independently selected from the group consisting of C.sub.1-6
alkyl, C.sub.1-6 haloalkyl and C.sub.3-6 cycloalkyl, wherein the
aliphatic portions of R.sup.f, R.sup.g and R.sup.h are optionally
further substituted with from one to three members selected from
the group consisting of --OH, --OR.sup.o, --OC(O)NHR.sup.o,
--OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o, --S(O)R.sup.o,
--S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0056] In a certain embodiment of the invention, in the compounds
having formulae Ia and Ib, R.sup.3a is a member independently
selected from the group consisting of hydrogen, halogen,
--OR.sup.f, --NR.sup.fR.sup.g, --C(O)R.sup.f, --C(O)OR.sup.f,
--S(O)R.sup.f, --S(O).sub.2R.sup.f, --S(O).sub.2NR.sup.fR.sup.g,
--R.sup.h, --CN, X.sup.3NR.sup.fR.sup.g, NR.sup.gC(O)R.sup.f,
X.sup.3N.sub.3 and --Y, wherein Y is a five to six-membered aryl, a
five or six-membered heteroaryl ring or a three to eight-membered
heterocycloalkyl ring selected from the group consisting of
homopiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl,
piperidinyl, azetidinyl, pyranyl, tetrahydrofuranyl, piperazinzyl,
phenyl, pyridyl, oxazolyl, pyrimidinyl, oxadiazolyl and thiazolyl,
optionally substituted with from one to three substituents selected
from the group consisting of halogen, --OR.sup.f,
--NR.sup.fR.sup.g, --R.sup.h, --CN, wherein each R.sup.f and
R.sup.g is independently selected from hydrogen, C.sub.1-6 alkyl,
C.sub.1-6 haloalkyl and C.sub.3-6 cycloalkyl, and each R.sup.h is
independently selected from the group consisting of C.sub.1-6
alkyl, C.sub.1-6haloalkyl and C.sub.3-6 cycloalkyl, wherein the
aliphatic portions of R.sup.f, R.sup.g and R.sup.h are optionally
further substituted with from one to three members selected from
the group consisting of --OH, --OR.sup.o, --OC(O)NHR.sup.o,
--OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o, --S(O)R.sup.o,
--S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl. The subcript m may be from 0 to 2; or
alternatively from 0-1.
[0057] In another embodiment of the invention, in the compounds
having formulae Ia or Ib, the symbol R.sup.3a moiety on the
pyrazole ring is hydrogen, halogen, chloro, fluoro, bromo,
oxazolyl, pyridyl,oxadiazolyl thiazolyl, --R.sup.h or cyano; and
optionally the symbol R.sup.1, when present, is selected from the
group consisting of --CO.sub.2H or C.sub.1-4 alkyl, optionally
substituted with --OH, --OR.sup.m, --S(O).sub.2R.sup.m, --CO.sub.2H
and --CO.sub.2R.sup.m. In yet another embodiment, R.sup.1, when
present, is hydrogen or C.sub.1-6 alkyl. m is an integer from
0-2.
[0058] In another embodiment of the invention, in compounds of
formulae Ia and Ib, the R.sup.3a substitutent is selected from the
group consisting of hydrogen, halogen, --OR.sup.f, NR.sup.fR.sup.g,
--R.sup.h, --Y, --CN, X.sup.3N.sub.3, --SO.sub.2R.sup.h,
X.sup.3NR.sup.fR.sup.g, X.sup.3Y, --S(O).sub.3R.sup.f,
--C(C.dbd.NOR.sup.f)NR.sup.fR.sup.g, --NO.sub.2, and
--NR.sup.gC(O)R.sup.f, wherein Y is an optionally substituted group
selected from the group consisting of phenyl, pyridyl, pyrimidinyl,
oxazolyl, thiazolyl, oxadiazolyl and morpholinyl, and R.sup.h is an
optionally substituted group selected from the group consisting of
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-8 cycloalkyl, and
R.sup.f and R.sup.g are each independently an optionally
substituted group selected from the group consisting of hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-8 cycloalkyl. In
certain aspects of this embodiment, the R.sup.3a substituent is
selected from the group consisting of hydrogen, fluoro, chloro,
bromo, iodo, amino, --CH.sub.3, oxazolyl, thiazolyl, pyridyl,
pyrimidinyl, morpholinyl, oxdiazolyl, --NHC(O)CH.sub.3, --CN,
CH.sub.2N.sub.3, CH.sub.2SO.sub.3H, NO.sub.2,
--(C.dbd.NOH)NH.sub.2, --S(O).sub.2CH.sub.3 and
CH.sub.2NH.sub.2.
[0059] In yet another embodiment of the invention, in the compounds
having formulae Ia or Ib, the subscript m is 0 or 1; R.sup.2a is
hydrogen, halogen or --CN ; R.sup.2c is selected from the group
consisting of F, Cl, Br, CN, NO.sub.2, --CO.sub.2CH.sub.3,
--C(O)CH.sub.3 and --S(O).sub.2CH.sub.3; R.sup.2d is selected from
the group consisting of --SR.sup.c, --O--X.sup.2--OR.sup.c,
--X.sup.2--OR.sup.c, --R.sup.e and --OR.sup.c; and R.sup.3a
substituents is selected from the group consisting of halogen,
C.sub.1-4 alkyl and C.sub.1-4 haloalkyl, wherein the aliphatic
portions of R.sup.3a are optionally substituted with from one to
three members selected from the group consisting of --OH,
--OR.sup.o, --OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2, --SH,
--SR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.o, --S(O).sub.2N(R.sup.o).sub.2,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2,
--C(O)NHR.sup.o, --C(O)N(R.sup.o).sub.2, --C(O)R.sup.o,
--NHC(O)R.sup.o, --NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2,
--NR.sup.oC(O)NH.sub.2, --NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein each R.sup.o is
independently an unsubstituted C.sub.1-6 alkyl.
[0060] In one preferred embodiment, in the compounds of the
invention having the formula Ib, when R.sup.2a is H, R.sup.2c is
chloro, R.sup.2d is methoxy, m is 0, a is N, c is N, and b and d
are CH, then R.sup.3a is other than hydrogen, methyl, unsubstituted
2-pyridyl, unsubstituted 2-pyrimidinyl or unsubstituted
2-oxazolyl.
[0061] In one specific embodiment, the present invention provides
compounds having formula Ia and Ib wherein the subscript m is an
integer of from 0 to 4. The symbol R.sup.1 is a substituent
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
--CO.sub.2R.sup.a, --X.sup.1CO.sub.2R.sup.a,
--X.sup.1SO.sub.2R.sup.a and --X.sup.1OR.sup.a, --COR.sup.a,
--CONR.sup.aR.sup.b, --X.sup.1NR.sup.aR.sup.b,
--X.sup.1NR.sup.aCOR.sup.b, --X.sup.1CONR.sup.aR.sup.b,
X.sup.1S(O).sub.2NR.sup.aR.sup.b and X.sup.1S(O).sub.2R.sup.a,
wherein X.sup.1 is C.sub.1-4 alkylene and each R.sup.a and R.sup.b
is independently selected from the group consisting of hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl; and
wherein the aliphatic portions of each of said R.sup.1 substituents
is optionally substituted with from one to three members selected
from the group consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(O)NHR.sup.m,
--NR.sup.mC(O)N(R.sup.m).sub.2, --NHC(O)N(R.sup.m).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.m, --NHCO.sub.2R.sup.m,
--NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2 and
--NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl. The symbols
R.sup.2a, R.sup.2c and R.sup.2d are each substituents independently
selected from the group consisting of hydrogen, halogen, cyano,
heteroaryl, --NO.sub.2, --CO.sub.2R.sup.c, --CONR.sup.cR.sup.d,
--C(O)R.sup.c, --S(O)R.sup.e, --S(O).sub.2R.sup.e, --R.sup.e,
--C(NOR.sup.c)R.sup.d, --C(NR.sup.cV).dbd.NV,
--N(V)C(R.sup.c).dbd.NV, --X.sup.2C(NOR.sup.c)R.sup.d,
--X.sup.2C(NR.sup.cV).dbd.NV, --X.sup.2N(V)C(R.sup.c).dbd.NV,
--X.sup.2NR.sup.cR.sup.d, --X.sup.2SR.sup.c, --X.sup.2CN,
--X.sup.2NO.sub.2, --X.sup.2CO.sub.2R.sup.c,
--X.sup.2CONR.sup.cR.sup.d, --X.sup.2C(O)R.sup.c,
--X.sup.2OC(O)NR.sup.cR.sup.d, --X.sup.2NR.sup.dC(O)R.sup.c,
--X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.e).dbd.NH, --X.sup.2S(O)R.sup.e,
--X.sup.2S(O).sub.2R.sup.e, --X.sup.2NR.sup.cS(O).sub.2R.sup.e,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d, --X.sup.2N.sub.3, --OR.sup.c,
--SR.sup.c, --NR.sup.dC(O)R.sup.e, --NR.sup.dC(O).sub.2R.sup.e,
--S(O).sub.2R.sup.e, --S(O).sub.2NR.sup.cR.sup.d,
--X.sup.2OR.sup.c, --O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d and --NR.sup.d--X.sup.2CO.sub.2R.sup.c.
Within each of R.sup.2a, R.sup.2c and R.sup.2d, X.sup.2 i.sub.s
C.sub.1-4 alkylene and each R.sup.c and R.sup.d is independently
selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, and
C.sub.3-6 cycloalkyl. Optionally, R.sup.c and R.sup.d when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members. The symbol R.sup.e is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl and heteroaryl, and each of R.sup.c, R.sup.d and
R.sup.e is optionally further substituted with from one to three
members selected from the group consisting of --OH, --OR.sup.n,
--OC(O)NHR.sup.n, --OC(O)N(R.sup.n).sub.2, --SH, --SR.sup.n,
--S(O)R.sup.n, --S(O).sub.2R.sup.n, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.n, --S(O).sub.2N(R.sup.n).sub.2,
--NHS(O).sub.2R.sup.n, --NR.sup.nS(O).sub.2R.sup.n, --C(O)NH.sub.2,
--C(O)NHR.sup.n, --C(O)N(R.sup.n).sub.2, --C(O)R.sup.n,
--NHC(O)R.sup.n, --NR.sup.nC(O)R.sup.n, --NHC(O)NH.sub.2,
--NR.sup.nC(O)NH.sub.2, --NR.sup.nC(O)NHR.sup.n, --NHC(O)NHR.sup.n,
--NR.sup.nC(O)N(R.sup.n).sub.2, --NHC(O)N(R.sup.n).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.n, --NHCO.sub.2R.sup.n,
--NR.sup.nCO.sub.2R.sup.n, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.n, --N(R.sup.n).sub.2, --NR.sup.nS(O)NH.sub.2 and
--NR.sup.nS(O).sub.2NHR.sup.n, wherein each R.sup.n is
independently an unsubstituted C.sub.1-6 alkyl; and wherein V is
independently selected from the group consisting of --R.sup.c,
--CN, --CO.sub.2R.sup.e and --NO.sub.2. Each of ring vertices a, b,
c and d in formulae Ia and Ib is independently selected from N and
C(R.sup.3a), and from one to two of said ring vertices is N. The
symbol R.sup.3a in formulae Ia and Ib is independently selected
from the group consisting of hydrogen, halogen, --OR.sup.f,
--OC(O)R.sup.f, --NR.sup.fR.sup.g, --SR.sup.f, --R.sup.h, --CN,
--NO.sub.2, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--OC(O)NR.sup.fR.sup.g, --NR.sup.gC(O)R.sup.f,
--NR.sup.gC(O).sub.2R.sup.h, --NR.sup.f--C(O)NR.sup.fR.sup.g,
--NH--C(NH.sub.2).dbd.NH, --NR.sup.hC(NH.sub.2).dbd.NH,
--NH--C(NH.sub.2).dbd.NR.sup.h, --NH--C(NHR.sup.h).dbd.NH,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --NR.sup.fS(O).sub.2R.sup.h,
--NR.sup.fS(O).sub.2NR.sup.fR.sup.g, --N.sub.3, --X.sup.3OR.sup.f,
--X.sup.3OC(O)R.sup.f, --X.sup.3NR.sup.fR.sup.g, --X.sup.3SR.sup.f,
--X.sup.3CN, --X.sup.3NO.sub.2, --X.sup.3CO.sub.2R.sup.f,
--X.sup.3CONR.sup.fR.sup.g, --X.sup.3C(O)R.sup.f,
--X.sup.3OC(O)NR.sup.fR.sup.g, --X.sup.3NR.sup.gC(O)R.sup.f,
--X.sup.3NR.sup.gC(O).sub.2R.sup.h,
--X.sup.3NR.sup.f--C(O)NR.sup.fR.sup.g,
--X.sup.3NH--C(NH.sub.2).dbd.NH,
--X.sup.3NR.sup.hC(NH.sub.2).dbd.NH,
--X.sup.3NH--C(NH.sub.2).dbd.NR.sup.h,
--X.sup.3NH--C(NHR.sup.h).dbd.NH, --X.sup.3S(O)R.sup.h,
--X.sup.3S(O).sub.2R.sup.h, --X.sup.3NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, --Y, --X.sup.3Y and
--X.sup.3N.sub.3. The symbol Y is a five to ten-membered aryl,
heteroaryl or heterocycloalkyl ring, optionally substituted with
from one to three substituents selected from the group consisting
of halogen, --OR.sup.f, --NR.sup.fR.sup.g, --R.sup.h, --SR.sup.f,
--CN, --NO.sub.2, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g,
--C(O)R.sup.f, --NR.sup.gC(O)R.sup.f, --S(O)R.sup.h,
--S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --X.sup.3OR.sup.f,
--X.sup.3NR.sup.fR.sup.g, --X.sup.3NR.sup.fS(O).sub.2R.sup.h and
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, and wherein each X.sup.3 is
independently selected from the group consisting of C.sub.1-4
alkylene, C.sub.2-4 alkenylene and C.sub.2-4 alkynylene; each
R.sup.f and R.sup.g is independently selected from hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
C.sub.2-8 alkenyl, C.sub.2-8 alkynyl, aryl, heteroaryl,
aryl-C.sub.1-4 alkyl, and aryloxy-C.sub.1-4 alkyl, or when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members; and each R.sup.h is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl, heteroaryl, aryl-C.sub.1-4 alkyl, and
aryloxy-C.sub.1-4 alkyl, wherein the aliphatic portions of X.sup.3,
R.sup.f, R.sup.g and R.sup.h are optionally further substituted
with from one to three members selected from the group consisting
of --OH, --OR.sup.o, --OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2,
--SH, --SR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0062] In another specific embodiment, in compounds having formula
Ia and Ib, R.sup.1 is independently selected from the group
consisting of C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6
cycloalkyl, --CO.sub.2R.sup.a, --X.sup.1CO.sub.2R.sup.a,
--X.sup.1SO.sub.2R.sup.a, --X.sup.1OR.sup.a, --COR.sup.a,
--CONR.sup.aR.sup.b, --X.sup.1NR.sup.aR.sup.b,
--X.sup.1NR.sup.aCOR.sup.b, --X.sup.1CONR.sup.aR.sup.b,
X.sup.1S(O).sub.2NR.sup.aR.sup.b and X.sup.1S(O).sub.2R.sup.a,
wherein X.sup.1 is C.sub.1-4 alkylene and each R.sup.a and R.sup.b
is independently selected from the group consisting of hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl and C.sub.3-6 cycloalkyl. The
aliphatic portions of each of said R.sup.1 substituents is
optionally substituted with from one to three members selected from
the group consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(O)NHR.sup.m,
--NR.sup.mC(O)N(R.sup.m).sub.2, --NHC(O)N(R.sup.m).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.m, --NHCO.sub.2R.sup.m,
--NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2 and
--NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl. The substituents
R.sup.2a, R.sup.2c and R.sup.2d are each independently selected
from the group consisting of hydrogen, halogen, cyano, heteroaryl,
--NO.sub.2, --CO.sub.2R.sup.c, --CONR.sup.cR.sup.d, --C(O)R.sup.c,
--S(O)R.sup.c, --S(O).sub.2R.sup.e, --R.sup.e,
--C(NOR.sup.c)R.sup.d, --C(NR.sup.cV).dbd.NV,
--N(V)C(R.sup.c).dbd.NV, --X.sup.2C(NOR.sup.c)R.sup.d,
--X.sup.2C(NR.sup.cV).dbd.NV, --X.sup.2N(V)C(R.sup.c).dbd.NV,
--X.sup.2NR.sup.cR.sup.d, --X.sup.2SR.sup.c, --X.sup.2CN,
--X.sup.2NO.sub.2, --X.sup.2CO.sub.2R.sup.c,
--X.sup.2CONR.sup.cR.sup.d, --X.sup.2C(O)R.sup.c,
--X.sup.2OC(O)NR.sup.cR.sup.d, --X.sup.2NR.sup.dC(O)R.sup.c,
--X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.e).dbd.NH, --X.sup.2S(O)R.sup.e,
--X.sup.2S(O).sub.2R.sup.e, --X.sup.2NR.sup.cS(O).sub.2R.sup.e,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d, --X.sup.2N.sub.3, --SR.sup.c,
--R.sup.e, --NR.sup.dC(O)R.sup.c, --NR.sup.dC(O).sub.2R.sup.e,
--S(O).sub.2R.sup.e, --S(O).sub.2NR.sup.cR.sup.d,
--X.sup.2OR.sup.c, --O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d and --NR.sup.d--X.sup.2CO.sub.2R.sup.c;
in which within each of R.sup.2a, R.sup.2c and R.sup.2d, X.sup.2 is
C.sub.1-4 alkylene and each R.sup.c and R.sup.d is independently
selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8 haloalkyl,
C.sub.3-6 cycloalkyl, or optionally, R.sup.c and R.sup.d when
attached to the same nitrogen atom can be combined with the
nitrogen atom to form a five or six-membered ring having from 0 to
2 additional heteroatoms as ring members; and each R.sup.e is
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8
alkenyl, C.sub.2-8 alkynyl, aryl and heteroaryl, and each of
R.sup.c, R.sup.d and R.sup.e is optionally further substituted with
from one to three members selected from the group consisting of
--OH, --OR.sup.n, --OC(O)NHR.sup.n, --OC(O)N(R.sup.n).sub.2, --SH,
--SR.sup.n, --S(O)R.sup.n, --S(O).sub.2R.sup.n, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.n, --S(O).sub.2N(R.sup.n).sub.2,
--NHS(O).sub.2R.sup.n, --NR.sup.nS(O).sub.2R.sup.n, --C(O)NH.sub.2,
--C(O)NHR.sup.n, --C(O)N(R.sup.n).sub.2, --C(O)R.sup.n,
--NHC(O)R.sup.n, --NR.sup.nC(O)R.sup.n, --NHC(O)NH.sub.2,
--NR.sup.nC(O)NH.sub.2, --NR.sup.nC(O)NHR.sup.n, --NHC(O)NHR.sup.n,
--NR.sup.nC(O)N(R.sup.n).sub.2, --NHC(O)N(R.sup.n).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.n, --NHCO.sub.2R.sup.n,
--NR.sup.nCO.sub.2R.sup.n, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.n, --N(R.sup.n).sub.2, --NR.sup.nS(O)NH.sub.2 and
--NR.sup.nS(O).sub.2NHR.sup.n. Each R.sup.n is independently an
unsubstituted C.sub.1-6 alkyl; and wherein V is independently
selected from the group consisting of --R.sup.c, --CN,
--CO.sub.2R.sup.e and --NO.sub.2. Each of ring vertices a, b, c and
d in formulae Ia and Ib is independently selected from N and
C(R.sup.3a), and from one to two of said ring vertices is N. The
substituent R.sup.3a is independently selected from the group
consisting of hydrogen, halogen, --OR.sup.f, --OC(O)R.sup.f,
--NR.sup.fR.sup.g, --SR.sup.f, --R.sup.h, --CN, --NO.sub.2,
--CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--OC(O)NR.sup.fR.sup.g, --NR.sup.gC(O)R.sup.f,
--NR.sup.gC(O).sub.2R.sup.h, --NR.sup.f--C(O)NR.sup.fR.sup.g,
--NH--C(NH.sub.2).dbd.NH, --NR.sup.hC(NH.sub.2).dbd.NH,
--NH--C(NH.sub.2).dbd.NR.sup.h, --NH--C(NHR.sup.h).dbd.NH,
--S(O)R.sup.h--S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --NR.sup.fS(O).sub.2R.sup.h,
--NR.sup.fS(O).sub.2NR.sup.fR.sup.g, --N.sub.3, --X.sup.3OR.sup.f,
--X.sup.3OC(O)R.sup.f, --X.sup.3NR.sup.fR.sup.g, --X.sup.3SR.sup.f,
--X.sup.3CN, --X.sup.3NO.sub.2, --X.sup.3CO.sub.2R.sup.f,
--X.sup.3CONR.sup.fR.sup.g, --X.sup.3C(O)R.sup.f,
--X.sup.3OC(O)NR.sup.fR.sup.g, --X.sup.3NR.sup.gC(O)R.sup.f,
--X.sup.3NR.sup.gC(O).sub.2R.sup.h,
--X.sup.3NR.sup.f--C(O)NR.sup.fR.sup.g,
--X.sup.3NH--C(NH.sub.2).dbd.NH,
--X.sup.3NR.sup.hC(NH.sub.2).dbd.NH,
--X.sup.3NH--C(NH.sub.2).dbd.NR.sup.h,
--X.sup.3NH--C(NHR.sup.h).dbd.NH, --X.sup.3S(O)R.sup.h,
--X.sup.3S(O).sub.2R.sup.h, --X.sup.3NR.sup.fS(O).sub.2R.sup.h,
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, --Y, --X.sup.3Y and
--X.sup.3N.sub.3. The symbol Y is a five to ten-membered aryl,
heteroaryl or heterocycloalkyl ring, optionally substituted with
from one to three substituents selected from the group consisting
of halogen, --OR.sup.f, --NR.sup.fR.sup.g, --R.sup.h, --SR.sup.f,
--CN, --NO.sub.2, --CO.sub.2R.sup.f, --CONR.sup.fR.sup.g,
--C(O)R.sup.f, --NR.sup.gC(O)R.sup.f, --S(O)R.sup.h,
--S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --X.sup.3OR.sup.f,
--X.sup.3NR.sup.fR.sup.g, --X.sup.3NR.sup.fS(O).sub.2R.sup.h and
--X.sup.3S(O).sub.2NR.sup.fR.sup.g, and wherein each X.sup.3 is
independently selected from the group consisting of C.sub.1-4
alkylene, C.sub.2-4 alkenylene and C.sub.2-4 alkynylene and each
R.sup.f and R.sup.g is independently selected from hydrogen,
C.sub.1-8 alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
C.sub.2-8 alkenyl, C.sub.2-8 alkynyl, aryl, heteroaryl,
aryl-C.sub.1-4 alkyl, and aryloxy-C.sub.1-4 alkyl, or when attached
to the same nitrogen atom can be combined with the nitrogen atom to
form a five or six-membered ring having from 0 to 2 additional
heteroatoms as ring members, and each R.sup.h is independently
selected from the group consisting of C.sub.1-8 alkyl, C.sub.1-8
haloalkyl, C.sub.3-6 cycloalkyl, C.sub.2-8 alkenyl, C.sub.2-8
alkynyl, aryl, heteroaryl, aryl-C.sub.1-4 alkyl, and
aryloxy-C.sub.1-4 alkyl, wherein the aliphatic portions of X.sup.3,
R.sup.f, R.sup.g and R.sup.h are optionally further substituted
with from one to three members selected from the group consisting
of --OH, --OR.sup.o, --OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2,
--SH, --SR.sup.o, --S(O)R.sup.o, --S(O).sub.2R.sup.o,
--SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.o,
--S(O).sub.2N(R.sup.o).sub.2, --NHS(O).sub.2R.sup.o,
--NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2, --C(O)NHR.sup.o,
--C(O)N(R.sup.o).sub.2, --C(O)R.sup.o, --NHC(O)R.sup.o,
--NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2, --NR.sup.oC(O)NH.sub.2,
--NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein R.sup.o is unsubstituted
C.sub.1-6 alkyl.
[0063] In another specific embodiment of the invention, in
compounds having formula Ia or Ib, each R.sup.1 is a substituent
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8 haloalkyl, C.sub.3-6 cycloalkyl,
--CO.sub.2R.sup.a, --X.sup.1CO.sub.2R.sup.a,
--X.sup.1SO.sub.2R.sup.a and --X.sup.1OR.sup.a, wherein the
aliphatic portions of each of said R.sup.1 substituents is
optionally substituted with from one to three members selected from
the group consisting of --OH, --OR.sup.m, --OC(O)NHR.sup.m,
--OC(O)N(R.sup.m).sub.2, --SH, --SR.sup.m, --S(O)R.sup.m,
--S(O).sub.2R.sup.m, --SO.sub.2NH.sub.2, --S(O).sub.2NHR.sup.m,
--S(O).sub.2N(R.sup.m).sub.2, --NHS(O).sub.2R.sup.m,
--NR.sup.mS(O).sub.2R.sup.m, --C(O)NH.sub.2, --C(O)NHR.sup.m,
--C(O)N(R.sup.m).sub.2, --C(O)R.sup.m, --NHC(O)R.sup.m,
--NR.sup.mC(O)R.sup.m, --NHC(O)NH.sub.2, --NR.sup.mC(O)NH.sub.2,
--NR.sup.mC(O)NHR.sup.m, --NHC(O)NHR.sup.m,
--NR.sup.mC(O)N(R.sup.m).sub.2, --NHC(O)N(R.sup.m).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.m, --NHCO.sub.2R.sup.m,
--NR.sup.mCO.sub.2R.sup.m, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.m, --N(R.sup.m).sub.2, --NR.sup.mS(O)NH.sub.2 and
--NR.sup.mS(O).sub.2NHR.sup.m, wherein each R.sup.m is
independently an unsubstituted C.sub.1-6 alkyl. The R.sup.2a
substituent is selected from the group consisting of hydrogen,
halogen, cyano, heteroaryl, --NO.sub.2, --CO.sub.2R.sup.c,
--CONR.sup.cR.sup.d, --C(O)R.sup.c, --S(O)R.sup.e,
--S(O).sub.2R.sup.e, --R.sup.e, --C(NOR.sup.c)R.sup.d,
--C(NR.sup.cV).dbd.NV, --N(V)C(R.sup.c).dbd.NV,
--X.sup.2C(NOR.sup.c)R.sup.d, --X.sup.2C(NR.sup.cV).dbd.NV,
--X.sup.2N(V)C(R.sup.c).dbd.NV, --X.sup.2NR.sup.cR.sup.d,
--X.sup.2SR.sup.c, --X.sup.2CN, --X.sup.2NO.sub.2,
--X.sup.2CO.sub.2R.sup.c, --X.sup.2CONR.sup.cR.sup.d,
--X.sup.2C(O)R.sup.c, --X.sup.2OC(O)NR.sup.cR.sup.d,
--X.sup.2NR.sup.dC(O)R.sup.c, --X.sup.2NR.sup.dC(O).sub.2R.sup.e,
--X.sup.2NR.sup.cC(O)NR.sup.cR.sup.d,
--X.sup.2NH--C(NH.sub.2).dbd.NH,
--X.sup.2NR.sup.eC(NH.sub.2).dbd.NH,
--X.sup.2NH--C(NH.sub.2).dbd.NR.sup.e,
--X.sup.2NH--C(NHR.sup.c).dbd.NH, --X.sup.2S(O)R.sup.c,
--X.sup.2S(O).sub.2R.sup.c, --X.sup.2NR.sup.cS(O).sub.2R.sup.c,
--X.sup.2S(O).sub.2NR.sup.cR.sup.d and --X.sup.2N.sub.3. The
R.sup.2c and R.sup.2d substituents are each independently selected
from the group consisting of halogen, --OR.sup.c, --SR.sup.c,
--R.sup.e, --CN, --NO.sub.2, --CO.sub.2R.sup.c, --C(O)R.sup.c,
--NR.sup.dC(O)R.sup.c, --NR.sup.dC(O).sub.2R.sup.e,
--S(O).sub.2R.sup.e, --S(O).sub.2NR.sup.cR.sup.d,
--X.sup.2OR.sup.c, --O--X.sup.2OR.sup.c, --X.sup.2NR.sup.cR.sup.d,
--O--X.sup.2NR.sup.cR.sup.d and --NR.sup.d--X.sup.2CO.sub.2R.sup.c.
Each R.sup.3a substituent is independently selected from the group
consisting of hydrogen, halogen, --OR.sup.f, --OC(O)R.sup.f,
--NR.sup.fR.sup.g, --SR.sup.f, --R.sup.h, --CN, --NO.sub.2,
--CO.sub.2R.sup.f, --CONR.sup.fR.sup.g, --C(O)R.sup.f,
--OC(O)NR.sup.fR.sup.g, --NR.sup.gC(O)R.sup.f,
--NR.sup.gC(O).sub.2R.sup.h, --NR.sup.f--C(O)NR.sup.fR.sup.g,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h,
--S(O).sub.2NR.sup.fR.sup.g, --NR.sup.fS(O).sub.2NR.sup.fR.sup.g,
--X.sup.3OR.sup.f, --X.sup.3NR.sup.fR.sup.g, --X.sup.3SR.sup.f,
--X.sup.3CN, --C(C.dbd.NOR.sup.f)NR.sup.fR.sup.g,
X.sup.3SO.sub.3R.sup.f, --X.sup.3CO.sub.2R.sup.f,
--X.sup.3CONR.sup.fR.sup.g, --X.sup.3C(O)R.sup.f,
--X.sup.3NR.sup.gC(O)R.sup.f, --X.sup.3NR.sup.gC(O).sub.2R.sup.h,
--Y, --X.sup.3Y, --X.sup.3N.sub.3, wherein Y is selected from the
group consisting of a five or six-membered aryl ring, a five or
six-membered heteroaryl ring and three to eight membered
heterocycloalkyl ring, wherein said Y group is optionally
substituted with from one to three substituents selected from the
group consisting of halogen, --OR.sup.f, --NR.sup.fR.sup.g,
--R.sup.h, --SR.sup.f, --CN, --NO.sub.2, --CO.sub.2R.sup.f,
--CONR.sup.fR.sup.g, --C(O)R.sup.f, --NR.sup.gC(O)R.sup.f,
--S(O)R.sup.h, --S(O).sub.2R.sup.h, --NR.sup.fS(O).sub.2R.sup.h and
--S(O).sub.2NR.sup.fR.sup.g, and wherein each X.sup.3 is
independently C.sub.1-4 alkylene, and each R.sup.f and R.sup.g is
independently selected from hydrogen, C.sub.1-8 alkyl, C.sub.1-8
haloalkyl and C.sub.3-6 cycloalkyl, and each R.sup.h is
independently selected from the group consisting of C.sub.1-8
alkyl, C.sub.1-8haloalkyl and C.sub.3-6 cycloalkyl, wherein the
aliphatic portions of X.sup.3, R.sup.f, R.sup.g and R.sup.h is
optionally further substituted with from one to three members
selected from the group consisting of --OH, --OR.sup.o,
--OC(O)NHR.sup.o, --OC(O)N(R.sup.o).sub.2, --SH, --SR.sup.o,
--S(O)R.sup.o, --S(O).sub.2R.sup.o, --SO.sub.2NH.sub.2,
--S(O).sub.2NHR.sup.o, --S(O).sub.2N(R.sup.o).sub.2,
--NHS(O).sub.2R.sup.o, --NR.sup.oS(O).sub.2R.sup.o, --C(O)NH.sub.2,
--C(O)NHR.sup.o, --C(O)N(R.sup.o).sub.2, --C(O)R.sup.o,
--NHC(O)R.sup.o, --NR.sup.oC(O)R.sup.o, --NHC(O)NH.sub.2,
--NR.sup.oC(O)NH.sub.2, --NR.sup.oC(O)NHR.sup.o, --NHC(O)NHR.sup.o,
--NR.sup.oC(O)N(R.sup.o).sub.2, --NHC(O)N(R.sup.o).sub.2,
--CO.sub.2H, --CO.sub.2R.sup.o, --NHCO.sub.2R.sup.o,
--NR.sup.oCO.sub.2R.sup.o, --CN, --NO.sub.2, --NH.sub.2,
--NHR.sup.o, --N(R.sup.o).sub.2, --NR.sup.oS(O)NH.sub.2 and
--NR.sup.oS(O).sub.2NHR.sup.o, wherein each R.sup.o is
independently an unsubstituted C.sub.1-6 alkyl.
[0064] In another embodiment of the invention, the compounds of the
invention having formula Ib is represented by formulae Ib.sup.1 and
Ib.sup.2:
##STR00003##
or a pharmaceutically acceptable salt, hydrate or N-oxide thereof.
In one embodiment, in formulae Ib.sup.1 and Ib.sup.2, R.sup.2c is
halogen, cyano or nitro; the symbol R.sup.2d is selected from
--SR.sup.c, --O--X.sup.2--OR.sup.c, --X.sup.2--OR.sup.c, --R.sup.e,
--OR.sup.c and --NR.sup.dC(O)R.sup.c; each of ring vertices a, b, c
and d is independently selected from N and C(R.sup.3a), and from
one to two of said ring vertices is N; and each R.sup.3a is
independently selected from the group consisting of hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.3-6
cycloalkyl, C.sub.3-6 heterocycloalkyl, --S(O).sub.2R.sup.h, amino,
phenyl, pyridyl, pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolyl
imiazolyl and thiazolyl. In one embodiment, the ring vertex a is N.
In another embodiment, the ring vertex b is N. In another
embodiment, the ring vertex c is N. In another embodiment, the ring
vertex d is N. In yet another embodiment, the ring vertices a and c
are each N; b is hydrogen; and d is C(R.sup.3a), wherein R.sup.3a
on ring vertex d is other than hydrogen. In another embodiment, the
ring vertex a is N; b is C(R.sup.3a) wherein R.sup.3a on ring
vertex b is other than hydrogen; and c and d are each hydrogen. In
another embodiment, the ring vertex a is N; b and c are each
hydrogen; and d is C(R.sup.3a), wherein R.sup.3a on ring vertex d
is other than hydrogen. In another embodiment, the ring vertex a is
C(R.sup.3a), wherein R.sup.3a on ring vertex a is other than
hydrogen; b is N; c and d are each hydrogen. In another embodiment,
the ring vertex a is N; b and d are each hydrogen; and c is
C(R.sup.3a); wherein R.sup.3a on ring vertex c is other than
hydrogen. In another embodiment, the ring vertices a and c are each
N; b is hydrogen; and d is C(R.sup.3a), wherein R.sup.3a on ring
vertex d is other than hydrogen.
[0065] In yet another embodiment of the invention, in formulae
Ib.sup.1 and Ib.sup.2, R.sup.2c and R.sup.2d are each a member
independently selected from the group consisting of halogen,
C.sub.1-4 alkoxy, C.sub.1-4 alkyl and C.sub.1-4 haloalkyl; R.sup.3a
on the pyrazole ring is an imidazolyl group optionally substituted
with C.sub.1-4 alkyl; and the ring vertex a is nitrogen and the
ring vertices b, c, and d are each C(H). Within certain aspects of
this embodiment, R.sup.2c is selected from the group consisting of
fluoro, chloro, bromo and iodo; and R.sup.2d is selected from the
group consisting of methoxy, ethoxy, trifluoromethoxy, methyl,
ethyl, trifluoromethyl and 2-fluoroethoxy.
[0066] In yet another embodiment of the invention, the compounds of
the invention having formula Ib is represented by formulae Ib.sup.3
and Ib.sup.4:
##STR00004##
or a pharmaceutically acceptable salt, hydrate and an N-oxide
thereof. In one embodiment, in formulae Ib.sup.3 and Ib.sup.4,
R.sup.2c is independently halogen, cyano or nitro; R.sup.2d is
selected from --SR.sup.c, --O--X.sup.2--OR.sup.c,
--X.sup.2--OR.sup.c, --R.sup.e, --OR.sup.c, --NR.sup.cR.sup.d,
--NR.sup.cS(O).sub.2R.sup.e and --NR.sup.dC(O)R.sup.c; R.sup.2a is
selected from the group consisting of F, Cl, Br, I, --CO.sub.2Me,
--CONH.sub.2, CN, oxazolyl, --CH.sub.2NH.sub.2, --CH.sub.2NHMe,
--CH.sub.2NMe.sub.2 and --CH.dbd.N--OH; each of ring vertices a, b,
c and d is independently selected from N and C(R.sup.3a), and from
one to two of said ring vertices is N; and each R.sup.3a is
independently selected from the group consisting of hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.3-6
cycloalkyl, C.sub.3-6 heterocycloalkyl, --S(O).sub.2R.sup.h, amino,
phenyl, pyridyl, pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolyl and
thiazolyl. In one embodiment the ring vertex a is N. In another
embodiment the ring vertex b is N. In another embodiment the ring
vertex c is N. In another embodiment the ring vertex d is N. In yet
another embodiment, the ring vertices a and c are each N; b is
hydrogen; and d is C(R.sup.3a), wherein R.sup.3a on ring vertex d
is other than hydrogen. In another embodiment, the ring vertex a is
N; b is C(R.sup.3a) wherein R.sup.3a on ring vertex b is other than
hydrogen; and c and d are each hydrogen. In another embodiment, the
ring vertex a is N; b and c are each hydrogen; and d is
C(R.sup.3a), wherein R.sup.3a on ring vertex d is other than
hydrogen. In another embodiment, the ring vertex a is C(R.sup.3a),
wherein R.sup.3a on ring vertex a is other than hydrogen; b is N; c
and d are each hydrogen. In another embodiment, the ring vertex a
is N; b and d are each hydrogen; and c is C(R.sup.3a); wherein
R.sup.3a on ring vertex c is other than hydrogen. In another
embodiment, the ring vertices a and c are each N; b is hydrogen;
and d is C(R.sup.3a), wherein R.sup.3a on ring vertex d is other
than hydrogen.
[0067] In yet another embodiment of the invention, in formulae
Ib.sup.3 and Ib.sup.4, R.sup.2a, R.sup.2c and R.sup.2d are each a
member independently selected from the group consisting of
hydrogen, halogen, C.sub.1-4 alkoxy, C.sub.1-4 alkyl and C.sub.1-4
haloalkyl; R.sup.3a on the pyrazole ring is an imidazolyl group
optionally substituted with C.sub.1-4 alkyl; and the ring vertex a
is nitrogen and the ring vertices b, c, and d are each C(H). Within
certain aspects of this embodiment, R.sup.2a and R.sup.2c in are
each independently selected from the group consisting of fluoro,
chloro, bromo and iodo; and R.sup.2d is selected from the group
consisting of methoxy, ethoxy, trifluoromethoxy, methyl, ethyl,
trifluoromethyl and 2-fluoroethoxy.
[0068] In yet another embodiment, the compounds of the invention
having formula Ia is represented by formulae Ia.sup.1 or
Ia.sup.2:
##STR00005##
or a pharmaceutically acceptable salt, hydrate or N-oxide thereof;
wherein the symbol R.sup.2c is halogen, cyano or nitro; the symbol
R.sup.2d is selected from --SR.sup.c, --O--X.sup.2--OR.sup.c,
--X.sup.2--OR.sup.c, --R.sup.e, --OR.sup.c and
--NR.sup.dC(O)R.sup.c; each of ring vertices a, b, c and d is
independently selected from N and C(R.sup.3a), and from one to two
of said ring vertices is N; and each R.sup.3a is independently
selected from the group consisting of hydrogen, halogen, C.sub.1-6
alkyl, C.sub.1-6 haloalkyl, C.sub.3-6 cycloalkyl, C.sub.3-6
heterocycloalkyl, --S(O).sub.2R.sup.h, amino, phenyl, pyridyl,
pyrimidinyl, oxazoylyl, oxadiazolyl, isoxazolyl and thiazolyl. In
one embodiment, the ring vertex d is N. In another embodiment the
ring vertex b is N. In another embodiment the ring vertex c is N.
In another embodiment the ring vertex d is N. In another
embodiment, the ring vertex a is N; b and d are each hydrogen; and
c is C(R.sup.3a) wherein R.sup.3a is other than hydrogen. In
another embodiment, the ring vertex a is C(R.sup.3a), wherein
R.sup.3a on ring vertex a is other than hydrogen; b is N; and c and
d are each hydrogen. In another embodiment, the ring vertex a is N;
b and c are each hydrogen; and d is C(R.sup.3a) wherein R.sup.3a on
ring vertex d is other than hydrogen. In another embodiment, the
ring vertex a is C(R.sup.3a), wherein R.sup.3a on ring vertex a is
other than hydrogen; b and c are each hydrogen; and d is N. In
another embodiment, the ring vertex a is C(R.sup.3a), wherein
R.sup.3a on ring vertex a is other than hydrogen; b and d are each
N; and c is hydrogen. In another embodiment, the ring vertices a
and b are each hydrogen; c is C(R.sup.3a), wherein R.sup.3a on ring
vertex c is other than hydrogen; and d is N.
[0069] In yet another embodiment, the compounds of the invention
having formula Ia is represented by formula Ia.sup.3 and
Ia.sup.4:
##STR00006##
or pharmaceutically acceptable salt, hydrate or N-oxide thereof;
wherein R.sup.2c is halogen, cyano or nitro; R.sup.2d is selected
from --SR.sup.c, --O--X.sup.2--OR.sup.c, --X.sup.2--OR.sup.c,
--R.sup.e, --OR.sup.c and --NR.sup.dC(O)R.sup.c; R.sup.2a is
selected from the group consisting of F, Cl, Br, I, --CO.sub.2Me,
--CONH.sub.2, CN, oxazolyl, --CH.sub.2NH.sub.2, --CH.sub.2NHMe, and
--CH.sub.2NMe.sub.2; each of ring vertices a, b, c and d is
independently selected from N and C(R.sup.3a), and from one to two
of said ring vertices is N; and each R.sup.3a is independently
selected from the group consisting of hydrogen, halogen, C.sub.1-6
alkyl, C.sub.1-6 haloalkyl, C.sub.3-6 cycloalkyl, C.sub.3-6
heterocycloalkyl, --S(O).sub.2R.sup.h, amino, phenyl, pyridyl,
pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolyl and thiazolyl. In
one embodiment, the ring vertex a is N. In another embodiment the
ring vertex b is N. In another embodiment the ring vertex c is N.
In another embodiment, the ring vertex d is N. In another
embodiment, the ring vertex a is C(R.sup.3a), wherein R.sup.3a on
ring vertex a is other than hydrogen; b is N; and c and d are each
hydrogen. In another embodiment, the ring vertex a is N; b and c
are each hydrogen; and d is C(R.sup.3a) wherein R.sup.3a on ring
vertex d is other than hydrogen. In another embodiment, the ring
vertex a is C(R.sup.3a), wherein R.sup.3a on ring vertex a is other
than hydrogen; b and c are each hydrogen; and d is N. In another
embodiment, the ring vertex a is C(R.sup.3a), wherein R.sup.3a on
ring vertex a is other than hydrogen; b and d are each N; and c is
hydrogen. In another embodiment, the ring vertices a and b are each
hydrogen; c is C(R.sup.3a), wherein R.sup.3a on ring vertex c is
other than hydrogen; and d is N.
[0070] A family of specific compound of particular interest having
formulae Ia and Ib consists of compounds, pharmaceutically
acceptable salts, hydrates or N-oxides thereof, as set forth in
Table 1.
TABLE-US-00001 TABLE 1 1.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyrid-
in-1-yl- ethanone 2.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[4,3-b]pyrid-
in-2-yl- ethanone 3.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3-
,4- b]pyridin-2-yl)-ethanone 4.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyra-
zin-1-yl-7- oxide)-ethanone 5.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyra-
zin-1-yl-7- oxide)-ethanone 6.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-py-
razolo[3,4- b]pyridin-1-yl-ethanone. 7.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-py-
razolo[3,4- b]pyridin-2-yl-ethanone. 8.
2-(3-Amino-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-metho-
xy-phenyl)- 2-methyl-piperazin-1-yl]-ethanone. 9.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-chloro-pyrazolo[3-
,4- b]pyridin-1-yl)-ethanone. 10.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-methyl- pyrazolo[3,4-b]pyridin-1-yl)-ethanone. 11.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyri-
din-2-yl- ethanone. 12.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-b]pyri-
din-1-yl- ethanone. 13.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-p-
yrazolo[4,3- c]pyridin-1-yl-ethanone. 14.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-p-
yrazolo[3,4- c]pyridin-2-yl-ethanone. 15.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-pyridin-2-yl-pyr-
azolo[3,4- b]pyridin-1-yl)-ethanone. 16.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyr-
azolo[3,4- b]pyridin-1-yl)-ethanone. 17.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3-
,4-b]pyridin- 1-yl-ethanone. 18.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3-
,4-b]pyridin- 2-yl-ethanone. 19.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone. 20.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyra-
zolo[3,4- b]pyridin-1-yl)-ethanone. 21.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-fluoro-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone. 22.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-oxazol-2- yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 23.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyri-
din-2-yl- ethanone 24.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyri-
din-1-yl- ethanone 25.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3-
,4-b]pyridin- 1-yl-ethanone 26.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-thiazol-2- yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 27.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-pyridin-2- yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 28.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone 29.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-methyl- pyrazolo[3,4-b]pyridin-1-yl)-ethanone 30.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-thiazol-2-yl-pyr-
azolo[3,4- b]pyridin-1-yl)-ethanone 31.
1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-py-
razolo[3,4- b]pyridine-3-carbonitrile 32.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-b]pyr-
idin-1-yl-2- oxide)-ethanone 33.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone 34.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[-
3,4- b]pyridin-2-yl)-ethanone 35.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-morpholin-4-yl-p-
yrazolo[3,4- b]pyridin-1-yl)-ethanone 36.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-p-
yrazolo[3,4- c]pyridin-1-yl-ethanone 37.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(pyrazolo[3,4-c]pyr-
idin-1-yl-6- oxide)-ethanone 38.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[-
4,3- c]pyridin-2-yl)-ethanone 39.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[3,-
4-b]pyridin- 2-yl)-ethanone 40.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[3,-
4-b]pyridin- 1-yl)-ethanone 41.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methanesulfonyl-
pyrazolo[3,4-b]pyridin-1-yl)-ethanone 42.
2-(3-Azidomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-
-phenyl)- piperazin-1-yl]-ethanone 43.
(1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-
pyrazolo[3,4-b]pyridin-3-yl)-methanesulfonic acid 44.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone 45.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[-
3,4- d]pyrimidin-2-yl)-ethanone 46.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[-
3,4- d]pyrimidin-1-yl)-ethanone 47.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methoxy-pyrazolo-
[3,4- d]pyrimidin-1-yl)-ethanone 48.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[-
3,4- b]pyridin-2-yl)-ethanone 49.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[-
3,4- b]pyridin-1-yl)-ethanone 50.
2-(6-Azido-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-pheny-
l)- piperazin-1-yl]-ethanone 51.
2-(6-Amino-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-pheny-
l)- piperazin-1-yl]-ethanone 52.
2-(7-Azido-pyrazolo[3,4-c]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-pheny-
l)- piperazin-1-yl]-ethanone 53.
2-(7-Amino-pyrazolo[3,4-c]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-pheny-
l)- piperazin-1-yl]-ethanone 54.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyra-
zolo[3,4- b]pyridin-2-yl)-ethanone 55.
2-(5-Amino-3-methyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-meth-
oxy- phenyl)-piperazin-1-yl]-ethanone 56.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methyl-5-nitro-p-
yrazolo[3,4- b]pyridin-1-yl)-ethanone 57.
2-(3-Amino-6-methyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-meth-
oxy- phenyl)-piperazin-1-yl]-ethanone 58.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3- [1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone 59.
1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-N-hyd-
roxy-1H- pyrazolo[3,4-b]pyridine-3-carboxamidine 60.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-[1,2,4]oxadiazol-
-3-yl- pyrazolo[3,4-b]pyridin-1-yl)-ethanone 61.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(5-methyl-[1,2,4-
]oxadiazol- 3-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 62.
N-(1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-
- pyrazolo[3,4-b]pyridin-6-yl)-acetamide 63.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methanesulfonyl-
pyrazolo[3,4-b]pyridin-1-yl)-ethanone 64.
2-(3-Aminomethyl-pyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-3-methoxy-
-phenyl)- piperazin-1-yl]-ethanone 65.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,-
4-b]pyridin- 1-yl)-ethanone 66.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-(-
3-iodo- pyrazolo[3,4-b]pyridin-1-yl)-ethanone 67.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol--
2-yl- pyrazolo[3,4-b]pyridin-1-yl)-ethanone 68.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-y-
l)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 69.
1-[4-(4-Chloro-3-ethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl-
)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 70.
1-{4-[4-Chloro-3-(2-fluoro-ethoxy)-phenyl]-piperazin-1-yl}-2-[3-(1H-im-
idazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 71.
1-[4-(4-Chloro-3-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 72.
1-[4-(4-Chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 73.
1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 74.
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 75.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 76.
1-{4-[4-Chloro-3-(2-fluoro-ethoxy)-phenyl]-2-methyl-piperazin-1-yl}-2--
[3-(1H- imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 77.
1-[4-(4-Chloro-3-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 78.
1-[4-(4-Chloro-5-ethoxy-2-fluoro-phenyl)-piperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 79.
1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 80.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[-
3-(1H- imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 81.
1-[4-(4-Chloro-3-ethoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 82.
1-{4-[4-Chloro-2-fluoro-5-(2-fluoro-ethoxy)-phenyl]-piperazin-1-yl}-2--
[3-(1H- imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 83.
1-[4-(4-Chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-[3-(1H-imi-
dazol-2-yl)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone
84.
1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl-
)- pyrazolo[3,4-b]pyridin-1-yl]-ethanone 85.
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-[-
3-(1H- imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone 86.
2-[4-Amino-3-(1H-imidazol-2-yl)-pyrazolo[3,4-d]pyrimidin-1-yl]-1-[4-(4-
-chloro-3- methoxy-phenyl)-piperazin-1-yl]-ethanone 87.
2-[5-Chloro-3-(1H-imidazol-2-yl)-indazol-1-yl]-1-[4-(4-chloro-3-methox-
y-phenyl)- piperazin-1-yl]-ethanone 88.
2-[6-Chloro-3-(1H-imidazol-2-yl)-indazol-1-yl]-1-[4-(4-chloro-3-methox-
y-phenyl)- piperazin-1-yl]-ethanone
[0071] Another family of specific compounds of particular interest
having formulae Ia and Ib consists of compounds, pharmaceutically
acceptable salts, hydrates or N-oxides thereof as set forth in
Table 2.
Preparation of Compounds
[0072] As provided in the examples below, the compounds and
intermediates of the present invention can be prepared by one of
skill in the art in a component assembly manner. Schemes 1A-1M
illustrate a variety of methods for the preparation of a variety of
azaindazole-type derivatives. In each of these schemes, X is
halogen; Nu is nucleophilic group; the symbol within an aryl ring
indicate the replacement of one to two carbon(s) of said aryl ring
vertex (vertices) with nitrogen atom(s); L is a ligand; and
non-interfering substituents are provided as --R, --R', --R'', and
--R'''.
##STR00007##
[0073] Scheme 1A shows the synthesis of azaindazole derivatives
from halo-pyridine-carbaldehyde or ketone.
##STR00008##
[0074] Scheme 1B shows the synthesis of azaindazole derivatives
from halo-cyanopyridines.
##STR00009##
[0075] Scheme 1C shows the synthesis of azaindazole derivatives
from amino-methyl-pyridine.
##STR00010##
[0076] Scheme 1D shows the reaction of azaindazole derivatives with
an .alpha.-haloacetate or .alpha.-haloacetamide.
##STR00011##
[0077] Scheme 1E shows the reaction of azaindazole derivatives with
an electrophilic halogen source (X+).
##STR00012##
[0078] Scheme 1F shows a metal-assisted coupling reaction of a
halo-azaindazole derivative.
##STR00013##
[0079] Scheme 1G shows a metal-assisted amination reaction of a
halo-azaindazole derivative.
##STR00014##
[0080] Scheme 1H shows the amination of an azaindazole
derivative.
##STR00015##
[0081] Scheme 1I shows the functionalization of an azaindazole
derivative.
##STR00016##
[0082] Scheme 1J shows the synthesis of a pyrazolopyrazine
derivative.
##STR00017##
[0083] Scheme 1K shows the synthesis of a pyrazolopyrimidine
derivative.
##STR00018##
[0084] Scheme 1L shows the synthesis of a pyrazolopyrimidine
derivative.
##STR00019##
[0085] Scheme 1M shows the synthesis of N-oxide derivatives of the
invention.
##STR00020##
[0086] Scheme 1N shows the synthesis of 3-imidazolyl-azaindazoles
derivatives.
[0087] Scheme 1N illustrates the synthesis of 3-imidazolyl
substituted pyrazolo[3,4-b]pyridines wherein R represents a
non-interfering substituent, such as, for example, a protecting
group, a carboxy ester or the remainder of the compound of formula
Ib. As shown in Scheme 1N, the reaction of NH.sub.2OH with 3-cyano
pyrazolo[3,4-b]pyridine provides a hydroxylamidine compound, which
upon reduction using hydrogen gas and a catalyst (e.g., Pd/C or
Pd(OH).sub.2 produces an the amidine product. Cyclization of the
amindine by treatment with chloroacetaldehyde will produce the
imidazole product.
##STR00021##
[0088] Scheme 1O shows the synthesis of 3-imidazolyl-azaindazoles
derivatives.
[0089] Scheme 1O illustrates the synthesis of 3-imidazolyl
substituted pyrazolo[3,4-b]pyridines. R' represents a
non-interfering substituent, such as, for example, a protecting
group, a carboxy ester, or the remainder of the compound of formula
Ib. As shown in Scheme 1O, the reaction of
3-cyano-pyrazolo[3,4-b]pyridine with ethylene diamine produces the
cyclized imidazoline product, which upon oxidation produces the
imidazole.
##STR00022##
[0090] Scheme 1P shows the synthesis of 3-imidazolyl-azaindazoles
derivatives. 3-imidazolyl-azaindazoles shown in Scheme 1P can be
produced in kilogram scale using the outlined synthetic route.
[0091] Scheme 1P illustrates the synthesis of 3-imidazolyl
substituted pyrazolo[3,4-b]pyridines, wherein R' represents a
non-interfering substituent, such as, for example, a protecting
group, or a carboxy ester, or the remainder of the compound of
formula I (see also the Examples). As shown in Scheme 1P, using a
transmetallation process, a 3-iodo-pyrazolo[3,4-b]pyridine
intermediate can be converted to a 3-formyl-pyrazolo[3,4-b]pyridine
intermediate, which upon treatment with glyoxal, is cyclized to
form 3-imidazolyl-pyrazolo[3,4]pyridine product.
IV. PHARMACEUTICAL COMPOSITIONS
[0092] In addition to the compounds provided above, compositions
for modulating CCR1 activity in humans and animals will typically
contain a pharmaceutical carrier or diluent.
[0093] The term "composition" as used herein is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly
or indirectly, from combination of the specified ingredients in the
specified amounts. By "pharmaceutically acceptable" it is meant the
carrier, diluent or excipient must be compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0094] The pharmaceutical compositions for the administration of
the compounds of this invention may conveniently be presented in
unit dosage form and may be prepared by any of the methods well
known in the art of pharmacy and drug delivery. All methods include
the step of bringing the active ingredient into association with
the carrier which constitutes one or more accessory ingredients. In
general, the pharmaceutical compositions are prepared by uniformly
and intimately bringing the active ingredient into association with
a liquid carrier or a finely divided solid carrier or both, and
then, if necessary, shaping the product into the desired
formulation. In the pharmaceutical composition the active object
compound is included in an amount sufficient to produce the desired
effect upon the process or condition of diseases.
[0095] The pharmaceutical compositions containing the active
ingredient may be in a form suitable for oral use, for example, as
tablets, troches, lozenges, aqueous or oily suspensions,
dispersible powders or granules, emulsions and self emulsifications
as described in U.S. Patent Application 2002-0012680, hard or soft
capsules, syrups, elixirs, solutions, buccal patch, oral gel,
chewing gum, chewable tablets, effervescent powder and effervescent
tablets. Compositions intended for oral use may be prepared
according to any method known to the art for the manufacture of
pharmaceutical compositions and such compositions may contain one
or more agents selected from the group consisting of sweetening
agents, flavoring agents, coloring agents, antioxidants and
preserving agents in order to provide pharmaceutically elegant and
palatable preparations. Tablets contain the active ingredient in
admixture with non-toxic pharmaceutically acceptable excipients
which are suitable for the manufacture of tablets. These excipients
may be for example, inert diluents, such as cellulose, silicon
dioxide, aluminum oxide, calcium carbonate, sodium carbonate,
glucose, mannitol, sorbitol, lactose, calcium phosphate or sodium
phosphate; granulating and disintegrating agents, for example, corn
starch, or alginic acid; binding agents, for example PVP,
cellulose, PEG, starch, gelatin or acacia, and lubricating agents,
for example magnesium stearate, stearic acid or talc. The tablets
may be uncoated or they may be coated, enterically or otherwise, by
known techniques to delay disintegration and absorption in the
gastrointestinal tract and thereby provide a sustained action over
a longer period. For example, a time delay material such as
glyceryl monostearate or glyceryl distearate may be employed. They
may also be coated by the techniques described in the U.S. Pat.
Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotic
therapeutic tablets for control release.
[0096] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin, or olive oil. Additionally, emulsions can be
prepared with a non-water miscible ingredient such as oils and
stabilized with surfactants such as mono-diglycerides, PEG esters
and the like.
[0097] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxy-propylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents may be a naturally-occurring phosphatide, for
example lecithin, or condensation products of an alkylene oxide
with fatty acids, for example polyoxy-ethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and a hexitol such as polyoxyethylene sorbitol monooleate, or
condensation products of ethylene oxide with partial esters derived
from fatty acids and hexitol anhydrides, for example polyethylene
sorbitan monooleate. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl,
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose or saccharin.
[0098] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0099] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0100] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, for example olive oil or arachis oil, or a mineral
oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally-occurring gums, for example gum
acacia or gum tragacanth, naturally-occurring phosphatides, for
example soy bean, lecithin, and esters or partial esters derived
from fatty acids and hexitol anhydrides, for example sorbitan
monooleate, and condensation products of the said partial esters
with ethylene oxide, for example polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening and flavoring
agents.
[0101] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative and
flavoring and coloring agents. Oral solutions can be prepared in
combination with, for example, cyclodextrin, PEG and
surfactants.
[0102] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous or oleagenous suspension. This
suspension may be formulated according to the known art using those
suitable dispersing or wetting agents and suspending agents which
have been mentioned above. The sterile injectable preparation may
also be a sterile injectable solution or suspension in a non-toxic
parenterally-acceptable diluent or solvent, for example as a
solution in 1,3-butane diol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as a solvent or suspending medium. For
this purpose any bland fixed oil may be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid find use in the preparation of injectables.
[0103] The compounds of the present invention may also be
administered in the form of suppositories for rectal administration
of the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient which is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
include cocoa butter and polyethylene glycols. Additionally, the
compounds can be administered via ocular delivery by means of
solutions or ointments. Still further, transdermal delivery of the
subject compounds can be accomplished by means of iontophoretic
patches and the like. 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.
[0104] In exemplary embodiment, the inhibitory agent of this
invention may be deposited within a medical device, such as a
stent, and delivered to the treatment site for treatment of a
portion of the body.
[0105] Stents have been used as delivery vehicles for therapeutic
agents (i.e., drugs). Intravascular stents are generally
permanently implanted in coronary or peripheral vessels. Stent
designs include those of U.S. Pat. No. 4,733,655 (Palmaz), U.S.
Pat. No. 4,800,882 (Gianturco), or U.S. Pat. No. 4,886,062
(Wiktor). Such designs include both metal and polymeric stents, as
well as self-expanding and balloon-expandable stents. Stents may
also used to deliver a drug at the site of contact with the
vasculature, as disclosed in U.S. Pat. No. 5,102,417 (Palmaz) and
in International Patent Application Nos. WO 91/12779 (Medtronic,
Inc.) and WO 90/13332 (Cedars-Sanai Medical Center), U.S. Pat. No.
5,419,760 (Narciso, Jr.) and U.S. Pat. No. 5,429,634 (Narciso,
Jr.), for example. Stents have also been used to deliver viruses to
the wall of a lumen for gene delivery, as disclosed in U.S. patent
application Ser. No. 08/746,404, filed Nov. 8, 1996 (Donovan et
al.).
[0106] The term "deposited" means that the inhibitory agent is
coated, adsorbed, placed, or otherwise incorporated into the device
by methods known in the art. For example, the inhibitory agent may
be embedded and released from within ("matrix type") or surrounded
by and released through ("reservoir type") polymer materials that
coat or span the medical device. In the later example, the
inhibitory agent may be entrapped within the polymer materials or
coupled to the polymer materials using one or more the techniques
for generating such materials known in the art. In other
formulations, the inhibitory agent may be linked to the surface of
the medical device without the need for a coating by means of
detachable bonds and release with time, can be removed by active
mechanical or chemical processes, or are in a permanently
immobilized form that presents the inhibitory agent at the
implantation site.
[0107] In one embodiment, the inhibitory agent may be incorporated
with polymer compositions during the formation of biocompatible
coatings for medical devices, such as stents. The coatings produced
from these components are typically homogeneous and are useful for
coating a number of devices designed for implantation.
[0108] The polymer may be either a biostable or a bioabsorbable
polymer depending on the desired rate of release or the desired
degree of polymer stability, but a bioabsorbable polymer is
preferred for this embodiment since, unlike a biostable polymer, it
will not be present long after implantation to cause any adverse,
chronic local response. Bioabsorbable polymers that could be used
include, but are not limited to, poly(L-lactic acid),
polycaprolactone, polyglycolide (PGA), poly(lactide-co-glycolide)
(PLLA/PGA), poly(hydroxybutyrate),
poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester,
polyanhydride, poly(glycolic acid), poly(D-lactic acid),
poly(L-lactic acid), poly(D,L-lactic acid), poly(D,L-lactide)
(PLA), poly (L-lactide) (PLLA), poly(glycolic acid-co-trimethylene
carbonate) (PGA/PTMC), polyethylene oxide (PEO), polydioxanone
(PDS), polyphosphoester, polyphosphoester urethane, poly(amino
acids), cyanoacrylates, poly(trimethylene carbonate),
poly(iminocarbonate), copoly(ether-esters) (e.g., PEO/PLA),
polyalkylene oxalates, polyphosphazenes and biomolecules such as
fibrin, fibrinogen, cellulose, starch, collagen and hyaluronic
acid, polyepsilon caprolactone, polyhydroxy butyric acid,
polyorthoesters, polyacetals, polydihydropyrans,
polycyanoacrylates, cross linked or amphipathic block copolymers of
hydrogels, and other suitable bioabsorbable poplymers known in the
art. Also, biostable polymers with a relatively low chronic tissue
response such as polyurethanes, silicones, and polyesters could be
used and other polymers could also be used if they can be dissolved
and cured or polymerized on the medical device such as polyolefins,
polyisobutylene and ethylene-alphaolefin copolymers; acrylic
polymers and copolymers, vinyl halide polymers and copolymers, such
as polyvinyl chloride; polyvinylpyrrolidone; polyvinyl ethers, such
as polyvinyl methyl ether; polyvinylidene halides, such as
polyvinylidene fluoride and polyvinylidene chloride;
polyacrylonitrile, polyvinyl ketones; polyvinyl aromatics, such as
polystyrene, polyvinyl esters, such as polyvinyl acetate;
copolymers of vinyl monomers with each other and olefins, such as
ethylene-methyl methacrylate copolymers, acrylonitrile-styrene
copolymers, ABS resins, and ethylene-vinyl acetate copolymers;
pyran copolymer; polyhydroxy-propyl-methacrylamide-phenol;
polyhydroxyethyl-aspartamide-phenol; polyethyleneoxide-polylysine
substituted with palmitoyl residues; polyamides, such as Nylon 66
and polycaprolactam; alkyd resins, polycarbonates;
polyoxymethylenes; polyimides; polyethers; epoxy resins,
polyurethanes; rayon; rayon-triacetate; cellulose, cellulose
acetate, cellulose butyrate; cellulose acetate butyrate;
cellophane; cellulose nitrate; cellulose propionate; cellulose
ethers; and carboxymethyl cellulose.
[0109] Polymers and semipermeable polymer matrices may be formed
into shaped articles, such as valves, stents, tubing, prostheses
and the like.
[0110] In one embodiment of the invention, the inhibitory agent of
the invention is coupled to a polymer or semipermeable polymer
matrix that is formed as a stent or stent-graft device.
[0111] Typically, polymers are applied to the surface of an
implantable device by spin coating, dipping or spraying. Additional
methods known in the art can also be utilized for this purpose.
Methods of spraying include traditional methods as well as
microdeposition techniques with an inkjet type of dispenser.
Additionally, a polymer can be deposited on an implantable device
using photo-patterning to place the polymer on only specific
portions of the device. This coating of the device provides a
uniform layer around the device which allows for improved diffusion
of various analytes through the device coating.
[0112] In preferred embodiments of the invention, the inhibitory
agent is formulated for release from the polymer coating into the
environment in which the medical device is placed. Preferably, the
inhibitory agent is released in a controlled manner over an
extended time frame (e.g., months) using at least one of several
well-known techniques involving polymer carriers or layers to
control elution. Some of these techniques were previously described
in U.S. Patent Application 20040243225A1, the entire disclosure of
which is incorporated in its entirety.
[0113] Moreover, as described for example in U.S. Pat. No.
6,770,729, which is incorporated herein in its entirety, the
reagents and reaction conditions of the polymer compositions can be
manipulated so that the release of the inhibitory agent from the
polymer coating can be controlled. For example, the diffusion
coefficient of the one or more polymer coatings can be modulated to
control the release of the inhibitory agent from the polymer
coating. In a variation on this theme, the diffusion coefficient of
the one or more polymer coatings can be controlled to modulate the
ability of an analyte that is present in the environment in which
the medical device is placed (e.g. an analyte that facilitates the
breakdown or hydrolysis of some portion of the polymer) to access
one or more components within the polymer composition (and for
example, thereby modulate the release of the inhibitory agent from
the polymer coating). Yet another embodiment of the invention
includes a device having a plurality of polymer coatings, each
having a plurality of diffusion coefficients. In such embodiments
of the invention, the release of the inhibitory agent from the
polymer coating can be modulated by the plurality of polymer
coatings.
[0114] In yet another embodiment of the invention, the release of
the inhibitory agent from the polymer coating is controlled by
modulating one or more of the properties of the polymer
composition, such as the presence of one or more endogenous or
exogenous compounds, or alternatively, the pH of the polymer
composition. For example, certain polymer compositions can be
designed to release a inhibitory agent in response to a decrease in
the pH of the polymer composition. Alternatively, certain polymer
compositions can be designed to release the inhibitory agent in
response to the presence of hydrogen peroxide.
V. METHODS OF TREATING DISEASES MODULATED BY CCR1, CCR2 AND/OR
CCR3
[0115] In yet another aspect, the present invention provides
methods of treating CCR1, CCR2 and/or CCR3-mediated conditions or
diseases by administering to a subject having such a disease or
condition, a therapeutically effective amount of a compound of
formula I above. The "subject" is defined herein to include animals
such as mammals, including, but not limited to, primates (e.g.,
humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats,
mice and the like.
[0116] CCR1 provides a target for interfering with or promoting
specific aspects of immune cell functions, or more generally, with
functions associated with CCR1 expression on a wide range of cell
types in a mammal, such as a human. Compounds that inhibit CCR1,
are particularly useful for modulating monocyte, macrophage,
lymphocyte, granulocyte, NK cell, mast cells, dendritic cell,
neutrophils, and certain immune derived cell (for example,
osteoclasts) function for therapeutic purposes. Accordingly, the
present invention is directed to compounds which are useful in the
prevention and/or treatment of a wide variety of inflammatory and
immunoregulatory disorders and diseases (see Saeki, et al., Current
Pharmaceutical Design 9:1201-1208 (2003)).
[0117] For example, an instant compound that inhibits one or more
functions of CCR1 may be administered to inhibit (i.e., reduce or
prevent) inflammation or cellular infiltration associated with an
immune disorder. As a result, one or more inflammatory processes,
such as leukocyte emigration or infiltration, chemotaxis,
exocytosis (e.g., of enzymes, histamine) or inflammatory mediator
release, can be inhibited. For example, monocyte infiltration to an
inflammatory site (e.g., an affected joint in arthritis, or into
the CNS in MS) can be inhibited according to the present
method.
[0118] Similarly, an instant compound that promotes one or more
functions of CCR1 is administered to stimulate (induce or enhance)
an inflammatory response, such as leukocyte emigration, chemotaxis,
exocytosis (e.g., of enzymes, histamine) or inflammatory mediator
release, resulting in the beneficial stimulation of inflammatory
processes. For example, monocytes can be recruited to combat
bacterial infections.
[0119] Diseases and conditions associated with inflammation, immune
disorders and infection can be treated using the method of the
present invention. In a preferred embodiment, the disease or
condition is one in which the actions of immune cells such
monocyte, macrophage, lymphocyte, granulocyte, NK cell, mast cell,
dendritic cell, or certain immune derived cell (for example,
osteoclasts) are to be inhibited or promoted, in order to modulate
the inflammatory or autoimmune response.
[0120] In one group of embodiments, diseases or conditions,
including chronic diseases, of humans or other species can treated
with modulators of CCR1, CCR2 or CCR3 function. These diseases or
conditions include: (1) allergic diseases such as systemic
anaphylaxis or hypersensitivity responses, drug allergies, insect
sting allergies and food allergies, (2) inflammatory bowel
diseases, such as Crohn's disease, ulcerative colitis, ileitis and
enteritis, (3) vaginitis, (4) psoriasis and inflammatory dermatoses
such as dermatitis, eczema, atopic dermatitis, allergic contact
dermatitis, urticaria and pruritus, (5) vasculitis, (6)
spondyloarthropathies, (7) scleroderma, (8) asthma and respiratory
allergic diseases such as allergic asthma, allergic rhinitis,
hypersensitivity lung diseases and the like, (9) autoimmune
diseases, such as fibromyalagia, scleroderma, ankylosing
spondylitis, juvenile RA, Still's disease, polyarticular juvenile
RA, pauciarticular juvenile RA, polymyalgia rheumatica, rheumatoid
arthritis, psoriatic arthritis, osteoarthritis, polyarticular
arthritis, multiple sclerosis, systemic lupus erythematosus, type I
diabetes, type II diabetes, glomerulonephritis, and the like, (10)
graft rejection (including allograft rejection and graft-v-host
disease), and (11) other diseases in which undesired inflammatory
responses or immune disorders are to be inhibited, such as
cardiovascular disease including atherosclerosis and restenosis,
myositis, neurodegenerative diseases (e.g., Alzheimer's disease),
encephalitis, meningitis, hepatitis, nephritis, sepsis,
sarcoidosis, allergic conjunctivitis, otitis, chronic obstructive
pulmonary disease, sinusitis, Behcet's syndrome and gout and (12)
immune mediated food allergies such as Celiac disease.
[0121] In another group of embodiments, diseases or conditions can
be treated with modulators of CCR1 function. Examples of diseases
to be treated with modulators of CCR1 function include cancers,
cardiovascular diseases, diseases in which angiogenesis or
neovascularization play a role (neoplastic diseases, retinopathy
and macular degeneration), infectious diseases (viral infections,
e.g., HIV infection, and bacterial infections) and
immunosuppressive diseases such as organ transplant conditions and
skin transplant conditions. The term "organ transplant conditions"
is meant to include bone marrow transplant conditions and solid
organ (e.g., kidney, liver, lung, heart, pancreas or combination
thereof) transplant conditions.
[0122] The compounds of the present invention are accordingly
useful in the prevention and treatment of a wide variety of
inflammatory and immunoregulatory disorders and diseases.
[0123] Depending on the disease to be treated and the subject's
condition, the compounds of the present invention may be
administered by oral, parenteral (e.g., intramuscular,
intraperitoneal, intravenous, ICV, intracisternal injection or
infusion, subcutaneous injection, or implant), by implantation
(e.g., as when the compound is coupled to a stent device), by
inhalation spray, nasal, vaginal, rectal, sublingual, or topical
routes of administration and may be formulated, alone or together,
in suitable dosage unit formulations containing conventional
non-toxic pharmaceutically acceptable carriers, adjuvants and
vehicles appropriate for each route of administration.
[0124] In the treatment or prevention of conditions which require
chemokine receptor modulation an appropriate dosage level will
generally be about 0.001 to 100 mg per kg patient body weight per
day which can be administered in single or multiple doses.
Preferably, the dosage level will be about 0.01 to about 25 mg/kg
per day; more preferably about 0.05 to about 10 mg/kg per day. A
suitable dosage level may be about 0.01 to 25 mg/kg per day, about
0.05 to 10 mg/kg per day, or about 0.1 to 5 mg/kg per day. Within
this range the dosage may be 0.005 to 0.05, 0.05 to 0.5 or 0.5 to
5.0 mg/kg per day. For oral administration, the compositions are
preferably provided in the form of tablets containing 1.0 to 1000
milligrams of the active ingredient, particularly 1.0, 5.0, 10.0,
15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,
400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of
the active ingredient for the symptomatic adjustment of the dosage
to the patient to be treated. The compounds may be administered on
a regimen of 1 to 4 times per day, preferably once or twice per
day.
[0125] It will be understood, however, that the specific dose level
and frequency of dosage for any particular patient may be varied
and will depend upon a variety of factors including the activity of
the specific compound employed, the metabolic stability and length
of action of that compound, the age, body weight, hereditary
characteristics, general health, sex and diet of the subject, as
well as the mode and time of administration, rate of excretion,
drug combination, and the severity of the particular condition for
the subject undergoing therapy.
[0126] Diseases and conditions associated with inflammation, immune
disorder, infection and cancer can be treated or prevented with the
present compounds, compositions, and methods.
[0127] The compounds and compositions of the present invention can
be combined with other compounds and compositions having related
utilities to prevent and treat the condition or disease of
interest, such as inflammatory or autoimmune disorders, conditions
and diseases, including inflammatory bowel disease, rheumatoid
arthritis, osteoarthritis, psoriatic arthritis, polyarticular
arthritis, multiple sclerosis, allergic diseases, psoriasis, atopic
dermatitis and asthma, and those pathologies noted above.
[0128] For example, in the treatment or prevention of inflammation
or autoimmunity or for example arthritis associated bone loss, the
present compounds and compositions may be used in conjunction with
an anti-inflammatory or analgesic agent such as an opiate agonist,
a lipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, a
cyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, an
interleukin inhibitor, such as an interleukin-1 inhibitor, an NMDA
antagonist, an inhibitor of nitric oxide or an inhibitor of the
synthesis of nitric oxide, a non steroidal anti-inflammatory agent,
or a cytokine-suppressing anti-inflammatory agent, for example with
a compound such as acetaminophen, aspirin, codeine, fentanyl,
ibuprofen, indomethacin, ketorolac, morphine, naproxen, phenacetin,
piroxicam, a steroidal analgesic, sufentanyl, sunlindac, tenidap,
and the like. Similarly, the instant compounds and compositions may
be administered with an analgesic listed above; a potentiator such
as caffeine, an H.sub.2 antagonist (e.g., ranitidine), simethicone,
aluminum or magnesium hydroxide; a decongestant such as
phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline,
ephinephrine, naphazoline, xylometazoline, propylhexedrine, or levo
desoxy ephedrine; an antitussive such as codeine, hydrocodone,
caramiphen, carbetapentane, or dextromethorphan; a diuretic; and a
sedating or non sedating antihistamine.
[0129] Likewise, compounds and compositions of the present
invention may be used in combination with other drugs that are used
in the treatment, prevention, suppression or amelioration of the
diseases or conditions for which compounds and compositions of the
present invention are useful. Such other drugs may be administered,
by a route and in an amount commonly used therefor,
contemporaneously or sequentially with a compound or composition of
the present invention. When a compound or composition of the
present invention is used contemporaneously with one or more other
drugs, a pharmaceutical composition containing such other drugs in
addition to the compound or composition of the present invention is
preferred. Accordingly, the pharmaceutical compositions of the
present invention include those that also contain one or more other
active ingredients or therapeutic agents, in addition to a compound
or composition of the present invention. Examples of other
therapeutic agents that may be combined with a compound or
composition of the present invention, either administered
separately or in the same pharmaceutical compositions, include, but
are not limited to: (a) VLA-4 antagonists, (b) corticosteroids,
such as beclomethasone, methylprednisolone, betamethasone,
prednisone, prenisolone, dexamethasone, fluticasone,
hydrocortisone, budesonide, triamcinolone, salmeterol, salmeterol,
salbutamol, formeterol; (c) immunosuppressants such as cyclosporine
(cyclosporine A, Sandimmune.RTM., Neoral.RTM.), tacrolirnus
(FK-506, Prograf.RTM.), rapamycin (sirolimus, Rapamune.RTM.) and
other FK-506 type immunosuppressants, and mycophenolate, e.g.,
mycophenolate mofetil (CellCept.RTM.); (d) antihistamines
(H1-histamine antagonists) such as bromopheniramine,
chlorpheniramine, dexchloipheniramine, triprolidine, clemastine,
diphenhydramine, diphenylpyraline, tripelennamine, hydroxyzine,
methdilazine, promethazine, trimeprazine, azatadine,
cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,
terfenadine, loratadine, cetirizine, fexofenadine,
descarboethoxyloratadine, and the like; (e) non steroidal anti
asthmatics (e.g., terbutaline, metaproterenol, fenoterol,
isoetharine, albuterol, bitolterol and pirbuterol), theophylline,
cromolyn sodium, atropine, ipratropium bromide, leukotriene
antagonists (e.g., zafmlukast, montelukast, pranlukast, iralukast,
pobilukast and SKB-106,203), leukotriene biosynthesis inhibitors
(zileuton, BAY-1005); (f) non steroidal anti-inflammatory agents
(NSAIDs) such as propionic acid derivatives (e.g., alminoprofen,
benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen,
fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen,
miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen,
tiaprofenic acid and tioxaprofen), acetic acid derivatives (e.g.,
indomethacin, acemetacin, alclofenac, clidanac, diclofenac,
fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,
isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin and
zomepirac), fenamic acid derivatives (e.g., flufenamic acid,
meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic
acid), biphenylcarboxylic acid derivatives (e.g., diflunisal and
flufenisal), oxicams (e.g., isoxicam, piroxicam, sudoxicam and
tenoxican), salicylates (e.g., acetyl salicylic acid and
sulfasalazine) and the pyrazolones (e.g., apazone, bezpiperylon,
feprazone, mofebutazone, oxyphenbutazone and phenylbutazone); (g)
cyclooxygenase-2 (COX-2) inhibitors such as celecoxib
(Celebrex.RTM.) and rofecoxib (Vioxx.RTM.); (h) inhibitors of
phosphodiesterase type IV (PDE IV); (i) gold compounds such as
auranofin and aurothioglucose, (j) etanercept (Enbrel.RTM.), (k)
antibody therapies such as orthoclone (OKT3), daclizumab
(Zenapax.RTM.), basiliximab (Simulect.RTM.) and infliximab
(Remicade.RTM.), (l) other antagonists of the chemokine receptors,
especially CCR5, CXCR2, CXCR3, CCR2, CCR3, CCR4, CCR7, CX.sub.3CR1
and CXCR6; (m) lubricants or emollients such as petrolatum and
lanolin, (n) keratolytic agents (e.g., tazarotene), (o) vitamin
D.sub.3 derivatives, e.g., calcipotriene or calcipotriol
(Dovonex.RTM.), (p) PUVA, (q) anthralin (Drithrocreme.RTM.), (r)
etretinate (Tegison.RTM.) and isotretinoin and (s) multiple
sclerosis therapeutic agents such as interferon .beta.-1.beta.
(Betaseron.RTM.), interferon (.beta.-1.alpha. (Avonex.RTM.),
azathioprine (Imurek.RTM., Imuran.RTM.), glatiramer acetate
(Capoxone.RTM.), a glucocorticoid (e.g., prednisolone) and
cyclophosphamide (t) DMARDS such as methotrexate (u) other
compounds such as 5-aminosalicylic acid and prodrugs thereof;
hydroxychloroquine; D-penicillamine; antimetabolites such as
azathioprine, 6-mercaptopurine and methotrexate; DNA synthesis
inhibitors such as hydroxyurea and microtubule disrupters such as
colchicine. The weight ratio of the compound of the present
invention to the second active ingredient may be varied and will
depend upon the effective dose of each ingredient. Generally, an
effective dose of each will be used. Thus, for example, when a
compound of the present invention is combined with an NSAID the
weight ratio of the compound of the present invention to the NSAID
will generally range from about 1000:1 to about 1:1000, preferably
about 200:1 to about 1:200. Combinations of a compound of the
present invention and other active ingredients will generally also
be within the aforementioned range, but in each case, an effective
dose of each active ingredient should be used.
VI. EXAMPLES
[0130] The following examples are offered to illustrate, but not to
limit the claimed invention.
[0131] Reagents and solvents used below can be obtained from
commercial sources such as Aldrich Chemical Co. (Milwaukee, Wis.,
USA). .sup.1H-NMR spectra were recorded on a Varian Mercury 400 MHz
NMR spectrometer. Significant peaks are provided relative to TMS
and are tabulated in the order: multiplicity (s, singlet; d,
doublet; t, triplet; q, quartet; m, multiplet) and number of
protons. Mass spectrometry results are reported as the ratio of
mass over charge, followed by the relative abundance of each ion
(in parenthesis). In the examples, a single m/e value is reported
for the M+H (or, as noted, M-H) ion containing the most common
atomic isotopes. Isotope patterns correspond to the expected
formula in all cases. Electrospray ionization (ESI) mass
spectrometry analysis was conducted on a Hewlett-Packard MSD
electrospray mass spectrometer using the HP1100 HPLC for sample
delivery. Normally the analyte was dissolved in methanol at 0.1
mg/mL and 1 microlitre was infused with the delivery solvent into
the mass spectrometer, which scanned from 100 to 1500 daltons. All
compounds could be analyzed in the positive ESI mode, using
acetonitrile/water with 1% formic acid as the delivery solvent. The
compounds provided below could also be analyzed in the negative ESI
mode, using 2 mM NH.sub.4OAc in acetonitrile/water as delivery
system.
[0132] Compounds within the scope of this invention can be
synthesized as described below, using a variety of reactions known
to the skilled artisan. A sample of useful routes to the
azaindazole derivatives and certain compounds of the invention are
provided below or elsewhere within the present application. In the
descriptions of the syntheses that follow, some of the
arylpiperazine and heteroaromatic subunit precursors were obtained
from commercial sources. These commercial sources include Aldrich
Chemical Co., Acros Organics, Ryan Scientific Incorporated, Oakwood
Products Incorporated, Lancaster Chemicals, Sigma Chemical Co.,
Lancaster Chemical Co., TCI-America, Alfa Aesar, Davos Chemicals,
and GFS Chemicals. Certain relevant arylpiperazine compounds can be
commercially obtained. Others could be prepared as described in
U.S. patent application Ser. No. 11/008,774, the contents of which
is hereby incorporated in its entirety for all purposes. Also,
standard chemistries have been employed to link the arylpiperazine
and heteroaromatic subunits (whether commercially obtained or
prepared by the methods below) using a suitably optimized linker,
such as the acetyl unit described in the body of this
invention.
[0133] One skilled in the art will also recognize that alternative
methods may be employed to synthesize the target compounds of this
invention, and that the approaches described within the body of
this document are not exhaustive, but do provide broadly applicable
and practical routes to compounds of interest.
[0134] Certain molecules claimed in this patent can exist in
different enantiomeric and diastereomeric forms and all such
variants of these compounds are claimed.
[0135] Regioisomerism is a common property in organic chemistry,
and is especially common with regards to certain structural types
provided herein. Those skilled in the art will recognize, with
respect to the compounds described herein, that the coupling
reactions with the heteroaromatic ring systems can lead to either
one of or a mixture of detectable regioisomers.
[0136] The detailed description of the experimental procedures used
to synthesize key compounds in this text lead to molecules that are
described by the physical data identifying them as well as by the
structural depictions associated with them.
[0137] Those skilled in the art will also recognize that during
standard work up procedures in organic chemistry, acids and bases
are frequently used. Salts of the parent compounds are sometimes
produced, if they possess the necessary intrinsic acidity or
basicity, during the experimental procedures described within this
patent.
Example 1
Synthesis of 1H-pyrazolo[3,4-b]pyridine
##STR00023##
[0139] 2-Chloro-3-formylpyridine (15.02 g, 106 mmol, 1 equiv),
hydrazine (10 mL, excess), and dioxane (90 mL) were combined in a
sealed tube and heated at 150.degree. C. for 16 hr. After cooling
to room temperature, the solvent was evaporated in vacuo to provide
a crude residue which was diluted with dichloromethane (600 mL).
The organic solution was washed with water (50 mL), brine (50 mL)
and dried over anhydrous sodium sulfate. The solvent was removed in
vacuo to provide 1H-pyrazolo[3,4-b]pyridine as a yellow powder
which was used without further purification: LCMS (ES) M+H 120.3,
R.sub.f 0.20 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20% to
100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile).
Example 2
Synthesis of 3-Thiazol-2-yl-1H-pyrazolo[3,4-b]pyridine
##STR00024##
[0141] To a suspension of
2-chloro-3-[(2-thiazolyl)carbonyl]pyridine (257.5 mg, 1.2 mmol, 1
equiv) in dioxane (3 mL) in a sealed tube was added hydrazine (2
mL). The mixture was heated at 150.degree. C. overnight, cooled to
room temperature and concentrated in vacuo to provide a crude
residue. The resultant residue was diluted with dichloromethane
(300 mL), washed with water (50 mL) and brine (50 mL). The organic
layer was separated, dried over sodium sulfate, filtered and
concentrated in vacuo to provide
3-thiazol-2-yl-1H-pyrazolo[3,4-b]pyridine (212.3 mg) as a yellow
powder which used without further purification: LCMS (ES) M+H
203.5, R.sub.f 2.68 min (Agilent Zorbax SB-C18, 2.1.times.50 mm,
5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20%
to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile).
Example 3
Synthesis of 1H-Pyrazolo[3,4-b]pyridin-3-ylamine
##STR00025##
[0143] 2-Chloro-3-cyanopyridine (2.77 g), hydrazine (5 mL), and
dioxane (100 mL) were combined in a sealed tube and heated at
150.degree. C. for 16 hr. The reaction mixture was cooled to room
temperature and concentrated in vacuo to provide a crude residue.
The resultant residue was dissolved in ethyl acetate (100 mL) and
washed with saturated NaCl solution (50 mL). The organic layer was
dried over sodium sulfate, filtered, and concentrated in vacuo to
afford 1H-pyrazolo[3,4-b]pyridin-3-ylamine as a yellow solid which
was used without further purification.
Example 4
Synthesis of 1H-pyrazolo[3,4-c]pyridine
##STR00026##
[0145] Preparation of 3-N-acetylamino-4-methylpyridine: To solution
of 3-amino-4-methylpyridine (540.2 mg, 5.0 mmol, 1 equiv) in
dichloromethane (20 mL) was added pyridine (0.8 mL, 10.0 mmol, 2
equiv) and acetic anhydride (0.57 mL, 6.0 mmol, 1.2 equiv). The
resultant solution was stirred at room temperature for 16 h and
concentrated in vacuo to provide a crude residue. The residue was
diluted with dichloromethane (200 mL), and washed with saturated
sodium bicarbonate aqueous solution (50 mL) and brine (50 mL). The
organic layer was separated, dried over sodium sulfate, and
concentrated in vacuo to yield 3-acetylamino-4-methylpyridine
(400.2 mg) as yellow solid which was used without further
purification.
[0146] Preparation of 1-pyrazolo[3,4-c]pyridin-1-yl-ethanone: To a
suspension of 3-acetylamino-4-methylpyridine (301.5 mg, 2.0 mmol, 1
equiv) in toluene (3 mL) was added tert-butyl nitrite (t-BuONO)
(420 .mu.L, 3.2 mmol, 1.6 equiv), acetic anhydride (560 .mu.L, 6.0
mmol, 3 equiv) and potassium acetate (235.2 mg, 2.4 mmol, 1.2
equiv). The resultant mixture was heated at 80.degree. C. for 2
hours, cooled to room temperature, and diluted with ethyl acetate
(200 mL). The mixture was washed with saturated sodium bicarbonate
solution(50 mL), water (50 mL) and brine (50 mL). The organic layer
was dried over sodium sulfate and concentrated in vacuo to provide
a crude residue. The residue was purified by flash chromatography
(silica, 15% ethyl acetate/hexane to 50% ethyl acetate/hexane) to
give 1-pyrazolo[3,4-c]pyridin-1-yl-ethanone (20.2 mg) which was
used without further purification.
[0147] Synthesis of 1H-pyrazolo[3,4-c]pyridine: To a solution of
1-pyrazolo[3,4-c]pyridin-1-yl-ethanone (20.2 mg, 0.17 mmol, 1
equiv) in tetrahydrofuran (2 mL) and methanol (0.5 mL) was added
sodium hydroxide aqueous solution (2M, 0.25 mL). The reaction
solution was stirred at room temperature for 1 hr and then
concentrated in vacuo to provide a crude residue. The crude residue
was diluted with water (20 mL) and extracted with dichloromethane
(2.times.100 mL). The combined organic layers were washed with
brine, dried over sodium sulfate, and concentrated in vacuo to
provide 1H-pyrazolo[3,4-c]pyridine as white powder, which used
without further purification: LCMS (ES) M+H 120.3, R.sub.f 0.22 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 5
Synthesis of 3-Iodo-1H-pyrazolo[3,4-b]pyridine
##STR00027##
[0149] To a solution of 1H-pyrazolo[3,4-b]pyridine (500.0 mg, 4.2
mmol, 1 equiv) in DMF (10 mL) at 0.degree. C., was added iodine
(2.13 g, 8.4 mmol, 2 equiv) and potassium hydroxide (943 mg, 16.8
mmol, 4 equiv). The resultant mixture was allowed to warm to room
temperature and stirred for 1 hour. The reaction solution was
slowly quenched with saturated sodium thiosulfate
(Na.sub.2S.sub.2O.sub.5) solution (10 mL), and extracted with ethyl
acetate (2.times.200 mL). The combined organic layers were washed
with water (3.times.50 mL), brined (50 mL), dried over sodium
sulfate and concentrated in vacuo to give
3-Iodo-1H-pyrazolo[3,4-b]pyridine (1.02 g) as a yellow powder which
was used without further purification: LCMS (ES) M+H 246.2, R.sub.f
2.17 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a
1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%
water, B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 6
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[3,4-b]pyridine-1-ylethanone and
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[3,4-b]pyridine-2-ylethanone
##STR00028##
[0151]
2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiper-
azin-1-yl]ethanone (arylpiperazine) (4.81 g, 14.32 mmol, 1 equiv),
1H-pyrazole[3,4-b]pyridine (2.27 g, 17.18 mmol, 1.2 equiv), and
potassium carbonate (20.00 g, 143.2 mmol, 10 equiv) were dissolved
in dimethylformamide (DMF) (10 mL) and heated at 80.degree. C. for
1 hour, then cooled to room temperature. The resultant mixture was
diluted with ethyl acetate (300 mL), and washed with water
(3.times.150 mL) and brine (100 mL). The organic layer was dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to provide a crude
residue. The crude residue was purified by flash chromatography
(silica, 100% ethyl acetate with 1% triethylamine to 100% acetone
with 1% triethylamine) provided
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[3,4-b]pyridine-1-ylethanone (2.3 g) and
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[3,4-b]pyridine-2-ylethanone (2.5 g).
[0152] For
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazi-
n-1-yl]-2-pyrazolo[3,4-b]pyridine-1-yl-ethanone; LCMS (ES) M+H
418.5, R.sub.f 2.34min (Agilent Zorbax SB-C18, 2.1.times.50mm,
5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20%
to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile): For
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[3,4-b]pyridine-2-ylethanone; LCMS (ES) M+H 418.5, R.sub.f
2.00 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C., 1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a
1.1 min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9%
water, B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 7
Synthesis of
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone
##STR00029##
[0154] Preparation of
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone: This compound was
synthesized according to the synthetic procedure outlined in
Example 6.
[0155] Synthesis of
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone: To a solution
of oxazole (40 .mu.L, 0.54 mmol, 3 equiv) in tetrahydrofuran (1 mL)
under nitrogen atmosphere, was added dropwise n-butyl lithium (2.5
M in Hexane, 220 .mu.L, 0.54 mmol, 3 equiv.). The resultant mixture
was stirred at -78.degree. C. for an additional 30 min followed by
the addition of ZnCl.sub.2 (0.5M in THF, 1.5 mL, 0.72 mmol, 4
equiv.). The reaction solution was allowed to warm to 0.degree. C.
and stirred 1 hr followed by the addition of
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone (100.2 mg, 0.18 mmol,
1 equiv) and palladium tetrakis(triphenylphosphine) (22.3 mg,
0.018, 0.1 equiv). The reaction mixture was then heated to reflux
for 48 hr, cooled to room temperature and diluted with ethyl
acetate (150 mL). The reaction mixture was washed with water (20
mL), brine (20 mL), dried over sodium sulfate, and concentrated in
vacuo to provide the crude product. Purification by preparative
HPLC provided the desired product
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-oxazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone as a white
powder (38.5 mg): LCMS (ES) M+H 485.5, R.sub.f 2.56 min (Agilent
Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1 ml/min flow
rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash at
100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile).
Example 8
Synthesis of
1-[4-(4-chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine--
1-ylethanone and
1-[4-(4-chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine--
2-ylethanone
##STR00030##
[0157] The two title compounds were synthesized according to the
synthetic procedure as outlined in Example 6: For
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine--
1-ylethanone; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.57 (dd,
1H), 8.11(s, 1H), 8.09 (dd, 1H), 7.22(d, 1H), 7.17 (dd, 1H), 6.49
(d, 1H), 6.42 (dd, 1H), 5.44 (s, 2H), 3.92 (s, 3H), 3.79 (m, 4H),
3.18 (m, 4H); MS (M+H).sup.+: 386.5: For
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-pyrazolo[3,4-b]pyridine--
2-ylethanone; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.65 (dd,
1H), 8.12(s, 1H), 8.02 (dd, 1H), 7.20(d, 1H), 7.03 (dd, 1H), 6.45
(d, 1H), 6.40 (dd, 1H), 5.35 (s, 2H), 3.88 (s, 3H), 3.87 (m, 2H),
3.79 (m, 2H), 3.15 (m, 4H); MS (M+H).sup.+: 386.5.
Example 9
Synthesis of 1H-Pyrazolo[4,3-c]pyridine
##STR00031##
[0159] 1H-Pyrazolo[4,3-c]pyridine was prepared according to the
procedure outlined in Example 1.
Example 10
Synthesis of
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[4,3-c]pyridine-1-yl-ethanone and
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[4,3-c]pyridine-2-yl-ethanone
##STR00032##
[0161] The two title compounds were synthesized according to the
procedure as outlined in Example 6. For
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[4,3-c]pyridine-1-yl-ethanone: LCMS (ES) M+H 418.5, R.sub.f
1.74 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile): For
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-yl]-2-py-
razolo[4,3-c]pyridine-2-yl-ethanone; LCMS (ES) M+H 418.5, R.sub.f
1.69 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 11
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-
e-1-yl-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-
e-2-yl-ethanone
##STR00033##
[0163] The two title compounds were synthesized according to the
procedure outlined in Example 6: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-
e-1-yl-ethanone; .sup.1H NMR (400 MHz, CDCl.sub.3) 9.02 (s, 1H),
8.34 (d, 1H), 8.09 (d, 1H), 7.63 (dd, 1H), 7.22 (d, 1H), 6.48 (d,
1H), 6.42 (dd, 1H), 5.38 (s, 2H), 3.88 (s, 3H), 3.79 (m, 4H), 3.14
(m, 4H) MS (M+H).sup.+, 386.5: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4-c]pyridin-
e-2-yl-ethanone; .sup.1H NMR (400 MHz, CDCl.sub.3)) 9.22 (s, 1H),
8.13 (d, 1H), 8.10 (d, 1H), 7.50 (dd, 1H), 7.19 (d, 1H), 6.45 (d,
1H), 6.39 (dd, 1H), 5.37 (s, 2H), 3.85 (s, 3H), 3.76 (m, 4H), 3.14
(m, 4H). MS (M+H).sup.+, 386.5.
Example 12
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4--
b]pyridin-1-yl-ethanone and
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4--
b]pyridin-2-yl-ethanone
##STR00034##
[0165] The two title compounds were synthesized according to the
procedure outlined in Example 6. For
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4--
b]pyridin-1-yl-ethanone: LCMS (ES) M+H, 404.5, R.sub.f 2.14 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C., 1 ml/min
flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash
at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile): For
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-pyrazolo[3,4--
b]pyridin-2-yl-ethanone; LCMS (ES) M+H, 404.5, R.sub.f 1.76 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C., 1 ml/min
flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash
at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile).
Example 13
Synthesis of
1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-thiazol-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone
##STR00035##
[0167] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 501.5, R.sub.f 2.82
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 14
Synthesis 3-pyrid-2-yl-1H-pyrazolo[3,4-b]pyridine
##STR00036##
[0169] The title compound was synthesized according to the
procedure outlined in Example 2.
Example 15
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
(3-pyridin-2-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone
##STR00037##
[0171] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 495.54, R.sub.f 2.73
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 16
Synthesis of 3-Chloro-1H-pyrazolo[3,4-b]pyridine
##STR00038##
[0173] 1H-pyrazolo[3,4-b]pyridine (89 mg) and N-chlorosuccinimide
(220 mg) were combined in CH.sub.2Cl.sub.2 (4 mL) and heated at
45.degree. C. for 16 hr, then cooled to room temperature. The
resultant mixture was purified by flash chromatography (silica gel,
50% hexane/ethyl acetate) to afford
3-chloro-1H-pyrazolo[3,4-b]pyridine.
Example 17
Synthesis of
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-chloropyrazolo[3,4-b]-
pyridin-1-yl)ethanone
##STR00039##
[0175] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 420.5, R.sub.f 2.37
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 18
Synthesis of 3-Methyl-1H-pyrazolo[3,4-b]pyridine
##STR00040##
[0177] To a solution of 2-chloro-3-cyanopyridine (139 mg) in
tetrahydrofuran (5 mL) at 0.degree. C. was added dropwise a
solution of MeMgBr (3M in ether, 0.67 mL). The resultant mixture
was warmed to room temperature and stirred for 3 hr. The reaction
solution was cooled to 0 .degree. C. and to it was added aqueous
HCl solution (2M, 5 mL). The reaction solution was then stirred an
additional 16 hr at room temperature and then neutralized by the
addition of saturated sodium bicarbonate (NaHCO.sub.3) solution.
The reaction solution was filtered to remove any precipitates and
the filtrate was washed with ethyl acetate (3.times.10 mL) and
aqueous brine (NaCl) solution (10 mL). The organic layer was dried
over sodium sulfate (Na.sub.2SO.sub.4), filtered and concentrated
in vacuo to give 3-acetyl-2-chloropyridine as a yellow powder which
was used without further purification. The title compound
(3-Methyl-1H-pyrazolo[3,4-b]pyridine) was synthesized from
3-acetyl-2-chloropyridine according to the procedure outlined in
Example 2.
Example 19
Synthesis of
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-methylpyrazolo[3,4-b]-
pyridin-1-yl)ethanone
##STR00041##
[0179] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 400.5, R.sub.f 2.12
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 20
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-(3-met-
hylpyrazolo[3,4-b]pyridin-1-yl)ethanone
##STR00042##
[0181] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 432.5, R.sub.f 2.42
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 21
Synthesis of
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-pyridin-2-yl-pyrazolo-
[3,4-b]pyridin-1-yl)ethanone
##STR00043##
[0183] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 463.5, R.sub.f 2.32
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 22
Synthesis of
1-[4-(4-Chloro-3-methoxyphenyl)piperazin-1-yl]-2-(3-thiazol-2-yl-pyrazolo-
[3,4-b]pyridin-1-yl)ethanone
##STR00044##
[0185] The title compound was synthesized according the procedure
outlined in Example 6: LCMS (ES) M+H 469.5, R.sub.f 2.43 min
(Agilent Zorbax SB-C18, 2.1 50 mm, 5.mu., 35.degree. C., 1 ml/min
flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash
at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile).
Example 23
Synthesis of
2-(3-Aminopyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5-methoxyp-
henyl)-2-methylpiperazin-1-yl]ethanone
##STR00045##
[0187] 1H-Pyrazolo[3,4-b]pyridin-3-ylamine (67 mg),
2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-y-
l]-ethanone (167 mg) and K.sub.2CO.sub.3 (414 mg) were combined in
DMF (1 mL) and heated at 80.degree. C. for 2 hr, then cooled to
room temperature. The resultant mixture was purified by preparative
HPLC to provide
2-(3-Aminopyrazolo[3,4-b]pyridin-1-yl)-1-[4-(4-chloro-2-fluoro-5--
methoxyphenyl)-2-methylpiperazin-1-yl]ethanone as a yellow powder.
LCMS (ES) M+H 433.5, R.sub.f 2.06 min (Agilent Zorbax SB-C18,
2.1.times.50 mm, 35.degree. C., 1 ml/min flow rate, a 2.5 min
gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile).
Example 24
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-yl-pyrazol-
o[3,4-b]pyridin-1-yl)-ethanone
##STR00046##
[0189] The title was synthesized according to the procedure
outlined in Example 7: LCMS (ES) M+H 453.5, R.sub.f 2.20 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 25
Synthesis of 3-Fluoro-1H-pyrazolo[3,4-b]pyridine
##STR00047##
[0191] The title was synthesized according to the procedure
outlined in Example 5 using SelectFluor.TM.
(1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane
bis(tetrafluoroborate)) as the electrophile.
Example 26
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-fluoro-pyrazolo[3,4-
-b]pyridin-1-yl)-ethanone
##STR00048##
[0193] The title compound was synthesized according to the
procedure outlined in Example 6: LCMS (ES) M+H 404.5, R.sub.f 2.27
min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.,
1 ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.08% formic acid/5% water/94.9% acetonitrile).
Example 27
Synthesis of
1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyraz-
olo[3,4-b]pyridine-3-carbonitrile
##STR00049##
[0195]
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo-
[3,4-b]pyridin-1-yl)-ethanone (128 mg) and CuCN (112 mg) were
combined in N-methylpyridone (NMP) (1 mL) and heated at 165.degree.
C. for 16 hr, then cooled to room temperature. The reaction mixture
was purified on preparative HPLC to afford
1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyraz-
olo[3,4-b]pyridine-3-carbonitrile as a white powder: CMS (ES) M+H
411.5, R.sub.f 2.33 min (Agilent Zorbax SB-C18, 2.1.times.50 mm,
5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20%
to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile).
Example 28
Synthesis of 1H-pyrazolo[4,3-b]pyridine
##STR00050##
[0197] 1H-pyrazolo[4,3-b]pyridine was synthesized according to the
procedure outlined in Example 4: LCMS (ES) M+H 120.3.
Example 29
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-
-1-yl-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-
-2-yl-ethanone
##STR00051##
[0199] The two title compounds were synthesized according to the
procedure outlined in Example 6: For
1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-
-1-yl-ethanone; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.71
(dd, 1H), 8.26 (s, 1H), 8.04 (dd, 1H), 7.36 (dd, 1H), 7.20 (d, 1H),
6.69 (d, 1H), 6.51 (dd, 1H), 5.57 (s, 2H), 3.82 (s, 3H), 3.73 (m,
2H), 3.59 (m, 2H), 3.31 (m, 2H), 3.19 (m, 2H). LCMS (ES) M+H 386.5,
R.sub.f 1.84 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20% to
100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile). For
1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-pyrazolo[4,3-b]pyridine-
-2-yl-ethanone; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.55 (d,
1H), 8.34 (s, 1H), 7.99 (d, 1H), 7.19 (m, 2H), 6.44 (d, 1H), 6.40
(dd, 1H), 5.34 (s, 2H), 3.86 (s, 3H), 3.77 (m, 2H), 3.72 (m, 2H),
3.13 (m, 4H). LCMS (ES) M+H 386.5, R.sub.f 1.69 min (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 35.degree. C., 1 ml/min flow rate, a 2.5
min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile).
Example 30
Synthesis of 2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-acetic
acid
##STR00052##
[0201] Preparation of pyrazolo[3,4-b]pyridin-2-yl-acetic acid ethyl
ester: This compound was synthesized according to the procedure
outlined in Example 6, using chloro-acetic acid ethyl ester in
place of
2-Chloro-1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-(S)-methylpiperazin-1-
-yl]ethanone.
[0202] Preparation of (3-Chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic
acid ethyl ester: To a solution of
pyrazolo[3,4-b]pyridin-2-yl-acetic acid ethyl ester 57 (40.2 mg,
0.2 mmol, 1 equiv) in 1 mL of dichloromethane was added NCS (32.7
mg, 1.2 mmol, 1.2 equiv). The resultant mixture was heated at
70.degree. C. for 30 min., cooled to room temperature, and diluted
with 100 mL of dichloromethane. The organic solution was washed
with 50 mL of saturated sodium bicarbonate aqueous solution, and 50
mL of brine. The organic layer was separated and dried over sodium
sulfate. Evaporation of solvent in vacuo gave 46.7 mg of
(3-chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic acid ethyl ester as
yellow solid.
[0203] Synthesis of
2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid:
(3-Chloro-pyrazolo[3,4-b]pyridin-2-yl)-acetic acid ethyl ester was
treated with 1N lithium hydroxide (LiOH) (1 equiv) in 1 mL of MeOH
to provide 2-(3-Chloro-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid,
which was used as directly in subsequent reactions without further
purification: LCMS(ES) M+H 212.0, R.sub.f 0.34 min (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1 ml/min flow rate,
a 2.5 min gradient of 20% to 100% B with a 1.1 min wash at 100% B;
A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic
acid/5% water/94.9% acetonitrile).
Example 31
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-2-(3-chloro-pyrazolo[3,4--
b]pyridine-2-yl)-ethanone
##STR00053##
[0205] The title compound was synthesized according to standard
amide formation conditions using
2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (HATU) as the coupling reagent: LCMS(ES) M+H
420.4, R.sub.f 2.17 min (Agilent Zorbax SB-C18, 2.1.times.50 mm,
5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20%
to 100% B with a 1.1 min wash at 100% B; A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile.
Example 32
Synthesis of 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic
acid
##STR00054##
[0207] Preparation of
2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethyl ester: To
a solution of pyrazolo[3,4-b]pyridin-1-yl-acetic acid ethyl ester
(205.4 mg, 1 mmol, 1 equiv) in 10 mL of dichloromethane at
0.degree. C., was added meta-chloroperoxybenzoic acid (mCPBA)
(345.3 mg, 1.5 mmol, 1.5 equiv). The resultant mixture was allowed
to warm to room temperature, and the reaction was stirred
overnight. 1 mL of pyridine was added to the reaction mixture, and
the mixture was stirred for another 30 min before the solvent was
removed to provide a residue. The residue was diluted with 200 mL
of dichloromethane, and washed with 1 N NaOH aqueous solution (10
mL.times.2), brine (20 mL). The organic layer was separated and
dried over sodium sulfate. Evaporation in vacuo gave
2-(pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethyl ester as
pale yellow solid, which was used without further purification:
LCMS(ES) M+H 222.4, R.sub.f 1.48 min (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5
min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile.
[0208] Synthesis of 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic
acid: 2-(pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid ethyl
ester was treated with 1N LiOH (1 equiv) in 1 mL of methanol (MeOH)
to provide 2-(Pyrazolo[3,4-b]pyridin-1-yl-7-oxide)-acetic acid:
LCMS(ES) M+H 194.2, R.sub.f 0.22 min (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C., 1 ml/min flow rate, a 2.5
min gradient of 20% to 100% B with a 1.1 min wash at 100% B; A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile
Example 33
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)piperazin-1-yl]-2-(pyrazolo[3,4-b]pyridin-
-1-yl-7-oxide)-ethanone
##STR00055##
[0210] The title compound was prepared according to standard amide
formation conditions as described in Example 43 using HATU as the
coupling reagent: LCMS(ES) M+H 402.5, R.sub.f 1.54 min (Agilent
Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1 ml/min flow
rate, a 2.5 min gradient of 20% to 100% B with a 1.1 min wash at
100% B; A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile.
Example 34
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-
-b]pyridin-1-yl)-ethanone.
##STR00056##
[0212] 2-chloro-3-cyano-6-picoline was reduced by
diisobutylaluminum hydride (DIBAL-H) following a literature
procedure (Baker et. al., J. Org. Chem., 1980, 45, 1354-1362.)
followed by the hydrazine condensation protocol as described in
Example 1 to provide the corresponding
6-Methyl-1H-pyrazolo[3,4-b]pyridine, which was then subjected to
the alkylation protocol in described in Example 6 to provide title
compound as a white powder: LCMS (ES) M+H 400.5, R.sub.f 2.161 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1
ml/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B, A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 35
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-
-b]pyridine-2-yl)-ethanone
##STR00057##
[0214] Preparation of
(6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid ethyl ester: To
a solution of 1H-6-methyl-pyrazolo[3,4-b]pyridine (1 mmol, 1 eq.)
in 3 mL of THF was added NaH (1.5 mmol, 1.5 eq.) portion by portion
at 0.degree. C. under nitrogen. The resultant mixture was stirred
at 0.degree. C. for 10 minutes followed by the slow addition of
2-chloro ethyl acetate (excess) at 0.degree. C. The resultant
mixture was slowly to warmed to rt, and stirred for another 2 h. To
the reaction mixture was added saturated NH.sub.4Cl aq. solution,
and aqueous mixture was extracted with 300 mL of EtOAc. The organic
extract was separated and washed with sat. sodium bicarbonate aq.
solution, brine solution, filtered and dried over sodium sulfate.
The organic solvent was removed in vacuo, and the crude residue was
purified by silica gel chromatography to provide 50.2 mg desired
product: HPLC retention time=0.78 minutes (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a
2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100%
B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic
acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=220.1,
found=220.4.
[0215] Preparation of
(6-methyl-pyrazolo[3,4-b]pyridine-2-yl)-acetic acid: This compound
was synthesized according to standard ester hydrolysis protocol as
described in Example 30 using 1N LiOH as the base. The isolated
product was used in the next step without purification.
[0216] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-methyl-pyrazolo[3,4-
-b]pyridine-2-yl)-ethanone: The title compound was synthesized
according to standard peptide coupling protocol using HATU as the
coupling reagent: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.04
(s, 1H), 7.96 (d, 1H), 7.20 (d, 1H), 6.95 (d, 1H), 6.45(d, 1H),
6.40 (dd, 1H), 5.29 (s, 2H), 3.92 (m, 2H), 3.88(s, 3H), 3.78 (m,
2H), 3.13 (m, 4H). LCMS observed for (M+H).sup.+: 400.5.
Example 36
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-morpholin-4-yl-pyra-
zolo[3,4-b]pyridin-1-yl)-ethanone
##STR00058##
[0218] A mixture of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b-
]pyridin-1-yl)-ethanone (102.4 mg), morpholine (0.20 mL), Xantphos
(35 mg), Pd.sub.2(dba).sub.3 (18.3 mg) and Cs.sub.2CO.sub.3 (97 mg)
in THF (1 mL) was heated to 80.degree. C. for 12 h. The reaction
mixture was allowed to cool to room temperature, diluted by EtOAc
(3 mL) and filtered. The filtrate was evaporated in vacuo. The
crude residue was purified by flash chromatography (silica,
Hexane/EtOAc) to provide the title compound as a white powder: LCMS
(ES) M+H 471.6, R.sub.f 2.043 min (Agilent Zorbax SB-C18,
2.1.times.50 mm, 35.degree. C., 1 ml/min flow rate, a 2.5 min
gradient of 20% to 100% B with a 1.1 min wash at 100% B, A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile).
Example 37
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-2--
pyrazolo[3,4-c]pyridin-1-yl-ethanone
##STR00059##
[0220] The title compound was synthesized following the alkylation
protocol as described in Example 6: LCMS (ES) M+H 418.4, R.sub.f
2.055 min (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C., 1 ml/min flow rate, a 2.5 min gradient of 20% to
100% B with a 1.1 min wash at 100% B, A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.08% formic acid/5% water/94.9%
acetonitrile).
Example 38
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-oxy-pyrazolo[3,4-c]-
pyridin-1-yl)-ethanone
##STR00060##
[0222] A mixture of 6-azaindazole (119 mg), H.sub.2O.sub.2 (0.2 mL)
in acetic acid (5 mL) was heated to 60.degree. C. for 2 h. The
resultant mixture was cooled to room temperature and concentrated
in vacuo. The crude residue was dissolved in EtOAc (10 mL), washed
with sat. aqueous NaHCO.sub.3 solution (3 mL), dried
(Na.sub.2SO.sub.4), filtered and evaporated in vacuo. The crude
product (the N-oxide) was subjected to the alkylation protocol as
described in Example 6 to provide the title compound as a white
powder: LCMS (ES) M+H 402.4, R.sub.f 2.147 min (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 35.degree. C., 1 ml/min flow rate, a 2.5
min gradient of 20% to 100% B with a 1.1 min wash at 100% B, A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile).
Example 39
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[4,3-
-c]pyridine-2-yl)-ethanone
##STR00061##
[0224] Preparation of 1H-4-chloro-pyrazolo[4,3-c]pyridine: To a
mixture of 2-chloro-4-iodopyridine-3-carbaldehyde (6.24 mmol, 1
eq.) and 5 mL of ethanol was added 4mL of hydrazine (excess), the
resultant mixture was stirred at rt for 6 h. The reaction solution
was concentrated in vacuo, and the crude residue was diluted with
50 mL of water, and extracted with 500 mL of dichloromethane. The
organic layer was then washed with brine, dried over anhydrous
sodium sulfate, filtered and concentrated to provide a crude
residue. To the crude residue was dissolved with 10 mL of
dichloromethane and stirred for 5 minutes. The precipitated solids
were isolated by filtration, washed with 2 mL of dichloromethane,
and dried in vacuo to provided 350.2 mg of
1H-4-chloro-pyrazolo[4,3,c]pyridine: HPLC retention time=0.44
minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100%
B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expected=154.0, found=154.3.
[0225] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[4,3-
-c]pyridine-2-yl)-ethanone: Using
1H-4-chloro-pyrazolo[4,3-c]pyridine, the title compound was
synthesized according to alkylation protocol in Example 6: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.18 (d, 1H), 7.31(dd, 1H), 7.21
(d, 1H), 6.47 (d, 1H), 6.42 (dd, 1H), 5.28 (s, 2H), 3.88 (s, 3H),
3.77 (m, 4H), 3.14 (m, 4 H). LCMS observed for (M+H).sup.+:
420.4.
Example 40
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-2-yl)-ethanone
##STR00062##
[0227] Preparation of 1H-4-iodo-pyrazolo[3,4,b]pyridine: To a
mixture of 2-chloro-4-iodopyridine-3-carbaldehyde (6.24 mmol, 1
eq.) and 5 mL of ethanol was added 4mL of hydrazine (excess), the
resultant mixture was stirred at rt for 6 h. The reaction mixture
was concentrated in vacuo and crude residue was diluted with 50 mL
of water, and extracted with 500 mL of dichloromethane. The organic
layer was washed with brine, dried over anhydrous sodium sulfate
and concentration in vacuo to provide a crude residue. To this
residue was added 10 mL of dichloromethane, the resultant mixture
was stirred for 5 minutes which resulted in the precipation of the
undesired cyclization isomer (1H-4-chloro-pyrazolo[4,3,c]pyridine)
which was removed by filtration. The filtrate was concentrated in
vacuo, and purified be by silica gel column (35% acetone in hexane
to 50% acetone in hexane) to provide 250.0 mg of
1H-4-iodo-pyrazolo[3,4,b]pyridine with a purity around 85%, which
was used without further purification: HPLC retention time=1.22
minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100%
B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=246.0, found=246.1.
[0228] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-2-yl)-ethanone: The title compounds were synthesized
according to the standard alkylation procedure described in Example
6. For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-1-yl)-ethanone: HPLC retention time=2.50 minutes (Agilent
Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1
ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1
minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9%
water, B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+H
expect=512.0, found=512.4; For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-2-yl)-ethanone: HPLC retention time=2.23 minutes (Agilent
Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C.) using 1 ml/min flow
rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash
at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=512.0,
found=512.4
Example 41
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methylsulfonyl-pyra-
zolo[4,3-c]pyridine-1-yl)-ethanone
##STR00063##
[0230] A mixture of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-iodo-pyrazolo[4,3-c-
]pyridine-1-yl)-ethanone (0.1 mmol, 1 eq.), CuI (0.3 mmol, 3 eq.)
and NaSO.sub.2Me (0.3 mmol, 1 eq.) in 1 mL of DMSO was heated at
80.degree. C. for 2 h. The reaction solution was cooled to rt, and
diluted with 20 mL of sat. NH.sub.4Cl aq. solution and 200 mL of
EtOAc. The diluted mixture was stirred vigorously for 2 h. The
organic layer was then separated, washed with brine, dried over
sodium sulfate, filtered and concentrated in vacuo to provide the
crude product. Purification by HPLC provided 40.2 mg desired
product: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.76 (d, 1H),
8.49(d, 1H), 7.65 (d, 1H), 7.23 (d, 1H), 6.50 (d, 1H), 6.44 (dd,
1H), 5.53 (s, 2H), 3.89 (s, 3H), 3.79 (m, 4H), 3.22 (m+s, 4H+3H).
LCMS observed for (M+H).sup.+: 464.4.
Example 42
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-amidomethyl-pyrazol-
o[3,4-b]pyridine-1-yl)-ethanone
##STR00064##
[0232] Preparation of (3-methyl-pyrazolo[3,4-b]pyridine-1-yl)acetic
acid ethyl ester: This compound was synthesized following the
alkylation protocol similar to the one described in Example 6: HPLC
retention time=2.06 minutes (Agilent Zorbax SB-C18, 2.1.times.50
mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a 2.5 minute
gradient of 0% to 100% B with a 1.1 minute wash at 100% B (A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile); MS (ES) M+H expect=220.1,
found=220.4.
[0233] Preparation of
[3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethyl
ester: A mixture of (3-methyl-pyrazolo[3,4-b]pyridine-1-yl)ethyl
acetate (2.5 mmol, 1 eq.), NBS (3.0 mmol, 1.2 eq.), and benzoly
peroxide (0.05 mmol, 0.02 eq.) in 10 mL of CCl.sub.4 was refluxed
for 1.5 h. The resultant mixture was cooled to rt, and diluted with
500 mL of EtOAc. The resultant solution was then washed with 100 mL
of sat. sodium bicarbonate aqueous solution, brine solution, dried
over anhydrous sodium sulfate, filtered and concentrated in vacuo.
The crude residue was purified by silica gel chromatography (20%
EtOAc in hexane to 35% EtOAc in hexane) to provide 450.2 mg of the
desired product: HPLC retention time=2.50 minutes (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow
rate, a 2.5 minute gradient of 0% to 100% B with a 1.1 minute wash
at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=298.0,
found=298.3.
[0234] Preparation of (3-azido-pyrazolo[3,4-b]pyridine-1-yl)acetic
acid ethyl ester: A mixture of
(3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethyl
ester (0.5 mmol, 1 eq.) and sodium azide (1 mmol, 2 eq.) in 1 mL of
DMF was heated at 80.degree. C. for 1 h. The resultant mixture was
cooled to rt, diluted with 150 mL of EtOAc, washed with water (40
mL.times.3), brine, dried over anhydrous sodium sulfate, filtered
and concentrated in vacuo. The solvent was removed in vacuo to
provide 135.2 mg desired product: HPLC retention time=1.84 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid /5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=261.1, found=261.4.
[0235] Preparation of
[3-(azidomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic acid: This
compound was synthesized according to standard hydrolysis protocol
as described in Example 30 using 1 N LiOH: HPLC retention time=1.94
minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C.) using 1 ml/min flow rate, a 2.5 minute gradient of 0% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=233.1, found=233.4.
[0236] Preparation of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-azidomethyl-pyrazol-
o[3,4-b]pyridine-1-yl)-ethanone: This compound was synthesized
according to standard peptide coupling procedure as described below
in Examle 43 using HATU as the coupling reagent: HPLC retention
time=2.36 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
20% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=441.2, found=441.5.
[0237] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-amidomethyl-pyrazol-
o[3,4-b]pyridine-1-yl)-ethanone: To a solution of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-azidomethyl-pyrazol-
o[3,4-b]pyridine-1-yl)-ethanone (0.21 mmol, 1 eq.) in 2 mL of THF
was added dropwise at rt a solution of
tris(2-carboxyethyl)phosphine HCl salt in 0.5 mL of water. The
resultant mixture was stirred at rt for 30 min. The reaction
solution was concentrated in vacuo, and the crude residue was
diluted with 150 mL of dichloromethane, washed with 25 mL of water,
brine, and dried over sodium sulfate. The solvent was removed in
vacuo, and the crude residue was purified by HPLC to provide 26.2
mg final product: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.17
(dd, 1H), 8.15(dd, 1H), 7.22 (d, 1H), 7.12 (dd, 1H), 6.50 (d, 1H),
6.44 (dd, 1H), 5.40 (s, 2H), 4.25 (s, 2H), 3.89 (s, 3H), 3.77 (m,
4H), 3.19 (m, 4H). LCMS observed for (M+H).sup.+: 416.4.
Example 43
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonic
acid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone
##STR00065##
[0239] Preparation of [3-(sulfonic
acid-methyl)-pyrazolo[3,4-b]pyridine-1-yl]acetic acid ethyl ester:
A mixture of (3-(bromomethyl)-pyrazolo[3,4-b]pyridine-1-yl)acetic
acid ethyl ester (0.13 mmol, 1 eq.) and sodium sulfite (1.8 mmol,
excess) in a mixture of 1 mL of DMF and 0.5 mL of water was heated
at 80.degree. C. for one hour. The resultant solution was cooled to
rt, and the solvent was removed in vacuo. The residue was extracted
with 1:1 MeOH:CH.sub.2Cl.sub.2 (30 mL.times.3). The combined
organic extracts were dried in vacuo, and the crude residue was
used without further purification: HPLC retention time=1.63 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 35.degree. C.) using 1
ml/min flow rate, a 2.5 minute gradient of 0% to 100% B with a 1.1
minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9%
water, B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+H
expect=300.1, found=300.5.
[0240] Preparation of [3-(sulfonic
acid-methyl)-pyrazolo[3,4-b]pyridine-1-yl]acetic acid: This
compound was synthesized according to standard hydrolysis protocol
as described in Example 30 using 1 N LiOH as the base. The crude
product was used without further purification.
[0241] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonic
acid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone: A mixture of
sulfonic acid (100.2 mg, contain lots of inorganic salt),
1H-4-(4-chloro-3-methoxy-phenyl)piperazine 2.times. HCl salt (0.37
mmol, excess), and HATU (0.37 excess) was suspended in 3 mL of
pyridine, stirred at rt for 3 h. The pyridine solvent was removed
in vacuo, and the crude residue was extracted with dichloromethane
(10 mL.times.3). The organic extracts were removed, and the crude
residue was purified by HPLC to provide 10 mg of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-sulfonic
acid-methyl-pyrazolo[3,4-b]pyridine-1-yl)-ethanone: HPLC retention
time=0.28 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
20% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=480.1, found=480.5.
Example 44
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[3,4-
-b]pyridine-1-yl)-ethanone
##STR00066##
[0243] Preparation of
(3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethyl
ester: To a solution of
(3-iodo-5-pyrazolo[3,4-b]pyridine-1-yl)acetic acid ethyl ester
(0.61, leq.) in 2 mL of DMF was added N-chlorosuccinimide (NCS)
(0.73, 1.2 eq.) as a solid. The resultant mixture was heated at
70.degree. C. for 3 h. The reaction mixture was cooled to rt, and
diluted with 250 mL of EtOAc. The diluted mixture was then washed
with water (100 mL.times.3), brine, dried over sodium sulfate,
filtered and concentrated in vacuo. The crude residue was purified
by silica gel chromatography (15% EtOAc to 75% EtOAc in hexane) to
provide 100.4 mg white solid as final product: HPLC retention
time=2.48 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
20% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=365.9, found=366.3.
[0244] Preparation of
(3-iodo-5-chloro-pyrazolo[3,4-b]pyridine-1-yl)acetic acid: This
compound was synthesized according to standard hydrolysis procedure
as described in Example 30 using 1N LiOH: HPLC retention time=1.78
minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100%
B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=337.9, found=337.9.
[0245] Preparation of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-5-chloro-pyraz-
olo[3,4-b]pyridine-1-yl)-ethanone: The title compound was
synthesized according to standard peptide coupling protocol as
described in Example 43 using HATU as the coupling reagent: HPLC
retention time=2.71 minutes (Agilent Zorbax SB-C18, 2.1.times.50
mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a 2.5 minute
gradient of 20% to 100% B with a 1.1 minute wash at 100% B (A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile); MS (ES) M+H expect=546.0,
found=546.4.
[0246] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-chloro-pyrazolo[3,4-
-b]pyridine-1-yl)-ethanone: To a solution of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3iodo-5-chloro-pyrazo-
lo[3,4-b]pyridine-1-yl)-ethanone (0.037 mmol, 1 eq.) in 1.5 mL of
dichloromethane under a nitrogen atmosphere cooled to -40.degree.
C., was added dropwise, 30 .mu.l of 2.0 M solution of isopropyl
magnesium chloride (0.056 mmol, 1.5 eq.) in THF. The resultant
mixture was for 30 minutes at -40.degree. C. followed by dropwise
addition of an ammonium chloride aqueous (aq) solution at low
temperature. The reaction solution was warmed to rt, diluted with
200 mL of EtOAc, washed with 50 mL of water, brine, dried over
sodium sulfate, filtered and concentrated in vacuo. The crude
residue was purified by HPLC to provide 5 mg final product: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.44 (d, 1H), 8.05(m, 1H), 7.23
(d, 1H), 6.50 (d, 1H), 6.43 (dd, 1H), 5.42 (s, 2H), 3.89 (s, 3H),
3.76 (m, 4H), 3.20 (m, 4H). LCMS observed for (M+H).sup.+:
421.1.
Example 45
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-2-yl)-ethanone
##STR00067##
[0248] Preparation of 1H-4-chloro-pyrazolo[3,4-d]pyrimidine: This
compound was synthesized according to standard hydrazine
cyclization protocol as described in Example 1: HPLC retention
time=0.36 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
0% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=155.0, found=155.0.
[0249] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-2-yl)-ethanone: These compounds were synthesized
using 1H-4-chloro-pyrazolo[3,4-d]pyrimidine following the
alkylation procedure as described in Example 6: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-1-yl)-ethanone; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.76 (s, 1H), 8.22 (s, 1H), 7.22 (d, 1H), 6.54 (d, 1H),
6.44 (dd, 1H), 5.41 (s, 2H), 3.88 (s, 3H), 3.77 (m, 4H), 3.23 (m,
4H), LCMS observed for (M+H).sup.+: 421.1: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-2-yl)-ethanone; HPLC retention time=1.70 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=421.1, found=421.1.
Example 46
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-methoxy-pyrazolo[3,-
4-d]pyrimidine-1-yl)-ethanone
##STR00068##
[0251] To a solution of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(4-chloro-pyrazolo[3,4-
-d]pyrimidine-1-yl)-ethanone (0.024 mmol, 1 eq.) in 1 mL of MeOH
was added solid potassium carbonate (excess), the resultant mixture
was heated at 70.degree. C. for 30 minutes, then filtered and dried
under vacuum. The crude product was purified by HPLC to provide the
desired product as a white powder: .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.90 (s, 1H), 8.07 (s, 1H), 7.22 (d, 1H), 6.50
(d, 1H), 6.44 (dd, 1H), 5.29 (s, 2H), 3.90 (s, 3H), 3.81 (m, 4H),
3.19 (m, 4H), 2.25 (s, 3H). LCMS observed for (M+H).sup.+:
418.9.
Example 47
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-2-yl)-ethanone
##STR00069##
[0253] Preparation of (2,6-dichloro-3-pyridinyl)methanol: To a
solution of 2,6-dichloro-3-nicotic acid (9 mmol, 1 eq.) in 10 mL of
dry THF at 0.degree. C., was added NaBH.sub.4 (27 mmol, 3 eq.)
portion by portion under nitrogen atmosphere. After the evolution
of hydrogen gas subsided (which is observed as bubbling in the
reaction mixture), BF.sub.3.OMe.sub.2 (27 mmol, 3 eq.) was added
dropwise to the reaction mixture at 0.degree. C. The resultant
mixture was stirred at 0.degree. C. for 20 minutes followed by the
slow addition of sat. NH.sub.4Cl aq. solution. The reaction
solution was then warmed to rt, and extracted with 300 mL of EtOAc,
and the organic layer was washed with brine, dried over sodium
sulfate, filtered and concentrated in vacuo to provide a white
solid, which was used in subsequent reaction without further
purification: HPLC retention time=0.71 minutes (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow
rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash
at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1%
formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=178.0,
found=178.0.
[0254] Preparation of 2,6-dichloro-3-formylpyridine: To a solution
of the above alcohol (2 mmol, 1 eq.) in 10 mL of dichloromethane
was added potassium carbonate (excess) as a solid, and Dess-Martin
periodinate (2 mmol, 1 eq.) at rt. The resultant mixture was
stirred at rt for 30 minutes. A 5% sodium thiosulfate aq. solution
was added to the reaction mixture and the resultant mixture was
stirred for another 10 minutes. The reaction mixture was extracted
with 300 mL of EtOAc, and the organic layer was washed with 50 ml
of 5% sodium thiosulfate aq. solution, Sat. sodium bicarbonate aq.
solution, brine, and dried over sodium sulfate. Evaporation of
solvent in vacuo to provide 200.1 mg of the desired product as a
white solid: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.37 (s,
1H), 8.17(d, 1H), 7.42 (d, 1H).
[0255] Preparation of 6-choloro-pyrazolo[3,4-b]pyridine: To a
solution of 2,6-dichloro-3-formylpyridine (0.89 mmol, 1 eq.) in 3
ml THF was added hydrazine (1.06 mmol, 1.2 eq.) at rt. The
resultant solution was heated at 120.degree. C. in sealed tube for
overnight. The solvent was removed in vacuo, and the residue was
dry loaded on silica gel column. Purification by silica gel
chromatography provide 29.5 mg final product: HPLC retention
time=2.17 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
0% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=154.0, found=154.0.
[0256] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-1-yl)-ethanone and
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-2-yl)-ethanone: The two title compounds were
synthesized according to the standard coupling procedure described
in Example 6: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-1-yl)-ethanone; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.08 (s, 1H), 8.01(d, 1H), 7.25 (d, 1H), 7.16 (d, 1H),
6.50(d, 1H), 6.45 (dd, 1H), 5.40 (s, 2H), 3.90 (s, 3H), 3.77 (m,
4H), 3.23 (m, 4H). LCMS observed for (M+H).sup.+: 420.5: For
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-chloro-pyrazolo[3,4-
-b]pyridine-2-yl)-ethanone: HPLC retention time=1.66 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile). LCMS observed for (M+H).sup.+: 420.5.
Example 48
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4--
d]pyridine-1-yl)-ethanone
##STR00070##
[0258] Preparation of 1H-6-hydrazo-pyrazolo[3,4-d]pyridine: To a
solution of 2,6-dichloro-3-pyridinecarbaldehyde in 2 mL of dioxane
was added excess amount of hydrazine. The resultant solution was
heated at 150.degree. C. overnight. Upon cooling to rt, the desired
product precipitated out of solution as a white solid. The crude
product was isolated by filtration, washed with a small amount of
dioxane, and dried in vacuo. The crude product was used without
further purification: HPLC retention time=1.78 minutes (Agilent
Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1
ml/min flow rate, a 2.5 minute gradient of 20% to 100% B with a 1.1
minute wash at 100% B (A=0.1% formic acid/5% acetonitrile/94.9%
water, B=0.1% formic acid/5% water/94.9% acetonitrile); MS (ES) M+H
expect=337.9, found=337.9.
[0259] Preparation of 1H-6-azido-pyrazolo[3,4-dlpyridine:
1H-6-hydrazo-pyrazolo[3,4-d]pyridine was suspended into a mixture
of 5 mL of concentrated HCl and 10 mL of water at 0.degree. C., and
to it was added dropwise a solution of sodium nitrate in 5 mL of
water. The resultant mixture was stirred at 0.degree. C. for 10 min
and warmed to rt. The reaction mixture was neutralized to pH=7-8,
and extracted with (200 mL.times.2) EtOAc. The combined organic
extract was washed with brine, dried over sodium sulfate, filtered
and concentrated in vacuo to provide the desired product which was
used without further purification: HPLC retention time=0.50 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=161.0, found=160.8.
[0260] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4--
d]pyridine-1-yl)-ethanone: The title compound was synthesized
according to alkylation protocol described in Example 6: HPLC
retention time=2.22 minutes (Agilent Zorbax SB-C18, 2.1.times.50
mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a 2.5 minute
gradient of 20% to 100% B with a 1.1 minute wash at 100% B (A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5%
water/94.9% acetonitrile); MS (ES) M+H expect=427.1,
found=427.1.
Example 49
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-amido-pyrazolo[3,4--
b]pyridine-1-yl)-ethanone
##STR00071##
[0262] To a solution of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-azido-pyrazolo[3,4--
b]pyridine-1-yl)-ethanone (0.071 mmol, 1 eq.) in 1 mL of EtOAc was
added SnCl.sub.2.2H.sub.2O as a solid. The resultant mixture was
heated at 40.degree. C. for 2 h. The resultant mixture was cooled
to rt and diluted with 200 mL of EtOAc and 50 mL of Sat. sodium
bicarbonate aq. solution. The diluted mixture was stirred for an
additional 1 h, before the organic layer was separated, washed with
brine, and dried over sodium sulfate. The solvent was removed in
vacuo, and the residue was purified by HPLC to provide 10 mg title
compound: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.86 (s, 1H),
7.74(d, 1H), 7.24 (d, 1H), 7.20 (d, 1H), 6.45(d, 1H), 6.39 (dd,
1H), 5.16 (s, 2H), 3.88 (s, 3H), 3.89 (m, 4H), 3.13 (m, 4H). LCMS
observed for (M+H).sup.+: 401.1.
Example 50
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-amino-pyrazolo[3,4--
c]pyridine-1-yl)-ethanone
##STR00072##
[0264] Preparation of 7-hydrazo-pyrazolo[3,4-c]pyridine: To a
solution of 2-chloro-3-fluoro-4-formylpyridine (5.75 mmol, 1 eq.)
in 20 mL of THF was added 1 mL of hydrazine (excess). The resultant
solution was heated at 110.degree. C. in sealed tube for 5 h. The
reaction was cooled to rt and solvent was removed in vacuo. The
crude residue was washed several times with hexane, EtOAc, and
dried in vacuo to provide a light yellow solid, which was used
without further purification: HPLC retention time=0.20 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=150.1, found=150.0.
[0265] Preparation of 7-azido-pyrazolo[3,4-c]pyridine: This
compound was synthesized according to protocol described in Example
48: HPLC retention time=0.26 minutes (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a
2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100%
B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic
acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=161.0,
found=160.9.
[0266] Preparation of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-azido-pyrazolo[3,4--
c]pyridine-2-yl)-ethanone: This compound was synthesized according
to protocol described in Example 6: HPLC retention time=2.43
minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree.
C.) using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100%
B with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=427.1, found=427.2.
[0267] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(7-amino-pyrazolo[3,4--
c]pyridine-1-yl)-ethanone: The title compound was synthesized
according to the procedure outlined in Example 49: .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.86 (s, 1H), 7.74(d, 1H), 7.24 (d, 1H),
7.20 (d, 1H), 6.45(d, 1H), 6.39 (dd, 1H), 5.16 (s, 2H), 3.88 (s,
3H), 3.89 (m, 4H), 3.13 (m, 4H). LCMS observed for (M+H).sup.+:
401.1.
Example 51
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(oxazole-2yl)-pyraz-
olo[3,4-b]pyridine-2-yl]-ethanone
##STR00073##
[0269] Preparation of (3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic
acid ethyl ester: To a solution of
3-Iodo-2H-pyrazolo[3,4-b]pyridine (4 mmol, 1 eq.) in 10 mL of dry
THF was added dropwise 0.5 M KHMDS (potassium hexamethyldisilazide)
in toluene (4.4 mmol, 1.1 eq.) at -78.degree. C., under nitrogen
atmosphere, and the resultant solution was stirred at for 30
minutes at -78.degree. C. Chloro ethyl acetate (8 mmol, 2eq.) was
added dropwise to the reaction solution and the reaction solution
was warmed to rt over 1.5 hour and stirred overnight. Following an
aqueous workup, the crude product was purified by silica gel
chromatography (20% EtOAc in hexane to 70% EtOAc in hexane) to
provide 70.2 mg of (3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic acid
ethyl ester: HPLC retention time=2.63 minutes (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow
rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash
at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.08%
formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=332.0,
found=332.1.
[0270] Preparation of (3-iodo-pyrazolo[3,4-b]pyridine-2-yl)acetic
acid: This compound was synthesized according to the standard ester
hydrolysis protocol as described in Example 30 using 1N LiOH as the
base. The crude product was used in the next step without
purification: HPLC retention time=1.02 minutes (Agilent Zorbax
SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow
rate, a 2.5 minute gradient of 20% to 100% B with a 1.1 minute wash
at 100% B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.08%
formic acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=303.0,
found=303.5.
[0271] Preparation of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b-
]pyridine-2-yl)-ethanone: This compound was synthesized according
to standard peptide coupling protocol using HATU as the coupling
reagent: HPLC retention time=297 minutes (Agilent Zorbax SB-C18,
2.1.times.50 mm, 35.degree. C.) using 1 ml/min flow rate, a 2.5
minute gradient of 20% to 100% B with a 1.1 minute wash at 100% B
(A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.08% formic
acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=512.0,
found=512.5.
[0272] Synthesis of 1-[4-(4-chloro-3-methoxy
-phenyl)-piperazin-1-yl]-2-[3-(oxazole-2yl)-pyrazolo[3,4-b]pyridine-2-yl]-
-ethanone: To a mixture of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b-
]pyridine-2-yl)-ethanone (0.071 mmol, 1 eq.) and tetrakis
triphenylphosphine palladium (0.025 mmol, 0.35 eq.) under nitrogen
atmosphere was added 0.5 mL of THF and 2-oxazole-(tri-n-butyl)Tin
(0.48 mmol, 6.7 eq.). The resultant mixture was heated in a sealed
tube at 80.degree. C. for 48 h. The reaction solution was cooled to
rt, diluted with 30 mL of NH.sub.4Cl sat. aq. solution, and
extracted with 300 mL of EtOAc. The organic layer was separated,
washed with brine, dried over sodium sulfate, filtered and
concentrated in vacuo. The crude residue was purified by silica gel
chromatography (0% to 15% MeOH in EtOAc) to provide 12.3 mg the
title compound: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.77 (dd,
1H), 8.51(dd, 1H), 7.81 (d, 1H), 7.22 (m, 3H), 6.52(d, 1H), 6.48
(dd, 1H), 6.01 (s, 2H), 3.90 (s, 3H), 3.80 (m, 4H), 3.27 (m, 4H).
LCMS observed for (M+H).sup.+: 453.5.
Example 52
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4--
b]pyridine-1-yl]ethanone
##STR00074##
[0274] Preparation of 3-methyl-5-nitro-pyrazolo[3,4-b]pyridine:
3-Methyl-pyrazolo[3,4-b]pyridine (1 mmol, 1 eq.) was suspended into
a mixture of 1:1 fuming nitric acid and concentrated sulfuric acid
(1 mL :1 mL), and the resultant mixture was heated at 90.degree. C.
for 30 minutes. The reaction mixture was then cooled to rt, and
poured into a mixture of sodium bicarbonate and ice. The resultant
solution was warmed up to rt and extracted with 300 mL of EtOAc.
The organic extract was separated, washed with brine, dried over
sodium sulfate, filtered, and concentrated in vacuo. The crude
residue was purified by silica gel chromatography to provide 70.2
mg of 3-methyl-5-nitro-pyrazolo[3,4-b]pyridine.
[0275] Preparation of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-nitro-pyrazolo[3,4--
b]pyridine-1-yl]-ethanone: This compound was synthesized from
3-methyl-5-nitro-pyrazolo[3,4-b]pyridine according to the
alkylation protocol described in Example 6: HPLC retention
time=1.46 minutes (Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu.,
35.degree. C.) using 1 ml/min flow rate, a 2.5 minute gradient of
20% to 100% B with a 1.1 minute wash at 100% B (A=0.1% formic
acid/5% acetonitrile/94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=445.1, found=445.1.
[0276] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4--
b]pyridine-1-yl]-ethanone:
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-nitro-pyrazolo[3,4--
b]pyridine-1-yl]-ethanone (15 mg) is combined with 200 mg of iron
powder in 2 mL of acetic acid at 100.degree. C. for 30 min. After
cooling to rt, the reaction solution was diluted with EtOAc and
filtered. The filtrate was evaporated in vacuo and purified by HPLC
to provide
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(5-amino-pyrazolo[3,4--
b]pyridine-1-yl]-ethanone: HPLC retention time=1.46 minutes
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C.)
using 1 ml/min flow rate, a 2.5 minute gradient of 20% to 100% B
with a 1.1 minute wash at 100% B (A=0.1% formic acid/5%
acetonitrile/ 94.9% water, B=0.1% formic acid/5% water/94.9%
acetonitrile); MS (ES) M+H expect=414.2, found=415.1.
Example 53
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-amino-6-methyl-pyra-
zolo[3,4-b]pyridine-1-yl]-ethanone
##STR00075##
[0278] Preparation of 1H-3-amino-6-methyl-pyrazolo[3,4-d]pyridine:
This compound was synthesized according to the cyclization
procedure using hydrazine described in Example 3 and the crude
product was used in the next step without further purification.
[0279] Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-amino-6-methyl-pyra-
zolo[3,4-b]pyridine-1-yl]-ethanone: This compound was synthesized
according to the standard coupling procedure described in Example
6: .sup.1H NMR (400 MHz, CDCl.sub.3) 7.75 (d, 1H), 7.22 (d, 1H),
6.86 (d, 1H), 6.48 (d, 1H), 6.42(dd, 1H), 5.18 (s, 2H), 3.89 (s,
3H), 3.75 (m, 4H), 3.16 (m, 4H), 2.62 (s, 3H). LCMS observed for
(M+H).sup.+: 415.5.
Example 54
Synthesis of
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl) ethanone
##STR00076##
[0281] Preparation of
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone: A mixture of
2-Chloro-1-[(S)-4-(4-chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-
-1-yl]-ethanone (1.37 g, 4.08 mmol, 1 eq),
3-Iodo-1H-pyrazolo[3,4-b]pyridine (1.0 g, 4.08 mmol, 1 eq),
potassium carbonate (2.26 g, 16.4 mmol, 4 eq), and DMF (15 ml) was
stirred overnight at 90.degree. C. The reaction solution was
diluted with ethyl acetate, washed with saturated aqueous
NaHCO.sub.3, and concentrated in vacuo. The crude product was
purified by flash chromatography to provide
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone (2.2 g).
[0282] Preparation of
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-
-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile: A mixture
of
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-ethanone (2.2 g, 4.0 mmol, 1
eq), CuCN (3.6 g, 40 mmol, 10 eq), and DMF (25 ml) was stirred at
175.degree. C. for 1 hrs. The reaction mixture was cooled to rt,
diluted with ethyl acetate and filtered. The filtrate was washed
with water, dried over Na.sub.2SO.sub.4, and purified by flash
chromatography to provide
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1-yl]-
-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile (1.6
g).
[0283] Preparation of
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-o-
xo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine: A
mixture of
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-(S)-methyl-piperazin-1--
yl]-2-oxo-ethyl}-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile (1.6 g,
3.6 mmol, 1 eq), NH.sub.2OH.HCl (0.84 g, 10.8 mmol, 3 eq), TEA (1.5
ml), and ethanol (10 ml) was stirred at 65.degree. C. overnight.
The reaction solution was concentrated in vacuo, and dissolved in
ethyl acetate, washed with brine, and concentrated to provide
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2-o-
xo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine (1.2
g).
[0284] Preparation of
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone: A
mixture of
1-{2-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]--
2-oxo-ethyl}-N-hydroxy-1H-pyrazolo[3,4-b]pyridine-3-carboxamidine
(1.2 g), trimethyl orthoformate (20 ml) and para-toluene sulfonic
acid (PTSA) (0.1 g) was stirred at 100.degree. C. overnight. The
reaction mixture was concentrated in vacuo to provide a crude
residue which was purified by flash chromatography to provide
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-2-methyl-piperazin-1-yl]-2--
(3-[1,2,4]oxadiazol-3-yl-pyrazolo[3,4-b]pyridin-1-yl)-ethanone (0.7
g). LCMS Retention time: 2.61 min (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a
2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100%
B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic
acid/5% water/94.9% acetonitrile). LCMS observed for (M+H).sup.+:
486.
Example 55
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4--
b]pyridine-1-yl)-ethanone
##STR00077##
[0286] A solution of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4--
b]pyridine-1-yl)-ethanone (0.15 mmol, 1 eq.) and hydroxyl amine HCl
salt (0.45 mmol, 3 eq.) in 2.5 mL of EtOH was heated at 60.degree.
C. for 1 h. The reaction mixture was cooled to rt, and concentrated
in vacuo. The crude residue was dissolved with 200 mL of
dichloromethane, washed with 50 mL of 5% K.sub.2CO.sub.3 aq.
solution, brine solution, dried over sodium sulfate, filtered and
concentrated in vacuo to provide the desired product as a white
solid: HPLC retention time=1.61 minutes (Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a
2.5 minute gradient of 20% to 100% B with a 1.1 minute wash at 100%
B (A=0.1% formic acid/5% acetonitrile/94.9% water, B=0.1% formic
acid/5% water/94.9% acetonitrile); MS (ES) M+H expect=444.1,
found=444.5.
Example 56
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(oxadiazole-3-yl)-p-
yrazolo[3,4-b]pyridine-1-yl]-ethanone
##STR00078##
[0288] To a suspension of (0.067 mmol, 1 eq.) of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-cyano-pyrazolo[3,4--
b]pyridine-1-yl)-ethanone in 2 mL of trimethylorthoformate was
added camphorsulfonic acid (CSA) (5.0 mg, catalytic amount). The
resultant mixture was heated at 50.degree. C. for 10 minutes and
cooled to rt. The reaction solution was concentration in vacuo to
provide a crude residue which was purified by HPLC chromatography
to provide 20.0 mg of the title compound: .sup.1H NMR (400 MHz,
CDCl.sub.3) 8.83 (s, 1H), 8.62(dd, 1H), 7.35 (dd, 1H), 7.22 (d,
1H), 6.51 (d, 1H), 6.44 (dd, 1H), 5.59 (s, 2H), 3.89 (s, 3H), 3.79
(m, 4H), 3.23 (m, 4H). LCMS observed for (M+H).sup.+: 454.5.
Example 57
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(5-methyl-oxadiazol-
e-3-yl)-pyrazolo[3,4-b]pyridine-1-yl]-ethanone
##STR00079##
[0290] The title compound was synthesized according to the
cyclization procedure using trimethylorthoacetate as described in
Example 56: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.61 (dd,
1H), 7.31(dd, 1H), 7.22 (d, 1H), 6.54 (d, 1H), 6.42 (dd, 1H), 5.57
(s, 2H), 3.89 (s, 3H), 3.77 (m, 4H), 3.21 (m, 4H), 2.69 (s, 1H).
LCMS observed for (M+H).sup.+: 468.5.
Example 58
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-acetimido-pyrazolo[-
3,4-b]pyridine-1-yl)-ethanone
##STR00080##
[0292]
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(6-amido-pyrazol-
o[3,4-b]pyridine-1-yl)-ethanone, acetic anhydride (1.2 equiv) and
pyridine (3 equiv) was combined in DCM at rt for 30 min: HPLC
retention time=1.82 minutes (Agilent Zorbax SB-C18, 2.1.times.50
mm, 5.mu., 35.degree. C.) using 1 ml/min flow rate, a 2.5 minute
gradient of 20% to 100% B with a 1.1 minute wash at 100% B (A=0.1%
formic acid/5% acetonitrile/94.9% water, B=0.08% formic acid/5%
water/94.9% acetonitrile); MS (ES) M+H expect=443.1,
found=442.8.
Example 59
Synthesis of
1-[4-(4-chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-methylsulfonyl-pyra-
zolo[4,3-c]pyridine-1-yl)-ethanone
##STR00081##
[0294] The title compound was synthesized from
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b-
]pyridin-1-yl)-ethanone according to the protocol described in
Example 41: .sup.1H NMR (400 MHz, CDCl.sub.3) 8.65 (d, 1H), 8.48
(d, 1H), 7.39 (dd, 1H), 7.22 (d, 1H), 6.51 (s, 1H), 6.44(d, 1H),
5.53 (s, 2H), 3.91 (s, 3H), 3.78 (m, 4H), 3.34 (s, 3H), 3.22 (m,
4H), LCMS observed for (M+H).sup.+: 415.0.
Example 60
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-y-
l-pyrazolo[3,4-b]pyridin-1-yl)-ethanone
##STR00082##
[0296] Preparation of Ethyl
(3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate: To a mixture of
3-iodo-1H-pyrazolo[3,4-b]pyridine (9.8 g, 40 mmol, 1 equiv) and
potassium carbonate (27.6 g, 5 equiv) in 15 mL of DMF at 90.degree.
C. was added ethyl chloroacetate (8.5 mL, 40 mmol, 1 equiv). Two
hours later, the reaction mixture was diluted with ethyl acetate
followed by washing with saturated aqueous NaHCO.sub.3. The organic
layer was dried and concentrated to provide the crude product.
Purification of the crude product by flash chromatography gave
ethyl (3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate (11g).
[0297] Preparation of (3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic
acid: Ethyl (3-Iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetate (11 g, 33
mmol, 1 equiv) was dissolved in 50 mL of THF and 50 mL of MeOH to
the solution was added 40 mL of 1N LiOH for 3 h. The organic
solvents were evaporated and the remaining aqueous phase was
neutralized with 1N HCl to a pH of about 1 which resulted in the
precipitation of the desired product as a white solid was filtered
and air dried to give (3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic
acid.
[0298] Preparation of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl--
pyrazolo[3,4-b]pyridin-1-yl)-ethanone: A mixture of
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid (3.03 g, 10 mmol,
1 equiv), 1-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazine (2.45
g, 1 equiv), BOP reagent (4.86 g, 1 equiv), triethylamine (4.2 mL,
3 equiv) in 10 mL of MF was stirred at rt overnight. To the
reaction mixture was then added water and the solid precipitates
were removed by filtration and air dried to give
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl--
pyrazolo[3,4-b]pyridin-1-yl)-ethanone. LCMS (ES) observed for M+H
530.0.
[0299] Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-y-
l-pyrazolo[3,4-b]pyridin-1-yl)-ethanone: To a solution of oxazole
(690 mg 10 mmol, 2.5 equiv) in tetrahydrofuran (5 mL) under
nitrogen atmosphere, was added dropwise n-butyl lithium (2.5 M in
Hexane, 4.8 mL, 3 equiv.). The resultant mixture was stirred at
-78.degree. C. for an additional 60 min followed by the addition of
ZnCl.sub.2 (0.5 M in THF, 32 mL, 4 equiv.). The reaction solution
was allowed to warm to 0.degree. C. and stirred 1 h followed by the
addition of
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-2-yl--
pyrazolo[3,4-b]pyridin-1-yl)-ethanone (2.12 g, 4 mmol, 1 equiv) and
palladium tetrakis(triphenylphosphine) (462 mg, 0.1 equiv). The
reaction mixture was then heated to reflux for 12 hr, cooled to
room temperature and diluted with ethyl acetate. The reaction
mixture was washed with water, brine, dried over sodium sulfate,
and concentrated in vacuo to provide the crude product.
Purification by flash chromatography provided of the desired
product
1-[4-(4-Chloro-2-fluoro-5-methoxy-phenyl)-piperazin-1-yl]-2-(3-oxazol-2-y-
l-pyrazolo[3,4-b]pyridin-1-yl)-ethanone as a white powder (1.03 g).
LCMS (ES) observed for M+H 471.1. HPLC retention time=2.4 min
(Agilent Zorbax SB-C18, 2.1.times.50 mm, 5.mu., 35.degree. C., 1
mL/min flow rate, a 2.5 min gradient of 20% to 100% B with a 1.1
min wash at 100% B; A=0.1% formic acid/5% acetonitrile/94.9% water,
B=0.1% formic acid/5% water/94.9% acetonitrile).
Example 61
Synthesis of
1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol--
2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00083##
[0301] Step 1: A mixture of 3-iodo-7-azaindazole (25.50 g) and
K.sub.2CO.sub.3 (41.4 g) in DMF (200 mL) was heated to 85.degree.
C. and t-butyl chloroacetate (14.3 mL) was slowly added. The
mixture was stirred at this temperature for 1 hour (h), cooled to
room temperature followed by the addition of water (300 mL).
Filtration of the reaction mixture provided
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl
ester.
[0302] Step 2: A 250 mL flask was charged with
(3-iodo-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester
(15.0 g), PdCl.sub.2(dppf) (3.0 g), Zn(CN).sub.2 (4.96 g), DMF (200
mL) and H.sub.2O (14 mL). The flask containing the resultant
suspension was degassed and backfilled with nitrogen gas repeatedly
for 5 minutes, followed by addition of Pd.sub.2(dba).sub.3 (3.85 g)
to the reaction mixture. The reaction mixture was heated under
N.sub.2 at 90.degree.C. for 16 h, cooled to room temperature,
diluted with H.sub.2O (800 mL) and filtered. The collected solid
was washed with toluene (10 mL) to provide
(3-cyano-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester
as a yellow solid.
[0303] Step 3: A mixture of
(3-cyano-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl ester,
hydroxylamine hydrochloride (8.28 g) and Et.sub.3N (22.6 mL) in
EtOH (120 mL) was heated over night under N.sub.2 at 65.degree.C.
The mixture was cooled to room temperature, filtered and the
collected solid was washed with H.sub.2O (100 mL) and Et.sub.2O (50
mL.times.2) to afforded
[3-(N-hydroxycarbamimidoyl)-pyrazolo[3,4-b]pyridin-1-yl]-acetic
acid tert-butyl ester.
[0304] Step 4:
[3-(N-Hydroxycarbamimidoyl)-pyrazolo[3,4-b]pyridin-1-yl]-acetic
acid tert-butyl ester (6.17 g) in a 100 mL vial was charged with
AcOH (45 mL) and Ac.sub.2O (4.3 mL). The resultant mixture was
stirred at room temperature for 1 h at this time the initial
suspension became a clear solution. To this solution was added Pd/C
(10%, 900 mg) and stirred under 1 atm H.sub.2 balloon, and
resultant mixture was stirred over night at room temperature.
Filtration from celite washed with DCM/MeOH and evaporation gave
(3-amidino-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid tert-butyl
ester which was used without further purification.
[0305] Step 5: (3-Amidino-pyrazolo[3,4-b]pyridin-1-yl)-acetic acid
tert-butyl ester obtained above in a 100 mL vial was charged with
chloroacetylaldehyde (5.72 mL), dioxane (50 mL) and K.sub.2CO.sub.3
(12.42 g). The resultant mixture was stirred at 80.degree. C. for
4h and more chloroacetylaldehyde (5.72 mL) and K.sub.2CO.sub.3
(12.42 g) were added. The mixture was stirred another 1 h at
80.degree. C. and stirred at 120.degree.C. for another 1 h, cooled
to room temperature, diluted with dichloromethane (DCM), washed
with brine, dried (Na.sub.2SO.sub.4), filtered and evaporated in
vacuo. Purification by flash chromatography provided
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
tert-butyl ester as a brown oil.
[0306] Step 6:
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
tert-butyl ester (977 mg) was dissolved in trifluoroacetic acid
(TFA) (10 mL) and stirred at room temperature for 1 hr. The mixture
was evaporated in vacuo to provide
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
as a brown oil, which was used without further purification.
[0307] Step 7: (Protocol A--the HBTU coupling procedure) A solution
of [3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]acetic acid
(0.30 M, 0.40 mL, 0.12 mmol) was transferred to a vial.
1-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazine dihydrochloride (48
mg, 0.14 mmol), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HBTU) (55 mg, 0.14 mmol) and i-Pr.sub.2NEt
(0.30 mL) were added to the vial and the mixture was stirred at
ambient temperature. After 30 minutes, LC/MS analysis indicated
formation of desired product and complete consumption of the
carboxylic acid starting material. The mixture was diluted with
EtOAc, washed with water (1.times.) and brine (1.times.), dried
over Na.sub.2SO.sub.4 and evaporated. The residue was purified by
silica gel chromatography (1% to 8% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[4-(4-chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol--
2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid: .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.79 (dd, 0.6 H, J=8.4, 1.6 Hz),
8.66 (dd, 0.4 H, J=8.0, 1.6 Hz), 8.57-8.55 (m, 1H), 7.29-7.26 (m,
1H), 7.22-7.16 (m, 1H), 7.09-7.05 (m, 2H), 6.50-6.45 (m, 2H), 5.45
(s, 0.6H), 5.43 (s, 1.4H), 4.07-4.01 (m, 2H), 3.81-3.69 (m, 4H),
3.17-3.13 (m, 1.6H), 3.08-3.02 (m, 2.4H), 1.50-1.42 (m, 3H); LC/MS
m/z (M+H).sup.+ 484.4.
Example 62
Synthesis of
1-{4-[4-Chloro-2-fluoro-5-(2-fluoroethoxy)-phenyl]piperazin-1-yl}-2-[3-(1-
H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00084##
[0309] The title compound was prepared following Protocol A.
1-[4-Chloro-2-fluoro-5-(2-fluoroethoxy)phenyl]-piperazine
dihydrochloride and
[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (2% to 3.5% MeOH in CH.sub.2Cl.sub.2) to
provide
1-{4-[4-chloro-2-fluoro-5-(2-fluoroethoxy)phenyl]piperazin-1-yl}-2-[3-(1H-
-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .quadrature..delta. 8.79 (dd, 0.6
H, J=8.0, 1.8 Hz), 8.67 (d, 0.4 H, J=6.4 Hz), 8.57-8.55 (m, 1H),
7.30-7.25 (m, 1H), 7.11-7.06 (m, 2H), 6.63 (d, 0.6H, J=7.6 Hz),
6.57 (d, 0.4H, J=7.6 Hz), 6.54 (d, 1H, J=7.6 Hz), 5.45 (s, 0.7H),
5.43 (s, 1.3H), 4.83-4.80 (m, 1H), 4.71-4.68 (m, 1H), 4.31-4.18 (m,
2H), 3.82-3.76 (m, 3H), 3.50 (t, 1H, J=5.2 Hz), 3.23-3.04 (m, 4H);
LC/MS m/z (M+H).sup.+ 502.4.
Example 63
Synthesis of
1-[4-(4-Chloro-3-ethoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyra-
zolo[3,4-b]pyridin-1-yl]ethanone
##STR00085##
[0311] The title compound was prepared following Protocol A.
1-(4-Chloro-3-ethoxyphenyl)piperazine dihydrochloride and
[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (4% to 15% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[4-(4-chloro-3-ethoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)pyra-
zolo[3,4-b]pyridin-1-yl]ethanone as a tan solid (25 mg): .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 8.79 (dd, 0.6H), 8.66 (dd, 0.4H),
8.57-8.54 (m, 1H), 7.29-7.19 (m, 4H), 6.49-6.40 (m, 2H), 5.45 (s,
0.7H), 5.43 (s, 1.3 H), 4.10-4.04 (m, 2H), 3.81-3.69 (m, 4H),
3.23-3.16 (m, 4H), 1.50-1.45 (m, 3H); LC/MS m/z (M+H).sup.+
466.4.
Example 64
Synthesis of
1-[4-(4-Chloro-2-fluoro-5-methoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-
-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00086##
[0313] The title compound was prepared following Protocol A.
1-(4-Chloro-2-fluoro-5-methoxyphenyl)piperazine dihydrochloride and
[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (1% to 10% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[4-(4-chloro-2-fluoro-5-methoxyphenyl)piperazin-1-yl]-2-[3-(1H-imidazol-
-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid (27 mg):
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.79 (dd, 0.6H), 8.67
(dd, 0.4H), 8.57-8.55 (m, 1H), 7.31-7.20 (m, 2H), 7.12-7.06 (m,
2H), 6.49-6.45 (m, 1H), 5.45 (s, 0.6H), 5.43 (s, 1.4H), 3.86 (s,
0.9H), 3.85 (s, 2.1 H), 3.81-3.75 (m, 4H), 3.15-3.08 (m, 4H); LC/MS
m/z (M+H).sup.+ 470.4.
Example 65
Synthesis of
1-[(S)-4-(4-Chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00087##
[0315] The title compound was prepared following Protocol A.
(S)-1-(4-Chloro-3-methoxyphenyl)-3-methylpiperazine dihydrochloride
and [3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid
were used as the coupling components. The crude product was
purified by silica gel chromatography (1% to 8% MeOH in
CH.sub.2Cl.sub.2) to provide
1-[(S)-4-(4-chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.79 (dd, 0.6H), 8.66
(dd, 0.4H), 8.57-8.55 (m, 1H), 7.29-7.19 (m, 4H), 6.44-6.39 (m,
2H), 5.42 (br. s, 2H), 4.83 (br. s, 0.3H), 4.49 (br. s, 0.3H), 4.30
(br. s, 0.3H), 3.89 (s, 1.2H), 3.88 (s, 1.8H), 3.83 (br. s, 0.3H),
3.72-3.69 (m, 1H), 3.54-3.52 (m, 1H), 3.38 (br. s, 1H), 3.19-3.15
(m, 1H), 3.00 (br. s, 1H), 2.80 (br. s, 1H), 1.50-1.43 (m, 3H);
LC/MS m/z (M+H).sup.+ 466.4.
Example 66
Synthesis of
1-[(S)-4-(4-Chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-
-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00088##
[0317] The title compound was prepared following protocol A.
(S)-1-(4-Chloro-2-fluoro-5-methoxy-phenyl)-3-methylpiperazine
dihydrochloride and
[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (2% to 3% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[(S)-4-(4-chloro-2-fluoro-5-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-
-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan
solid (29 mg): .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.79 (dd,
1H), 8.55 (dd, 1H), 7.27-7.21 (m, 3H), 7.07 (d, 1H), 6.43 (br. d,
1H), 5.46-5.37 (m, 2H), 4.83 (br. s, 0.3H), 4.51-4.48 (m, 0.6H),
4.28-4.21 (m, 0.6H), 3.86 (s, 0.9H), 3.85 (s, 2.1H), 3.79 (br. s,
0.3H), 3.67 (br. s, 0.3H), 3.33-3.21 (m, 2.5H), 2.95-2.93 (m,
0.9H), 2.83-2.76 (m, 1.6H), 1.48-1.40 (m, 3H); LC/MS m/z
(M+H).sup.+ 484.4.
Example 67
Synthesis of
1-[(R)-4-(4-Chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00089##
[0319] The title compound was prepared following Protocol A.
(R)-1-(4-Chloro-3-methoxyphenyl)-2-methylpiperazine and
[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (1% to 7.5% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[(R)-4-(4-chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.76 (d, 0.6H), 8.66 (dd,
0.3H), 8.57-8.54 (m, 1H), 7.29-7.19 (m, 4H), 6.48-6.40 (m, 2H),
5.53-5.40 (m, 2H), 4.26 (br. d, 0.6H), 4.00 (br. d, 0.6H), 3.88 (s,
1.3H), 3.86 (s, 1.7H), 3.80-3.49 (m, 3.2H), 3.33 (br. s, 0.6H),
3.17-3.14 (m, 2H), 1.51-1.42 (m, 3H); LC/MS m/z (M+H).sup.+
466.4.
Example 68
Synthesis of
1-[(S)-4-(4-Chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00090##
[0321] The title compound was prepared following protocol A.
(S)-1-(4-Chloro-3-methoxyphenyl)-2-methylpiperazine and
[3-(1H-imidazol-2-yl)pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (1% to 7% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[(S)-4-(4-chloro-3-methoxyphenyl)-3-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.77 (d, 0.6H), 8.66 (d,
0.3H), 8.57-8.54 (m, 1H), 7.30-7.19 (m, 4H), 6.48-6.40 (m, 2H),
5.54-5.36 (m, 2H), 4.25 (br. d, 0.6H), 4.00 (br. d, 0.6H), 3.88 (s,
1.3H), 3.86 (s, 1.7H), 3.82-3.48 (m, 3.2H), 3.36-3.29 (m, 0.6H),
3.17-3.13 (m, 2H), 1.51-1.43 (m, 3H); LC/MS m/z (M+H).sup.+
466.4.
Example 69
Synthesis of
1-[(R)-4-(4-Chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00091##
[0323] The title compound was prepared following Protocol A.
(R)-1-(4-Chloro-3-methoxyphenyl)-3-methylpiperazine and
[3-(1H-imidazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]acetic acid were
used as the coupling components. The crude product was purified by
silica gel chromatography (1% to 7.5% MeOH in CH.sub.2Cl.sub.2) to
provide
1-[(R)-4-(4-chloro-3-methoxyphenyl)-2-methylpiperazin-1-yl]-2-[3-(1H-imid-
azol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as a tan solid:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. .quadrature.8.75 (d,
0.6H), 8.66 (dd, 0.4H), 8.57-8.54 (m, 1H), 7.29-7.18 (m, 4H),
6.44-6.39 (m, 2H), 5.42 (br. s, 2H), 4.82 (br. s, 0.3H), 4.45 (br.
s, 0.3H), 4.33 (br. s, 0.3H), 3.88 (s, 1.2H), 3.87 (s, 1.8H), 3.83
(br. s, 0.3H), 3.73-3.67 (m, 1H), 3.54-3.52 (m, 1H), 3.38 (br. s,
1H), 3.17-3.13 (m, 1H), 2.99 (br. s, 1H), 2.80 (br. s, 1H),
1.50-1.42 (m, 3H); LC/MS m/z (M+H).sup.+ 466.4.
Example 70
Synthesis of
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-((S)-4-(4-chlor-
o-3-ethoxyphenyl)-2-methylpiperazin-1-yl)ethanone
##STR00092##
[0325] The title compound was prepared following Protocol A. To a
vial containing (S)-1-(4-chloro-3-ethoxyphenyl)-3-methylpiperazine
dihydrochloride (70 mg, 0.21 mmol) was added
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
(51 mg, 0.21 mmol), HBTU (81 mg, 0.21 mmol), DMF (0.7 mL), and
DIPEA (0.15 mL, 0.87 mmol). The reaction mixture was maintained at
30.degree. C. for 24 h. The solution was diluted with EtOAc (30 mL)
and washed with 1N HCl (2.times.10 mL) and sat. aq. NaCl
(2.times.10 mL). The organic phase was dried over MgSO.sub.4 and
concentrated in vacuo. The resulting residue was purified by
preparative HPLC (20.fwdarw.95% gradient of MeCN--H.sub.2O with
0.1% TFA) and the pure fractions lyophilized to afford the
indicated compound (11 mg, 11% yield): MS (ES) [M+H].sup.+ expected
480.2, found 480.5; .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.94
(br s, 1H), 8.79 (dd, J=1.6, 8.0, 1H), 8.55 (dd, J=1.6, 4.4, 1H),
7.19-7.26 (m, 4H), 6.37-6.43 (m, 2H), 5.40 (br s, 2H), 2.78-4.81
(m, 10H), 4.07 (q, J=6.8, 2H), 1.46 (t, J=6.8, 3H).
Example 71
Synthesis of
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(3-(2-fluoro-
ethoxy)-4-chlorophenyl)piperazin-1-yl)ethanone
##STR00093##
[0327] This compound was prepared by Protocol A. To a vial
containing 1-(3-(2-fluoroethoxy)-4-chlorophenyl)piperazine
dihydrochloride (70 mg, 0.21 mmol) was added
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
(51 mg, 0.21 mmol), HBTU (83 mg, 0.22 mmol), DMF (0.7 mL), and
DIPEA (0.20 mL, 1.2 mmol). The reaction mixture was maintained at
20.degree. C. for 24 h. The solution was diluted with EtOAc (30 mL)
and washed with 1N HCl (2.times.10 mL) and sat. aq. NaCl
(2.times.10 mL). The organic phase was dried over MgSO.sub.4 and
concentrated in vacuo. The resultant residue was purified by
preparative HPLC (20.fwdarw.95% gradient of MeCN--H.sub.2O with
0.1% TFA) and the pure fractions lyophilized to afford the
indicated compound (20 mg, 20% yield): MS (ES) [M+H].sup.+ expected
484.2, found 484.4. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.94
(br s, 1H), 8.78 (dd, J=1.2, 8, 1H), 8.55 (dd, J=1.2, 4.6, 1H),
7.16-7.26 (m, 4H), 6.45-6.53 (m, 2H), 5.44 (s, 2H), 4.77 (dt,
J=4.0, 46.8, 2H), 4.26 (dt, J=4.0, 26.8, 2H), 3.69-3.78 (m, 4H),
3.10-3.21 (m, 4H).
Example 72
Synthesis of
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-((S)-4-(3-(2-fl-
uoroethoxy)-4-chlorophenyl)-2-methylpiperazin-1-yl)ethanone
##STR00094##
[0329] This compound was prepared by Protocol A. To a vial
containing
(S)-1-(3-(2-fluoroethoxy)-4-chlorophenyl)-3-methylpiperazine
dihydrochloride (80 mg, 0.23 mmol) was added
2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic acid
(51 mg, 0.21 mmol), HBTU (81 mg, 0.21 mmol), DMF (0.7 mL), and
DIPEA (0.2 mL, 1.2 mmol). The reaction mixture was maintained at
30.degree. C. for 24 h. The solution was diluted with EtOAc (30 mL)
and washed with 1N HCl (2.times.10 mL) and sat. aq. NaCl
(2.times.10 mL). The organic phase was dried over MgSO.sub.4 and
concentrated in vacuo. The resultant residue was purified by
preparative HPLC (20.fwdarw.95% gradient of MeCN--H.sub.2O with
0.1% TFA) and the pure fractions lyophilized to afford the
indicated compound (14 mg, 13% yield): MS (ES) [M+H].sup.+ expected
498.2, found 498.4. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.95
(br s, 1H), 8.79 (dd, J=1.6, 8.2, 1H), 8.55 (dd, J=1.6, 4.4, 1H),
7.19-7.26 (m, 4H), 6.44-6.50 (m, 2H), 5.40 (br s, 2H), 4.77 (dt,
J=4.2, 47.2, 2H), 4.26 (dt, J=4.2, 27.2, 2H), 2.78-4.42 (m,
10H).
Example 73
Synthesis of
1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-p-
yrazolo[3,4-b]pyridin-1-yl]-ethanone
##STR00095##
[0331] This compound was prepared by Protocol A. A vial was charged
with 2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic
acid (55 mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),
1-(4-Chloro-3-methyl-phenyl)-piperazine dihydrochloride (142 mg,
0.50 mmol), anhydrous DMF (2.0 mL), and DIPEA (0.5 mL). The vial
was capped, heated to 45.degree. C., and stirred overnight. The
following day, the volatiles were removed in vacuo and separation
by preparative hplc (reverse phase, acetonitrile-water gradient)
gave
1-[4-(4-Chloro-3-methyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)-p-
yrazolo[3,4-b]pyridin-1-yl]-ethanone: MS (ES) [M+H].sup.+ found:
436.4
Example 74
Synthesis of
1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazo-
l-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone
##STR00096##
[0333] This compound was prepared by Protocol A. A vial was charged
with 2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic
acid (55 mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),
1-(4-Chloro-3-trifluoromethyl-phenyl)-piperazine dihydrochloride
(170 mg, 0.50 mmol), anhydrous DMF (2.0 mL), and DIPEA (0.5 mL).
The vial was capped, heated to 45.degree. C., and stirred
overnight. The following day, the volatiles were removed in vacuo
and separation by preparative hplc (reverse phase,
acetonitrile-water gradient) gave
1-[4-(4-Chloro-3-trifluoromethyl-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazo-
l-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone: MS (ES) [M+H].sup.+
found: 490.4
Example 75
Synthesis of
1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidaz-
ol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone
##STR00097##
[0335] This compound was prepared by Protocol A. A vial was charged
with 2-(3-(1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)acetic
acid (55 mg, 0.226 mmol), HBTU (125 mg, 0.33 mmol),
1-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazine dihydrochloride
(177 mg, 0.50 mmol), anhydrous DMF (2.0 mL), and DIPEA (0.4 mL).
The vial was capped, heated to 45.degree. C., and stirred
overnight. The following day, the volatiles were removed in vacuo
and separation by preparative hplc (reverse phase,
acetonitrile-water gradient) gave
1-[4-(4-Chloro-3-trifluoromethoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidaz-
ol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone. MS (ES) [M+H].sup.+
found: 506.4
Example 76
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)--
pyrazolo[3,4-b]pyridin-1-yl]-ethanone
##STR00098##
[0337] The title compound was prepared from
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-iodo-pyrazolo[3,4-b-
]pyridin-1-yl]-ethanone according to the procedure similar to those
described from step 2 to step 5 in the synthesis of Example 61:
.sup.1H NMR (CDCl.sub.3, 400 MHz) .quadrature..delta. 10.22 (br,
1H), 8.82 (dd, 1H), 8.56 (dd, 1H), 7.20-7.30 (m, 3H), 7.11 (s, 1H),
6.47 (d, 1H), 6.42 (dd, 1H), 5.44 (s, 2H), 3.88 (s, 3H), 3.80 (m,
4H), 3.19 (m, 4H); MS (ES) M+H expect 452.2.
Example 77
Synthesis of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imidaz-
ol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone
##STR00099##
[0339] To the solution of
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1H-imidazol-2-yl)--
pyrazolo[3,4-b]pyridin-1-yl]-ethanone (50 mg, 0.11 mmol, 1 eq) in
THF was added 60% sodium hydride (5.7 mg, 0.14 mmol, 1.3 eq) and
stirred for 1 hr, followed by the addition of iodomethane (25.8 mg,
0.16 mmol, 1.5 eq). 2 hrs later, LCMS indicated that the major peak
is the desired product. preparative hplc (reverse phase,
acetonitrile-water gradient) gave
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-[3-(1-methyl-1H-imidaz-
ol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]-ethanone. MS (ES) [M+H].sup.+
found: 465.2.
Example 78
##STR00100##
[0340] Step 1: Tert-butyl
4-(4-chloro-3-methoxyphenyl)piperazine-1-carboxylate
##STR00101##
[0342] To a 3-necked, 5-L Morton flask equipped with a mechanical
stirrer, gas adapter, heating mantle and thermometer was added
rac-BINAP (4.24 g, 0.005 equiv) and Pd.sub.2(dba).sub.3 (3.20 g,
0.0025 equiv). The flask was evacuated, and back-filled with
nitrogen. Toluene (100 mL) was added by cannula. The mixture was
stirred at room temperature for 15 min to give a purple solution.
Toluene (2.0 L) was then added. 2-Chloro-5-bromoanisole (300.3 g,
1.356 mol, 1 equiv) was added in one portion. Boc-piperazine (252.4
g, 1 equiv) was added in one portion. Sodium tert-butoxide (183.0
g, 1.4 equiv) was added in one portion. The flask was evacuated and
back-filled with nitrogen. The mixture was then heated to an
internal temperature of 60.degree. C. A heterogeneous light-orange
slurry was obtained. After 1 h, the mixture becomes a homogeneous
brown solution. After an additional 15 h, the mixture was cooled to
room temperature. EtOAc (2.0 L) was added to the stirring mixture.
The solid was filtered. The filtrate was washed with EtOAc (100
mL). The combined filtrate was washed with 10% aq. K.sub.2CO.sub.3
solution (1.times.1 L), water (1.times.1 L), and dried over
MgSO.sub.4. The solvent was removed in vacuo to afford the product
as an orange solid (410.3 g, 93% yield).
Step 2: 1-(4-chloro-3-methoxyphenyl)piperazine dihydrochloride
##STR00102##
[0344] A 4-L beaker equipped with a mechanical stirrer was charged
with tert-butyl
4-(4-chloro-3-methoxyphenyl)piperazine-1-carboxylate (500 g, 1.53
mol, 1 equiv) and MeOH (1.50 L). While stirring at room temperature
conc. 37% HCl (500 mL, 4 equiv) was added over 5 min. The internal
temperature rose to 40.degree. C., and the solution became thick
with precipitate. After 15 min, the mixture was heated to an
internal temperature of 60.degree. C. on a hotplate. (Foaming begun
at approximately 50.degree. C. as the mixture warms.) After 2 h at
60.degree. C., the solution was cooled to room temperature, and
subsequently to 5.degree. C. in a refrigerator. The product was
collected by filtration in two batches. Each batch of the red-brown
filtrate was washed with EtOAc (2.times.500 mL) to give a light
yellow solid. The two batches were combined to afford the product
(391.3 g). The filtrate was concentrated to a volume of 300 mL in
vacuo and treated with hot (50.degree. C.) MeOH (500 mL). The
mixture was cooled to 5.degree. C. in a refrigerator for 24 h. The
resulting precipitate was collected by filtration and washed with
EtOAc (2.times.200 mL) to afford an additional 44.3g of product
(total of 435.6 g, 95% yield).
Step 3:
2-chloro-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanone
##STR00103##
[0346] To a 3 L flask equipped with a mechanical stirrer was added
1-(4-chloro-3-methoxyphenyl)piperazine dihydrochloride (220 g, 0.73
mol, 1 equiv), CH.sub.2Cl.sub.2 (1000 mL), and water (1000 mL). The
biphasic mixture was cooled to 5.degree. C. with an ice-water bath.
K.sub.2CO.sub.3 (506 g, 5 equiv) was added to the vigorously
stirring solution in portions to minimize foaming. A solution of
chloroacetyl chloride (124.4 g, 1.5 equiv) in CH.sub.2Cl.sub.2 (100
mL) was added dropwise from an addition funnel, while maintaining
an internal temperature below 8.degree. C. After 1 h, the cooling
bath was removed, and the reaction warmed to room temperature.
After an additional 1 h, the layers were partitioned. The aqueous
phase was extracted with CH.sub.2Cl.sub.2 (2.times.300 mL), and the
combined organic layers dried over 3:1
Na.sub.2SO.sub.4/K.sub.2CO.sub.3 (addition of K.sub.2CO.sub.3 helps
the solution phase to become clear). After filtration, the filtrate
was concentrated in vacuo, and the residue was dried for 16 h under
vacuum to afford the product as an off-white solid (410 g, 92%
yield).
Step 6: 7-azaindazole-3-carboxaldehyde
##STR00104##
[0348] A 5L 3-necked flask equipped with a digital thermometer, a 1
L addition funnel and mechanical stirrer (all glassware dried in
oven and cooled in air for 30 min before use) was charged with
3-iodo-7-azaindaozle (196.0 g, 0.80 mol) and 1 L of anhydrous THF
(in SureSeal bottle from Aldrich and used as is). The solids
completely dissolved in THF at room temperature to form a dark
brown solution. The flask was then cooled to -5.degree. C. with an
ice/NaCl bath and moderate stirring and o-tolylmagnesium chloride
(1 M solution in THF, 880 mL, 1.1 equiv) was added dropwise to keep
the internal temperature between -5.degree. C. to -3.degree. C.
(after .about.820 mL of o-tolylmagnesium chloride solution was
added, the temperature no longer rose). The whole addition process
took 2 hr and 25 min. At the end of the addition, the mixture was a
homogeneous dark brown solution.
[0349] After an additional 1 hr, isopropylmagnesium chloride
solution (2 M in THF, 480 mL, 1.2 equiv) was added dropwise to keep
internal temperature <4.degree. C. After 25 min and about 200 mL
isopropylmagnesium chloride solution was added, brown precipitate
started to form. After a total of 380 mL of isopropylmagnesium
chloride solution was added, the mixture became homogeneous again.
The whole addition process was done in 45 min. After another 1 hr
25 min, a small amount of sample was taken out and quenched with
D.sub.2O. LCMS analysis of this sample indicated the complete
Iodo-Mg exchange.
[0350] 1-Formylpiperdine (120 mL, 1.3 equiv) was then added
dropwise to keep the internal temperature between <2.degree. C.
After about 30 mL 1-formylpiperidine was added, the internal
temperature did not go up anymore and rest of the
1-formylpiperidine was added relatively quickly. The whole addition
process took 20 min. At the end of the addition, the mixture was
still a dark homogeneous solution and was allowed to slowly warm up
to room temperature and moderately stirred for 18 hr.
[0351] The mixture was re-cooled to 0.degree. C. with an ice/NaCl
bath and quenched by slow addition of a mixture of saturated
NH.sub.4Cl solution (750 mL)/concentrated HCl solution (250 mL) to
keep the internal temperature at <35.degree. C. After the
addition was complete, stirring was allowed to continue for 1 hr
and a yellow precipitate appeared. The mixture was filtered and the
solid was washed with THF (100 mL). The collected filtrate was
transferred to a separation funnel and the pH of the aqueous layer
was adjusted to between 5 and 6 with the addition of NaHCO.sub.3
(around 5 g). The THF layer was separated and washed with sat. NaCl
solution (2.times.100 mL). The combined aqueous layers (including
the NaCl wash and quenched aqueous layer) was extracted with EtOAc
(3.times.250 mL). The combined organic layers was dried
(Na.sub.2SO.sub.4), filtered and evaporated in vacuo (bath
temperature <30.degree. C.) to give a brownish solid. This solid
was triturated with Et.sub.2O (600 mL) and filtered. The collected
solid was washed with Et.sub.2O (2.times.100 mL) to give
7-azaindazole-3-carboxaldehyde as a yellowish solid (86.6 g,
73%).
Step 4:
1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl-
-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00105##
[0353] A mixture of 7-azaindazole-3-carboxaldehyde (86.6 g, 0.59
mol, 1 equiv), NaI (8.8 g, 0.1 equiv) and K.sub.2CO.sub.3 (162.5 g,
2 equiv) in DMF (0.5 L) in a 5 L flask was heated to 85.degree. C.
(the heating process took around 1.5 h).
2-chloro-1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)ethanone
(175 g, 1 equiv) was added in small portions to the reaction
mixture. The whole addition process took about 30 min. The mixture
was then stirred at 85.degree. C. for 30 min and LCMS confirmed
that the reaction was complete. After cooling down to room
temperature, the mixture was transferred to a 4 L flask with 2 L
ice. The reaction flask was rinsed with small amount of acetone (30
mL) and transferred to the DMF/ice mixture in the 4 L flask also. A
lot of brownish solids precipitated out. After the ice completely
melted, the mixture was filtered. The collected solid was washed
with water (1 L), blended, and then washed with water (1 L) to get
rid of some residual DMF. The collected solid contained a lot of
water, so it was dissolved in CH.sub.2Cl.sub.2 (4 L) and the
mixture was transferred to a 5 L separation funnel. The bottom
CH.sub.2Cl.sub.2 layer was separated and the top aq. layer was
washed with CH.sub.2Cl.sub.2 (2.times.100 mL). The combined
CH.sub.2Cl.sub.2 layers were dried (Na.sub.2SO.sub.4), filtered and
evaporated in vacuo to give
1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl-pyrazo-
lo[3,4-b]pyridin-1-yl]ethanone as a brownish solid (236.4 g, 97%)
which was used without purification.
Step 5:
1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-im-
idazol-2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone
##STR00106##
[0355]
1-[4-(4-Chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-formyl--
pyrazolo[3,4-b]pyridin-1-yl]ethanone_(300 g, 723 mmol, 1 equiv),
glyoxal trimer dihydrate (60.6 g, 0.4 equiv), and ammonium acetate
(222.9 g, 4 equiv) were suspended in a mixture of THF (720 mL) and
MeOH (720 mL) in a 5 L round-bottomed flask fitted with a magnetic
stir bar and nitrogen inlet. Acetic acid (84 mL, 2 equiv) was added
and the mixture was heated in a 45.degree. C. oil bath (solids
dissolved upon heating). After 12 hours, LC/MS analysis indicated
complete consumption of aldehyde starting material and formation of
desired product (LC/MS m/z (M+H).sup.+ 452.1). The MeOH/THF were
removed by rotary evaporation. The residue was dissolved in 10%
MeOH in CH.sub.2Cl.sub.2 (ca. 1.5 L) and the mixture was shaken
vigorously with aqueous potassium carbonate (ca. 210 g potassium
carbonate in ca. 1.5 L water, pH of aqueous=8-9). The layers were
separated, and the aqueous layer was extracted with 10% MeOH in
CH.sub.2Cl.sub.2 (2.times.100 mL). The combined organic layers were
concentrated to give a brown oily solid. The crude product was
suspended in 10% MeOH in EtOAc (ca. 1 L). Anhydrous
Na.sub.2SO.sub.4 (ca. 60 g) and silica gel (ca. 100 g) were added
and the slurry was heated gently with a heat gun to dissolve the
crude product. The slurry was transferred to a 2 L fritted glass
filter funnel containing silica gel (ca. 100 g, pre-equilibrated
with 10% MeOH in EtOAc), and the product was eluted through the
silica gel plug with 10% MeOH in EtOAc (ca. 6 L) and 1% Et.sub.3N,
10% MeOH in EtOAc (ca. 10 L). (Note: Incomplete dissolution of the
product and/or precipitation of product in the presence of silica
gel complicated the filtration). The solvents were removed by
rotary evaporation. The residue was triturated with MeCN
(1.times.300 mL) and dried (rotary evaporation followed by high
vacuum) to provide
1-[4-(4-chloro-5-ethoxy-2-fluorophenyl)piperazin-1-yl]-2-[3-(1H-imidazol--
2-yl)-pyrazolo[3,4-b]pyridin-1-yl]ethanone as an off-white solid
(190 g, 58%, LC/MS purity >98%).
Step 7:
2-(3-1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(4-chl-
oro-3-methoxyphenyl)piperazin-1-yl)ethanone hydrochloride salt
##STR00107##
[0357] A 2 L flask with a magnetic stirrer was charged with the
product of step 6 (5.1 g, 11.28 mmol) and EtOAc (900 mL). The
resulting suspension was heated to form a clear solution and cooled
to room temperature under moderate stirring. HCl in Et.sub.2O (2M,
6.2 mL, 12.42 mmol) was added dropwise to the resulting solution at
room temperature over 5 min. After the addition, the resultant
suspension was stirred at room temperature for another 1 h. The
solid was collected by filtration and washed with Et.sub.2O (150
mL.times.2) and dried in vacuo to afford 5.4 g of an off-white
powder. A 250 mL flask with a magnetic stirrer was charged with the
powder obtained above (5.4 g), acetone (100 mL) and deionized water
(16 mL). The resulting suspension was heated to form a clear
solution and stirred to cool. When the solution became cloudy
(crystal seeds appeared), acetone (540 mL) was added slowly to the
suspension over 20 min. The resulting suspension was heated to
50.degree. C. and stirred for 2 h. Filtration while hot, washing
with hot acetone (50 mL.times.2) and drying in vacuo gave 3.3 g
(60%) of the product as an off-white solid: m. p. 164-165.degree.
C. The crystals appear as prisms under a polarizing microscope.
Example 79
##STR00108##
[0358] Step 1: Synthesis of
1-{2-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-1H-pyraz-
olo[3,4-b]pyridine-3-carbonitrile
##STR00109##
[0360] To a 2000 ml flask was charged with
1-[4-(4-Chloro-3-methoxy-phenyl)-piperazin-1-yl]-2-(3-iodo-pyrazolo[3,4-b-
]pyridin-1-yl)-ethanone(40 g, 78.1 mmol), dppf (3.86 g, 6.96 mmol),
Zn(CN).sub.2 (9.6 g, 81.6 mmol), DMF (360 ml) and H.sub.2O (20 ml).
The resulting suspension was degassed using N.sub.2 for 5 min,
followed by addition of Pd.sub.2(dba).sub.3 (4.24 g, 4.64 mmol).
The reaction mixture was heated under N.sub.2 at 90.degree. C. for
2 h (monitor by TLC and LC-MS). After cooling to room temperature,
diluted with EtOAc (1500 ml), filtered to remove the precipitate
and washed with H.sub.2O (1000.times.2 ml), saturated EDTA.4Na (800
ml.times.2), brine and dried over Na.sub.2SO.sub.4. After
evaporation of the solvent, ether (150 mL) was added and stirred
for 2 h. The resulting solid was filtered to give the desired
product 30 g (93%) as light yellow powder. Recrystallization from
refluxing CH.sub.3CN (160 mL) afforded 26 g (80%) light yellow
crystals: mp 183-185.degree. C.; R.sub.t=2.38 min; MS (ES) M+H
expect 411.1, found 411.1.
Step 2: Synthesis of
1-(4-(4-chloro-3-methoxyphenyl)piperazin-1-yl)-2-(3-(4,5-dihydro-1H-imida-
zol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)ethanone
##STR00110##
[0362] A 250 mL flask was charged with
1-(2-(4-chloro-3-methoxyphenyl)piperazin-1-yl)-2-oxoethyl)-1H-pyrazol[3,4-
-b]pyridine-3-carbonitrile (15.3 g, 37.2 mmol), EtOH (40 mL,
.about.1 M). Under ice-bath and stirring, AcOH (6.75 mL, 112 mmol)
was added, followed by ethylenediamine (25 mL, 372 mmol). The
resulting mixture was heated at 120.degree. C. (bath) under N.sub.2
(observed mixture starting refluxing) for 1.5 h. TLC and LC-MS
indicated the disappearance of starting material and formation of
imidazoline. After cooling to room temperature, mixture was diluted
with DCM (700 mL) and washed with H.sub.2O (350 mL). The H.sub.2O
layer was back extracted with DCM (150 mL), and the combined
organic layer was washed brine (350 mL) and dried over MgSO.sub.4.
After evaporating of the solvent under reduced pressure, the
residue was suspended in hot EtOAc (80 mL). After cooling to room
temperature, the solid was collected by filtration and washed with
EtOAc (30 mL) to afford the title compound as white powders (16 g,
95%) which was used directly for next step: mp 133-135.degree. C.;
R.sub.t=1.369 min. MS (ES) M+H expect 454.2, found 454.4.
Step 3:
2-(3-1H-imidazol-2-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-1-(4-(4-chl-
oro-3-methoxyphenyl)piperazin-1-yl)ethanone
##STR00111##
[0364] The above imidazoline (12.3 g, 27.1 mmol) in a 500 mL flask
was charged with anhydrous DMSO (108 mL, .about.0.25 M). DMP (17.2
g, 40.6 mmol) was added in portions under stirring. The resulting
mixture was stirred at 45.degree. C. under N.sub.2 for 2 h (monitor
by TLC and LC-MS). After cooling to room temperature, quenching the
reaction with sat Na.sub.2S.sub.2O.sub.3 (100 mL) (ice-bath),
followed 3 N NaOH (100 mL) (pH 12 to 13) and H.sub.2O (300 mL) and
extracted with DCM (600 mL+300 mL). The combined organic layer was
washed with sat NaHCO.sub.3 (300 mL), brine (300 mL) and dried
(MgSO.sub.4, 120 g). After evaporation of the organic solvent, the
residue yellow solid (.about.11 g) was dissolved in hot CH.sub.3CN
(20 mL). After cooling to room temperature, the resulting solid was
collected by filtration to afford 6.7 g (55%) of title compound as
light tan crystals : mp 149-152.degree. C.; R.sub.t=1.309 min. MS
(ES) M+H expect 452.2, found 452.4. Mother liquor was concentrated
and afforded another 0.6 g (total isolated yield 60%).
Example 80
[0365] This example illustrates the evaluation of the biological
activity associated with compounds of interest (candidate
compounds) of the invention.
Materials and Methods
[0366] A. Cells
[0367] 1. CCR1 Expressing Cells
[0368] a) THP-1 Cells
[0369] THP-1 cells were obtained from ATCC (TIB-202) and cultured
as a suspension in RPMI-1640 medium supplemented with 2 mM
L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM
HEPES, 1 mM sodium pyruvate, 0.05% 2-mercaptoethanol and 10% FBS.
Cells were grown under 5% CO.sub.2/95% air, 100% humidity at
37.degree. C. and subcultured twice weekly at 1:5 (cells were
cultured at a density range of 2.times.10.sup.5 to 2.times.10.sup.6
cells/mL) and harvested at 1.times.10.sup.6 cells/mL. THP-1 cells
express CCR1 and can be used in CCR1 binding and functional
assays.
[0370] b) Isolated Human Monocytes
[0371] Monocytes were isolated from human buffy coats using the
Miltenyi bead isolation system (Miltenyi, Auburn, Calif.). Briefly,
following a Ficoll gradient separation to isolate peripheral blood
mononuclear cells, cells were washed with PBS and the red blood
cells lysed using standard procedures. Remaining cells were labeled
with anti-CD14 antibodies coupled to magnetic beads (Miltenyi
Biotech, Auburn, Calif.). Labeled cells were passed through
AutoMACS (Miltenyi, Auburn, Calif.) and positive fraction
collected. Monocytes express CCR1 and can be used in CCR1 binding
and functional assays.
[0372] B. Assays
[0373] 1. Inhibition of CCR1 Ligand Binding
[0374] CCR1 expressing cells were centrifuged and resuspended in
assay buffer (20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl.sub.2, 5
mM MgCl.sub.2, and with 0.2% bovine serum albumin) to a
concentration of 5.times.10.sup.6 cells/mL for THP-1 cells and
5.times.10.sup.5 for monocytes. Binding assays were set up as
follows. 0.1 mL of cells (5.times.10.sup.5 THP-1 cells/well or
5.times.10.sup.4 monocytes) was added to the assay plates
containing the compounds, giving a final concentration of
.about.2-10 .mu.M each compound for screening (or part of a dose
response for compound IC.sub.50 determinations). Then 0.1 mL of
.sup.125I labeled MIP-1.alpha. (obtained from Perkin Elmer Life
Sciences, Boston, Mass.) or 0.1 mL of .sup.125I labeled
CCL15/leukotactin (obtained as a custom radiolabeling by Perkin
Elmer Life Sciences, Boston, Mass.) diluted in assay buffer to a
final concentration of .about.50 .mu.M, yielding .about.30,000 cpm
per well, was added (using .sup.125I labeled MIP-1.alpha. with
THP-1 cells and .sup.125I labeled CCL15/leukotactin with
monocytes), the plates sealed and incubated for approximately 3
hours at 4.degree. C. on a shaker platform. Reactions were
aspirated onto GF/B glass filters pre-soaked in 0.3%
polyethyleneimine (PEI) solution, on a vacuum cell harvester
(Packard Instruments; Meriden, Conn.). Scintillation fluid (40
.mu.I; Microscint 20, Packard Instruments) was added to each well,
the plates were sealed and radioactivity measured in a Topcount
scintillation counter (Packard Instruments). Control wells
containing either diluent only (for total counts) or excess
MIP-1.alpha. or MIP-1.beta. (1 .mu.g/mL, for non-specific binding)
were used to calculate the percent of total inhibition for
compound. The computer program Prism from GraphPad, Inc. (San
Diego, Calif.) was used to calculate IC.sub.50 values. IC.sub.50
values are those concentrations required to reduce the binding of
labeled MIP-1.alpha. to the receptor by 50%. . (For further
descriptions of ligand binding and other functional assays, see
Dairaghi, et al., J. Biol. Chem. 274:21569-21574 (1999), Penfold,
et al., Proc. Natl. Acad. Sci. USA. 96:9839-9844 (1999), and
Dairaghi, et al,. J. Biol. Chem. 272:28206-28209 (1997)).
[0375] 2. Calcium Mobilization
[0376] To detect the release of intracellular stores of calcium,
cells (THP-1 or monocytes) were incubated with 3 .mu.M of INDO-1AM
dye (Molecular Probes; Eugene, Oreg.) in cell media for 45 minutes
at room temperature and washed with phosphate buffered saline
(PBS). After INDO-1AM loading, the cells were resuspended in flux
buffer (Hank's balanced salt solution (HBSS) and 1% FBS). Calcium
mobilization was measured using a Photon Technology International
spectrophotometer (Photon Technology International; New Jersey)
with excitation at 350 nm and dual simultaneous recording of
fluorescence emission at 400 nm and 490 nm. Relative intracellular
calcium levels were expressed as the 400 nm/490 nm emission ratio.
Experiments were performed at 37.degree. C. with constant mixing in
cuvettes each containing 10.sup.6 cells in 2 mL of flux buffer. The
chemokine ligands may be used over a range from 1 to 100 nM. The
emission ratio was plotted over time (typically 2-3 minutes).
Candidate ligand blocking compounds (up to 10 .mu.M) were added at
10 seconds, followed by chemokines at 60 seconds (i.e.,
MIP-1.alpha.; R&D Systems; Minneapolis, Minn.) and control
chemokine (i.e., SDF-1.alpha.; R&D Systems; Minneapolis, Minn.)
at 150 seconds.
[0377] 3. Chemotaxis Assays
[0378] Chemotaxis assays were performed using 5 m pore
polycarbonate, polyvinylpyrrolidone-coated filters in 96-well
chemotaxis chambers (Neuroprobe; Gaithersburg, Md.) using
chemotaxis buffer (Hank's balanced salt solution (HBSS) and 1%
FBS). CCR1 chemokine ligands (i.e., MIP-1.alpha.,
CCL15/Leukotactin; R&D Systems; Minneapolis, Minn.) are use to
evaluate compound mediated inhibition of CCR1 mediated migration.
Other chemokines (i.e., SDF-1a; R&D Systems; Minneapolis,
Minn.) are used as specificity controls. The lower chamber was
loaded with 29 .mu.l of chemokine (i.e., 0.1 nM CCL15/Leukotactin)
and varying amounts of compound; the top chamber contained 100,000
THP-1 or monocyte cells in 20 .mu.l. The chambers were incubated
1-2 hours at 37.degree. C., and the number of cells in the lower
chamber quantified either by direct cell counts in five high
powered fields per well or by the CyQuant assay (Molecular Probes),
a fluorescent dye method that measures nucleic acid content and
microscopic observation.
[0379] C. Identification of Inhibitors of CCR1
[0380] 1. Assay
[0381] To evaluate small organic molecules that prevent the
receptor CCR1 from binding ligand, an assay was employed that
detected radioactive ligand (i.e, MIP-1.alpha. or
CCL15/Leukotactin) binding to cells expressing CCR1 on the cell
surface (for example, THP-1 cells or isolated human monocytes). For
compounds that inhibited binding, whether competitive or not, fewer
radioactive counts are observed when compared to uninhibited
controls.
[0382] THP-1 cells and monocytes lack other chemokine receptors
that bind the same set of chemokine ligands as CCR1 (i.e.,
MIP-1.alpha., MPIF-1, Leukotactin, etc.). Equal numbers of cells
were added to each well in the plate. The cells were then incubated
with radiolabeled MIP-1.alpha.. Unbound ligand was removed by
washing the cells, and bound ligand was determined by quantifying
radioactive counts. Cells that were incubated without any organic
compound gave total counts; non-specific binding was determined by
incubating the cells with unlabeled ligand and labeled ligand.
Percent inhibition was determined by the equation:
% inhibition=(1-[(sample cpm)-(nonspecific cpm)]/[(total
cpm)-(nonspecific cpm)]).times.100.
[0383] 2. Dose Response Curves
[0384] To ascertain a candidate compound's affinity for CCR1 as
well as confirm its ability to inhibit ligand binding, inhibitory
activity was titered over a 1.times.10.sup.-10 to 1.times.10.sup.4
M range of compound concentrations. In the assay, the amount of
compound was varied; while cell number and ligand concentration
were held constant.
[0385] 3. CCR1 Functional Assays
[0386] CCR1 is a seven transmembrane, G-protein linked receptor. A
hallmark of signaling cascades induced by the ligation of some such
receptors is the pulse-like release of calcium ions from
intracellular stores. Calcium mobilization assays were performed to
determine if the candidate CCR1 inhibitory compounds were able to
also block aspects of CCR1 signaling. Candidate compounds able to
inhibit ligand binding and signaling with an enhanced specificity
over other chemokine and non-chemokine receptors were desired.
[0387] Calcium ion release in response to CCR1 chemokine ligands
(i.e., MIP-1.alpha., MPIF-1, Leukotactin, etc.) was measured using
the calcium indicator INDO-1. THP-1 cells or monocytes were loaded
with INDO-1/AM and assayed for calcium release in response to CCR1
chemokine ligand (i.e., MIP-1.alpha.) addition. To control for
specificity, non-CCR1 ligands, specifically bradykinin, was added,
which also signals via a seven transmembrane receptor. Without
compound, a pulse of fluorescent signal will be seen upon
MIP-1.alpha. addition. If a compound specifically inhibits
CCR1-MIP-1.alpha. signaling, then little or no signal pulse will be
seen upon MIP-1.alpha. addition, but a pulse will be observed upon
bradykinin addition. However, if a compound non-specifically
inhibits signaling, then no pulse will be seen upon both
MIP-1.alpha. and bradykinin addition.
[0388] One of the primary functions of chemokines is their ability
to mediate the migration of chemokine receptor-expressing cells,
such as white blood cells. To confirm that a candidate compound
inhibited not only CCR1 specific binding and signaling (at least as
determined by calcium mobilization assays), but also CCR1 mediated
migration, a chemotaxis assay was employed. THP-1 myelomonocytic
leukemia cells, which resemble monocytes, as wells as freshly
isolated monocytes, were used as targets for chemoattraction by
CCR1 chemokine ligands (i.e., MIP-1.alpha., CCL15/leukotactin).
Cells were place in the top compartment of a microwell migration
chamber, while MIP-1.alpha. (or other potent CCR1 chemokine ligand)
and increasing concentrations of a candidate compound was loaded in
the lower chamber. In the absence of inhibitor, cells will migrate
to the lower chamber in response to the chemokine agonist; if a
compound inhibited CCR1 function, then the majority of cells will
remain in the upper chamber. To ascertain a candidate compound's
affinity for CCR1 as well as to confirm its ability to inhibit CCR1
mediated cell migration, inhibitory activity was titered over a
1.times.10.sup.-10 to 1.times.10.sup.-4 M range of compound
concentrations in this chemotaxis assay. In this assay, the amount
of compound was varied; while cell number and chemokine agonist
concentrations were held constant. After the chemotaxis chambers
were incubated 1-2 hours at 37.degree. C., the responding cells in
the lower chamber were quantified by labeling with the CyQuant
assay (Molecular Probes), a fluorescent dye method that measures
nucleic acid content, and by measuring with a Spectrafluor Plus
(Tecan). The computer program Prism from GraphPad, Inc. (San Diego,
Calif.) was used to calculate IC.sub.50 values. IC.sub.50 values
are those compound concentrations required to inhibit the number of
cells responding to a CCR1 agonist by 50%.
[0389] 4. In Vivo Efficacy
[0390] a) Rabbit Model of Destructive Joint Inflammation
[0391] To study the effects of candidate compounds on inhibiting
the inflammatory response of rabbits to an intra-articular
injection of the bacterial membrane component lipopolysaccharide
(LPS), a rabbit model of destructive joint inflammation is used.
This study design mimics the destructive joint inflammation seen in
arthritis. Intra-articular injection of LPS causes an acute
inflammatory response characterized by the release of cytokines and
chemokines, many of which have been identified in rheumatoid
arthritic joints. Marked increases in leukocytes occur in synovial
fluid and in synovium in response to elevation of these chemotactic
mediators. Selective antagonists of chemokine receptors have shown
efficacy in this model (see Podolin, et al., J. Immunol.
169(11):6435-6444 (2002)).
[0392] A rabbit LPS study is conducted essentially as described in
Podolin, et al. ibid., female New Zealand rabbits (approximately 2
kilograms) are treated intra-articularly in one knee with LPS (10
ng) together with either vehicle only (phosphate buffered saline
with 1% DMSO) or with addition of CCX-105 (dose 1=50 .mu.M or dose
2=100 .mu.M) in a total volume of 1.0 mL. Sixteen hours after the
LPS injection, knees are lavaged and cells counts are performed.
Beneficial effects of treatment were determined by histopathologic
evaluation of synovial inflammation. Inflammation scores are used
for the histopathologic evaluation: 1--minimal, 2--mild,
3--moderate, 4--moderate-marked.
[0393] b) Evaluation of a Candidate Compound in a Rat Model of
Collagen Induced Arthritis
[0394] A 17 day developing type II collagen arthritis study is
conducted to evaluate the effects of a candidate compound on
arthritis induced clinical ankle swelling. Rat collagen arthritis
is an experimental model of polyarthritis that has been widely used
for preclinical testing of numerous anti-arthritic agents (see
Trentham, et al., J. Exp. Med. 146(3):857-868 (1977), Bendele, et
al., Toxicologic Pathol. 27:134-142 (1999), Bendele, et al.,
Arthritis Rheum. 42:498-506 (1999)). The hallmarks of this model
are reliable onset and progression of robust, easily measurable
polyarticular inflammation, marked cartilage destruction in
association with pannus formation and mild to moderate bone
resorption and periosteal bone proliferation.
[0395] Female Lewis rats (approximately 0.2 kilograms) are
anesthetized with isoflurane and injected with Freund's Incomplete
Adjuvant containing 2 mg/mL bovine type II collagen at the base of
the tail and two sites on the back on days 0 and 6 of this 17 day
study. A candidate compound is dosed daily in a sub-cutaneous
manner from day 0 till day 17 at a efficacious dose. Caliper
measurements of the ankle joint diameter were taken, and reducing
joint swelling is taken as a measure of efficacy.
[0396] In the table below, structures and activity are provided for
representative compounds described herein. Activity is provided as
follows for either the chemotaxis assay or binding assay as
described above: +, IC.sub.50>12.5 uM; ++, 2500
nM<IC.sub.50<12.5 uM; +++, 1000 nM<IC.sub.50<2500 nM;
and ++++, IC.sub.50<1000 nM.
TABLE-US-00002 TABLE 2 Structure Structure ##STR00112## 1.001/++++
##STR00113## 1.002/++++ ##STR00114## 1.003/++++ ##STR00115##
1.004/++++ ##STR00116## 1.005/++++ ##STR00117## 1.006/++++
##STR00118## 1.007/++++ ##STR00119## 1.008/++++ ##STR00120##
1.009/++++ ##STR00121## 1.010/++++ ##STR00122## 1.011/++
##STR00123## 1.012/++++ ##STR00124## 1.013/++++ ##STR00125##
1.014/+++ ##STR00126## 1.015/++++ ##STR00127## 1.016/++++
##STR00128## 1.017/++++ ##STR00129## 1.018/++++ ##STR00130##
1.019/++++ ##STR00131## 1.020/++++ ##STR00132## 1.021/++++
##STR00133## 1.022/++++ ##STR00134## 1.023/++++ ##STR00135##
1.024/++++ ##STR00136## 1.025/++++ ##STR00137## 1.026/++++
##STR00138## 1.027/++++ ##STR00139## 1.028/ ++++ ##STR00140##
1.029/++++ ##STR00141## 1.030/++++ ##STR00142## 1.031/++++
##STR00143## 1.032/++++ ##STR00144## 1.033/++++ ##STR00145##
1.034/++++ ##STR00146## 1.035/++++ ##STR00147## 1.036/++++
##STR00148## 1.037/++++ ##STR00149## 1.038/++++ ##STR00150##
1.039/++++ ##STR00151## 1.040/++++ ##STR00152## 1.041/+++
##STR00153## 1.042/++++ ##STR00154## 1.043/++++ ##STR00155##
1.044/++++ ##STR00156## 1.045/++++ ##STR00157## 1.046/++++
##STR00158## 1.047/++++ ##STR00159## 1.048/++++ ##STR00160##
1.049/++++ ##STR00161## 1.050/++++ ##STR00162## 1.051/++++
##STR00163## 1.052/++++ ##STR00164## 1.053/++++ ##STR00165##
1.054/++++ ##STR00166## 1.055/++++ ##STR00167## 1.056/++++
##STR00168## 1.057/++++ ##STR00169## 1.058/++++ ##STR00170##
1.059/++++ ##STR00171## 1.060/++++ ##STR00172## 1.061/++++
##STR00173## 1.062/+++ ##STR00174## 1.063/++++ ##STR00175##
1.064/++++ ##STR00176## 1.065/++++ ##STR00177## 1.066/++++
##STR00178## 1.067/++++ ##STR00179## 1.068/++++ ##STR00180##
1.069/++++ ##STR00181## 1.070/++++ ##STR00182## 1.071/++++
##STR00183## 1.072/++++ ##STR00184## 1.073/++++ ##STR00185##
1.074/++++ ##STR00186## 1.075/++++ ##STR00187## 1.076/++++
##STR00188## 1.077/++++ ##STR00189## 1.078/++ ##STR00190##
1.079/++++ ##STR00191## 1.080/++++ ##STR00192## 1.081/++++
* * * * *