WO2002060900A2 - Antagonists of mcp-1 function and methods of use thereof - Google Patents

Antagonists of mcp-1 function and methods of use thereof Download PDF

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Publication number
WO2002060900A2
WO2002060900A2 PCT/US2002/003016 US0203016W WO02060900A2 WO 2002060900 A2 WO2002060900 A2 WO 2002060900A2 US 0203016 W US0203016 W US 0203016W WO 02060900 A2 WO02060900 A2 WO 02060900A2
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Prior art keywords
optionally substituted
lower alkyl
amino
carbonyl
pyridin
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PCT/US2002/003016
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French (fr)
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WO2002060900A3 (en
WO2002060900A8 (en
Inventor
Edgardo Laborde
Louise Robinson
Fanying Meng
Brian T. Peterson
Hugo O. Villar
Steven E. Anuskiewicz
Yoshiro Ishiwata
Shoji Yokochi
Yukiharu Matsumoto
Takuji Kakigami
Hideaki Inagaki
Takahito Jomori
Kouji Matsushima
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Telik, Inc.
Sanwa Kaguku Kenkyusho Co., Ltd.
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Priority to JP2002561468A priority Critical patent/JP4290984B2/en
Priority to EP02707672A priority patent/EP1358188A2/en
Priority to MXPA03006850A priority patent/MXPA03006850A/en
Priority to AU2002242065A priority patent/AU2002242065B2/en
Priority to CA002432997A priority patent/CA2432997A1/en
Priority to KR10-2003-7010170A priority patent/KR20030079976A/en
Priority to BR0206839-7A priority patent/BR0206839A/en
Publication of WO2002060900A2 publication Critical patent/WO2002060900A2/en
Publication of WO2002060900A3 publication Critical patent/WO2002060900A3/en
Publication of WO2002060900A8 publication Critical patent/WO2002060900A8/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A61K31/5513—1,4-Benzodiazepines, e.g. diazepam or clozapine
    • A—HUMAN NECESSITIES
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    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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Definitions

  • the present invention relates to chemical compounds, pharmaceutical compositions comprising said compounds, uses of said compounds and compositions, methods of treatment employing said compounds and compositions, and processes for preparing said compounds.
  • this invention relates to novel compounds which are antagonists of MCP-1 function and are useful in the prevention or treatment of chronic or acute inflammatory or autoimmune diseases, especially those associated with aberrant lymphocyte or monocyte accumulation such as arthritis, asthma, atherosclerosis, diabetic nephropathy, inflammatory bowel disease, Crohn's disease, multiple sclerosis, nephritis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, and transplant rejection. More specifically, the invention is related to pharmaceutical compositions comprising these compounds and the use of these compounds and compositions in the prevention or treatment of such diseases.
  • Chemokines Structure and Function
  • leukocytes The migration of leukocytes from blood vessels into diseased tissues is an important process in the initiation of normal inflammatory responses to certain stimuli or insults to the immune system. However, this process is also involved in the onset and progression of life-threatening inflammatory and autoimmune diseases; blocking leukocyte recruitment in these disease states, therefore, can be an effective therapeutic strategy.
  • the mechanism by which leukocytes leave the bloodstream and accumulate at inflammatory sites involves three distinct steps: (1 ) rolling, (2) arrest and firm adhesion, and (3) transendothelial migration [Springer, Nature 346:425-433 (1990); Lawrence and Springer, Cell 65:859-873 (1991 ); Butcher, Cell 67:1033-1036 (1991 )].
  • the second step is mediated at the molecular level by chemoattractant receptors on the surface of leukocytes which bind chemoattractant cytokines secreted by proinflammatory cells at the site of damage or infection.
  • Receptor binding activates leukocytes, increases their adhesiveness to the endothelium, and promotes their transmigration into the affected tissue, where they can secrete inflammatory and chemoattractant cytokines and degradative proteases that act on the subendothelial matrix, facilitating the migration of additional leukocytes to the site of injury.
  • chemokines are a large family of low molecular weight (8 to 10 kD) proteins that share the ability to stimulate directed cell migration (“chemotaxis”) [Schall, Cytokine 3:165-183 (1991 ); Murphy, Rev Immun 12:593-633 (1994)].
  • Chemokines are characterized by the presence of four conserved cysteine residues and are grouped into two main subfamilies based on whether the two amino-terminal cysteines are separated by one amino acid (CXC subfamily, also known as ⁇ -chemokines) or immediately adjacent to each other (CC subfamily, also referred to as ⁇ -chemokines) [Baggiolini et al., Adv Immunol 55:97-179 (1994); Baggiolini et al., Annu Rev Immunol 15:675-705 (1997); Deng et al., Nature 381 :661-666 (1996); Luster, New Engl J Med 338:436445 (1998); Saunders and Tarby, Drug Discovery Today 4:80-92 (1999)].
  • CXC subfamily also known as ⁇ -chemokines
  • CC subfamily also referred to as ⁇ -chemokines
  • the chemokines of the CXC subfamily are produced by a wide range of cells and act predominantly on neutrophils as mediators of acute inflammation.
  • the CC chemokines which include MCP-1 , RANTES, MIP-1 ⁇ , and MIP-1 ⁇ , are also produced by a variety of cells, but these molecules act mainly on monocytes and lymphocytes in chronic inflammation.
  • chemokines Like many cytokines and growth factors, chemokines utilize both high and low affinity interactions to elicit full biological activity.
  • Studies performed with labeled ligands have identified chemokine binding sites ("receptors") on the surface of neutrophils, monocytes, T cells, and eosinophils with affinities in the 500 pM to 10 nM range [Kelvin et al., J Leukoc Biol 54:604-612 (1993); Murphy, Annu Rev Immunol 12:593-633 (1994); Raport et al., J Leukoc Biol 59:18-23 (1996); Premack and Schall, Nature Med 2:1174-1178 (1996)].
  • the cloning of these receptors has revealed that cell surface high-affinity chemokine receptors belong to the seven transmembrane (“serpentine”) G-protein-coupled receptor (GPCR) superfamily.
  • Chemokine receptors are expressed on different cell types, including non-leukocyte cells. Some receptors are restricted to certain cells (e.g., the
  • CXCR1 receptor is predominantly restricted to neutrophils), whereas others are more widely expressed (e.g., the CCR2 receptor is expressed on monocytes, T cells, natural killer cells, dendritic cells, and basophils).
  • chemokines are rather promiscuous with regard to their binding partners.
  • MIP-1 ⁇ and RANTES bind to the CCR1 and CCR4 receptors
  • MCP-1 binds to the CCR2 and CCR4 receptors.
  • most chemokines receptors bind more than one chemokine
  • CC receptors bind only CC chemokines
  • CXC receptors bind only CXC chemokines.
  • This ligand-receptor restriction may be related to the structural differences between CC and CXC chemokines, which have similar primary, secondary, and tertiary structures, but different quaternary structures [Lodi et al., Science 263:1762-1767 (1994)].
  • chemokines The binding of chemokines to their serpentine receptors is transduced into a variety of biochemical and physiological changes, including inhibition of cAMP synthesis, stimulation of cytosolic calcium influx, upregulation or activation of adhesion proteins, receptor desensitization and internalization, and cytoskeletal rearrangements leading to chemotaxis [Vaddi et al., J Immunol 153:4721-4732 (1994); Szabo et al., Eur J Immunol 27:1061-1068 (1997); Campbell et al., Science 279:381-384 (1998); Aragay et al., Proc Natl Acad Sci USA 95:2985- 2990 (1998); Franci et al., J Immunol 157:5606-5612 (1996); Aramori et al., EMBO J 16:4606-4616 (1997); Haribabu et al., J Biol Chem 272:28726-28731 (1997); Newton e
  • chemokine binding also triggers cellular activation, resulting in lysozomal enzyme release and generation of toxic products from the respiratory burst [Newton et al., Methods Enzymol 287:174- 186 (1997); Zachariae et al., J Exp Med 171 :2177-2182 (1990); Vaddi et al., J Leukocyte Biol 55:756-762 (1994)].
  • the molecular details of the chemokine- receptor interactions responsible for inducing signal transduction, as well as the specific pathways that link binding to the above-mentioned physiological changes, are still being elucidated.
  • chemokines of both subfamilies bind to various extracellular matrix proteins such as the glycosaminoglycans (GAGs) heparin, chondroitin sulfate, heparan sulfate, and dermatan sulfate with affinities in the middle nanomolar to millimolar range.
  • GAGs glycosaminoglycans
  • Chemokines have been implicated as important mediators of allergic, inflammatory and autoimmune disorders and diseases, such as asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, and transplant rejection. Accordingly, it has been postulated that the use of antagonists of chemokine function may help reverse or halt the progression of these disorders and diseases.
  • MCP-1 appears to play a significant role during the early stages of allergic responses because of its ability to induce mast cell activation and LTC4 release into the airway, which directly induces AHR (airways hyper-responsiveness) [Campbell et al., J Immunol 163:2160-2167 (1999)].
  • MCP-1 has been found in the lungs of patients with idiopathic pulmonary fibrosis and is thought to be responsible for the influx of mononuclear phagocytes and the production of growth factors that stimulate mesenchymal cells and subsequent fibrosis [Antoniades et al., Proc Natl Acad Sci USA 89:5371-5375 (1992)].
  • MCP-1 is also involved in the accumulation of monocytes in pleural effusions which is associated with both Mycobacterium tuberculosis infection and malignancy [Strieter et al., J Lab Clin Med 123:183-197 (1994)].
  • MCP-1 has also been shown to be constitutively expressed by synovial fibroblasts from rheumatoid arthritis patients, and its levels are higher in rheumatoid arthritis joints compared to normal joints or those from other arthritic diseases [Koch et al., J Clin Invest 90:772-779 (1992)]. These elevated levels of MCP-1 are probably responsible for the monocyte infiltration into the synovial tissue. Increased levels of synovial MIP-1 ⁇ and RANTES have also been detected in patients with rheumatoid arthritis [Kundel et al., J Leukocyte Biol 59:6-12 (1996)].
  • MCP-1 also plays a critical role in the initiation and development of atherosclerotic lesions. MCP-1 is responsible for the recruitment of monocytes into atherosclerotic areas, as shown by immunohistochemistry of macrophage-rich arterial wall [Yla-Herttuala et al., Proc Natl Acad Sci USA 88:5252-5256 (1991 ); Nelken et al., J Clin Invest 88:1 121 -1 127 (1991 )] and anti-MCP-1 antibody detection [Takeya et al., Human Pathol 24:534-539 (1993)].
  • LDL-receptor/MCP-1 -deficient and apoB-transgenic/MCP-1 -deficient mice show significantly less lipid deposition and macrophage accumulation throughout their aortas compared with wild-type MCP-1 strains [Alcami et al., J Immunol 160:624- 633 (1998); Gosling et al., J Clin Invest 103:773-778 (1999); Gu et al., Mol. Cell. 2:275-281 (1998); Boring et al., Nature 394:894-897 (1998).
  • MCP-1 multiple sclerosis
  • stroke Other inflammatory diseases marked by specific site elevations of MCP-1 include multiple sclerosis (MS), glomerulonephritis, and stroke.
  • chemokine antagonists reported to date are either neutralizing antibodies to specific chemokines or receptor-ligand antagonists, that is, agents that compete with specific chemokines for binding to their cognate serpentine receptors but, unlike the chemokines themselves, do not activate these receptors towards eliciting a functional response [Howard et al., Trend Biotechnol ! 4:46-51 (1996)].
  • anti-chemokine antibodies have been shown to curtail inflammation in a number of animal models (e.g., anti-MIP-1 ⁇ in bleomycin- induced pulmonary fibrosis [Smith et al., Leukocyte Biol 57:782-787 (1994)]; anti-IL-8 in reperfusion injury [Sekido et al., Nature 365:654-657 (1995)], and anti-MCP-1 in a rat model of glomerulonephritis [Wada et al., FASEB J 10:1418-1425 (1996)]).
  • anti-MIP-1 ⁇ in bleomycin- induced pulmonary fibrosis
  • anti-IL-8 in reperfusion injury
  • anti-MCP-1 in a rat model of glomerulonephritis
  • MRL-/pr mouse arthritis model In the MRL-/pr mouse arthritis model, administration of an MCP-1 antagonist significantly reduced the overall histopathological score after the early onset of the disease [Gong et al., J Exp Med 186: 131 -137 (1997)].
  • a major problem associated with using antibodies to antagonize chemokine function is that they must be humanized before use in chronic human diseases.
  • the ability of multiple chemokines to bind and activate a single receptor forces the development of a multiple antibody strategy or the use of cross-reactive antibodies in order to completely block or prevent pathological conditions.
  • chemokine receptor antagonists suggest that inflammatory disorders characterized by multiple or redundant chemokine expression profiles will be relatively more refractory to treatment by these agents.
  • a different approach to target chemokine function would involve the use of compounds that disrupt the chemokine-GAG interaction.
  • One class of such agents with potential therapeutic application would consist of small organic molecules that bind to the chemokine low affinity GAG-binding domain. Compounds of this class might not inhibit binding of the chemokine to its high-affinity receptor per se, but would disrupt chemokine localization within the extracellular matrix and provide an effective block for directed leukocyte-taxis within tissues.
  • the heparinoids suramin and pentosan polysulphate both inhibit angiogenesis under conditions where heparin is either ineffective or even stimulatory [Wellstein and Czubayko, Breast Cancer Res Treat 38:109-119 (1996)].
  • the anti-angiogenic capacity of the drug has also been shown to be targeted against VEGF [Waltenberger et al., J Mol Cell Cardiol 28:1523-1529 (1996)] which, like FGF, possesses heparin-binding domains similar to those of the chemokines.
  • Heparin or heparin sulphate has also been shown to directly compete for GAG interactions critical for T-cell adhesion mediated by MIP-1 ⁇ in vitro [Tanaka et al., Nature 361:79-82 (1993)].
  • the present invention relates to compounds that inhibit MCP-1 -induced chemotaxis of human monocytic cells both in vitro and in vivo.
  • novel MCP-1 antagonists are useful for the treatment of inflammatory diseases, especially those associated with lymphocyte and/or monocyte accumulation, such as atherosclerosis, diabetic nephropathy, inflammatory bowel disease, Crohn's disease, multiple sclerosis, nephritis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, and other chronic or acute autoimmune disorders.
  • these compounds can be used in the treatment of allergic hypersensitivity disorders, such as asthma and allergic rhinitis, characterized by basophil activation and eosinophil recruitment.
  • a first embodiment of the present invention provides compounds of
  • Y is O, S or N-R 7 ,
  • Z is N or C-R 8 .
  • R 1 R 2 , R 3 , and R 8 are independently, hydrogen, or optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), -CF 3 , halogen, nitro, -CN, -OR 9 , -SR 9 ,
  • R a and R 1 ⁇ 0 ⁇ are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(d- 2 alkyl) 2 , lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl(lower alkyl), aryl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 9 and R 10 together are -(CH 2 ) 4 -6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ . 2 alkyl) group,
  • R 10 together are -(CH 2 ) 4 -6- optionally interrupted by one O, S, NH,
  • R 4 and R 5 are independently, hydrogen, lower alkyl optionally substituted lower alkyl, optionally substituted aryl, or optionally substituted aryl(lower alkyl), or, together, are -(CH 2 ) 2 . 4 -,
  • R 11 and R 12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 11 and
  • R 12 together are -(CH 2 ) 4 . 6 -, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
  • the compounds of this invention may possess one or more chiral centers, and can therefore be produced as individual stereoisomers or as mixtures of stereoisomers, depending on whether individual stereoisomers or mixtures of stereoisomers of the starting materials are used.
  • some of the compounds of the invention are capable of further forming pharmaceutically acceptable salts and esters.
  • the compounds of this invention may further exist in tautomeric forms and can therefore be produced as individual tautomeric forms or as mixtures of tautomeric forms. Unless indicated otherwise, the description or naming of a compound or groups of compounds is intended to include both the individual isomers or mixtures (racemic or otherwise) of stereoisomers and their tautomeric forms.
  • a second embodiment of the present invention provides compounds of Formula (la) or Formula (lla):
  • Y is O, S or N-R 7 ,
  • Z is N or C-R 8 .
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 7 and R 8 are as defined in the first embodiment
  • a third embodiment of the present invention provides pharmaceutical compositions comprising pharmaceutically acceptable excipients and a therapeutically effective amount of at least one compound of this invention.
  • a fourth embodiment of the present invention provides methods for treating chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection in mammals in need thereof, comprising the administration to such mammal of a therapeutically effective amount of at least one compound of this invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.
  • chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection in mammals in need thereof, comprising the
  • a fifth embodiment of this invention provides uses of the compounds of the invention in the preparation of medicaments for the treatment of chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection.
  • chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection.
  • a sixth embodiment of this invention provides a process for the preparation of the compounds of the invention or pharmaceutically acceptable salts thereof.
  • Alkyl is a linear or branched saturated hydrocarbon radical having from 1 to 20 carbon atoms.
  • alkyl radicals are: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, dodecyl, etc.
  • Lower alkyl as in “lower alkyl,” “lower alkoxy,” “cycloalkyl(lower alkyl),” “aryl(lower alkyl)”, or “heteroaryl(lower alkyl)", means a C- ⁇ - 10 alkyl radical.
  • Preferred lower alkyl radicals are those having from 1 to 6 carbon atoms.
  • alkenyl is a linear or branched hydrocarbon radical having from 2 to 20 carbon atoms and at least one carbon-carbon double bond. Examples of alkenyl radicals are: vinyl, 1-propenyl, isobutenyl, etc.
  • Alkynyl is a linear or branched hydrocarbon radical having from 2 to 20 carbon atoms and at least one carbon-carbon triple bond. Examples of alkynyl radicals are: propargyl, 1-butynyl, etc.
  • Cycloalkyl is a monovalent cyclic hydrocarbon radical having from 3 to 12 carbon atoms. Examples of cycloalkyl radicals are: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.
  • Cycloalkyl(lower alkyl) is a lower alkyl radical which is substituted with a cycloalkyl, as previously defined.
  • Examples of cycloalkyl(lower alkyl) radicals are cyclopropylmethyl, cyclobutylethyl, cyclopentylmethyl, cyclohexyl methyl, etc.
  • Heterocycloalkyl is a monovalent cyclic hydrocarbon radical having 3 to
  • Halo(lower alkyl) is a radical derived from lower alkyl containing at least one halogen substituent.
  • Non-limiting examples of halo(lower alkyl) radicals include: -CF 3 , C 2 F 5 , etc..
  • Aryl as in “aryl”, “aryloxy”, and “aryl(lower alkyl)", is a radical derived from an aromatic hydrocarbon containing 6 to 16 ring carbon atoms, having a single ring (e.g., phenyl), or two or more condensed rings, preferably 2 to 3 condensed rings (e.g., naphthyl), or two or more aromatic rings, preferably 2 to 3 aromatic rings, which are linked by a single bond (e.g., biphenyl).
  • Preferred aryl radicals are those containing from 6 to 14 carbon atoms.
  • Heteroaryl as in “heteroaryl” and “heteroaryl(lower alkyl)", is a radical derived from an aromatic hydrocarbon containing 5 to 14 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic aromatic rings (e.g., thienyl, furyl, pyrrolyl, pyrimidinyl, isoxazolyl, oxazolyl, indolyl, isobenzofuranyl, purinyl, isoquinolyl, pteridinyl, imidazolyl, pyridyl, pyrazolyl, pyrazinyl, quinolyl, etc.).
  • substituents on the substituted heteroaryl moiety include lower alkyl, substituted lower alkyl, halo(lower alkyl), halogen, nitro, -CN, -OR, -SR, and -NRR ' .
  • Aryl(lower alkyl) is a lower alkyl radical which is substituted with an aryl, as previously defined.
  • aryl(lower alkyl) is an aryl(lower alkyl) radical having one to three substituents on either or both of the aryl and the alkyl portion of the radical.
  • Heteroaryl(lower alkyl) is a lower alkyl radical which is substituted with a heteroaryl, as previously defined.
  • Substituted heteroaryl(lower alkyl) is a heteroaryl(lower alkyl) radical having one to three substituents on the heteroaryl portion or the alkyl portion of the radical, or both.
  • Lower alkoxy is an -OR radical, where R is a lower alkyl or cycloalkyl.
  • Halogen means fluoro, chloro, bromo, or iodo.
  • Steps are compounds that have the same sequence of covalent bonds and differ in the relative disposition of their atoms in space.
  • Inner salts or “Zwitterions” can be formed by transferring a proton from the carboxyl group onto the lone pair of electrons of the nitrogen atom in the amino group.
  • Tautomers are isomeric compounds that differ from one another by interchanged positions of ⁇ and ⁇ bonds. The compounds are in equilibrium with one another. They may also differ from one another in the position at which a hydrogen atom is attached.
  • “Pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients may be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.
  • “Pharmaceutically acceptable salts and esters” means any salt and ester that is pharmaceutically acceptable and has the desired pharmacological properties.
  • Such salts include salts that may be derived from an inorganic or organic acid, or an inorganic or organic base, including amino acids, which is not toxic or undesirable in any way.
  • Suitable inorganic salts include those formed with the alkali metals, e.g. sodium and potassium, magnesium, calcium, and aluminum.
  • Suitable organic salts include those formed with organic bases such as the amine bases, e.g. ethanolamine, diethanolamine, triethanolamine, tromethamine, ⁇ /-methylglucamine, and the like.
  • Such salts also include acid addition salts formed with inorganic acids (e.g.
  • esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the compounds, e.g. C1-6 alkyl esters.
  • a pharmaceutically acceptable salt or ester may be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly, where there are more than two acidic groups present, some or all of such groups can be salified or esterified.
  • Disease includes any unhealthy condition of an animal (which includes human and non-human mammals), including particularly various forms of inflammatory illnesses or diseases, such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, immune disorders, and transplant rejection.
  • inflammatory illnesses or diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, immune disorders, and transplant rejection.
  • the first embodiment of the present invention provides compounds of Formula (I) and Formula (II):
  • R and R >10 . together are -(CH 2 ) 4 . 6 - optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N- (optionally substituted C ⁇ - 2 alkyl) group
  • examples include piperazinyl, 4- methylpiperazinyl, 4-morpholyl, and hexahydropyhmidyl.
  • Y is O or N-R 7 .
  • R 1 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR 9 , -NR 9 R 10 , wherein R 9 and R 10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C ⁇ - 2 alkyl) 2 , lower alkyl(optionally substituted heterocycloalkyl), aryl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl).
  • 6 - optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ - 2 alkyl) group.
  • R 9 and R 10 together are -(CH 2 ) . 6 - optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ - 2 alkyl) group.
  • alkyl 2 alkyl 2 , alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl).
  • R 4 and R 5 are independently, hydrogen or lower alkyl, or together are -(CH 2 ) 2 . 4 -. More preferably, R 4 and R 5 are independently, hydrogen or lower alkyl.
  • a particular preferred "substituted aryl” is a phenyl group substituted with a R 13 and optionally substituted with up to four R 14 s, where R 13 and R 14 are defined with respect to formulae la and lla.
  • R 13 and R 14 are defined with respect to formulae la and lla.
  • Y is N-R 7 and Z is N
  • R 1 is lower alkyl
  • R 4 and R 5 are hydrogen
  • R 11 and R 12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 11 and R 12 together are -(CH 2 ) 4 . 6 -, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
  • R 2 is -NR 9 R 10 , wherein R 9 and R 10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C ⁇ - 2 alkyl) 2 , lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), benzyl, optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 9 and R 10 together are -(CH 2 ) .
  • 6 - optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ - 2 alkyl) group.
  • Y is N-R 7 and Z are N
  • R 1 and R 8 are lower alkyl
  • R 2 is -NR 9 R 10 ,
  • R 4 and R 5 are hydrogen
  • R ,1"1 and R 12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 11 and R 12 together are -(CH 2 ) 4 . 6 -, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
  • the second embodiment of the present invention provides compounds of Formula (la) or Formula (lla):
  • Y is O, S or N-R 7 , Z is N or C-R 8 , R 1 , R 2 , R 3 , R 4 , R 5 , R 7 and R 8 are as defined in the first embodiment,
  • R 13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl), -CF 3 , halogen, nitro, -CN, -OR 15 , -SR 15 , -NR 15 R 16 ,
  • R 15 and R 16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, -CF 3 , cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), or, together, are -(CH 2 ) 4 .6- optionally interrupted by one O, S, NH or N-(C ⁇ - 2 alkyl) group, each R 14 is independently selected from optionally substituted lower alkyl, optionally
  • R 9 and R 10 together are -(CH 2 ) 4 -6- optionally interrupted by one O, S, NH or N-(C ⁇ - 2 alkyl) group
  • examples include piperazinyl, 4-methylpiperazinyl, morpholyl, and hexahydropyrimidyl.
  • n is a stereocompatible integer of 0 to 4. The term "stereocompatible" limits the number of substituents permissible by available valences in accordance with space requirements of substituents, among other.
  • n is an integer of 1 to 2. More preferably, where R 13 is not hydrogen, n is 1.
  • Y is N-R 7 and Z is N, R 1 is lower alkyl,
  • R 2 is -NR 9 R 10 , wherein R 9 and R 10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C ⁇ - 2 alkyl) 2 , lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), benzyl, optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R 9 and R 10 together are -(CH 2 ) - 6 - optionally interrupted by one O, S, NH, N-(aryl),
  • N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ . 2 alkyl) group such as piperazinyl
  • R 13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl),
  • R 15 and R 16 are independently, hydrogen, lower alkyl, alkenyl, alkynyl, -CF 3 , cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally .
  • substituted heteroaryl optionally substituted heteroaryl(lower alkyl) or R 15 and R 16 together are -(CH 2 ) 4 . 6 - optionally interrupted by one O, S, NH or N-(C-
  • R 4 and R 5 are hydrogen. Most preferably, independently,
  • Y is O, and R 1 is lower alkyl.
  • Y is N-R 7 , R 7 are hydrogen or lower alkyl, and R 1 is lower alkyl.
  • Z is C-R 8 , and R 8 is hydrogen.
  • R 2 and R 3 are independently selected from hydrogen, lower alkyl, halogen, OR 9 , -NR 9 R 10 , where R 9 and R 10 are independently lower alkyl, substituted lower alkyl, or substituted aryl, or R 9 and R 10 together are -(CH 2 ) . 6 - optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-COOH, N-(carboxy(lower alkyl)) or N-(optionally substituted C ⁇ - 2 alkyl) group.
  • R 17 and R 18 are, independently, lower alkyl, substituted lower alkyl, or substituted aryl, or, together, are -(CH 2 ) 4 . 6 - optionally interrupted by one O, S, NH or N-(C-
  • Z is N
  • R 2 is 4-methylpiperazinyl
  • R 13 is 3-CF 3
  • R 14 is 4-F.
  • the preferred compounds of the invention are listed in Tables 1-4 below.
  • the compounds of this invention may possess one or more chiral centers, and can therefore exist as individual stereoisomers or as mixtures of stereoisomers. In such cases, all stereoisomers also fall within the scope of this invention.
  • the compounds of this invention may also exist in various tautomeric forms, and in such cases, all tautomers also fall within the scope of this invention.
  • the invention compounds include the individually isolated stereoisomers and tautomers as well as mixtures of such stereoisomers and their tautomers.
  • Pharmaceutically acceptable base addition salts of the compounds of Formula (I) and Formula (II) include salts which may be formed when acidic protons present in the parent compound are capable of reacting with inorganic or organic bases.
  • the parent compound is treated with an excess of an alkaline reagent, such as hydroxide, carbonate, or alkoxide, containing an appropriate cation.
  • Cations such as Na ⁇ K + , Ca 2+ , and NH 4 + are examples of cations present in pharmaceutically acceptable salts.
  • the Na + salts are especially useful.
  • Acceptable inorganic bases therefore, include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide. Salts may also be prepared using organic bases, such as choline, dicyclohexylamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, procaine, ⁇ /-methylglucamine, and the like [for a nonexclusive list see, for example, Berge et al., "Pharmaceutical Salts," J. Pharma. Sci. 66:1 (1977)].
  • the free acid form may be regenerated by contacting the base addition salt with an acid and isolating the free acid in the conventional manner.
  • the free acid forms can differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents.
  • Pharmaceutically acceptable acid addition salts of the compounds of Formula (I) and Formula (II) include salts which may be formed when the parent compound contains a basic group.
  • Acid addition salts of the compounds are prepared in a suitable solvent from the parent compound and an excess of a non-toxic inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid (giving the sulfate and bisulfate salts), nitric acid, phosphoric acid and the like, or a non-toxic organic acid such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, salicylic acid, p-toluenesulfonic acid, hexanoic acid, hepta
  • the free base form may be regenerated by contacting the acid addition salt with a base and isolating the free base in the conventional manner.
  • the free base forms can differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents.
  • salts of amino acids such as arginate and the like, gluconate, and galacturonate [see Berge, supra (1977)].
  • Some of the compounds of the invention may form inner salts or Zwitterions.
  • Certain of the compounds of the present invention may also exist in one or more solid or crystalline phases or polymorphs, the variable biological activities of such polymorphs or mixtures of such polymorphs are also included in the scope of this invention.
  • a third embodiment of the present invention provides pharmaceutical compositions comprising pharmaceutically acceptable excipients and a therapeutically effective amount of at least one compound of this invention.
  • Pharmaceutical compositions of the compounds of this invention, or derivatives thereof may be formulated as solutions or lyophilized powders for parenteral administration. Powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use.
  • the liquid formulation is generally a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, 5% dextrose in water or buffered sodium or ammonium acetate solution.
  • Such formulations are especially suitable for parenteral administration but may also be used for oral administration. It may be desirable to add excipients such as polyvinylpyrrolidinone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride, or sodium citrate.
  • these compounds may be encapsulated, tableted, or prepared in an emulsion or syrup for oral administration.
  • Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to facilitate preparation of the composition.
  • Liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohols, or water.
  • Solid carriers include starch, lactose, calcium sulfate, dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar, or gelatin.
  • the carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax.
  • the amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage unit.
  • the pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulation, and compressing, when necessary, for tablet forms; or milling, mixing, and filling for hard gelatin capsule forms.
  • a liquid carrier When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion, or an aqueous or non-aqueous suspension.
  • Such a liquid formulation may be administered directly p.o. or filled into a soft gelatin capsule.
  • a pharmaceutical composition of the present invention is packaged in a container with a label indicating the use of the pharmaceutical composition in the treatment of a disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, and transplant rejection, or a chronic or acute immune disorder, or a combination of any of these disease conditions.
  • a disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, and transplant rejection, or a chronic or acute immune disorder, or a combination of any of these disease conditions.
  • a fourth embodiment of the present invention provides a method for treating a chronic or acute inflammatory disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a chronic or acute immune disorder, or a transplant rejection in mammals in need thereof, comprising the administration to such mammal of a therapeutically effective amount of at least one compound of general Formula (I) and Formula (II) or pharmaceutically acceptable salts and esters thereof.
  • a chronic or acute inflammatory disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a
  • the compounds of the present invention inhibit chemotaxis of a human monocytic cell line (THP-1 cells) induced by human MCP-1 in vitro. This inhibitory effect has also been observed in vivo. Indeed, the compounds have been shown to reduce monocyte infiltration in a thioglycollate-induced inflammation model in mice.
  • the compounds of the present invention have been found to prevent the onset or ameliorate symptoms in several animal models of inflammation.
  • the compounds inhibited recruitment of monocytes into the glomeruli in an anti-Thy-1 antibody-induced model of nephritis; reduced paw swelling in a rat model of adjuvant arthritis; inhibited neointimal hyperplasia after balloon injury in a rat model of restenosis, and reduced the amount of lesion of the aortic sinus in an apoE-deficient mouse model of atherosclerosis.
  • the use of the compounds of the invention for treating inflammatory and autoimmune disease by combination therapy may also comprise the administration of the compound of the invention to a mammal in combination with common anti-inflammatory drugs, cytokines, or immunomodulators.
  • the compounds of this invention are thus used to inhibit leukocyte migration in patients which require such treatment.
  • the method of treatment comprises the administration, orally or parenterally, of an effective quantity of the chosen compound of the invention, preferably dispersed in a pharmaceutical carrier.
  • Dosage units of the active ingredient are generally selected from the range of 0.01 to 1000 mg/kg, preferably 0.01 to 100 mg/kg, and more preferably 0.1 to 50 mg/kg, but the range will be readily determined by one skilled in the art depending on the route of administration, age, and condition of the patient. These dosage units may be administered one to ten times daily for acute or chronic disease. No unacceptable toxicological effects are expected when compounds of the invention are used in accordance with the present invention.
  • the invention compounds may be administered by any route suitable to the subject being treated and the nature of the subject's condition.
  • Routes of administration include, but are not limited to, administration by injection, including intravenous, intraperitoneal, intramuscular, and subcutaneous injection, by transmucosal or transdermal delivery, through topical applications, nasal spray, suppository and the like, or may be administered orally.
  • Formulations may optionally be liposomal formulations, emulsions, formulations designed to administer the drug across mucosal membranes or transdermal formulations. Suitable formulations for each of these methods of administration may be found in, for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.
  • the following general procedures may be employed for the preparation of the compounds of the present invention.
  • the starting materials and reagents used in preparing these compounds are either available from commercial suppliers such as the Aldrich Chemical Company (Milwaukee, WI), Bachem (Torrance, CA), Sigma (St. Louis, MO), or are prepared by methods well known to a person of ordinary skill in the art, following procedures described in such references as Fieser and Fieser's Reagents for Organic Synthesis, vols. 1-17, John Wiley and Sons, New York, NY, 1991 ; Rodd's Chemistry of Carbon Compounds, vols. 1-5 and supps., Elsevier Science Publishers, 1989; Organic Reactions, vols.
  • protective groups for amino, hydroxy, and carboxy groups are described in Greene et al., Protective Groups in Organic Synthesis, Second Edition, John Wiley and Sons, New York, 1991. Activation of carboxylic acids can be achieved by using a number of different reagents as described in Larock: Comprehensive Organic Transformations, VCH Publishers, New York, 1989.
  • the starting materials, intermediates, and compounds of this invention may be isolated and purified using conventional techniques, including precipitation, filtration, distillation, crystallization, chromatography, and the like.
  • the compounds may be characterized using conventional methods, including physical constants and spectroscopic methods.
  • Step (h) resolving a racemic mixture of any proportions of a compound of formula I or formula II to yield a stereoisomer thereof.
  • Step (a), above may be carried out in the presence of an organic solvent or a mixture of solvents at elevated temperatures.
  • Said organic solvent may be toluene, and the reaction may be carried out under refluxing conditions.
  • Step (b), above, may be carried out using the salt of the compound of the formula R 2 -H in an inorganic solvent.
  • Said salt may be the lithium, sodium, or potassium salt.
  • Step (c), above may be carried out in an organic solvent or a mixture of solvents at elevated temperatures.
  • the haloformylation reagent may be a compound of the formula A-(CO)-B wherein A and B are, independently, suitable leaving groups such as halogens, -COCI, -COBr and the like.
  • the haloformylation agent and organic solvent employed in step (c) may be oxalyl chloride and THF, respectively, and the ensuing reaction may be heated to above 50°C.
  • the compounds of the invention can further be synthesized as shown in the following examples. These examples are merely illustrative of some methods by which the compounds of this invention can be synthesized, and various modifications to these examples can be made and will be suggested to a person of ordinary skill in the art having regard to this disclosure.
  • Acylurea compounds of the present invention may be prepared starting with an aryl carboxamide and an isocyanate.
  • Carboxamide and isocyanate starting materials may be purchased from various different commercial sources, such as, for example the Aldrich Chemical Company, supra, or they may be prepared from standard procedures known in the art for preparing these compounds, such as the procedures described in the above-cited references.
  • the isocyanates may also be prepared according to the procedures described in the example below.
  • an aryl carboxamide is treated with an aryl isocyanate in an organic solvent or mixtures of suitable organic solvents.
  • the organic solvent is toluene.
  • the carboxamide and the isocyanate may be combined as solutions or suspensions, depending on the solubilities of the compounds in the selected solvent.
  • the carboxamide and the isocyanate may be added in a stoichiomethc ratio (1 :1), or a slight excess of the isocyanate may be used, for example between 1.01 fold and 2 fold excess, but typically about 1.01 to about 1.2 fold excess.
  • the isocyanate is added to a suspension of the carboxamide in toluene, and the resulting mixture is heated until the reaction is determined to be complete.
  • the reaction mixture may be heated at about 10 °C to about 150 °C, preferably at about 40 °C to about 120 °C under an inert atmosphere such as nitrogen, or the reaction mixture may be maintained at the refluxing temperature of the mixture.
  • the reaction may be allowed to proceed to completion in about 10 minutes to 24 hours.
  • the reaction is heated to reflux until the reaction is complete, over about 6 to 24 hours.
  • the resulting precipitated acylureas may be isolated by conventional techniques.
  • the product is isolated by filtration.
  • the precipitated solid may be filtered, washed with a solvent or a series of solvents, and isolated without further purification.
  • the precipitated acylureas may be washed with a combination of toluene, methanol and then with ether, and the product may be dried under vacuum.
  • the acylureas may be further purified using conventional techniques, such as by crystallization using conventional methods known in the art.
  • acylureas prepared according to this procedure may be converted to the corresponding salts prior to isolation and/or purification, or after crystallization.
  • Acylureas may also be prepared from the condensation of an aryl carboxamide with an amine.
  • the carboxamide may be prepared from the corresponding carboxylic acid or may be obtained from commercial sources.
  • the amine may be substituted where one substituted group is an amine protecting group such that the protecting group may be removed in a subsequent step if desired.
  • a carboxamide mixture in a suitable aprotic solvent is treated with a haloformylation reagent to form the corresponding carboxamide carbonylchloride derivative.
  • the aprotic solvent is dichloromethane, toluene, 2-methyltetrahydrofuran or THF
  • the haloformylation reagent is oxalyl chloride.
  • the aprotic solvent is THF.
  • Oxalyl chloride is preferably present in an excess, for example between 1.1 to 3.0 equivalents, typically about 1.5 equivalent over the carboxamide.
  • the reaction is generally performed under an inert atmosphere where the mixture is heated to 50 °C to 175 °C for 15 minutes to 24 hours until the reaction is deemed complete. Typically, the reaction is heated to reflux over 2 to 16 hours under nitrogen, and then cooled to room temperature.
  • the solvent is removed in vacuo by rotoevaporation or distillation, and the resulting carboxamide carbonylchloride is then condensed with a primary or secondary amine.
  • Condensation with the amine may be performed by the addition of a solution of the amine in an aprotic solvent, such as THF, under an inert atmosphere, at a temperature between 0 °C and 20 °C, preferably between 0 °C and 5 °C. If the chloroformylation and the subsequent condensation reaction is performed in the same solvent, the solvent removal step may be eliminated. Preferably, the reaction is performed at 0 °C to 5 °C for 1 to 24 hours, until the reaction is complete.
  • the solvent is removed by concentration under reduced pressure, and the acylurea can be isolated by conventional techniques such as filtration and washing of the crude product with a solvent, followed by drying under vacuum.
  • the preparation of the acylureas may also be performed starting with an amine or aniline derivative through a condensation reaction with a phosgene equivalent, followed by a condensation reaction with the carboxamide.
  • a solution of an aniline or aniline derivative and triphosgene in tetrachloroethane or other suitable organic solvent is combined and stirred at 25 °C to 80 °C under an inert atmosphere for 2 to 12 hours until the reaction is complete.
  • the solvent is removed under reduced pressure and the residue is dissolved in an aprotic solvent, such as toluene, and the resulting mixture is treated with a carboxamide.
  • the mixture is heated to about 50 °C to 150 °C, preferably from about 75 °C to 115 °C.
  • the reaction mixture is heated to reflux for 2 to 24 hours until the reaction is complete, and allowed to cool to room temperature.
  • the precipitated solid is isolated by conventional techniques such as filtration.
  • the filtered solid is then washed with a suitable solvent or mixtures of solvents.
  • the solid is washed with toluene, methanol and then ether, and the washed product is dried in vacuo to give the corresponding acylurea.
  • Amine substituted aryl acyl ureas of the present invention may be prepared starting from the corresponding aryl halide acyl ureas by the reaction of the aryl halide with an amine.
  • the aryl halide is an aryl chloride, which can be prepared from one or more of the above described procedures or from standard procedures known in the art for preparing these compounds.
  • the acyl urea is dissolved in an organic solvent or a mixture of suitable organic solvents.
  • the organic solvent is tetrahydrofuran.
  • the acyl urea and the amine may be combined as solutions or suspensions, depending on the solubilities of the compounds in the selected solvent or solvent mixtures.
  • the acyl urea and the amine may be added in a stoichiomethc ratio (1 :1 ) , or a slight excess of the amine may be used, for example, between 1.01 fold and 20 fold excess, but typically about 1.01 to about 10 fold excess.
  • the amine is added to the acyl urea in tetrahydrofuran and the resulting mixture is stirred at about 0 °C to refluxing temperatures of the solvent, preferably at about 10 °C to about 50 °C, most preferably at about room temperatures under an inert atmosphere such as nitrogen.
  • the reaction mixture is maintained at the reaction temperatures until the reaction proceeds to completion.
  • the reaction may be allowed to proceed to completion in about 10 minutes to 48 hours.
  • the reaction is stirred at room temperature for about 5 hours.
  • the resulting product may be isolated by conventional techniques.
  • the solvent and excess amine may be removed by evaporation under reduced pressure, and the residue is suspended in a solvent.
  • the solvent is water. The suspension or solid may be filtered and washed with water or a suitable solvent, and then isolated and dried using conventional methods.
  • Cyclic acyl ureas of the present invention may be prepared according to methods known in the art.
  • One method comprises the alkylation of the acyl urea nitrogens with an alkylating agent gene cally represented above as X-(CH 2 ) 2 . 4 -Y, where X and Y are leaving groups, and may be the same or different.
  • Leaving groups known in the art include halides, methanesulfonates, thfluoromethanesulfonates, p-toluenesulfonates, p-bromotoluenesulfonate, p- nitrobenzenesulfonates and the like.
  • Representative alkylating agents include
  • the acyl urea is treated with a base in an organic solvent or mixtures of solvents.
  • the base is an inorganic base such as sodium hydride, or an organic base such as dimethyl sulfoxide and sodium hydride.
  • the solvent is a polar, aprotic solvent such as tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, glycols, or mixtures of such solvents.
  • a solution or suspension of the acyl urea is slowly added to the base in an organic solvent at about 0 °C to about 25 °C, and the resulting mixture is stirred for about 10 minutes to about 5 hours, preferably about 30 minutes.
  • the alkylating agent is added and the mixture is stirred until the reaction is deemed complete.
  • Alkylation of both urea nitrogens may be accomplished in a single step, or may be accomplished sequentially in a two step procedure be exposing the partially alkylated product with the same or different base.
  • the reaction is then quenched with a solvent, preferably water, and the mixture is extracted multiple times with an organic solvent.
  • the extracting solvent is dichloromethane.
  • the combined organic extracts are washed with water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to afford the product, which may be purified using standard conditions known in the art. Purification may be performed by silica gel chromatography in a mixture of organic solvents, such as ethyl acetate and petroleum ether.
  • Example 1 Synthesis of ⁇ /-(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- ⁇ /- ⁇ [(4-chlorophenyl)amino]carbonyl ⁇ carboxamide (39).
  • Oxalyl chloride (4.5 mL of a 2M solution in dichloromethane) was added dropwise to a stirred suspension of 4-chloro-1 ,3-dimethylpyrazolo [5,4-b]pyridine-5-carboxylic acid (0.98 g) in anhydrous dichloromethane (14 mL), followed by 4 drops of ⁇ /, ⁇ /-dimethylformamide (DMF). After the evolution of gas subsided, 4 more drops of DMF were added, and this operation was repeated three more times. The mixture was stirred at room temperature for 45 min, filtered to remove trace amounts of insoluble material, and the solvent evaporated under reduced pressure.
  • Oxalyl chloride (1.4 mL of a 2M solution in dichloromethane) was added dropwise to a suspension of 4-chloro-1 ,3-dimethylpyrazolo[5,4-b] pyridine-5-carboxamide (0.41 g) in anhydrous dichloromethane (5 L). The mixture was heated at reflux for 15 h, and cooled to room temperature. The solvent was evaporated under reduced pressure and the residue was dissolved in anhydrous THF (6 mL). An aliquot of this solution (1 .5 mL) was added dropwise to an ice-cooled solution of 4-chloroaniline (57 mg) in anhydrous THF (2 mL).
  • Example 2 Preparation of ⁇ /- ⁇ [(3,4-Dichlorophenyl)amino]carbonyl ⁇ (1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl) carboxamide (2).
  • the solid was re-suspended in anhydrous dichloromethane (20 mL) and treated with a 2M solution of oxalyl chloride in dichloromethane (3.6 mL), followed by anhydrous DMF (0.1 mL). The mixture was stirred at room temperature for 1 h, filtered, and the filtrate concentrated under reduced pressure. The residue was cooled in an ice-bath and slowly treated with 28% ammonium hydroxide solution. The suspension was removed from the ice-bath, stirred at room temperature for 1 h, and filtered.
  • Example 3 Ethyl 2-[4-( ⁇ [(1 ,3-dimethypyrazolo[5,4-b]pyridin-5- yl)carbonylamino]carbonyl ⁇ amino)-2-(trifluoromethyl)phenoxy]acetate (34).
  • the precipitated solid was collected by filtration, washed with dichloromethane and dried under high vacuum.
  • the solid was re-suspended in acetone-DMF (8 mL:0.8 mL) and treated with potassium carbonate (0.18 g) and ethyl bromoacetate (0.15 mL).
  • the suspension was heated at reflux for 2 h, cooled to room temperature, and concentrated under reduced pressure.
  • the residue was treated with water (3 mL), and the resulting solids were filtered, washed with water and acetone, and dried under high vacuum to give the title compound as a white solid.
  • Example 4 Sodium 2-[4-( ⁇ [(1 ,3-Dimethypyrazolo[5,4-b]pyridin-5-yl)carbonyl- amino]carbonyl ⁇ amino)-2-(trifluoromethyl)phenoxy]acetate (35).
  • Example 7 (4- ⁇ [2-(Dimethylamino)ethyl]amino ⁇ -1 ,3-dimethylpyrazolo[5,4- b]pyridin-5-yl)-N-( ⁇ [4-fluoro-3- (trifluoromethyl)phenyl]amino ⁇ carbonyl)carboxamide (45).
  • Example 8 (6- ⁇ [2-(Dimethylamino)ethyl]methylamino ⁇ -1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-( ⁇ [4-fluoro-3-(trifluoromethyl) phenyl]amino ⁇ carbonyl)carboxamide hydrochloride (116).
  • this material (2.18 g) was dissolved in phenylphosphonic dichloride (10 mL) and stirred at 150 °C for 17 hrs. The solution was cooled to room temperature and poured into water, extracted with ethyl acetate, washed with saturated NaHCO 3 solution, and dried over Na 2 SO 4 . Evaporation of the solvent under reduced pressure produced a white solid.
  • This material (2.16 g) was dissolved in tetrahydrofuran (50 mL), dimethylaminoethylmethylamine (10.6 mL) was added at room temperature, and the mixture was heated at reflux for 20 hrs. The reaction mixture was cooled to room temperature, extracted with ethyl acetate.
  • Example 9 ⁇ 6-[2-(Diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5- yl ⁇ -N-( ⁇ [4-fluoro-3-(trifluoromethyl)phenyl]amino ⁇ carbonyl)carboxamide hydrochloride (120).
  • 6-Hydroxy-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carbonitrile (0.91 1 g) was dissolved in concentrated H 2 SO (3.0 mL) and stirred at 40 °C for 24 hrs. The solution was poured into ice-water and the precipitated solid was filtered, washed with water, and dried under high vacuum to give 6-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]py din-5-carboxamide as a white solid. A portion of this material (0.122 g) was dissolved in DMF (2.0 mL). 2-Chlorotriethylamine hydrochloride (0.192 g) and potassium carbonate (0.304 g) were added to the solution.
  • the reaction mixture was stirred at 60 °C for 18 h.
  • the reaction mixture was poured into water, extracted with ethyl acetate.
  • the extract was washed with saturated NaHCO 3 solution and the brine, and dried over Na 2 SO 4 .
  • the extract was evaporated under reduced pressure to give 6-[2- (diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carboxamide as a white solid.
  • a portion of this material (0.059 g) was dissolved in hot toluene (10 L) and azeotroped for 2 hrs. The solution was cooled to room temperature.
  • Example 10 N-( ⁇ [4-Fluoro-3-(trifluoromethyl)phenyl]amino ⁇ carbonyl)(1-methyl- 3-phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (123).
  • the solid was dissolved in acetic acid (20 mL), and reacted with zinc (2.00g) at 80 °C for 60 min. Zinc was removed by the filtration, and the filtrate was concentrated under reduced pressure.
  • the residue was dissolved in ethanol (30 mL), and treated with 5N sodium hydroxide solution (30 mL). The mixture was heated at reflux for 6 hrs, and then solvent was evaporated under reduced pressure. The residue was poured into water (40 mL), filtered, and the filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid.
  • the precipitated solid was collected by the filtration, washed with water, and dried under high vacuum to give 1-methyl-3-phenylpyrazolo[5,4- b]pyridine-5-carboxylic acid as a white powder.
  • the acid was suspended in anhydrous dichloromethane (50 mL) under an argon atmosphere and added oxalyl chloride (5.0 mL), followed by anhydrous DMF (0.5 mL). The mixture was stirred at room temperature for 6 hrs, and then concentrated under reduced pressure. The residue was cooled to ice-bath temperature and a saturated solution of ammonia in tetrahydrofuran (50 mL) was slowly added to the residue.
  • Example 11 N-( ⁇ [4-Fluoro-3-(trifluoromethyl)phenyl]amino ⁇ carbonyl)[4-(4- methylpiperazin yl)-1 ,3-diphenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide (126).
  • Ethyl 4-chloro-1 ,3-diphenylpyrazolo[5,4-b]pyhdine-5-carboxylate (900 mg) and methylpiperazine (4.50 g) were dissolved in tetrahydrofuran (90 mL) and stirred at room temperature for 6 hrs.
  • the reagents were removed under reduced pressure, the residue was dissolved in the mixture of ethanol (27 mL) and 5N sodium hydroxide solution (27 mL), and heated at reflux for 6 hrs.
  • the solvent was evaporated under reduced pressure, the residue was dissolved in water (36 mL), insoluble solids were removed by the filtration.
  • the filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum.
  • Example 13 Ethyl 1 - ⁇ 1 ,3-dimethyl-5-[N-( ⁇ [3-(1 -methylethoxy)phenyl]amino ⁇ carbonyl) carbamoyl]pyrazolo[5,4-b]pyridin-4-yl ⁇ piperidine-4-carboxylate (131). coo ⁇ t
  • Example 15 N-((r4-fluoro-3-(trifluoromethvl)phenvnamino ⁇ carbonyl)[3-methvl-4- (4-methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide (134).
  • reaction mixture was treated with 5N HCl and purified by ODS column (VARIAN MEGABOND ELUT C18) to give 3-methyl-4-(4-methyl-piperazin-1-yl) isoxazolo[5,4-b]pyridine-5-carboxylic acid as an amorphous solid.
  • a portion of this amorphous solid (417 mg) was dissolved in thionyl chloride (40 ml) and heated at 50 °C for 3.5 hrs. The mixture was concentrated under reduced pressure, the residue was dissolved in THF (20 ml), and reacted with ammonia gas at room temperature for 3 hrs.
  • a 96-well microchemotaxis chamber with a 5 ⁇ m-pore size, PVP-coated polycarbonate filter membrane (Neuro Probe Inc., Cabin John, MD) was used for testing.
  • Compounds were prepared as 10 mM stock solution in DMSO.
  • THP-1 cells (2x10 6 cells/mL) were labeled with 5 ⁇ M Calcein AM containing 0.1% F127 (Molecular Probe, Eugene, OR) at 37 °C for 30 min, and then pretreated with compound at room temperature for an additional 30 min.
  • the filter membrane was placed over the lower chamber, followed by a silicon gasket and the upper chamber.
  • the pretreated THP-1 cells (4x10 5 cells/50 ⁇ L of RPMI1640 medium per well) were added to the upper chamber and incubated in 5% CO 2 at 37 °C for 2 h.
  • the migrated cells were determined with a fluorescent plate reader (LJL BioSystems, Sunnyvale, CA).
  • Table 7 shows the IC 50 (concentration of compound that inhibited migration of 50% of the cells relative to control) for several compounds of the present invention.
  • Example 18 Thioglycollate-lnduced Inflammation Model 3% Brewer's thioglycollate broth (Difco, Detroit, Ml) was injected into the peritoneal cavity of ICR male mice, followed by subcutaneous administration of the test compound. The same dose of test compound was again administered immediately after the broth injection and 3h later. After 72 h, the number of total elicited cells and MOMA2- positive cells in the peritoneal cavity was analyzed using an EPICS XL Beckman Coulter. The results are shown in Table 9.
  • Example 19 Anti-Thy-1 Antibody-Induced Nephritis Model The efficacy of the compounds of the present invention was also evaluated in an animal model of nephritis. This model very closely simulates the conditions found in human mesangial proliferative glomerulonephritis.
  • Anti-Thy-1 nephritis was induced by intravenous injection of anti-Thy-1 -antibody to male Wistar rats.
  • the test compound was subcutaneously administered 2 h before, immediately after, and 5 h after the anti-Thy-1 antibody treatment, and then twice a day for the following 2 days.
  • Anti-MCP-1 antibody was intraperitoneally injected once a day for 3 days. Seven days after the anti-Thy-1 antibody treatment, the rats were sacrificed. The kidneys were perfused with 10% formaldehyde in PBS, surgically removed, and immersed in 10% formaldehyde.
  • the kidneys were then embedded in paraffin for glomerular histopathology or in OCT compound (Miles Inc., Elkhart, IN) in liquid nitrogen after immersion in 30% sucrose overnight. Immunohistochemical staining was performed with a mouse anti-rat ED-1 monoclonal antibody. Briefly, 5 ⁇ m renal sections were prepared and endogenous peroxidase blocked with 0.3% hydrogen peroxide. The sections were then blocked with Protein Block (DAKO, Japan) and stained with anti-ED-1 antibody for 45 min. The ED-1 antigen was visualized by peroxidase-labeled anti-mouse IgG and diaminobenzidine. The amount of urinary protein was determined with the DC protein assay kit (Bio-Rad, Hercules, CA). The effect of a representative test compound on the level of urinary protein excretion and the infiltration of ED-1 positive cells into the glomeruli is shown in Table 10.
  • Apolipoprotein E is a component of several plasma lipoproteins, including chylomicrons, VLDL, and HDL. Receptor-mediated catabolism of these lipoprotein particles is mediated through the interaction of apoE with the LDL receptor (LDLR) or with LDLR-related protein (LRP). ApoE-deficient mice exhibit hypercholesterolemia and develop complex atheromatous lesions similar to those seen in humans. The efficacy of the compounds of the present invention was also evaluated using this animal model.
  • LDLR LDL receptor
  • LRP LDLR-related protein
  • mice Male, 4 week-old apoE-deficient mice were fed on high-fat diet (15% fat, 1.25% cholesterol). The test compound was administered as food admixture for 8 weeks. At 12 week-old, the mice were fasted for 4 hours and then sacrificed under ether anesthesia. Blood was collected in the presence of heparin, and the hearts were perfused in situ with PBS (pH 7.4), followed by 4% paraformaldehyde for 5 min.
  • PBS pH 7.4
  • the hearts were embedded in OCT compound and sectioned at 10 ⁇ m using cryostat.
  • the sections were stained with oil red O.
  • Each section of the aortic valve was evaluated for oil red O staining by capturing images directly from a RGB camera attached to a light microscope; image analysis was performed with the IPAP-WIN software (Sumika Tekuno, Japan).
  • Five sections were examined for each animal, and the sum of the lesion areas was calculated and expressed as the percent of the total cross-sectional wall area.
  • Total cholesterol was determined with a Determiner assay kit (Kyowa medex, Japan).
  • Compound H reduced the percent of atherosclerotic lesion area and this reduction was statistically significant. There were no significant differences in plasma total cholesterol level and body weight between the control and the test compound-treated groups (data not shown).
  • a 48-well microchemotaxis chamber with a 5 ⁇ m-pore size, PVP-coated polycarbonate filter membrane (Neuro Probe Inc., Cabin John, MD) was used for testing.
  • Compounds were prepared as 10 mM stock solution in DMSO.
  • THP-1 cells were washed with RPMI 1640 medium supplemented with 0.5% BSA and 25 mM HEPES, pH 7.4, and suspended at a density of 4x10 6 cells/mL in the same medium.
  • a 150 ⁇ L aliquot of this suspension was treated with an equal volume of test compound solution and the mixture incubated at 37 °C for 15 min.
  • the lower chamber was loaded with 26 ⁇ L of a 2.5 nM solution of hMCP-1 (PeproTech) in medium.
  • the filter membrane was placed over the lower chamber, followed by a silicon rubber gasket and the upper chamber.
  • a 50 ⁇ L aliquot of the THP-1 cell suspension containing the test compound was added to the upper chamber and the assembly incubated in a 5% CO 2 atmosphere at 37 °C for 2 hr.
  • the chamber was then disassembled and the cells remaining on the upper surface of the filter were scrapped off with a rubber scrapper.
  • the filter was fixed with methanol, stained with Diff-Quik solution, and mounted on a slide glass. The cells that had migrated across the filter onto the lower surface were then counted by microscopic observation.
  • Table 12 shows the IC 50 (concentration of compound that inhibited migration of 50% of the cells relative to control) for several compounds of the present invention.
  • Table 12 Effect of Selected Compounds on MCP-1-lnduced Chemotaxis
  • CFA Complete Freunt's adjuvant
  • Collagen-induced arthritis was induced in female Lewis rats as follows. Rats were injected subcutaneously with the emulsion of bovine collagen type II (3.3 mg/kg at day 0 and 1.67 mg/kg at day 7) and complete Freund's adjuvant (CFA) at the base of the tail at day 0 and immunized additionally at day 7. The test compounds were administered i.p. at day 0 and then once a day for 18 days or p.o. (methotrexate) at day 0 and then 3 times a week until day 18. Paw swelling was measured by the increased volume of the legs using a plethysmometer. Table 14 shows the change in the swelling expressed as percent swelling relative to the volume of legs on day 0. Table 14. Effect of Compounds on the Collagen-induced Arthritis Model
  • Collagen-induced arthritis was induced in female Lewis rats as follows. Rats were injected subcutaneously with the emulsion of bovine collagen type II (3.3mg/kg at day 0 and 1.67mg/kg at day 7) and complete Freund's adjuvant (CFA) at the base of the tail at day 0 and immunized additionally at day 7. The test compounds were administered p.o. at day 0 and then twice a day for 21 days or 3 times a week until day 21 was also administered p.o. from day 13 to day 21 in another group. Paw swelling was measured by the increased volume of the legs using a plethysmometer. Table 15 shows the change in the swelling expressed as percent swelling relative to the volume of legs on day 0.
  • Example 25 Restenosis Model in Rat Effects of MCP-1 antagonists on rat carotid artery responses to the balloon catheter injury.
  • Anti-Thy-1 nephritis was induced by intravenous injection of anti-Thy-1 -antibody to male Wistar rats.
  • the test compound was per os administered 2 h before, immediately after, and 5 h after the anti-Thy-1 antibody treatment, and then twice a day for the following 2 days. Seven days after the anti-Thy-1 antibody treatment, the rats were sacrificed.
  • the amount of urinary protein was determined with the DC protein assay kit (Bio-Rad, Hercules, CA). The effect of a representative test compound on the level of urinary protein excretion is shown in Table 17.
  • Example 27 Oral pharmaceutical composition - Solid dosage formulation
  • a pharmaceutical composition for oral administration may be prepared by combining the following:
  • the mixture may be compressed to tablets, or filled into hard gelatin capsules.
  • the tablet may be coated by applying a suspension of film former
  • the film coat can comprise 2.0% to 6.0% of the tablet weight, preferably about 3.0%.
  • Example 28 Oral pharmaceutical composition preparation - Capsule
  • a pharmaceutical composition of a compound of the invention suitable for oral administration may also be prepared by combining the following:
  • Example 29 Pharmaceutical composition for parenteral administration
  • a pharmaceutical composition for parenteral administration may be prepared by combining the following:
  • the solution is sterilized and sealed in sterile containers.

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Abstract

The present invention relates to chemical compounds of Formula (I) or (II) (wherein Y, Z and R1 - R6 are defined herein), pharmaceutical compositions comprising said compounds, uses of said compounds and compositions, methods of treatment employing said compounds and compositions, and processes for preparing said compounds. Specifically, this invention relates to novel compounds which are antagonists of MCP-1 function and are useful in the prevention or treatment of chronic or acute inflammatory or autoimmune diseases. More specifically, the invention is related to pharmaceutical compositions comprising these compounds and the use of these compounds and compositions in the prevention or treatment of such diseases.

Description

ANTAGONISTS OF MCP-1 FUNCTION AND METHODS OF USE THEREOF
FIELD OF THE INVENTION The present invention relates to chemical compounds, pharmaceutical compositions comprising said compounds, uses of said compounds and compositions, methods of treatment employing said compounds and compositions, and processes for preparing said compounds. Specifically, this invention relates to novel compounds which are antagonists of MCP-1 function and are useful in the prevention or treatment of chronic or acute inflammatory or autoimmune diseases, especially those associated with aberrant lymphocyte or monocyte accumulation such as arthritis, asthma, atherosclerosis, diabetic nephropathy, inflammatory bowel disease, Crohn's disease, multiple sclerosis, nephritis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, and transplant rejection. More specifically, the invention is related to pharmaceutical compositions comprising these compounds and the use of these compounds and compositions in the prevention or treatment of such diseases.
BACKGROUND OF THE INVENTION
Chemokines: Structure and Function
The migration of leukocytes from blood vessels into diseased tissues is an important process in the initiation of normal inflammatory responses to certain stimuli or insults to the immune system. However, this process is also involved in the onset and progression of life-threatening inflammatory and autoimmune diseases; blocking leukocyte recruitment in these disease states, therefore, can be an effective therapeutic strategy.
The mechanism by which leukocytes leave the bloodstream and accumulate at inflammatory sites involves three distinct steps: (1 ) rolling, (2) arrest and firm adhesion, and (3) transendothelial migration [Springer, Nature 346:425-433 (1990); Lawrence and Springer, Cell 65:859-873 (1991 ); Butcher, Cell 67:1033-1036 (1991 )]. The second step is mediated at the molecular level by chemoattractant receptors on the surface of leukocytes which bind chemoattractant cytokines secreted by proinflammatory cells at the site of damage or infection. Receptor binding activates leukocytes, increases their adhesiveness to the endothelium, and promotes their transmigration into the affected tissue, where they can secrete inflammatory and chemoattractant cytokines and degradative proteases that act on the subendothelial matrix, facilitating the migration of additional leukocytes to the site of injury.
The chemoattractant cytokines, collectively known as "chemokines," are a large family of low molecular weight (8 to 10 kD) proteins that share the ability to stimulate directed cell migration ("chemotaxis") [Schall, Cytokine 3:165-183 (1991 ); Murphy, Rev Immun 12:593-633 (1994)].
Chemokines are characterized by the presence of four conserved cysteine residues and are grouped into two main subfamilies based on whether the two amino-terminal cysteines are separated by one amino acid (CXC subfamily, also known as α-chemokines) or immediately adjacent to each other (CC subfamily, also referred to as β-chemokines) [Baggiolini et al., Adv Immunol 55:97-179 (1994); Baggiolini et al., Annu Rev Immunol 15:675-705 (1997); Deng et al., Nature 381 :661-666 (1996); Luster, New Engl J Med 338:436445 (1998); Saunders and Tarby, Drug Discovery Today 4:80-92 (1999)].
The chemokines of the CXC subfamily, represented by IL-8, are produced by a wide range of cells and act predominantly on neutrophils as mediators of acute inflammation. The CC chemokines, which include MCP-1 , RANTES, MIP-1 α, and MIP-1 β, are also produced by a variety of cells, but these molecules act mainly on monocytes and lymphocytes in chronic inflammation.
Like many cytokines and growth factors, chemokines utilize both high and low affinity interactions to elicit full biological activity. Studies performed with labeled ligands have identified chemokine binding sites ("receptors") on the surface of neutrophils, monocytes, T cells, and eosinophils with affinities in the 500 pM to 10 nM range [Kelvin et al., J Leukoc Biol 54:604-612 (1993); Murphy, Annu Rev Immunol 12:593-633 (1994); Raport et al., J Leukoc Biol 59:18-23 (1996); Premack and Schall, Nature Med 2:1174-1178 (1996)]. The cloning of these receptors has revealed that cell surface high-affinity chemokine receptors belong to the seven transmembrane ("serpentine") G-protein-coupled receptor (GPCR) superfamily.
Chemokine receptors are expressed on different cell types, including non-leukocyte cells. Some receptors are restricted to certain cells (e.g., the
CXCR1 receptor is predominantly restricted to neutrophils), whereas others are more widely expressed (e.g., the CCR2 receptor is expressed on monocytes, T cells, natural killer cells, dendritic cells, and basophils).
Given that at least twice as many chemokines have been reported to date as there are receptors, there is a high degree of redundancy in the ligands and, not surprisingly, most chemokine receptors are rather promiscuous with regard to their binding partners. For example, both MIP-1α and RANTES bind to the CCR1 and CCR4 receptors, while MCP-1 binds to the CCR2 and CCR4 receptors. Although most chemokines receptors bind more than one chemokine, CC receptors bind only CC chemokines, and CXC receptors bind only CXC chemokines. This ligand-receptor restriction may be related to the structural differences between CC and CXC chemokines, which have similar primary, secondary, and tertiary structures, but different quaternary structures [Lodi et al., Science 263:1762-1767 (1994)]. The binding of chemokines to their serpentine receptors is transduced into a variety of biochemical and physiological changes, including inhibition of cAMP synthesis, stimulation of cytosolic calcium influx, upregulation or activation of adhesion proteins, receptor desensitization and internalization, and cytoskeletal rearrangements leading to chemotaxis [Vaddi et al., J Immunol 153:4721-4732 (1994); Szabo et al., Eur J Immunol 27:1061-1068 (1997); Campbell et al., Science 279:381-384 (1998); Aragay et al., Proc Natl Acad Sci USA 95:2985- 2990 (1998); Franci et al., J Immunol 157:5606-5612 (1996); Aramori et al., EMBO J 16:4606-4616 (1997); Haribabu et al., J Biol Chem 272:28726-28731 (1997); Newton et al., Methods Enzymol 287:174-186 (1997)]. In the case of macrophages and neutrophils, chemokine binding also triggers cellular activation, resulting in lysozomal enzyme release and generation of toxic products from the respiratory burst [Newton et al., Methods Enzymol 287:174- 186 (1997); Zachariae et al., J Exp Med 171 :2177-2182 (1990); Vaddi et al., J Leukocyte Biol 55:756-762 (1994)]. The molecular details of the chemokine- receptor interactions responsible for inducing signal transduction, as well as the specific pathways that link binding to the above-mentioned physiological changes, are still being elucidated. Notwithstanding the complexity of these events, it has been shown that in the case of the MCP-1 /CCR2 interaction, specific molecular features of MCP-1 can induce different conformations in CCR2 that are coupled to separate post-receptor pathways [Jarnagin et al., Biochemistry 38:16167-16177 (1999)]. Thus, it should be possible to identify ligands that inhibit chemotaxis without affecting other signaling events.
In addition to their high-affinity seven transmembrane GPCRs, chemokines of both subfamilies bind to various extracellular matrix proteins such as the glycosaminoglycans (GAGs) heparin, chondroitin sulfate, heparan sulfate, and dermatan sulfate with affinities in the middle nanomolar to millimolar range. These low-affinity chemokine-GAG interactions are believed to be critical not only for conformational activation of the ligands and presentation to their high-affinity serpentine receptors, but also for the induction of stable chemokine gradients that may function to stimulate haptotaxis (i.e., the migration of specific cell subtypes in response to a ligand gradient that is affixed upon the surface of endothelial cells or embedded within the extracellular matrix) [Witt and Lander, Curr Biol 4:394-400 (1994); Rot, Eur J Immunol 23:303-306 (1993); Webb et al., Proc Natl Acad Sci USA 90:7158-7162 (1993); Tanaka et al, Nature 361 :79-82 (1993); Gilat et al., J Immunol 153:4899-4906 (1994)]. Similar ligand-GAG interactions have been described for a variety of cytokines and growth factors, including the various members of the FGF family, hepatocyte growth factor, IL-3 and IL-7, GM-CSF, and VEGF [Roberts et al., Nature 332:376-378 (1988); Gilat et al., Immunol Today 17:16-20 (1996); Clarke et al., Cytokine 7:325-330 (1995); Miao et al., J Biol Chem 271 :4879-4886 (1996); Vlodavsky et al., Cancer Metastasis Rev 15:177-186 (1996)]. MCP-1 and Diseases
Chemokines have been implicated as important mediators of allergic, inflammatory and autoimmune disorders and diseases, such as asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, and transplant rejection. Accordingly, it has been postulated that the use of antagonists of chemokine function may help reverse or halt the progression of these disorders and diseases.
In particular, elevated expression of MCP-1 has been observed in a number of chronic inflammatory diseases [Proost et al., Int J Clin Lab Res 26:211-223 (1996); Taub, D. D. Cytokine Growth Factor Rev 7:355-376 (1996)] including rheumatoid arthritis [Robinson et al., Clin Exp Immunol 101 :398-407 (1995); Hosaka et al., ibid. 97:451-457 (1994); Koch et al., J Clin Invest 90:772- 779 (1992); Villiger et al., J Immunol 149:722-727 (1992)], asthma [Hsieh et al., J Allergy Clin Immunol 98:580-587 (1996); Alam et al., Am J Respir Crit Care Med 153:1398-1404 (1996); Kurashima et al., J Leukocyte Biol 59:313-316 (1996); Sugiyama et al., Eur Respir J 8:1084-1090 (1995)], and atherosclerosis [Yla-Herttuala et al., Proc Natl Acad Sci USA 88:5252-5256 (1991 ); Nelken et al., J Clin Invest 88:1121-1127 (1991 )].
MCP-1 appears to play a significant role during the early stages of allergic responses because of its ability to induce mast cell activation and LTC4 release into the airway, which directly induces AHR (airways hyper-responsiveness) [Campbell et al., J Immunol 163:2160-2167 (1999)].
MCP-1 has been found in the lungs of patients with idiopathic pulmonary fibrosis and is thought to be responsible for the influx of mononuclear phagocytes and the production of growth factors that stimulate mesenchymal cells and subsequent fibrosis [Antoniades et al., Proc Natl Acad Sci USA 89:5371-5375 (1992)]. In addition, MCP-1 is also involved in the accumulation of monocytes in pleural effusions which is associated with both Mycobacterium tuberculosis infection and malignancy [Strieter et al., J Lab Clin Med 123:183-197 (1994)].
MCP-1 has also been shown to be constitutively expressed by synovial fibroblasts from rheumatoid arthritis patients, and its levels are higher in rheumatoid arthritis joints compared to normal joints or those from other arthritic diseases [Koch et al., J Clin Invest 90:772-779 (1992)]. These elevated levels of MCP-1 are probably responsible for the monocyte infiltration into the synovial tissue. Increased levels of synovial MIP-1α and RANTES have also been detected in patients with rheumatoid arthritis [Kundel et al., J Leukocyte Biol 59:6-12 (1996)].
MCP-1 also plays a critical role in the initiation and development of atherosclerotic lesions. MCP-1 is responsible for the recruitment of monocytes into atherosclerotic areas, as shown by immunohistochemistry of macrophage-rich arterial wall [Yla-Herttuala et al., Proc Natl Acad Sci USA 88:5252-5256 (1991 ); Nelken et al., J Clin Invest 88:1 121 -1 127 (1991 )] and anti-MCP-1 antibody detection [Takeya et al., Human Pathol 24:534-539 (1993)]. LDL-receptor/MCP-1 -deficient and apoB-transgenic/MCP-1 -deficient mice show significantly less lipid deposition and macrophage accumulation throughout their aortas compared with wild-type MCP-1 strains [Alcami et al., J Immunol 160:624- 633 (1998); Gosling et al., J Clin Invest 103:773-778 (1999); Gu et al., Mol. Cell. 2:275-281 (1998); Boring et al., Nature 394:894-897 (1998).
Other inflammatory diseases marked by specific site elevations of MCP-1 include multiple sclerosis (MS), glomerulonephritis, and stroke. These findings suggest that the discovery of compounds that block MCP-
1 activity would be beneficial in treating inflammatory diseases.
Antagonists of Chemokine Function
Most chemokine antagonists reported to date are either neutralizing antibodies to specific chemokines or receptor-ligand antagonists, that is, agents that compete with specific chemokines for binding to their cognate serpentine receptors but, unlike the chemokines themselves, do not activate these receptors towards eliciting a functional response [Howard et al., Trend Biotechnol ! 4:46-51 (1996)].
The use of specific anti-chemokine antibodies has been shown to curtail inflammation in a number of animal models (e.g., anti-MIP-1α in bleomycin- induced pulmonary fibrosis [Smith et al., Leukocyte Biol 57:782-787 (1994)]; anti-IL-8 in reperfusion injury [Sekido et al., Nature 365:654-657 (1995)], and anti-MCP-1 in a rat model of glomerulonephritis [Wada et al., FASEB J 10:1418-1425 (1996)]). In the MRL-/pr mouse arthritis model, administration of an MCP-1 antagonist significantly reduced the overall histopathological score after the early onset of the disease [Gong et al., J Exp Med 186: 131 -137 (1997)]. A major problem associated with using antibodies to antagonize chemokine function is that they must be humanized before use in chronic human diseases. Furthermore, the ability of multiple chemokines to bind and activate a single receptor forces the development of a multiple antibody strategy or the use of cross-reactive antibodies in order to completely block or prevent pathological conditions.
Several small molecule antagonists of chemokine receptor function have been reported in the scientific and patent literature [White, J. Biol Chem 273:10095-10098 (1998); Hesselgesser, J. Biol Chem 273:15687-15692 (1998); Bright et al., Bioorg Med Chem Lett 8:771-774 (1998); Lapierre, 26th Natl Med Chem Symposium, June 14-18, Richmond (VA), USA (1998); Forbes et al., Bioorg Med Chem Lett 10:1803-18064 (2000); Kato et al., WO Patent 97/24325; Shiota et al., WO Patent 97/44329; Naya et al., WO Patent 98/04554; Takeda Industries, JP Patent 0955572 (1998); Schwender et al., WO Patent 98/02151 ; Hagmann et al., WO Patent 98/27815; Connor et al., WO Patent 98/06703; Wellington et al., US Patent 6,288,103 B1 (2001 )].
The specificity of the chemokine receptor antagonists, however, suggests that inflammatory disorders characterized by multiple or redundant chemokine expression profiles will be relatively more refractory to treatment by these agents.
A different approach to target chemokine function would involve the use of compounds that disrupt the chemokine-GAG interaction. One class of such agents with potential therapeutic application would consist of small organic molecules that bind to the chemokine low affinity GAG-binding domain. Compounds of this class might not inhibit binding of the chemokine to its high-affinity receptor per se, but would disrupt chemokine localization within the extracellular matrix and provide an effective block for directed leukocyte-taxis within tissues. An advantage of this strategy is the fact that most CC and CXC chemokines possess similar C-terminal protein folding domains that define the GAG-binding site, and, hence, such compounds would be more useful for the treatment of inflammatory disorders induced by multiple, functionally redundant chemokines [McFadden and Kelvin, Biochem Pharmacol 54: 1271 -1280 (1997)]. The use of small molecule drugs to bind cytokine ligands and disrupt interactions with extracellular GAGs has been reported with FGF-dependent angiogenesis [Folkman and Shing, Adv Exp Med Biol 313:355-364 (1992)]. For example, the heparinoids suramin and pentosan polysulphate both inhibit angiogenesis under conditions where heparin is either ineffective or even stimulatory [Wellstein and Czubayko, Breast Cancer Res Treat 38:109-119 (1996)]. In the case of suramin, the anti-angiogenic capacity of the drug has also been shown to be targeted against VEGF [Waltenberger et al., J Mol Cell Cardiol 28:1523-1529 (1996)] which, like FGF, possesses heparin-binding domains similar to those of the chemokines. Heparin or heparin sulphate has also been shown to directly compete for GAG interactions critical for T-cell adhesion mediated by MIP-1β in vitro [Tanaka et al., Nature 361:79-82 (1993)].
SUMMARY OF THE INVENTION The present invention relates to compounds that inhibit MCP-1 -induced chemotaxis of human monocytic cells both in vitro and in vivo. These novel MCP-1 antagonists are useful for the treatment of inflammatory diseases, especially those associated with lymphocyte and/or monocyte accumulation, such as atherosclerosis, diabetic nephropathy, inflammatory bowel disease, Crohn's disease, multiple sclerosis, nephritis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, and other chronic or acute autoimmune disorders. In addition, these compounds can be used in the treatment of allergic hypersensitivity disorders, such as asthma and allergic rhinitis, characterized by basophil activation and eosinophil recruitment. A first embodiment of the present invention provides compounds of
Formula (I) or Formula (II):
Figure imgf000010_0001
(") (II) wherein
Y is O, S or N-R7,
Z is N or C-R8,
R1 R2, R3, and R8 are independently, hydrogen, or optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR9, -SR9,
-NR9R10, -NR9(carboxy(lower alkyl)), -C(=O)R9, -C(=O)OR9
C(=O)NR ι9a DR1,0u, -OC(=O)Ra, -SO2Ra, -OSO2Ra -SO2NR9R10,
-NR9SO2R10 or
Figure imgf000010_0002
wherein Ra and R 1ι0υ are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(d-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl(lower alkyl), aryl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι.2 alkyl) group,
R' is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -SO2R9, or -SO2NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl (lower alkyl), aryl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and
R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH,
N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-
(optionally substituted Cι-2 alkyl) group,
R4 and R5 are independently, hydrogen, lower alkyl optionally substituted lower alkyl, optionally substituted aryl, or optionally substituted aryl(lower alkyl), or, together, are -(CH2)2.4-,
R6 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), -C(=O)R11, -C(=O)OR11,
-C(=O)NR11R12, -SO2R11, or -SO2NR11R12, wherein R11 and R12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R11 and
R12 together are -(CH2)4.6-, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
The compounds of this invention may possess one or more chiral centers, and can therefore be produced as individual stereoisomers or as mixtures of stereoisomers, depending on whether individual stereoisomers or mixtures of stereoisomers of the starting materials are used. In addition, some of the compounds of the invention are capable of further forming pharmaceutically acceptable salts and esters. The compounds of this invention may further exist in tautomeric forms and can therefore be produced as individual tautomeric forms or as mixtures of tautomeric forms. Unless indicated otherwise, the description or naming of a compound or groups of compounds is intended to include both the individual isomers or mixtures (racemic or otherwise) of stereoisomers and their tautomeric forms. Methods for the determination of stereochemistry and the separation of stereoisomers are well known to a person of ordinary skill in the art [see the discussion in Chapter 4 of March J.: Advanced Organic Chemistry, 4th ed. John Wiley and Sons, New York, NY, 1992]. All of these stereoisomers and pharmaceutical forms are intended to be included within the scope of the present invention.
A second embodiment of the present invention provides compounds of Formula (la) or Formula (lla):
Figure imgf000012_0001
(la)
(lla)
where:
Y is O, S or N-R7,
Z is N or C-R8,
R1, R2, R3, R4, R5, R7 and R8 are as defined in the first embodiment,
R13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, -NR15SO2R16 or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl) or, together, are -(CH2)4-6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group, each R14 is independently selected from optionally substituted lower alkyl, optionally substituted aryl, optionally substituted heteroaryl, hydroxy, halogen, -CF3, -OR17, -NR17R18, -C(=O)R18, -C(=O)OR18, -C(=O)NR17R18, wherein R17 and R18 are independently, hydrogen, lower alkyl, alkenyl, alkynyl, -CF3, optionally substituted heterocycloalkyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or, together, are -(CH2) .6-, optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group, and where n is an integer of 0 to 4, or a pharmaceutically acceptable salt thereof as a single stereoisomer or mixture of stereoisomers. A third embodiment of the present invention provides pharmaceutical compositions comprising pharmaceutically acceptable excipients and a therapeutically effective amount of at least one compound of this invention. A fourth embodiment of the present invention provides methods for treating chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection in mammals in need thereof, comprising the administration to such mammal of a therapeutically effective amount of at least one compound of this invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.
A fifth embodiment of this invention provides uses of the compounds of the invention in the preparation of medicaments for the treatment of chronic or acute inflammatory or autoimmune diseases such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a transplant rejection.
A sixth embodiment of this invention provides a process for the preparation of the compounds of the invention or pharmaceutically acceptable salts thereof.
DETAILED DESCRIPTION OF THE INVENTION
Definitions and General Parameters
The following definitions apply to the description of compounds of the present invention: "Alkyl" is a linear or branched saturated hydrocarbon radical having from 1 to 20 carbon atoms. Examples of alkyl radicals are: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, dodecyl, etc.
"Lower alkyl", as in "lower alkyl," "lower alkoxy," "cycloalkyl(lower alkyl)," "aryl(lower alkyl)", or "heteroaryl(lower alkyl)", means a C-ι-10 alkyl radical. Preferred lower alkyl radicals are those having from 1 to 6 carbon atoms.
"Alkenyl" is a linear or branched hydrocarbon radical having from 2 to 20 carbon atoms and at least one carbon-carbon double bond. Examples of alkenyl radicals are: vinyl, 1-propenyl, isobutenyl, etc.
"Alkynyl" is a linear or branched hydrocarbon radical having from 2 to 20 carbon atoms and at least one carbon-carbon triple bond. Examples of alkynyl radicals are: propargyl, 1-butynyl, etc.
"Cycloalkyl" is a monovalent cyclic hydrocarbon radical having from 3 to 12 carbon atoms. Examples of cycloalkyl radicals are: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. "Substituted cycloalkyl" is a monovalent cyclic hydrocarbon radical having from 3 to 12 carbon atoms, which is substituted with one, two, or three substituents each independently selected from aryl, substituted aryl, heteroaryl, halogen, -CF3, nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR\ -NRSO2R', -C(=O)NRR', -NRC(=O)R' or -PO3HR, wherein R and R' are, independently, hydrogen, lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl (lower alkyl), and having 3 to 12 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic rings (e.g. piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolizinyl, 1 ,4- diazaperhydroepinyl, etc.).
"Cycloalkyl(lower alkyl)" is a lower alkyl radical which is substituted with a cycloalkyl, as previously defined. Examples of cycloalkyl(lower alkyl) radicals are cyclopropylmethyl, cyclobutylethyl, cyclopentylmethyl, cyclohexyl methyl, etc. "Heterocycloalkyl" is a monovalent cyclic hydrocarbon radical having 3 to
12 carbon ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic rings (e.g. piperidyl, 4-morpholyl, 4- piperazinyl, pyrrolidinyl, perhydropyrrolizinyl, 1 ,4-diazaperhydroepinyl, etc.). "Substituted heterocycloalkyl" is a monovalent cyclic hydrocarbon radical having from 3 to 12 carbon atoms, which is substituted with one, two, or three substituents each independently selected from aryl, substituted aryl, heteroaryl, halogen, -CF3 nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -NRSO2R', -C(=O)NRR', -NRC(=O)R' or -PO3HR, wherein R and R' are, independently, hydrogen, lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl (lower alkyl), and having 3 to 12 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic rings (e.g. piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolizinyl, 1 ,4- diazaperhydroepinyl, etc.).
"Substituted heterocycloalkyl(lower alkyl)" is a lower alkyl radical which is substituted with a monovalent cyclic hydrocarbon radical having from 3 to 12 carbon atoms, which is substituted with one, two, or three substituents each independently selected from aryl, substituted aryl, heteroaryl, halogen, -CF3 nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -NRSO2R', -C(=O)NRR', -NRC(=O)R' or -PO3HR, wherein R and R' are, independently, hydrogen, lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl (lower alkyl), and having 3 to 12 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic rings (e.g. piperidyl, 4- morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolizinyl, 1 ,4- diazaperhydroepinyl, etc.).
"Substituted alkyl" or "substituted lower alkyl," is an alkyl or lower alkyl radical, respectively, which is substituted with one, two, or three substituents each independently selected from aryl, substituted aryl, heteroaryl, halogen, -CF3 nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -NRSO2R', -C(=O)NRR', -NRC(=O)R', or -PO3HR, wherein R and R' are, independently, hydrogen, lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl(lower alkyl).
"Halo(lower alkyl)" is a radical derived from lower alkyl containing at least one halogen substituent. Non-limiting examples of halo(lower alkyl) radicals include: -CF3, C2F5, etc..
"Aryl", as in "aryl", "aryloxy", and "aryl(lower alkyl)", is a radical derived from an aromatic hydrocarbon containing 6 to 16 ring carbon atoms, having a single ring (e.g., phenyl), or two or more condensed rings, preferably 2 to 3 condensed rings (e.g., naphthyl), or two or more aromatic rings, preferably 2 to 3 aromatic rings, which are linked by a single bond (e.g., biphenyl). Preferred aryl radicals are those containing from 6 to 14 carbon atoms. "Substituted aryl" is an aryl radical which is substituted with one, two, or three substituents each independently selected from alkyl, substituted alkyl, halo(lower alkyl), halogen, nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -PO3H2, -NRSO2R', -C(=O)NRR' or -NRC(=O)R', wherein R and R' are, independently, hydrogen, lower alkyl, substituted lower alkyl, cycloalkyl, aryl, substituted aryl, optionally substituted aryl(lower alkyl), heteroaryl, or heteroaryl(lower alkyl). Preferred substituted aryl radicals are those substituted with one, two, or three substituents each independently selected from the group consisting of lower alkyl, halogen, -CF3, nitro, -CN, -OR, -NRR', -C(=O)NRR', -SO2OR, -SO2NRR', -PO3H2, -NRSO2R' or -NRC(=O)R\
"Substituted aryloxy" is an -OR radical where R is a radical derived from an aromatic hydrocarbon containing 6 to 16 ring carbon atoms, having a single ring (e.g., phenyl), or two or more condensed rings, preferably 2 to 3 condensed rings (e.g., naphthyl), or two or more aromatic rings, preferably 2 to 3 aromatic rings, which are linked by a single bond (e.g., biphenyl), and which is substituted with one, two, or three substituents each independently selected from aryl, substituted aryl, heteroaryl, halogen, -CF3, nitro, -CN, -OR, -SR, -NRR', -C(=O)R, -OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -NRSO2R', -C(=O)NRR', -NRC(=O)R' or -PO3HR, wherein R and R' are, independently, hydrogen, lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl (lower alkyl). Preferred aryl radicals are those containing from 6 to 14 carbon atoms.
"Heteroaryl", as in "heteroaryl" and "heteroaryl(lower alkyl)", is a radical derived from an aromatic hydrocarbon containing 5 to 14 ring atoms, 1 to 5 of which are heteroatoms chosen, independently, from N, O, or S, and includes monocyclic, condensed heterocyclic, and condensed carbocyclic and heterocyclic aromatic rings (e.g., thienyl, furyl, pyrrolyl, pyrimidinyl, isoxazolyl, oxazolyl, indolyl, isobenzofuranyl, purinyl, isoquinolyl, pteridinyl, imidazolyl, pyridyl, pyrazolyl, pyrazinyl, quinolyl, etc.).
"Substituted heteroaryl" is a heteroaryl radical which is substituted with one, two, or three substituents each independently selected from alkyl, substituted alkyl, halogen, CF3, nitro, -CN, -OR, -SR, -NRR', -C(=O)R, OC(=O)R, -C(=O)OR, -SO2OR, -OSO2R, -SO2NRR', -NRSO2R', -C(=O)NRR', or -NRC(=O)R', wherein R and R' are, independently, hydrogen, lower alkyl, substituted lower alkyl, cycloalkyl, aryl, substituted aryl, aryl(lower alkyl), substituted aryl(lower alkyl), heteroaryl, or heteroaryl(lower alkyl). Particularly preferred substituents on the substituted heteroaryl moiety include lower alkyl, substituted lower alkyl, halo(lower alkyl), halogen, nitro, -CN, -OR, -SR, and -NRR'. "Aryl(lower alkyl)" is a lower alkyl radical which is substituted with an aryl, as previously defined.
"Substituted aryl(lower alkyl)" is an aryl(lower alkyl) radical having one to three substituents on either or both of the aryl and the alkyl portion of the radical. "Heteroaryl(lower alkyl)" is a lower alkyl radical which is substituted with a heteroaryl, as previously defined.
"Substituted heteroaryl(lower alkyl)" is a heteroaryl(lower alkyl) radical having one to three substituents on the heteroaryl portion or the alkyl portion of the radical, or both. "Lower alkoxy" is an -OR radical, where R is a lower alkyl or cycloalkyl.
"Halogen" means fluoro, chloro, bromo, or iodo.
"Stereoisomers" are compounds that have the same sequence of covalent bonds and differ in the relative disposition of their atoms in space.
"Inner salts" or "Zwitterions" can be formed by transferring a proton from the carboxyl group onto the lone pair of electrons of the nitrogen atom in the amino group.
"Tautomers" are isomeric compounds that differ from one another by interchanged positions of σ and π bonds. The compounds are in equilibrium with one another. They may also differ from one another in the position at which a hydrogen atom is attached.
"Pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients may be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.
"Pharmaceutically acceptable salts and esters" means any salt and ester that is pharmaceutically acceptable and has the desired pharmacological properties. Such salts include salts that may be derived from an inorganic or organic acid, or an inorganic or organic base, including amino acids, which is not toxic or undesirable in any way. Suitable inorganic salts include those formed with the alkali metals, e.g. sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include those formed with organic bases such as the amine bases, e.g. ethanolamine, diethanolamine, triethanolamine, tromethamine, Λ/-methylglucamine, and the like. Such salts also include acid addition salts formed with inorganic acids (e.g. hydrochloric and hydrobromic acids) and organic acids (e.g. acetic acid, citric acid, maleic acid, and the alkane and arene-sulfonic acids such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the compounds, e.g. C1-6 alkyl esters. When there are two acidic groups present, a pharmaceutically acceptable salt or ester may be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly, where there are more than two acidic groups present, some or all of such groups can be salified or esterified.
"Therapeutically effective amount" means that amount which, when administered to a mammal for treating a disease, is sufficient to effect such treatment for the disease. "Treating" or "treatment" of a disease in a mammal includes:
(1 ) Preventing the disease from occurring in a mammal which may be predisposed to the disease but does not yet experience or display symptoms of the disease;
(2) Inhibiting the disease, i.e., arresting its development, or (3) Relieving the disease, i.e., causing regression of the disease.
"Disease" includes any unhealthy condition of an animal (which includes human and non-human mammals), including particularly various forms of inflammatory illnesses or diseases, such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, immune disorders, and transplant rejection. The Compounds and Their Pharmaceutically Acceptable Salts
The first embodiment of the present invention provides compounds of Formula (I) and Formula (II):
Figure imgf000020_0001
(I) (II) wherein Y, Z, and R1 to R 2 are as defined above.
Where R and R >10 . together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N- (optionally substituted Cι-2 alkyl) group, examples include piperazinyl, 4- methylpiperazinyl, 4-morpholyl, and hexahydropyhmidyl.
Preferably, Y is O or N-R7.
Preferably, R1 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9R10,
Figure imgf000020_0002
wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), aryl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl).
More preferably, R1 is optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), aryl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl). Preferably, R2 is hydrogen, optionally substituted lower alkyl, cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9(carboxy(lower alkyl)), -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl (lower alkyl), or R9 and R10 together are -(CH2) .6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
More preferably, R2 is optionally substituted lower alkyl, cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9(carboxy(lower alkyl)), -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C1.2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl (lower alkyl), or R9 and R10 together are -(CH2) .6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
Preferably, R3 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, or -C(=O)NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl (lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2) -6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group. More preferably, R3 is optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, or -C(=O)NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
Preferably, R7 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -SO2R9, or -SO2NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι..2 alkyl)2, alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl).
Preferably, R8 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), -CF3, halogen, -OR9, -NR9R10, -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -OC(=O)R9, -SO2R9, -SO2NR9R10, -NR9SO2R10 or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, optionally substituted cycloalkyl, cycloalkyl (lower alkyl), optionally substituted aryl, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
Preferably, R4 and R5 are independently, hydrogen or lower alkyl, or together are -(CH2)2.4-. More preferably, R4 and R5 are independently, hydrogen or lower alkyl.
Preferably, R6 is hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, cycloalkyl (lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), -C(=O)R11, -C(=O)OR11, -C(=O)NR11R12, -SO2R11, or -SO2NR11R12, wherein R11 and R12 are independently, hydrogen, optionally substituted lower alkyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, heteroaryl, heteroaryl (lower alkyl), or R11 and R12 together are -(CH2)4-6-.
A particular preferred "substituted aryl" is a phenyl group substituted with a R13 and optionally substituted with up to four R14s, where R13 and R14 are defined with respect to formulae la and lla. The above-listed preferences equally apply for the compounds of
Formulae la and lla, below.
In a more preferred version of the first embodiment of the invention, Y is N-R7 and Z is N, R1 is lower alkyl, R4 and R5 are hydrogen, and
R6 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl), halo(lower alkyl), -C(=O)R11, -C(=O)OR11,
-C(=O)NR 1R12, -SO2R11, or -SO2NR11R12, wherein R11 and R12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R11 and R12 together are -(CH2)4.6-, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
More preferably still, R2 is -NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), benzyl, optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2) .6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
In another more preferred version of the first embodiment of the invention,
Y is N-R7 and Z are N,
R1 and R8 are lower alkyl,
R2 is -NR9R10,
R4 and R5 are hydrogen, and
R6 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl), halo(lower alkyl), -C(=O)R11, -C(=O)OR11,
-C(=O)NR >1"1RD1,2, -SO2R ,1"1, or -SO2NR ,1"1Rr->1"2, wherein R ,1"1 and R12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R11 and R12 together are -(CH2)4.6-, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
The second embodiment of the present invention provides compounds of Formula (la) or Formula (lla):
Figure imgf000024_0001
(la)
(lla) wherein:
Y is O, S or N-R7, Z is N or C-R8, R1, R2, R3, R4, R5, R7 and R8 are as defined in the first embodiment,
R13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16,
-C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, -NR15SO2R16 or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), or, together, are -(CH2)4.6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group, each R14 is independently selected from optionally substituted lower alkyl, optionally substituted aryl, optionally substituted heteroaryl, hydroxy, halogen, -CF3, -OR17 -NR17R18, -C(=O)R18, -C(=O)OR18, -C(=O)NR17R18, wherein R17 and R18 are independently, hydrogen, lower alkyl, alkenyl, alkynyl, -CF3, optionally substituted heterocycloalkyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or, together, are -(CH2)4-6-, optionally interrupted by one O, S, NH or N-(d-2 alkyl) group, and where n is an integer of 0 to 4, or a pharmaceutically acceptable salt thereof as a single stereoisomer or mixture of stereoisomers. Wherein R13 is -OR15, and R15 is optionally substituted lower alkyl, it may, for example, be optionally substituted with -C(=O)OR19, wherein R19 is hydrogen or lower alkyl.
Where R9 and R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group, examples include piperazinyl, 4-methylpiperazinyl, morpholyl, and hexahydropyrimidyl. n is a stereocompatible integer of 0 to 4. The term "stereocompatible" limits the number of substituents permissible by available valences in accordance with space requirements of substituents, among other. Preferably, R13 is hydrogen, optionally substituted lower alkyl, alkenyl, heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl) or, together, are -(CH2)4-6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group. More preferably, R13 is optionally substituted lower alkyl, alkenyl, heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl) or, together, are -(CH2) .6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group. Preferably, R14 is independently selected from optionally substituted lower alkyl, optionally substituted aryl, optionally substituted heteroaryl, hydroxy, halogen, -CF3, -OR17 -NR17R18, -C(=O)R18, -C(=O)OR18, -C(=O)NR17R18, wherein R17 and R18 are, independently, hydrogen, lower alkyl, alkenyl, or optionally substituted aryl.
Preferably, where R13 is not hydrogen, n is an integer of 1 to 2. More preferably, where R13 is not hydrogen, n is 1.
In another more preferred version of the second embodiment of the invention, Y is N-R7 and Z is N, R1 is lower alkyl,
R2 is -NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), benzyl, optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2) -6- optionally interrupted by one O, S, NH, N-(aryl),
N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι.2 alkyl) group such as piperazinyl,
4-methylpiperazinyl, 4-morpholyl, hexahydropyrimidyl, and
R13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl),
-CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15,
-C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, -NR15SO2R16 or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally . substituted heteroaryl, optionally substituted heteroaryl(lower alkyl) or R15 and R16 together are -(CH2)4.6- optionally interrupted by one O, S, NH or N-(C-|.2 alkyl) group.
More preferably still, R4 and R5 are hydrogen. Most preferably, independently,
1. Y is O, and R1 is lower alkyl.
2. Y is N-R7, R7 are hydrogen or lower alkyl, and R1 is lower alkyl.
3. Y -s N-R7, and R7 is methyl. 4. Z is N.
5. Z is C-R8, and R8 is hydrogen.
6. R2 and R3 are independently selected from hydrogen, lower alkyl, halogen, OR9, -NR9R10, where R9 and R10 are independently lower alkyl, substituted lower alkyl, or substituted aryl, or R9 and R10 together are -(CH2) .6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-COOH, N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
7. R13 is independently selected from halogen, optionally substituted aryl, -CF3, -CH3, -CN, -OR15, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16 , or -CO2H.
8. R14 is independently selected from halogen, optionally substituted lower alkyl, -CF3, -OR17, aryl, heteroaryl, -NR17R18, -C(=O)R17, -C(=O)OR17,
-C(=O)NR17R18, or -CO2H, where R17 and R18 are, independently, lower alkyl, substituted lower alkyl, or substituted aryl, or, together, are -(CH2)4.6- optionally interrupted by one O, S, NH or N-(C-|.2 alkyl) group.
9. Z is N, R2 is 4-methylpiperazinyl, R13 is 3-CF3, and R14 is 4-F. The preferred compounds of the invention are listed in Tables 1-4 below.
The compounds of this invention may possess one or more chiral centers, and can therefore exist as individual stereoisomers or as mixtures of stereoisomers. In such cases, all stereoisomers also fall within the scope of this invention. The compounds of this invention may also exist in various tautomeric forms, and in such cases, all tautomers also fall within the scope of this invention. The invention compounds include the individually isolated stereoisomers and tautomers as well as mixtures of such stereoisomers and their tautomers.
Some of the compounds of Formula (I) and Formula (II) are capable of further forming pharmaceutically acceptable salts and esters. All of these forms are included within the scope of the present invention. Pharmaceutically acceptable base addition salts of the compounds of Formula (I) and Formula (II) include salts which may be formed when acidic protons present in the parent compound are capable of reacting with inorganic or organic bases. Typically, the parent compound is treated with an excess of an alkaline reagent, such as hydroxide, carbonate, or alkoxide, containing an appropriate cation. Cations such as Na\ K+, Ca2+, and NH4 + are examples of cations present in pharmaceutically acceptable salts. The Na+ salts are especially useful. Acceptable inorganic bases, therefore, include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide. Salts may also be prepared using organic bases, such as choline, dicyclohexylamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, procaine, Λ/-methylglucamine, and the like [for a nonexclusive list see, for example, Berge et al., "Pharmaceutical Salts," J. Pharma. Sci. 66:1 (1977)]. The free acid form may be regenerated by contacting the base addition salt with an acid and isolating the free acid in the conventional manner. The free acid forms can differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents.
Pharmaceutically acceptable acid addition salts of the compounds of Formula (I) and Formula (II) include salts which may be formed when the parent compound contains a basic group. Acid addition salts of the compounds are prepared in a suitable solvent from the parent compound and an excess of a non-toxic inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid (giving the sulfate and bisulfate salts), nitric acid, phosphoric acid and the like, or a non-toxic organic acid such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, salicylic acid, p-toluenesulfonic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, lactic acid, o-(4-hydroxy-benzoyl)benzoic acid, 1 ,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid,
4-methyl-bicyclo[2.2.2.]oct-2-ene-1 -carboxylic acid, glucoheptonic acid, gluconic acid, 4,4'-methylenebis(3-hydroxy-2-naphthoic)acid, 3-phenylpropionic acid, trimethylacetic acid, fert-butylacetic acid, laurylsulfuric acid, glucuronic acid, glutamic acid, 3-hydroxy-2-naphthoic acid, stearic acid, muconic acid, and the like. The free base form may be regenerated by contacting the acid addition salt with a base and isolating the free base in the conventional manner. The free base forms can differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents.
Also included in the embodiment of the present invention are salts of amino acids such as arginate and the like, gluconate, and galacturonate [see Berge, supra (1977)].
Some of the compounds of the invention may form inner salts or Zwitterions.
Certain of the compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms, and are intended to be encompassed within the scope of the present invention.
Certain of the compounds of the present invention may also exist in one or more solid or crystalline phases or polymorphs, the variable biological activities of such polymorphs or mixtures of such polymorphs are also included in the scope of this invention.
Pharmaceutical Compositions
A third embodiment of the present invention provides pharmaceutical compositions comprising pharmaceutically acceptable excipients and a therapeutically effective amount of at least one compound of this invention. Pharmaceutical compositions of the compounds of this invention, or derivatives thereof, may be formulated as solutions or lyophilized powders for parenteral administration. Powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use. The liquid formulation is generally a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline solution, 5% dextrose in water or buffered sodium or ammonium acetate solution. Such formulations are especially suitable for parenteral administration but may also be used for oral administration. It may be desirable to add excipients such as polyvinylpyrrolidinone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride, or sodium citrate.
Alternatively, these compounds may be encapsulated, tableted, or prepared in an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to facilitate preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohols, or water. Solid carriers include starch, lactose, calcium sulfate, dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar, or gelatin. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage unit. The pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulation, and compressing, when necessary, for tablet forms; or milling, mixing, and filling for hard gelatin capsule forms. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion, or an aqueous or non-aqueous suspension. Such a liquid formulation may be administered directly p.o. or filled into a soft gelatin capsule.
Some specific examples of suitable pharmaceutical compositions are described in Examples 27-29.
Typically, a pharmaceutical composition of the present invention is packaged in a container with a label indicating the use of the pharmaceutical composition in the treatment of a disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, and transplant rejection, or a chronic or acute immune disorder, or a combination of any of these disease conditions. Methods of Use:
A fourth embodiment of the present invention provides a method for treating a chronic or acute inflammatory disease such as asthma, atherosclerosis, diabetic nephropathy, glomerulonephritis, inflammatory bowel disease, Crohn's disease, multiple sclerosis, pancreatitis, pulmonary fibrosis, psoriasis, restenosis, rheumatoid arthritis, or a chronic or acute immune disorder, or a transplant rejection in mammals in need thereof, comprising the administration to such mammal of a therapeutically effective amount of at least one compound of general Formula (I) and Formula (II) or pharmaceutically acceptable salts and esters thereof.
The compounds of the present invention inhibit chemotaxis of a human monocytic cell line (THP-1 cells) induced by human MCP-1 in vitro. This inhibitory effect has also been observed in vivo. Indeed, the compounds have been shown to reduce monocyte infiltration in a thioglycollate-induced inflammation model in mice.
The compounds of the present invention have been found to prevent the onset or ameliorate symptoms in several animal models of inflammation. For example, the compounds inhibited recruitment of monocytes into the glomeruli in an anti-Thy-1 antibody-induced model of nephritis; reduced paw swelling in a rat model of adjuvant arthritis; inhibited neointimal hyperplasia after balloon injury in a rat model of restenosis, and reduced the amount of lesion of the aortic sinus in an apoE-deficient mouse model of atherosclerosis.
The ability of the compounds of this invention to block the migration of monocytes and prevent or ameliorate inflammation, which is demonstrated in the specific examples, indicates their usefulness in the treatment and management of disease states associated with aberrant leukocyte recruitment.
The use of the compounds of the invention for treating inflammatory and autoimmune disease by combination therapy may also comprise the administration of the compound of the invention to a mammal in combination with common anti-inflammatory drugs, cytokines, or immunomodulators.
The compounds of this invention are thus used to inhibit leukocyte migration in patients which require such treatment. The method of treatment comprises the administration, orally or parenterally, of an effective quantity of the chosen compound of the invention, preferably dispersed in a pharmaceutical carrier. Dosage units of the active ingredient are generally selected from the range of 0.01 to 1000 mg/kg, preferably 0.01 to 100 mg/kg, and more preferably 0.1 to 50 mg/kg, but the range will be readily determined by one skilled in the art depending on the route of administration, age, and condition of the patient. These dosage units may be administered one to ten times daily for acute or chronic disease. No unacceptable toxicological effects are expected when compounds of the invention are used in accordance with the present invention. The invention compounds may be administered by any route suitable to the subject being treated and the nature of the subject's condition. Routes of administration include, but are not limited to, administration by injection, including intravenous, intraperitoneal, intramuscular, and subcutaneous injection, by transmucosal or transdermal delivery, through topical applications, nasal spray, suppository and the like, or may be administered orally. Formulations may optionally be liposomal formulations, emulsions, formulations designed to administer the drug across mucosal membranes or transdermal formulations. Suitable formulations for each of these methods of administration may be found in, for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.
Examples:
The following Examples serve to illustrate the preparation, properties, and therapeutic applications of the compounds of this invention. These Examples are not intended to limit the scope of this invention, but rather to show how to prepare and use the compounds of this invention.
Preparation of the Compounds of the Invention: General Procedures
The following general procedures may be employed for the preparation of the compounds of the present invention. The starting materials and reagents used in preparing these compounds are either available from commercial suppliers such as the Aldrich Chemical Company (Milwaukee, WI), Bachem (Torrance, CA), Sigma (St. Louis, MO), or are prepared by methods well known to a person of ordinary skill in the art, following procedures described in such references as Fieser and Fieser's Reagents for Organic Synthesis, vols. 1-17, John Wiley and Sons, New York, NY, 1991 ; Rodd's Chemistry of Carbon Compounds, vols. 1-5 and supps., Elsevier Science Publishers, 1989; Organic Reactions, vols. 1-40, John Wiley and Sons, New York, NY, 1991 ; March J.: Advanced Organic Chemistry, 4th ed., John Wiley and Sons, New York, NY; and Larock: Comprehensive Organic Transformations, VCH Publishers, New York, 1989. In some cases, protective groups may be introduced and finally removed.
For example, suitable protective groups for amino, hydroxy, and carboxy groups are described in Greene et al., Protective Groups in Organic Synthesis, Second Edition, John Wiley and Sons, New York, 1991. Activation of carboxylic acids can be achieved by using a number of different reagents as described in Larock: Comprehensive Organic Transformations, VCH Publishers, New York, 1989.
The starting materials, intermediates, and compounds of this invention may be isolated and purified using conventional techniques, including precipitation, filtration, distillation, crystallization, chromatography, and the like. The compounds may be characterized using conventional methods, including physical constants and spectroscopic methods.
Generally, a compound of formula I or formula II may be prepared as
Figure imgf000034_0001
(I) (II) follows: wherein Y, Z, and R >1 - n R6 are as defined in the first embodiment, said process comprising: (a) contacting a compound of formula lb or Mb
Figure imgf000035_0001
(lb) (lib) with a compound of the formula
RD — N=C=O
under conditions sufficient to produce a compounds of formula I or formula II, wherein R4 and R5 are both H; or
(b) contacting a compound of formula lc or formula lie
Figure imgf000035_0002
(lc) (lie) wherein X is halogen, nitro, -CN, or -OR9, with a compound of the formula
R2-H, under conditions sufficient to produce a compound of formula I or formula or
(c) contacting a compound of formula lb or formula lib
Figure imgf000035_0003
(lb) (lib) with a haloformylation reagent and a compound of the formula
Figure imgf000036_0001
under conditions sufficient to produce a compound of formula I or formula II, wherein R4 is H; or (d) elaborating substituents of a compound of formula I or formula II in a manner known perse; or
(e) reacting the free base of a compound of formula I or formula II with an acid to give a pharmaceutically acceptable addition salt; or
(f) reacting an acid addition salt of a compound of formula I or formula II with a base to form the corresponding free base; or
(g) converting a salt of a compound of formula I or formula II to another pharmaceutically acceptable salt of a compound of Formula I or II; or
(h) resolving a racemic mixture of any proportions of a compound of formula I or formula II to yield a stereoisomer thereof. Step (a), above, may be carried out in the presence of an organic solvent or a mixture of solvents at elevated temperatures. Said organic solvent may be toluene, and the reaction may be carried out under refluxing conditions.
Step (b), above, may be carried out using the salt of the compound of the formula R2-H in an inorganic solvent. Said salt may be the lithium, sodium, or potassium salt.
Step (c), above, may be carried out in an organic solvent or a mixture of solvents at elevated temperatures. The haloformylation reagent may be a compound of the formula A-(CO)-B wherein A and B are, independently, suitable leaving groups such as halogens, -COCI, -COBr and the like. The haloformylation agent and organic solvent employed in step (c) may be oxalyl chloride and THF, respectively, and the ensuing reaction may be heated to above 50°C. The compounds of the invention can further be synthesized as shown in the following examples. These examples are merely illustrative of some methods by which the compounds of this invention can be synthesized, and various modifications to these examples can be made and will be suggested to a person of ordinary skill in the art having regard to this disclosure.
Procedure A:
Figure imgf000037_0001
Acylurea compounds of the present invention may be prepared starting with an aryl carboxamide and an isocyanate. Carboxamide and isocyanate starting materials may be purchased from various different commercial sources, such as, for example the Aldrich Chemical Company, supra, or they may be prepared from standard procedures known in the art for preparing these compounds, such as the procedures described in the above-cited references. The isocyanates may also be prepared according to the procedures described in the example below. Typically, an aryl carboxamide is treated with an aryl isocyanate in an organic solvent or mixtures of suitable organic solvents. Preferably, the organic solvent is toluene. The carboxamide and the isocyanate may be combined as solutions or suspensions, depending on the solubilities of the compounds in the selected solvent. The carboxamide and the isocyanate may be added in a stoichiomethc ratio (1 :1), or a slight excess of the isocyanate may be used, for example between 1.01 fold and 2 fold excess, but typically about 1.01 to about 1.2 fold excess. Typically, the isocyanate is added to a suspension of the carboxamide in toluene, and the resulting mixture is heated until the reaction is determined to be complete. The reaction mixture may be heated at about 10 °C to about 150 °C, preferably at about 40 °C to about 120 °C under an inert atmosphere such as nitrogen, or the reaction mixture may be maintained at the refluxing temperature of the mixture. The reaction may be allowed to proceed to completion in about 10 minutes to 24 hours. Preferably, the reaction is heated to reflux until the reaction is complete, over about 6 to 24 hours.
Upon cooling of the reaction mixture, the resulting precipitated acylureas may be isolated by conventional techniques. Typically, the product is isolated by filtration. The precipitated solid may be filtered, washed with a solvent or a series of solvents, and isolated without further purification. Preferably, the precipitated acylureas may be washed with a combination of toluene, methanol and then with ether, and the product may be dried under vacuum. If desired, the acylureas may be further purified using conventional techniques, such as by crystallization using conventional methods known in the art. Optionally, acylureas prepared according to this procedure may be converted to the corresponding salts prior to isolation and/or purification, or after crystallization.
Procedure B:
Figure imgf000039_0001
Acylureas may also be prepared from the condensation of an aryl carboxamide with an amine. The carboxamide may be prepared from the corresponding carboxylic acid or may be obtained from commercial sources. Depending on the desired substitution of the amine, optionally, the amine may be substituted where one substituted group is an amine protecting group such that the protecting group may be removed in a subsequent step if desired. In the first step of the process, a carboxamide mixture in a suitable aprotic solvent is treated with a haloformylation reagent to form the corresponding carboxamide carbonylchloride derivative. Typically, the aprotic solvent is dichloromethane, toluene, 2-methyltetrahydrofuran or THF, and the haloformylation reagent is oxalyl chloride. Preferably, the aprotic solvent is THF. Oxalyl chloride is preferably present in an excess, for example between 1.1 to 3.0 equivalents, typically about 1.5 equivalent over the carboxamide. The reaction is generally performed under an inert atmosphere where the mixture is heated to 50 °C to 175 °C for 15 minutes to 24 hours until the reaction is deemed complete. Typically, the reaction is heated to reflux over 2 to 16 hours under nitrogen, and then cooled to room temperature. The solvent is removed in vacuo by rotoevaporation or distillation, and the resulting carboxamide carbonylchloride is then condensed with a primary or secondary amine. Condensation with the amine may be performed by the addition of a solution of the amine in an aprotic solvent, such as THF, under an inert atmosphere, at a temperature between 0 °C and 20 °C, preferably between 0 °C and 5 °C. If the chloroformylation and the subsequent condensation reaction is performed in the same solvent, the solvent removal step may be eliminated. Preferably, the reaction is performed at 0 °C to 5 °C for 1 to 24 hours, until the reaction is complete. The solvent is removed by concentration under reduced pressure, and the acylurea can be isolated by conventional techniques such as filtration and washing of the crude product with a solvent, followed by drying under vacuum.
Procedure C:
Figure imgf000040_0001
The preparation of the acylureas may also be performed starting with an amine or aniline derivative through a condensation reaction with a phosgene equivalent, followed by a condensation reaction with the carboxamide. Typically, a solution of an aniline or aniline derivative and triphosgene in tetrachloroethane or other suitable organic solvent is combined and stirred at 25 °C to 80 °C under an inert atmosphere for 2 to 12 hours until the reaction is complete. The solvent is removed under reduced pressure and the residue is dissolved in an aprotic solvent, such as toluene, and the resulting mixture is treated with a carboxamide. The mixture is heated to about 50 °C to 150 °C, preferably from about 75 °C to 115 °C. Preferably, the reaction mixture is heated to reflux for 2 to 24 hours until the reaction is complete, and allowed to cool to room temperature. The precipitated solid is isolated by conventional techniques such as filtration. The filtered solid is then washed with a suitable solvent or mixtures of solvents. Typically, the solid is washed with toluene, methanol and then ether, and the washed product is dried in vacuo to give the corresponding acylurea.
Procedure D:
Figure imgf000041_0001
Amine substituted aryl acyl ureas of the present invention may be prepared starting from the corresponding aryl halide acyl ureas by the reaction of the aryl halide with an amine. Preferably, the aryl halide is an aryl chloride, which can be prepared from one or more of the above described procedures or from standard procedures known in the art for preparing these compounds. Typically, the acyl urea is dissolved in an organic solvent or a mixture of suitable organic solvents. Preferably, the organic solvent is tetrahydrofuran. The acyl urea and the amine may be combined as solutions or suspensions, depending on the solubilities of the compounds in the selected solvent or solvent mixtures. The acyl urea and the amine may be added in a stoichiomethc ratio (1 :1 ) , or a slight excess of the amine may be used, for example, between 1.01 fold and 20 fold excess, but typically about 1.01 to about 10 fold excess. Typically, the amine is added to the acyl urea in tetrahydrofuran and the resulting mixture is stirred at about 0 °C to refluxing temperatures of the solvent, preferably at about 10 °C to about 50 °C, most preferably at about room temperatures under an inert atmosphere such as nitrogen. The reaction mixture is maintained at the reaction temperatures until the reaction proceeds to completion. The reaction may be allowed to proceed to completion in about 10 minutes to 48 hours. Preferably, the reaction is stirred at room temperature for about 5 hours. When the reaction is deemed complete, the resulting product may be isolated by conventional techniques. Typically, the solvent and excess amine may be removed by evaporation under reduced pressure, and the residue is suspended in a solvent. Preferably, the solvent is water. The suspension or solid may be filtered and washed with water or a suitable solvent, and then isolated and dried using conventional methods.
Procedure E:
Figure imgf000042_0001
Cyclic acyl ureas of the present invention may be prepared according to methods known in the art. One method comprises the alkylation of the acyl urea nitrogens with an alkylating agent gene cally represented above as X-(CH2)2.4-Y, where X and Y are leaving groups, and may be the same or different. Leaving groups known in the art include halides, methanesulfonates, thfluoromethanesulfonates, p-toluenesulfonates, p-bromotoluenesulfonate, p- nitrobenzenesulfonates and the like. Representative alkylating agents include
1 ,2-dibromoethane, 1 ,3-dibromoethane, 1 ,3-dibromopropane, and the corresponding sulfonates and mixed halosulfonates. Typically, the acyl urea is treated with a base in an organic solvent or mixtures of solvents. Preferably, the base is an inorganic base such as sodium hydride, or an organic base such as dimethyl sulfoxide and sodium hydride. Preferably, the solvent is a polar, aprotic solvent such as tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, glycols, or mixtures of such solvents. Typically, a solution or suspension of the acyl urea is slowly added to the base in an organic solvent at about 0 °C to about 25 °C, and the resulting mixture is stirred for about 10 minutes to about 5 hours, preferably about 30 minutes. The alkylating agent is added and the mixture is stirred until the reaction is deemed complete. Alkylation of both urea nitrogens may be accomplished in a single step, or may be accomplished sequentially in a two step procedure be exposing the partially alkylated product with the same or different base. The reaction is then quenched with a solvent, preferably water, and the mixture is extracted multiple times with an organic solvent. Preferably, the extracting solvent is dichloromethane. The combined organic extracts are washed with water, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to afford the product, which may be purified using standard conditions known in the art. Purification may be performed by silica gel chromatography in a mixture of organic solvents, such as ethyl acetate and petroleum ether.
Example 1 : Synthesis of Λ/-(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- Λ/-{[(4-chlorophenyl)amino]carbonyl}carboxamide (39).
Figure imgf000043_0001
Oxalyl chloride (4.5 mL of a 2M solution in dichloromethane) was added dropwise to a stirred suspension of 4-chloro-1 ,3-dimethylpyrazolo [5,4-b]pyridine-5-carboxylic acid (0.98 g) in anhydrous dichloromethane (14 mL), followed by 4 drops of Λ/,Λ/-dimethylformamide (DMF). After the evolution of gas subsided, 4 more drops of DMF were added, and this operation was repeated three more times. The mixture was stirred at room temperature for 45 min, filtered to remove trace amounts of insoluble material, and the solvent evaporated under reduced pressure. The residue was dried under high vacuum for 1 h, cooled in an ice-bath, and treated with ammonium hydroxide (28% NH3 in water, 20 mL) (Caution! exothermic reaction). The mixture was stirred at 0 °C for 30 min, and at room temperature for an additional 1 h. The solid was collected by filtration, washed with water, and dried under vacuum to give the carboxamide as a white powder.
Oxalyl chloride (1.4 mL of a 2M solution in dichloromethane) was added dropwise to a suspension of 4-chloro-1 ,3-dimethylpyrazolo[5,4-b] pyridine-5-carboxamide (0.41 g) in anhydrous dichloromethane (5 L). The mixture was heated at reflux for 15 h, and cooled to room temperature. The solvent was evaporated under reduced pressure and the residue was dissolved in anhydrous THF (6 mL). An aliquot of this solution (1 .5 mL) was added dropwise to an ice-cooled solution of 4-chloroaniline (57 mg) in anhydrous THF (2 mL). The ice-bath was removed and the mixture was stirred at room temperature for 1.5 h. The precipitated solid was collected by filtration, washed with dichloromethane and methanol, and dried under high vacuum to afford Λ/-(4-Chloro-1 ,3-dimethyl-pyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(4-chlorophenyl) amino]carbonyl}carboxamide as a white powder. 1H NMR (300 MHz, DMSO-dβ) δ 2.68 (s, 3H), 4.02 (s, 3H), 7.42 (d, 2H, J=8.7 Hz), 7.64 (d, 2H, J=8J Hz), 8.69 (s, 1 H), 10.47 (s, 1 H), 11.37 (s, 1 H). MS (API-CI) m/z 340, 342, 344.
Example 2: Preparation of Λ/-{[(3,4-Dichlorophenyl)amino]carbonyl}(1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl) carboxamide (2).
(i) Zn, AcOH
Figure imgf000044_0001
A suspension of 4-chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carboxylic acid (2.00 g) and zinc dust (1.74 g) in acetic acid (40 mL) was heated at 80 °C for 1.5 h. The mixture was cooled to room temperature, slowly poured into an ice-cooled solution of 5N sodium hydroxide (160 mL) and filtered. The filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid. The precipitate was filtered, washed with water, and dried under high vacuum. The solid was re-suspended in anhydrous dichloromethane (20 mL) and treated with a 2M solution of oxalyl chloride in dichloromethane (3.6 mL), followed by anhydrous DMF (0.1 mL). The mixture was stirred at room temperature for 1 h, filtered, and the filtrate concentrated under reduced pressure. The residue was cooled in an ice-bath and slowly treated with 28% ammonium hydroxide solution. The suspension was removed from the ice-bath, stirred at room temperature for 1 h, and filtered. The solid was washed with water and dried under high vacuum to produce 1 ,3-dimethylpyrazolo[5,4-b]pyri- dine-5-carboxamide as an off-white solid. A suspension of this carboxamide (0.13 g) in anhydrous dichloromethane (5 mL) was treated with a 2M solution of oxalyl chloride also in dichloromethane (0.5 mL) and the mixture heated at gentle reflux for 15 h. The solvent was removed under reduced pressure and the residue dissolved in anhydrous THF (1.2 mL). An aliquot of this solution (0.4 mL) was added to an ice-cooled solution of 3,4-dichloroaniline (36 mg) in anhydrous THF (0.7 mL). The ice-bath was removed and the mixture stirred at room temperature for 1 h. The precipitated solid was collected by filtration, washed with dichloromethane and MeOH, and dried under high vacuum to give the title compound as a white solid. 1H NMR (DMSO-d6) δ 2.56 (s, 3H), 4.02 (s, 3H), 7.56-7.63 (m, 2H), 8.03 (d, J=2.2 Hz, 1 H), 8.99 (d, J=2.0 Hz, 1 H), 9.08 (d, j=2.0 Hz, 1 H), 10.92 (s, 1 H), 1 1.28 (s, 1 H). MS (ESI) m/z 376, 378, 380. Example 3: Ethyl 2-[4-({[(1 ,3-dimethypyrazolo[5,4-b]pyridin-5- yl)carbonylamino]carbonyl} amino)-2-(trifluoromethyl)phenoxy]acetate (34).
(i) (COCI)2
Figure imgf000045_0001
1 ,3-Dimethylpyrazolo[5,4-b]pyridine-5-carboxamide (0.35 g) was suspended in anhydrous dichloromethane (10 mL) and treated with a 2M solution of oxalyl chloride in dichloromethane (1.8 mL). The suspension was heated at gentle reflux for 15 h, cooled to room temperature, and the solvent removed under reduced pressure. The residue was dissolved in anhydrous THF (7 mL). An aliquot of this solution (2 mL) was added to an ice-cooled solution of 4-amino-2-(trifluoromethyl)phenol (93 mg) in THF (0.5 mL). The ice-bath was removed and the reaction mixture stirred at 25°C for 1 h. The precipitated solid was collected by filtration, washed with dichloromethane and dried under high vacuum. The solid was re-suspended in acetone-DMF (8 mL:0.8 mL) and treated with potassium carbonate (0.18 g) and ethyl bromoacetate (0.15 mL). The suspension was heated at reflux for 2 h, cooled to room temperature, and concentrated under reduced pressure. The residue was treated with water (3 mL), and the resulting solids were filtered, washed with water and acetone, and dried under high vacuum to give the title compound as a white solid. 1H NMR (DMSO-d6) δ 1 .19-1.24 (t, J=7.1 Hz, 3H), 2.58 (s, 3H), 3.99 (s, 3H), 4.14-4.21 (q, J=7.1 Hz, 2H), 4.86 (s, 2H), 7.03 (d, J=9J Hz, 1 H), 7.52-7.60 (dd, J=2.4, 8.8 Hz, 1 H), 8.09 (d, J=2.4 Hz, 1 H), 8J5 (d, J=1.6 Hz, 1 H), 9.22 (d, J=1.6 Hz, 1 H), 12.12 (s, 1 H). MS (ESI) m/z 480.
Example 4: Sodium 2-[4-({[(1 ,3-Dimethypyrazolo[5,4-b]pyridin-5-yl)carbonyl- amino]carbonyl}amino)-2-(trifluoromethyl)phenoxy]acetate (35).
Figure imgf000046_0001
A solution of ethyl 2-[4-({[(1 ,3-dimethypyrazolo[5,4-b]pyridin-5-yl)carbonyl- amino]carbonyl}amino)-2-(trifluoromethyl)phenoxy]acetate (0.1 1 g) in DMF (3 mL) was treated with 5N sodium hydroxide solution (1.6 mL). The mixture was stirred at room temperature for 1.25 h, and then concentrated under reduced pressure. The residue was taken up in water (4 mL), filtered, and the filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid. The precipitated solid was collected by filtration, washed with water, and dried under high vacuum. A portion of this material was suspended in water (5 mL) and treated with 5N sodium hydroxide solution (27 μL). The mixture was sonicated, filtered, and the filtrate lyophilized to yield the title compound. 1H NMR (DMSO-d6) δ 2.53 (s, 3H), 3.99 (s, 3H), 4.17 (s, 2H), 6.85 (d, J=8.8 Hz, 1 H), 7.44-7.47 (br d, J=8.8 Hz, 1 H), 8.03 (d, J=2.1 Hz, 1 H), 8.52 (s, 1 H) 8.79 (d, J=1.6 Hz, 1 H), 9.23 (d, J=1.6 Hz, 1 H), 11.50 (br s, 1 H). MS (ESI) m/z 450.
Example 5: N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(4-hydroxy- 1,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (43).
DIPEA-H20
Figure imgf000047_0001
Figure imgf000047_0002
A suspension of (4-chloro-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl)-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (0.050 g) in THF (5 mL) was dissolved in a 1 :3 mixture of Λ/,Λ/-diisopropylethylamine (DIPEA) and water (4 mL). The solution was stirred at room temperature for 24 h, concentrated, and the residue purified by column chromatography on silica-gel eluting with 3:1 EtOAc-MeOH to give the title compound as a white powder. 1H NMR (DMSO-de) δ 2.46 (s, 3H), 3.75 (s, 3H), 7.45 (dd, J=9.9, 9.9 Hz, 1 H), 7.81 (m, 1 H), 8.11 (dd, J=6.6, 2.6 Hz, 1 H), 8.58 (s, 1 H), 11.51 (br s, 1 H). MS (ESI) m/z 410. Example 6: (1 ,3-Dimethyl-4-morpholin-4-ylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4- fluoro -3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide (65).
Figure imgf000047_0003
65
A solution of 4-chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carboxylic acid (450 mg) and morpholine (870 mg) in DMF (15 mL) was stirred at room temperature for 5 h, and then concentrated under reduced pressure. The residue was treated with water (10 mL), and the resulting solid was filtered, washed with water, and dried under high vacuum. The solid was suspended in anhydrous dichloromethane (20 mL) and treated with oxalyl chloride (1.0 mL) and anhydrous DMF (0.1 mL). The mixture was stirred at room temperature for 1 h, filtered, and the filtrate was concentrated under reduced pressure. The residue was cooled in an ice-bath and slowly treated with a saturated solution of ammonia in THF (20 mL). The suspension was removed from the ice-bath, stirred at room temperature for 1 h, and filtered. The solid was washed with water and dried under high vacuum to give 1 ,3-dimethyl-4-morpholin-4- ylpyrazolo[5,4-b]pyridine-5-carboxamide as a white powder. A portion of this material (0.33 g) was dissolved in hot toluene (30 mL) and azeotroped for 1 h. The solution was cooled to room temperature and treated with 4-fluoro-3- (trifluoromethyl)phenylisocyanate (0.27 g). The mixture was refluxed for 16 h and then cooled to room temperature. The precipitated solid was filtered, washed with toluene, and dried under high vacuum to give the title compound as a white powder. 1H NMR (DMSO-d6) δ 2.62 (s, 3H), 3.27 (m, 4H), 3.80 (m, 4H), 3.93 (s, 3H), 7.50 (dd, J=9.9, 9.9 Hz, 1 H), 7.92 (m, 1H), 8.13 (dd, J=6.6, 2.6 Hz, 1 H), 8.38 (s, 1 H), 10J2 (s, 1 H), 11.34 (s, 1 H). MS (ESI) m/z 479.
Example 7: (4-{[2-(Dimethylamino)ethyl]amino}-1 ,3-dimethylpyrazolo[5,4- b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (45).
Figure imgf000048_0001
4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carboxamide (0.10 g) was dissolved in hot toluene (30 mL) and azeotroped for 1 h. The solution was cooled to room temperature and reacted with 4-fluoro-3- (trifluoromethyl)phenylisocyanate (0.10 g). The mixture was refluxed for 16 h and then cooled to room temperature. The precipitated solid was filtered, washed with methanol, and dried under high vacuum to give (4-chloro-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3-(trifluoromethyl) phenyl]amino}carbonyl)carboxamide as a white powder. A portion of this material (0.050 g) was dissolved in THF (10 mL) and reacted with (2- aminoethyl)dimethylamine (0.10 g). The mixture was stirred at room temperature for 5 h, and then concentrated under reduced pressure. The residue was treated with water (10 mL), and the resulting solid was filtered, washed with water, and dried under high vacuum. A portion of this material was suspended in water (5 mL) and treated with 1 N hydrochloric acid solution (100 μL). The mixture was sonicated, filtered, and the filtrate lyophilized to give (4-{[2- (dimethylamino)ethyl]amino}-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl)-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride as a white powder. 1H NMR (DMSO-d6) δ 2.22 (s, 6H), 2.53 (t, J=5.8 Hz, 2H), 2.61 (s, 3H), 3.62 (t, J=5.8 Hz, 2H), 3.86 (s, 3H), 7.50 (dd, J=10.3, 9.5 Hz, 1 H), 7J6 (br s, 1 H), 7.86 (m, 1 H), 8.09 (dd, J=6.6, 2.6 Hz, 1 H), 8.40 (s, 1 H), 10.74 (s, 1 H), 10.97 (br s, 1 H). MS (ESI) m/z 480. The following additional compounds were prepared by the foregoing procedures:
[4-(Dimethylamino)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (44).
1H NMR (DMSO-de) δ 2.59 (s, 3H), 3.04 (s, 6H), 3.90 (s, 3H), 7.50 (dd, J=9.9, 9.9 Hz, 1 H), 7.90 (m, 1 H), 8.1 1 (dd, j=3.6, 2.6 Hz, 1 H), 8.29 (s, 1 H), 10.79 (s, 1 H), 11.18 (s, 1 H). MS (ESI) m/z 437.
(4-{[2-(Dimethylamino)ethyl]amino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-N- ({[4-fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (46). 1H NMR (DMSO-de) δ 2.69 (s, 3H), 2J9 (d, J=4.4 Hz, 6H), 3.40 (m, 2H), 3.83 (m, 2H), 3.89 (s, 3H), 7.52 (dd, J=9.9, 9.9 Hz, 1 H), 7.65 (br s, 1 H), 7.88 (m, 1 H), 8.08 (dd, j=6.6, 2.6 Hz, 1 H), 8.47 (s, 1 H), 10.67 (br s, 1 H), 10.73 (s, 1 H), 11.13 (s, 1 H).
(4-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5- yl)-N-({[4-fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide (47). 1H NMR (DMSO-de) δ 2.01 (s, 6H), 2.53 (t, J=5.8 Hz, 2H), 2.59 (s, 3H), 3.04 (s, 3H), 3.63 (t, J=5.8 Hz, 2H), 3.92 (s, 3H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.90 (m, 1 H), 8.14 (dd, J=3.6, 2.6 Hz, 1 H), 8.38 (s, 1 H), 10.99 (s, 1 H), 1 1.99 (s, 1 H). MS (ESI) m/z 494. (4-{[3-(Dimethylamino)propyl]amino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (49). 1H NMR (DMSO-de) δ 1.77 (m, 2H), 2.14 (s, 6H), 2.36 (t, J=6.6 Hz, 2H), 2.59 (s, 3H), 3.45 (t, J=6.6 Hz, 2H), 3.86 (s, 3H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.85 (m, 1 H), 8.08 (m, 1 H), 8.41 (s, 1 H), 10J8 (br s, 1 H). MS (ESI) m/z 494. {4-[Bis(2-hydroxyethyl)amino]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (51). 1H NMR (DMSO-de) δ 2.59 (s, 3H), 3.52 (m, 4H), 3.62 (m, 4H), 3.92 (s, 3H), 5.12 (br s, 2H), 7.53 (dd, j=9.9, 8.9 Hz, 1 H), 7.90 (m, 1 H), 8.20 (dd, J=6.6, 2.6 Hz, 1 H), 8.36 (s, 1 H), 10.86 (s, 1 H), 1 1.31 (br s, 1 H). MS (ESI) m/z 497. {1 ,3-Dimethyl-4-[methyl(1 -methylpyrrolidin-3-yl)amino]pyrazolo[5,4-b]pyridin-5- yl}-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (53). 1H NMR (DMSO-de) δ 1 .97 (m, 2H), 2.22 (s, 3H), 2.28 (m, 1 H), 2.54 (m, 1 H), 2.58 (s, 3H), 2.60-2.80 (m, 2H), 2.90 (s, 3H), 3.93 (s, 3H), 4.24 (m, 1 H), 7.50 (dd, j=10.3, 9.5 Hz, 1 H), 7.90 (m, 1 H), 8.12 (dd, J=6.6, 2.6 Hz, 1 H), 8.41 (s, 1 H), 10.7 (br s, 1 H), 11.4 (br s, 1 H). MS (ESI) m/z 506.
{4-[(4-Aminocyclohexyl)amino]-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (55). 1H NMR (DMSO-de) δ 1.64-1.83 (m, 8H), 2.63 (s, 3H), 3.18 (m, 1 H), 3.88 (s, 3H), 4.08 (m, 1 H), 7.52 (m, 1 H), 7.88 (m, 1 H), 8.10 (m, 1 H), 8.57 (s, 1 H), 10.6 (br s, 1 H), 10.9 (br s, 1 H). MS (ESI) m/z 506.
{1 ,3-Dimethyl-4-[(perhydropyrrolizin-7a-ylmethyl)amino]pyrazolo[5,4-b]pyridin-5- yl}-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (57). 1H NMR (DMSO-de) δ 1.59-1.81 (m, 8H), 2.59 (s, 3H) , 2.85-3.03 (m, 2H), 2.93 (d, J=6.2 Hz, 2H), 3.85 (s, 3H), 7.48 (dd, J=9.9, 9.5 Hz, 1 H), 7.59 (br s, 1 H), 7.82 (m, 1 H), 8.10 (dd, J=6.2, 2.6 Hz, 1 H), 8.50 (s, 1 H), 10.9 (br s, 1 H). MS (ESI) m/z 532.
{1 ,3-Dimethyl-4-[(2-perhydropyrrolizin-7a-ylethyl)amino]pyrazolo[5,4-b]pyridin-5- yl}-N-({[4-fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide (59). 1H NMR (DMSO-de) δ 1.57-1.78 (m, 10H), 2.57 (m, 2H), 2.60 (s, 3H) , 2.90 (m, 2H), 3.33 (t, J=6.2 Hz, 2H), 3.84 (s, 3H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.87 (m, 1 H), 8.12 (dd, J=6.2, 2.6 Hz, 1 H), 8.28 (s, 1 H), 10.82 (br s, 1 H), 11.52 (s, 1 H). MS (ESI) m/z 546.
{1 ,3-Dimethyl-4-[(2-perhydropyrrolizin-7a-ylethyl)amino]pyrazolo[5,4-b]pyridin-5- yl}-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (60).
1H NMR (DMSO-de) δ 1.89-2.18 (m, 10H), 2.65 (s, 3H) , 3.08 (m, 2H), 3.59 (m, 2H), 3.90 (s, 3H), 7.51 (dd, J=9.9, 9.5 Hz, 1 H), 7.87 (m, 1 H), 8.01 (br s, 1 H), 8.07 (dd, J=6.6, 2.6 Hz, 1 H), 8.50 (s, 1 H), 10.58 (br s, 1 H), 1 1.71 (s, 1 H), 1 1.1 1 (s, 1 H).
(1 ,3-Dimethyl-4-piperidylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (64).
1H NMR (DMSO-de) δ 1.62 (m, 2H), 1.71 (m, 4H), 2.62 (s, 3H), 3.24 (m, 4H), 3.92 (s, 3H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.92 (m, 1 H), 8.12 (dd, J=6.2, 2.6 Hz, 1 H), 8.35 (s, 1 H), 10.71 (s, 1 H), 1 1.31 (s, 1 H). MS (ESI) m/z 477.
(1 ,3-Dimethyl-4-piperazinylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (thfluoromethyl)phenyl]amino}carbonyl)carboxamide (66).
1H NMR (DMSO-de) δ 2.62 (s, 3H), 2.93 (m, 4H), 3.23 (m, 4H), 3.92 (s, 3H), 7.51 (dd, J=9.5, 9.5 Hz, 1 H), 7.90 (m, 1 H), 8.12 (dd, J=6.2, 2.6 Hz, 1 H), 8.36 (s, 1 H), 10J (br s, 1 H), 1 1.3 (br s, 1 H). MS (ESI) m/z 478.
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (84).
1H NMR (DMSO-de) δ 2.32 (s, 3H), 2.55 (m, 4H), 2.62 (s, 3H), 3.29 (m, 4H), 3.93 (s, 3H), 7.51 (dd, J=9.9, 9.9 Hz, 1 H), 7.91 (m, 1 H), 8.12 (dd, J=6.2, 2.6 Hz, 1 H), 8.37 (s, 1 H), 10.71 (s, 1 H), 11.32 (s, 1 H). MS (ESI) m/z 492. [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (85). 1H NMR (DMSO-de) δ 2.65 (s, 3H), 2.86 (d, J=4.4 Hz, 3H), 3.31 (m, 2H), 3.51 (m, 6H), 3.95 (s, 3H), 7.52 (dd, J=9.9, 9.9 Hz, 1 H), 7.88 (m, 1 H), 8.11 (m, 1 H), 8.44 (s, 1 H), 10.48 (br s, 1 H), 10.74 (s, 1 H), 11.38 (s, 1 H).
[4-(4-Ethylpiperazinyl)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (99).
1H NMR (DMSO-de) δ 1.03 (t, J=6.9 Hz, 3H), 2.41 (q, J=6.9 Hz, 2H), 2.57 (m, 4H), 2.62 (s, 3H), 3.31 (m, 4H), 3.93 (s, 3H), 7.50 (dd, J=9.9, 9.9 Hz, 1 H), 7.91 (m, 1 H), 8.13 (dd, J=6.2, 2.6 Hz, 1 H), 8.35 (s, 1 H), 10.72 (s, 1 H), 1 1.31 (s, 1 H). MS (ESI) m/z 506.
[4-(4-Ethylpiperazinyl)-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl]-N-({[4-fluoro-3- (thfluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (100). 1H NMR (DMSO-d6) δ 1.28 (d, J=7.3 Hz, 3H), 2.65 (s, 3H), 3.17-3.35 (m, 4H), 3.45-3.61 (m, 6H), 3.95 (s, 3H), 7.52 (dd, J=9.9, 9.5 Hz, 1 H), 7.89 (m, 1 H), 8.11 (dd, J=6.2, 2.6 Hz, 1 H), 8.44 (s, 1 H), 10.29 (br s, 1 H), 10.74 (s, 1 H), 1 1.39 (s, 1 H).
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl){4-[4-(2- hydroxyethyl)piperazinyl]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}carboxamide (101).
1H NMR (DMSO-de) δ 2.47 (t, J=5.9Hz, 2H), 2.62 (s, 3H), 2.63 (m, 4H), 3.28 (m, 4H), 3.53 (m, 2H), 3.92 (s, 3H), 4.38 (br s, 1 H), 7.51 (dd, J=9.9, 9.5 Hz, 1 H), 7.92 (m, 1 H), 8.13 (dd, J=6.2, 2.6 Hz, 1 H), 8.35 (s, 1 H), 10.7 (br s, 1 H), 1 1.3 (br s, 1 H). MS (ESI) m/z 522. [1 ,3-Dimethyl-4-(4-phenylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (103). 1H NMR (DMSO-de) δ 2.66 (s, 3H), 3.36 (m, 4H), 3.43 (m, 4H), 3.94 (s, 3H), 6.81 (t, J=7.3 Hz, 1 H), 7.01 (d, J=8.1 Hz, 1 H), 7.23 (dd, J=8.1 , 7.3 Hz, 1 H), 7.49 (dd, J=9.9, 9.5 Hz, 1 H), 7.91 (m, 1 H), 8.12 (dd, J=6.3, 2.6 Hz, 1 H), 8.40 (s, 1 H), 10J2 (s, 1 H), 1 1.38 (br s, 1 H). MS (ESI) m/z 554. {1 ,3-Dimethyl-4-[4-benzylpiperazinyl]pyrazolo[5,4-b]pyhdin-5-yl}-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (104). 1H NMR (DMSO-de) δ 2.59 (m, 3H), 2.61 (s, 4H), 3.29 (m, 4H), 3.55 (s, 2H), 3.92 (s, 3H), 7.24-7.33 (m, 5H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.92 (m, 1 H), 8.13 (dd, J=6.2, 2.6 Hz, 1 H), 8.35 (s, 1 H), 10.69 (s, 1 H), 11.29 (s, 1 H). MS (ESI) m/z 568.
4-Amino-1 -{5-[N-({[4-fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carbamoyl]- 1 ,3-dimethylpyrazolo[5,4-b]pyhdin-4-yl}pyhdinium chloride (106). 1H NMR (DMSO-de) δ 2.17 (s, 3H), 4.10 (s, 3H), 7.04 (d, J=7.3 Hz, 2H), 7.48 (dd, J=9.9, 9.5 Hz, 1 H), 7.82 (m, 1 H), 8.01 (dd, J=6.6, 2.6 Hz, 1 H), 8.36 (d, J=7.3 Hz, 2H), 8.80 (br s, 2H), 9.06 (s, 1 H), 10.4 (br s, 1 H), 11.4 (br s, 1 H). MS (ESI) m/z 486.
[1 ,3-Dimethyl-4-(4-methyl(1 ,4-diazaperhydroepinyl))pyrazolo[5,4-b]pyridin-5-yl]- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (107). 1H NMR (DMSO-de) δ 1.92 (m, 2H), 2.30 (s, 3H), 2.63 (s, 3H), 2J0 (m, 4H), 3.54 (m, 4H), 3.91 (s, 3H), 7.50 (dd, J=9.9, 9.5 Hz, 1 H), 7.91 (m, 1 H), 8.12 (m, 1 H), 8.36 (s, 1 H), 10.75 (s, 1 H), 1 1.22 (s, 1 H). MS (ESI) m/z 506.
[1 ,3-Dimethyl-4-(4-methyl(1 ,4-diazaperhydroepinyl))pyrazolo[5,4-b]pyridin-5-yl]- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (108). 1H NMR (DMSO-de) δ 2.15-2.32 (m, 2H), 2.65 (s, 3H), 2.83 (d, J=4.8Hz, 3H), 3.22-3.30(m, 1 H), 3.52-3.69 (m, 3H), 3.91-3.97 (m, 1 H), 3.94 (s, 3H), 7.52 (dd, j=9.9, 9.5 Hz, 1 H), 7.92 (m, 1 H), 8.13 (dd, J=6.2, 2.6 Hz, 1 H), 8.45 (s, 1 H), 10J4 (br s, 1 H), 10J6 (s, 1 H), 1 1.30 (s, 1 H).
Example 8: (6-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3-(trifluoromethyl) phenyl]amino}carbonyl)carboxamide hydrochloride (116).
Figure imgf000054_0001
Figure imgf000054_0002
A solution of 5-amino-1 ,3-dimethylpyrazole-4-carbaldehyde (6.15 g) in ethyl cyanoacetate (10 mL) was stirred at 185 °C for 3 hrs. The reaction mixture was allowed to cool to room temperature. The precipitated solid was filtered, washed with ethyl acetate, and dried under high vacuum to give 6-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]pyhdin-5-carbonitrile as a white powder.
A portion of this material (2.18 g) was dissolved in phenylphosphonic dichloride (10 mL) and stirred at 150 °C for 17 hrs. The solution was cooled to room temperature and poured into water, extracted with ethyl acetate, washed with saturated NaHCO3 solution, and dried over Na2SO4. Evaporation of the solvent under reduced pressure produced a white solid. This material (2.16 g) was dissolved in tetrahydrofuran (50 mL), dimethylaminoethylmethylamine (10.6 mL) was added at room temperature, and the mixture was heated at reflux for 20 hrs. The reaction mixture was cooled to room temperature, extracted with ethyl acetate. The extract was dried over Na2SO4, and evaporated under reduced pressure to give 6-{[2-(dimethylamino)ethyl]methylamino}-1 ,3- dimethylpyrazolo[5,4-b]pyridine-5-carbonithle as a brown oil. This material (2.96 g) was dissolved in concentrated H2SO (10 mL). The mixture was stirred at 60 °C for 15 hrs. The reaction mixture was neutralized with saturated NaHCO3 solution and extracted with dichloromethane. The extract was washed with NaCl solution, dried over Na2SO4, and evaporated under reduced pressure. The residue was purified by the recrystallization from the mixture of ethyl acetate and hexane to give 6-{[2-(dimethylamino)ethyl]methylamino}-1 ,3- dimethylpyrazolo[5,4-b]pyridine-5-carboxamide as a white solid. A portion of this material (0.150 g) was dissolved in hot toluene (20 mL) and azeotroped for 2 hrs. The solution was cooled to room temperature and reacted with 4-fluoro-3- (trifluoromethyl)phenylisocyanate (0.180 g). The mixture was refluxed for 2 days and then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was purified with the column chromatography (ethyl acetate) to give the acylurea as a white solid. A portion of this material (52mg) was dissolved in tetrahydrofuran (1.0 mL) and treated with 1 N hydrochloric acid solution (1.0 mL). This mixture was lyophilized to give (6-{[2- (dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride as a white solid. 1H NMR (DMSO-d6) δ 2.17 (s, 6H), 2.41 (s, 3H), 2.53 (t, J=6.4 Hz, 2H), 2.98 (s, 3H), 3.64 (t, J=6.4 Hz, 2H), 3.82 (s, 3H), 7.49 (m, 1 H), 7.87 (m, 1 H), 8.10 (m, 1 H), 8.26 (s, 1 H), 10.82 (br s, 1 H).
Example 9: {6-[2-(Diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5- yl}-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (120).
Figure imgf000055_0001
Figure imgf000055_0002
6-Hydroxy-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carbonitrile (0.91 1 g) was dissolved in concentrated H2SO (3.0 mL) and stirred at 40 °C for 24 hrs. The solution was poured into ice-water and the precipitated solid was filtered, washed with water, and dried under high vacuum to give 6-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]py din-5-carboxamide as a white solid. A portion of this material (0.122 g) was dissolved in DMF (2.0 mL). 2-Chlorotriethylamine hydrochloride (0.192 g) and potassium carbonate (0.304 g) were added to the solution. The reaction mixture was stirred at 60 °C for 18 h. The reaction mixture was poured into water, extracted with ethyl acetate. The extract was washed with saturated NaHCO3 solution and the brine, and dried over Na2SO4. The extract was evaporated under reduced pressure to give 6-[2- (diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridine-5-carboxamide as a white solid. A portion of this material (0.059 g) was dissolved in hot toluene (10 L) and azeotroped for 2 hrs. The solution was cooled to room temperature. 4- Fluoro-3-(trifluoromethyl) phenylisocyanate (0.098 g) was added to the solution. The mixture was heated at reflux for 23 hrs, and then, cooled to room temperature. The solvent was evaporated under reduced pressure, and the residue was purified with the column chromatography (ethyl acetate). A portion of this material (20mg) was dissolved in tetrahydrofuran (1.0 mL) and treated with 1 N hydrochloric acid solution (1.0 mL). The mixture was lyophilized to give {6-[2-(diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl}-N-({[4-fluoro- 3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride as a white solid. 1H NMR (DMSO-d6) δ 1.25 (m, 8H), 2.49 (s, 3H), 3.61 (m, 2H), 3.93 (s, 3H), 4.85 (m, 2H), 7.51 (m, 1 H), 7.86 (m, 1 H), 8.08 (m, 1 H), 8.52 (s, 1 H), 10.24 (br s, 1 H), 10.80 (d, J=4.0 Hz, 2H). MS (ESI) m/z 51 1 (M+H), 509 (M-H). N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(6-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (109).
1H NMR (DMSO-de) δ 2.33 (s, 3H), 3.69 (s, 3H), 7.69 (t, J=10.0 Hz, 1 H), 7.81 (m, 1 H), 8.10 (m, 1 H), 8.50 (s, 1 H), 11.27 (s, 1 H), 14.60 (br s, 1 H). MS (ESI) m/z 412 (M+H), 410 (M-H). [1 ,3-Dimethyl-6-(1 -methylethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (110). 1H NMR (DMSO-de) δ 1.45 (d, J=6.0 Hz, 6H), 2.48 (s, 3H), 3.90 (s, 3H), 5.53 (m, 1 H), 7.50 (t, J=9.6 Hz, 1 H), 7.87 (m, 1 H), 8.1 1 (m, 1 H), 8.62 (s, 1 H), 10.48 (br s, 1 H), 10.75 (br s, 1 H). MS (ESI) m/z 454 (M+H), 452 (M-H). [1 ,3-Dimethyl-6-(phenylmethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (thfluoromethyl)phenyl]amino}carbonyl)carboxamide (HI). 1H NMR (DMSO-de) δ 2.48 (s, 3H), 3.93 (s, 3H), 5.60 (s, 2H), 7.31-7.41 (m, 3H), 7.49 (t, J=9.6 Hz, 1 H), 7.58 (d, J=6.8 Hz, 2H), 7.85 (m, 1 H), 8.07 (m, 1 H), 8.58 (s, 1 H), 10.63 (br s, 1 H), 10J1 (br s, 1 H). MS (ESI) m/z 502 (M+H), 500 (M-H).
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(6-methoxy-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (112).
1H NMR (DMSO-de) δ 2.48 (s, 3H), 3.91 (s, 3H), 4.10(s, 3H), 7.50 (t, j=9.6 Hz, 1 H), 7.88 (m, 1 H), 8.09 (m, 1H), 8.55 (s, 1 H), 10.62 (br s, 1 H), 10.75 (br s, 1 H). MS (ESI) m/z 426 (M+H), 424 (M-H).
(1 ,3-Dimethyl-6-morpholin-4-ylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (113).
1H NMR (DMSO-de) δ 2.45 (s, 3H), 3.34 (m, 4H), 3J5 (m, 4H), 3.87 (s, 3H), 7.50 (t, J=9.6 Hz, 1 H), 7.87 (m, 1H), 8.10 (m, 1 H), 8.39 (s, 1 H), 10.69 (br s, 1 H), 1 1.14 (br s, 1 H). MS (ESI) m/z 481 (M+H), 479 (M-H).
[6-(Dimethylamino)-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (114).
1H NMR (DMSO-de) δ 2.40 (s, 3H), 3.03 (s, 6H), 3.82 (m, 4H), 2.98 (s, 3H), 3.64 (t, J=6.4 Hz, 2H), 3.82 (s, 3H), 7.49 (t, j=9.6 Hz, 1 H), 7.87 (m, 1 H), 8.10 (m, 1 H), 8.22 (s, 1 H), 10.76 (br s, 1 H), 11.05 (br s, 1 H). MS (ESI) m/z 439 (M+H), 437 (M-H). [1 ,3-Dimethyl-6-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (115). 1H NMR (DMSO-de) δ 2.45 (s, 3H), 2.80 (m, 3H), 3.12 (m, 2H), 3.48 (t, j=11.2 Hz, 1 H), 3.89 (s, 3H), 7.51 (t, J=9.6 Hz, 1 H), 7.87 (m, 1 H), 8.09 (m, 1 H), 8.41 (s, 1 H), 10.74 (br s, 1 H), 11.00 (br s, 1 H). {6-[2-(Dimethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
(117).
1H NMR (DMSO-de) δ 2.49 (s, 3H), 2.88 (s, 6H), 3.60 (m, 2H), 3.93 (s, 3H), 4.86
(m, 2H), 7.51 (t, J=9.6 Hz, 1 H), 7.87 (m, 1 H), 8.10 (m, 1 H), 8.55 (s, 1 H), 10.42 (br s, 1 H), 10.70 (br s, 1 H), 10J8 (br s, 1 H). MS (ESI) m/z 483 (M+H), 481 (M- H).
{6-[2-(Dimethylamino)-isopropoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N- ({[4-fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (118).
1H NMR (DMSO-de) δ 1.44 (d, J=6.4 Hz, 3H), 2.49 (s, 3H), 2.90 (s, 6H), 3.48- 3.59 (m, 2H), 3.93 (s, 3H), 5.84 (m, 2H), 7.51 (t, J=9.6 Hz, 1 H), 7.86 (m, 1 H), 8.10 (m, 1 H), 8.55 (s, 1 H), 10.02 (br s, 1 H), 10.65 (s, 1 H), 10.73 (s, 1 H). MS (ESI) m/z 497 (M+H), 495 (M-H). {6-[3-(Dimethylamino)propoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyhdin-5-yl}-N-({[4- fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
(119).
1H NMR (DMSO-de) δ 2.24 (m, 2H), 2.49 (s, 3H), 2J3 (s, 1 H), 2J8 (s, 5H), 3.48- 3.59 (m, 2H), 3.92 (s, 3H), 4.58 (t, J=5.0 Hz, 2H), 7.51 (t, J=9.6 Hz, 1 H), 7.89 (m, 1 H), 8.10 (m, 1 H), 8.55 (s, 1 H), 10.11 (br s, 1 H), 10.62 (s, 1 H), 10.79 (s, 1 H). MS (ESI) m/z 497 (M+H), 495 (M-H).
[1 ,3-Dimethyl-6-(2-pyrrolidinylethoxy)pyrazolo[5,4-b]pyhdin-5-yl]-N-({[4-fluoro-3- (thfluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (121). 1H NMR (DMSO-de) δ 1.88-2.05 (m, 4H), 2.49 (s, 3H), 3.12-3.18 (m, 2H), 3.68 (m, 4H), 3.93 (s, 3H), 4.85 (t, j=4.8 Hz, 2H), 7.51 (t, J=9.6 Hz, 1 H), 7.86 (m, 1 H), 8.09 (m, 1 H), 8.53 (s, 1 H), 10.63 (br s, 1 H), 10.62 (d, J=3.6 Hz, 2H). MS (ESI) m/z 509 (M+H), 507 (M-H).
[1 ,3-Dimethyl-6-(2-piperidylethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (122). 1H NMR (DMSO-de) δ 1.38 (br s, 1 H), 1.67-1.78 (m, 5H), 2.49 (s, 3H), 3.04 (br s, 2H), 3.57 (br s, 4H), 3.93 (s, 3H), 4.87 (br s, 2H), 7.53 (t, j=9.6 Hz, 1 H), 7.88 (m, 1 H), 8.09 (m, 1 H), 8.54 (s, 1 H), 10.17 (br s, 1 H), 10.78 (s, 1 H), 10.82 (s, 1 H). MS (ESI) m/z 523 (M+H), 521 (M-H).
Example 10: N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(1-methyl- 3-phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (123).
Figure imgf000059_0001
A mixture of 1-methyl-3-phenylpyrazol-5-ylamine (3.46 g) and diethyl 2- (ethoxymethylene)propan-l ,3-dioate (4.33 g) was heated at 120 °C for 3 hrs with stirring, and then ethanol formed in the reaction was evaporated under reduce pressure. The residue was treated with hexane (50 mL), and the precipitated solid was filtered, washed with hexane, and dried under high vacuum. The solid was dissolved in phosphorus oxychloride (20 mL), heated at reflux for 14 hrs, and the reaction mixture was concentrated under reduced pressure. The residue was treated with water (50 mL), and the resulting solid was filtered, washed with water, and dried under high vacuum to give ethyl 1-methyl-3- phenyl-4-chloro-pyrazolo[5,4-b]pyridin-5-carboxylate.
The solid was dissolved in acetic acid (20 mL), and reacted with zinc (2.00g) at 80 °C for 60 min. Zinc was removed by the filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethanol (30 mL), and treated with 5N sodium hydroxide solution (30 mL). The mixture was heated at reflux for 6 hrs, and then solvent was evaporated under reduced pressure. The residue was poured into water (40 mL), filtered, and the filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum to give 1-methyl-3-phenylpyrazolo[5,4- b]pyridine-5-carboxylic acid as a white powder. The acid was suspended in anhydrous dichloromethane (50 mL) under an argon atmosphere and added oxalyl chloride (5.0 mL), followed by anhydrous DMF (0.5 mL). The mixture was stirred at room temperature for 6 hrs, and then concentrated under reduced pressure. The residue was cooled to ice-bath temperature and a saturated solution of ammonia in tetrahydrofuran (50 mL) was slowly added to the residue. The ice-bath was removed, the reaction mixture was stirred at room temperature for 6 hrs, and the precipitated solid was filtered. The solid was washed with water and dried under high vacuum to give 1-methyl-3-phenylpyrazolo[5,4- b]pyridine-5-carboxamide as a white powder.
A portion of this material (50 mg) was dissolved in hot toluene (5.0 mL) and azeotroped for 6 hrs. The solution was cooled to room temperature, 4-fluoro- 3-(trifluoromethyl)phenylisocyanate (45 mg) was added, and refluxed for 16 hrs. The reaction mixture was cooled to room temperature and the precipitated solid was collected by the filtration, washed with methanol, and dried under high vacuum to give N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(1-methyl- 3-phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide as a white powder (45mg). 1H NMR (DMSO-de) δ 4.17 (s, 3H), 7.49-7.62 (m, 4H), 7.93 (m, 1 H), 8.12-8.15 (m, 3H), 9.15 (d, J=2.2 Hz, 1 H), 9.27 (d, J=2.2 Hz, 1 H), 10.98 (s, 1 H), 1 1.49 (s, 1 H). MS (ESI) m/z 456 (M-H), 458 (M+H).
N-({[4-Fluoro-3-(thfluoromethyl)phenyl]amino}carbonyl)(1-methylpyrazolo[5,4- b]pyridin-5-yl)carboxamide (124). 1H NMR (DMSO-de) δ 4.11 (s, 3H), 7.51 (dd, J=9.9, 9.9 Hz, 1 H), 7.91 (m, 1 H), 8.12 (dd, J=6.6, 2.6 Hz, 1 H), 8.37 (s, 1 H), 8.93 (d, J=2.2 Hz, 1 H), 9.12 (d, J=2.2 Hz, 1 H), 10.90 (s, 1 H), 1 1.30 (s, 1 H). MS (ESI) m/z 380 (M-H), 382 (M+H).
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(3-methyl-1- phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide (125). 1H NMR (DMSO-de) δ 2.68 (s, 3H), 7.37 (m, 1 H), 7.50-7.60 (m, 3H), 7.92 (m, 1 H), 8.13 (dd, J=6.6, 2.6 Hz, 1 H), 8.25 (dd, J=8.8, 1.1 Hz, 2H), 9.11 (d, J=2.2 Hz, 1 H), 9.19 (d, J=2.2 Hz, 1 H), 10.91 (s, 1 H), 1 1.32 (s, 1 H). MS (ESI) m/z 456 (M-H), 458 (M+H).
Example 11 : N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[4-(4- methylpiperazin yl)-1 ,3-diphenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide (126).
Figure imgf000060_0001
Ethyl 4-chloro-1 ,3-diphenylpyrazolo[5,4-b]pyhdine-5-carboxylate (900 mg) and methylpiperazine (4.50 g) were dissolved in tetrahydrofuran (90 mL) and stirred at room temperature for 6 hrs. The reagents were removed under reduced pressure, the residue was dissolved in the mixture of ethanol (27 mL) and 5N sodium hydroxide solution (27 mL), and heated at reflux for 6 hrs. The solvent was evaporated under reduced pressure, the residue was dissolved in water (36 mL), insoluble solids were removed by the filtration. The filtrate was cooled in an ice-bath and acidified to pH 4 with concentrated hydrochloric acid. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum.
The solid was suspended in anhydrous dichloromethane (50 mL) under an argon atmosphere. Oxalyl chloride (5.0 mL) and anhydrous DMF (0.5 mL) was added to the suspension, and the reaction mixture was stirred at room temperature for 6 hrs. The oxalyl chloride was evaporated under reduced pressure, the residue was cooled in an ice bath, and a saturated solution of ammonia in tetrahydrofuran (50 mL) was slowly added. The ice bath was removed, the suspension was stirred at room temperature for 6 hrs. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum to give 1 ,3-diphenylpyrazolo[5,4-b]pyridine-5-carboxamide as a white powder.
A portion of this material (82 mg) was dissolved in hot toluene (8.0 mL) and azeotroped for 6 hrs. The solution was cooled to room temperature, added 4-fluoro-3-(trifluoromethyl)phenylisocyanate (49 mg), and heated at reflux for 16 hrs. The reaction mixture was cooled to room temperature, the precipitated solid was filtered, washed with methanol, and dried under high vacuum to give N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[4-(4-methylpiperazinyl)-1 ,3- diphenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide as a white powder.
1H NMR (DMSO-de) δ 1.91 (m, 4H), 2.01 (s, 3H), 3.10 (m, 4H), 7.37 (m, 1 H), 7.48-7.60 (m, 6H), 7.68 (m, 2H), 7.91 (m, 1 H), 8.12 (m, 1 H), 8.18 (dd, J=8.8, 1.1 Hz, 2H), 8.50 (s, 1 H), 10.65 (s, 1 H), 11.38 (s, 1 H). MS (ESI) m/z 616 (M-H), 618 (M+H). N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[4-(4-methylpiperazinyl)- 1 ,3-diphenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide hydrochloride (127) MS (ESI) m/z 616 (M-H), 618 (M+H).
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[1-methyl-4-(4- methylpiperazinyl)-3-phenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide (128) 1H NMR (DMSO-de) δ 1.92 (m, 4H), 2.00 (s, 3H), 3.06 (m, 4H), 4.06 (s, 3H), 7.48-7.59 (m, 6H), 7.91 (m, 1 H), 8.12 (m, 1 H), 8.41 (s, 1 H), 10.67 (s, 1 H), 11.31 (s, 1 H). MS (ESI) m/z 554 (M-H), 556 (M+H).
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[1-methyl-4-(4- methylpiperazinyl)-3-phenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide hydrochloride (129) MS (ESI) m/z 554 (M-H), 556 (M+H).
Example 12: N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)
(1 ,3,6-thmethylpyrazo lo[5,4-b]pyridin-5-yl)carboxamide (130).
Figure imgf000062_0001
A solution of 5-amino-1 ,3-dimethylpyrazol-4-carbaldehyde (400 mg) and ethyl 3-oxobutanoate (600 mg) in piperidine (5 mL) was reflux for 3 hrs, the reagents were evaporated under reduced pressure, and the residue was washed with ether (5.0 mL) to give ethyl 1 ,3,6-trimethylpyrazolo[5,4-b]pyridin-5- yl)carboxylate as a white powder. The solid was dissolved in the mixture of ethanol (5.0 mL) and 5N sodium hydroxide solution (5.0 mL), the solution was heated at reflux for 6 hrs, and then concentrated under reduced pressure. The residue was poured into water (4 mL), insoluble solid was removed by the filtration, the filtrate was cooled in an ice bath, and acidified to pH 4 with concentrated hydrochloric acid. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum. The solid was suspended in anhydrous dichloromethane (5 mL) under an argon atmosphere. Oxalyl chloride (0.5 mL) and anhydrous DMF (0.05 mL) were added to the suspension, and the mixture was stirred at room temperature for 6 hrs. The reaction mixture was concentrated under reduced pressure, the residue was cooled in an ice bath, and a saturated solution of ammonia in tetrahydrofuran (5 mL) was slowly added into the residue. The ice-bath was removed and the suspension was stirred at room temperature for 1 hr. The precipitated solid was filtrated, washed with water, and dried under high vacuum to give 1 ,3,6-thmethylpyrazolo[5,4-b]pyhdin-5-carboxamide as a white powder. A portion of this material (50 mg) was dissolved in hot toluene (5.0 mL) and azeotroped for 6 hrs. The solution was cooled to room temperature, 4-fluoro-3- (t fluoromethyl)phenylisocyanate (55 mg) was added, and refluxed for 16 hrs. The reaction mixture was cooled to room temperature, the precipitated solid was filtered, washed with methanol, and dried under high vacuum to give N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(1 ,3,6-trimethylpyrazolo[5,4- b]pyridin-5-yl)carboxamide as a white powder.
1H NMR (DMSO-de) δ 2.51 (s, 3H), 2J3 (s, 3H), 3.97 (s, 3H), 7.51 (dd, J=9.9, 9.9 Hz, 1 H), 7.91 (m, 1 H), 8.12 (dd, j=6.2, 2.6 Hz, 1 H), 8.49 (s, 1 H), 10.75 (s, 1 H), 11.16 (s, 1 H). MS (ESI) m/z 408 (M-H), 410 (M+H).
Example 13: Ethyl 1 -{1 ,3-dimethyl-5-[N-({[3-(1 -methylethoxy)phenyl]amino} carbonyl) carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}piperidine-4-carboxylate (131). cooεt
Figure imgf000064_0002
Figure imgf000064_0001
(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[3-(1 - methylethoxy)phenyl]amino}carbonyl)carboxamide (40 mg) was dissolved in tetrahydrofuran (5.0 mL), ethyl pipehdin-4-carboxylate (160 mg) was added , and the mixture was stirred at room temperature for 5 hrs. The solvent was evaporated under reduced pressure, the residue was suspended in water (10 mL), the resulting solid was filtered, washed with water, and dried under high vacuum to give ethyl 1-{1 ,3-dimethyl-5-[N-({[3-(1- methylethoxy)phenyl]amino}carbonyl)carbamoyl]pyrazolo[5,4-b]pyridin-4- yl}piperidin-4-carboxylate as a white powder. 1H NMR (DMSO-de) δ 1.19 (t, J=7.3 Hz, 3H), 1.27 (d, J=5.9 Hz, 6H), 1.79 (m, 2H), 1.98 (m, 2H), 2.57 (m, 1 H), 2.61 (s, 3H), 3.14 (m, 2H), 3.41 (m, 2H), 3.92 (s, 3H), 4.09 (q, J=7.3 Hz, 2H), 4.59 (m, 1 H), 6.67 (dd, J=8.1 , 1.1 Hz, 1 H), 7.05 (dd, J=8.1 , 2.6 Hz, 1 H), 7.25 (m, 2H), 8.36 (s, 1 H), 10.54 (s, 1 H), 11.18 (s, 1 H). MS (ESI) m/z 521 (M-H), 523 (M+H). tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1 -methylethoxy)phenyl]amino}carbonyl) carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}amino)acetate carboxamide (132). 1H NMR (DMSO-de) δ 1.28 (d, J=6.2 Hz, 3H), 1.42 (s, 9H), 2.63 (s, 3H), 3.87 (s, 3H), 4.21 (d, J=5.9 Hz, 2H), 4.59 (m, 1 H), 6.67 (dd, J=7.4, 1.8 Hz, 1 H), 7.02 (dd, J=7.7, 2.6 Hz, 1 H), 7.24 (m, 2H), 7.99 (br, 1 H), 8.44 (s, 1 H), 10.57 (s, 1 H), 10.91 (s, 1 H). MS (ESI) m/z 495 (M-H), 497 (M+H). tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1 -methylethoxy)phenyl]amino}carbonyl) carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}methylamino)acetate (133). 1H NMR (DMSO-de) δ 1.27 (d, J=5.9 Hz, 3H), 1.42 (s, 9H), 2.64 (s, 3H), 3.06 (s, 3H), 3.88 (s, 2H), 3.93 (s, 3H), 4.60 (m, 1 H), 6.67 (dd, J=8.0, 1.8 Hz, 1 H), 7.06 (dd, J=8.1 , 1 .1 Hz, 1 H), 7.25 (m, 2H), 8.41 (s, 1 H), 10.57 (s, 1 H), 1 1.02 (s, 1 H). MS (ESI) m/z 509 (M-H), 511 (M+H).
Example 14: 3-methylisoxazolo[5,4-b]pyridine-5-carboxamide.
Figure imgf000065_0001
Figure imgf000065_0002
A mixture of 5-amino-3-methyl-isoxazole (5.26 g) and diethyl ethoxymethylenemalonate (10.9 ml) was heated at 120 °C for 3 hrs. The mixture was cooled to room temperature. The precipitated solid was collected, washed with n-hexane and dried under high vacuum. A portion of this solid (1 1.37 g) was dissolved in phosphoryl chloride (55 ml), heated at reflux for 1 1 hrs, and then cooled to room temperature. The mixture was concentrated under reduced pressure and the residue was purified with the column chromatography (ethyl acetate: n-hexane = 1 : 9) to give ethyl 3-methylisoxazolo[5,4-b]pyridine-5- carboxylate as a white solid. Zinc powder (2.75 g) was added to a solution of a portion of the solid (2.43 g) in acetic acid (24 ml), the mixture was stirred at 80 °C for 1.5 hrs, and then filtered. The filtrate was concentrated under reduced pressure. The residue was dissolved in a mixture of 5N NaOH (10 ml) and ethanol (10 ml), the solution was refluxed for 1.5 hrs, cooled to room temperature, and acidified with concentrated HCl. The precipitated solid was collected by the filtration, washed with water, and dried under high vacuum.
The solid was suspended in thionyl chloride (50 ml) and heated at 50 °C for 5 hrs. The thionyl chloride was evaporated under reduced pressure, the residue was dissolved with dichloromethane (5 ml), and reacted with ammonia gas at room temperature for 1 day. The reaction mixture was concentrated under reduced pressure, and the residue was treated with water. The precipitated solid was filtered, washed with water, and dried under high vacuum to give 3-methylisoxazolo[5,4-b]pyhdine-5-carboxamide. Such a carboxamide may be reacted with an isocyanate or an amine to produce an acylurea compound of the invention according to Procedure A or B, respectively, above. 1H NMR (DMSO-de) δ 2.62 (s, 3H), 7.72 (br.s, 1 H), 8.27 (br.s, 1 H), 8.87 (d, J=2.0Hz, 1 H), 9.11 (d, J=2.0Hz, 1 H). MS (ESI) m/z 176 (M-H).
Example 15: N-((r4-fluoro-3-(trifluoromethvl)phenvnamino}carbonyl)[3-methvl-4- (4-methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide (134).
Figure imgf000066_0001
A solution of ethyl 4-chloro-3-methylisoxazolo[5,4-b]pyridin-5-carboxylate (1.02 g) and 1 -methylpiperazine (1.12 ml) in tetrahydrofuran (10 ml) was stirred at room temperature for 1 hr and then concentrated under reduced pressure. The residue was treated with water, the precipitated solid was filtered, washed with water, and dried under high vacuum. This solid was suspended in ethanol (1 1 ml), added 5N NaOH (1 1 ml), and the mixture was stirred at 60 °C for 1 hr. The reaction mixture was treated with 5N HCl and purified by ODS column (VARIAN MEGABOND ELUT C18) to give 3-methyl-4-(4-methyl-piperazin-1-yl) isoxazolo[5,4-b]pyridine-5-carboxylic acid as an amorphous solid. A portion of this amorphous solid (417 mg) was dissolved in thionyl chloride (40 ml) and heated at 50 °C for 3.5 hrs. The mixture was concentrated under reduced pressure, the residue was dissolved in THF (20 ml), and reacted with ammonia gas at room temperature for 3 hrs. The reaction mixture was concentrated under reduced pressure, the residue was treated with water, the resulting solid was filtered, washed with water, and dried under high vacuum to give the 3-methyl-4- (4-methyl-piperazin-1 -yl)-isoxazolo[5,4-b]pyridine-5-carboxylic acid amide.
A portion of this material (50 mg) was dissolved in toluene (5 ml) and azeotroped for 3 hrs. The solution was cooled to room temperature and then reacted with 4-fluoro-3-(trifluoromethyl)phenyl isocyanate (0.031 ml). The mixture was refluxed for 3 hrs and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure and the residue was purified with p-TLC (methanol) to give N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)[3-methyl-4-(4- methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide (134).
1H NMR (DMSO-de) delta ppm: 2.23 (s, 3H), 2.64 (s, 3H), 3.16-3.51 (m, 8H), 7.45 (t, J=9JHz, 1 H), 7.84 (m, 1 H), 8.13 (m, 1 H), 8.39 (s, 1 H). MS (ESI) m/z 481 (M+H), 479 (M-H).
N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)[3-methyl-4-(4- methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide hydrochloride (135). 1H NMR (DMSO-de) δ 2.50 (m, 3H), 2.69 (s, 3H), 3.25-3.42 (m, 8H), 7.53 (t, J=9.8Hz, 1 H), 7.89 (m, 1 H), 8.1 1 (m, 1 H), 8.56 (br.s, 1 H), 10.61 (br.s, 1 H), 11.48 (br.s, 1 H). MS (ESI) m/z 481 (M+H), 479 (M-H).
[4-(dimethylamino)-3-methylisoxazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide (136).
1H NMR (DMSO-de) δ 2.64 (s, 3H), 3.10 (s, 6H), 7.53 (t, J=9.9Hz, 1 H), 7.91 (m, 1 H), 8.1 1 (m, 1 H), 8.36 (s, 1 H), 10.68 (br.s, 1 H), 11.32(br.s, 1 H). MS (ESI) m/z 426 (M+H), 424 (M-H).
(4-{[2-(dimethylamino)ethyl]methylamino}-3-methylisoxazolo[5,4-b]pyridin-5-yl)- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide (137).
1H NMR (DMSO-de) δ 2.50 (s, 6H), 2.65 (s, 3H), 3.08 (s, 3H), 3.35 (m, 2H), 3.58 (m, 2H), 7.52 (t, J=9JHz, 1 H), 7.91 (m, 1 H), 8.14 (m, 1 H), 8.46 (s, 1 H),10.80 (br.s 1 H). MS (ESI) m/z 483 (M+H), 481 (M-H).
(4-{[2-(dimethylamino)ethyl]methylamino}-3-methylisoxazolo[5,4-b]pyridin-5-yl)- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride (138). 1H NMR (DMSO-de) δ 2.69 (s, 3H), 2J5 (br.d, J=4Hz, 6H), 3.17 (br.s, 3H), 3.37 (m, 2H), 3.76 (m, 2H), 7.53 (t, J=9.5Hz, 1 H), 7.88 (m, 1 H), 8.10 (m, 1 H), 8.52 (s, 1 H),10.44 (br.s, 1 H), 10.67 (br.s, 1 H), 1 1.39 (br.s, 1 H). MS (ESI) m/z 483 (M+H), 481 (M-H).
N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(3-methyl-4-morpholin-4- ylisoxazolo[5,4-b]pyridin-5-yl)carboxamide (139).
1H NMR (DMSO-de) δ 2.66 (s, 3H), 3.79 (m, 4H), 7.52 (t, j=9.7 Hz, 1 H), 7.91 (1 Hrs), 8.12 (m, 1 H), 8.50 (s, 1 H), 10.63 (br.s, 1 H), 1 1.48 (br.s, 1 H). MS (ESI) m/z 468 (M+H), 466 (M-H).
Compounds of General Formulae III to VI:
The compounds shown in Tables 1 to 4 were prepared either by the procedures described above or by modifications of these procedures familiar to those skilled in the art.
Figure imgf000068_0001
III Table 1
Figure imgf000068_0002
Figure imgf000069_0001
Figure imgf000070_0002
Figure imgf000070_0001
Figure imgf000070_0003
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0002
Figure imgf000074_0001
V
Table 3
Figure imgf000074_0003
Figure imgf000075_0001
Table 4
Figure imgf000075_0002
Figure imgf000076_0001
The names of the compounds shown in Tables 1 to 4 are given in Tables 5 and 6. These names were generated with the Chemistry 4-D Draw™ software from Chemlnnovation Software, Inc. (San Diego, CA).
Table 5
Figure imgf000077_0001
Figure imgf000078_0001
Cmpd Name
4-({[(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-
41 yl)carbonylamino]carbonyl}amino)benzoic acid
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[3-
42 (trifluoromethyl)phenyl]amino}carbonyl)carboxamide
Table 6
Figure imgf000079_0001
Figure imgf000080_0001
(trifluoromethoxy)phenyl]amino}carbonyl)carboxamide, hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-
78 (1 ,3-thiazol-2-yl)phenyl)amino]carbonyl}carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-
79 (1 ,3-thiazol-2-yl)phenyl)amino]carbonyl}carboxamide hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-
80 phenylphenyl)amino]carbonyl}carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-
81 phenylphenyl)amino]carbonyl}carboxamide hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-
82 phenoxyphenyl)amino]carbonyl}carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4
83 methyl-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-
84 fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-
85 fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3-
86 fluoro-4-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-
87 hydroxy-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-
88 hydroxy-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
89 [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- hydroxy-5-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3-
90 hydroxy-5-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
91 [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3,5- bis(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride Methylethyl 5-[({[1 ,3-dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-
92 b]pyridin-5-yl]carbonylamino}carbonyl)amino]-2-chlorobenzoate hydrochloride
93 [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- (phenylcarbonyl)phenyl]amino}carbonyl)carboxamide hydrochloride
94 [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- chloro-4-(morpholin-4-ylcarbonyl)phenyl]amino}carbonyl)carboxamide
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-
95 chloro-3-(morpholin-4-ylcarbonyl)phenyl]amino}carbonyl)carboxamide hydrochloride
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-[({4-
96 chloro-3-[(4- methylpiperazinyl)carbonyl]phenyl}amino)carbonyl]carboxamide
Figure imgf000082_0001
Figure imgf000083_0001
pyrazolo[5,4-b]pyridin-4-yl}piperidine-4-carboxylate tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1 -
132 methylethoxy)phenyl]amino}carbonyl)- carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}amino)acetate carboxamide tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1-
133 methylethoxy)phenyl]amino}carbonyl)- carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}methylamino)acetate
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-
134 [3-methyl-4-(4-methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-
135 [3-methyl-4-(4-methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide hydrochloride
[4-(Dimethylamino)-3-methylisoxazolo[5,4-b]pyridin-5-yl]-
136 N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
(4-{[2-(Dimethylamino)ethyl]methylamino}-3-methylisoxazolo[5,4-
137 b]pyridin-5-yl)-
N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
(4-{[2-(Dimethylamino)ethyl]methylamino}-3-methylisoxazolo[5,4-
138 b]pyridin-5-yl)-
N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide hydrochloride
139 N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)- (3-methyl-4-morpholin-4-ylisoxazolo[5,4-b]pyridin-5-yl)carboxamide
(6-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4-
140 b]pyridin-5-yl)-N-({[3-(1- methylethoxy)phenyl]amino}carbonyl)carboxamide
141 (1,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[3-chloro-4-(morpholin-4- ylcarbonyl)phenyl]amino}carbonyl)carboxamide
142 (1,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-[({3-chloro-4-[(4- methylpiperazinyl)carbonyl]phenyl}amino)carbonyl]carboxamide
Example 16: Inhibition of MCP-1 -Induced Chemotaxis
A 96-well microchemotaxis chamber with a 5 μm-pore size, PVP-coated polycarbonate filter membrane (Neuro Probe Inc., Cabin John, MD) was used for testing. Compounds were prepared as 10 mM stock solution in DMSO. THP-1 cells (2x106 cells/mL) were labeled with 5 μM Calcein AM containing 0.1% F127 (Molecular Probe, Eugene, OR) at 37 °C for 30 min, and then pretreated with compound at room temperature for an additional 30 min. The lower chamber was loaded with medium containing 12.5 nM hMCP-1. The filter membrane was placed over the lower chamber, followed by a silicon gasket and the upper chamber. The pretreated THP-1 cells (4x105 cells/50μL of RPMI1640 medium per well) were added to the upper chamber and incubated in 5% CO2 at 37 °C for 2 h. The migrated cells were determined with a fluorescent plate reader (LJL BioSystems, Sunnyvale, CA). Table 7 shows the IC50 (concentration of compound that inhibited migration of 50% of the cells relative to control) for several compounds of the present invention.
Table 7: Effect of Selected Compounds on MCP-1 Induced Chemotaxis
Figure imgf000085_0001
Example 17: Thioglycollate-lnduced Inflammation Model
3% Brewer's thioglycollate broth (Difco, Detroit, Ml) was injected into the peritoneal cavity of ICR male mice, followed by subcutaneous administration of the test compound. The same dose of test compound was again administered immediately after the broth injection and 3 h later. After 72 h, the number of total elicited cells and MOMA2- positive cells in the peritoneal cavity was analyzed using an EPICS XL Beckman Coulter. The results are shown in Table 8.
Table 8a: Effect of Compound ll on a Thioglycollate-lnduced Inflammation Model
Figure imgf000085_0002
Figure imgf000086_0001
aAnti-MCP-1 Ab was intraperitoneally injected. Significant difference from control group: *P<0.05, **P<0.01 (ANOVA).
Example 18: Thioglycollate-lnduced Inflammation Model 3% Brewer's thioglycollate broth (Difco, Detroit, Ml) was injected into the peritoneal cavity of ICR male mice, followed by subcutaneous administration of the test compound. The same dose of test compound was again administered immediately after the broth injection and 3h later. After 72 h, the number of total elicited cells and MOMA2- positive cells in the peritoneal cavity was analyzed using an EPICS XL Beckman Coulter. The results are shown in Table 9.
Table 9.a Effect of Compounds on a Thioglycollate-lnduced
Inflammation Model
Figure imgf000086_0002
Significant difference from control group: *P<0.05, **P<0.01 (Least Significant Difference
Method).
Example 19: Anti-Thy-1 Antibody-Induced Nephritis Model The efficacy of the compounds of the present invention was also evaluated in an animal model of nephritis. This model very closely simulates the conditions found in human mesangial proliferative glomerulonephritis.
Anti-Thy-1 nephritis was induced by intravenous injection of anti-Thy-1 -antibody to male Wistar rats. The test compound was subcutaneously administered 2 h before, immediately after, and 5 h after the anti-Thy-1 antibody treatment, and then twice a day for the following 2 days. Anti-MCP-1 antibody was intraperitoneally injected once a day for 3 days. Seven days after the anti-Thy-1 antibody treatment, the rats were sacrificed. The kidneys were perfused with 10% formaldehyde in PBS, surgically removed, and immersed in 10% formaldehyde. The kidneys were then embedded in paraffin for glomerular histopathology or in OCT compound (Miles Inc., Elkhart, IN) in liquid nitrogen after immersion in 30% sucrose overnight. Immunohistochemical staining was performed with a mouse anti-rat ED-1 monoclonal antibody. Briefly, 5 μm renal sections were prepared and endogenous peroxidase blocked with 0.3% hydrogen peroxide. The sections were then blocked with Protein Block (DAKO, Japan) and stained with anti-ED-1 antibody for 45 min. The ED-1 antigen was visualized by peroxidase-labeled anti-mouse IgG and diaminobenzidine. The amount of urinary protein was determined with the DC protein assay kit (Bio-Rad, Hercules, CA). The effect of a representative test compound on the level of urinary protein excretion and the infiltration of ED-1 positive cells into the glomeruli is shown in Table 10.
Table 10a: Effect of Compound H on Anti-Thy-1 Antibody Induced Nephritis
Figure imgf000087_0001
Figure imgf000088_0001
Significant difference from control group: **P<0.01 (ANOVA).
Example 20: Apolipoprotein E-Deficient Mouse Model
Apolipoprotein E (apoE) is a component of several plasma lipoproteins, including chylomicrons, VLDL, and HDL. Receptor-mediated catabolism of these lipoprotein particles is mediated through the interaction of apoE with the LDL receptor (LDLR) or with LDLR-related protein (LRP). ApoE-deficient mice exhibit hypercholesterolemia and develop complex atheromatous lesions similar to those seen in humans. The efficacy of the compounds of the present invention was also evaluated using this animal model.
Male, 4 week-old apoE-deficient mice were fed on high-fat diet (15% fat, 1.25% cholesterol). The test compound was administered as food admixture for 8 weeks. At 12 week-old, the mice were fasted for 4 hours and then sacrificed under ether anesthesia. Blood was collected in the presence of heparin, and the hearts were perfused in situ with PBS (pH 7.4), followed by 4% paraformaldehyde for 5 min.
To determine cross-sectional lesion areas, the hearts were embedded in OCT compound and sectioned at 10 μm using cryostat. The sections were stained with oil red O. Each section of the aortic valve was evaluated for oil red O staining by capturing images directly from a RGB camera attached to a light microscope; image analysis was performed with the IPAP-WIN software (Sumika Tekuno, Japan). Five sections were examined for each animal, and the sum of the lesion areas was calculated and expressed as the percent of the total cross-sectional wall area. Total cholesterol was determined with a Determiner assay kit (Kyowa medex, Japan).
The effect of a representative test compound in this animal model of atherosclerosis is shown in Table 1 1.
Table 11a: Effect of Compound H on the ApoE-Deficient Mouse Model of Atherosclerosis
Figure imgf000089_0001
Significant difference from control group: *P<0.05 (t-test).
Compound H reduced the percent of atherosclerotic lesion area and this reduction was statistically significant. There were no significant differences in plasma total cholesterol level and body weight between the control and the test compound-treated groups (data not shown).
Example 21 : Inhibition of MCP-1 -Induced Chemotaxis
A 48-well microchemotaxis chamber with a 5 μm-pore size, PVP-coated polycarbonate filter membrane (Neuro Probe Inc., Cabin John, MD) was used for testing. Compounds were prepared as 10 mM stock solution in DMSO. THP-1 cells were washed with RPMI 1640 medium supplemented with 0.5% BSA and 25 mM HEPES, pH 7.4, and suspended at a density of 4x106 cells/mL in the same medium. A 150 μL aliquot of this suspension was treated with an equal volume of test compound solution and the mixture incubated at 37 °C for 15 min. The lower chamber was loaded with 26 μL of a 2.5 nM solution of hMCP-1 (PeproTech) in medium. The filter membrane was placed over the lower chamber, followed by a silicon rubber gasket and the upper chamber. A 50 μL aliquot of the THP-1 cell suspension containing the test compound was added to the upper chamber and the assembly incubated in a 5% CO2 atmosphere at 37 °C for 2 hr. The chamber was then disassembled and the cells remaining on the upper surface of the filter were scrapped off with a rubber scrapper. The filter was fixed with methanol, stained with Diff-Quik solution, and mounted on a slide glass. The cells that had migrated across the filter onto the lower surface were then counted by microscopic observation. Table 12 shows the IC50 (concentration of compound that inhibited migration of 50% of the cells relative to control) for several compounds of the present invention. Table 12: Effect of Selected Compounds on MCP-1-lnduced Chemotaxis
Figure imgf000090_0001
Example 22: Adjuvant Arthritis Model in Rat
Adjuvant arthritis was induced in male Lewis rats as follows. Complete Freunt's adjuvant (CFA) was mixed with PBS and 0.1 ml of the resulting emulsion was injected subcutaneously into the foot pad of the right hind paw (day=0). The test compound was administered p.o. at day 0 and then once a day for 26 days. Paw swelling was measured by the increased thickness of the foot pad. Table 13 shows the change in the foot pad edema expressed as percent swelling relative to the pad thickness on day 0.
Table 13. Effect of Compound 85 on the Adjuvant Arthritis Model
Figure imgf000091_0001
Example 23: Collagen-induced Arthritis Model in Rat
Collagen-induced arthritis was induced in female Lewis rats as follows. Rats were injected subcutaneously with the emulsion of bovine collagen type II (3.3 mg/kg at day 0 and 1.67 mg/kg at day 7) and complete Freund's adjuvant (CFA) at the base of the tail at day 0 and immunized additionally at day 7. The test compounds were administered i.p. at day 0 and then once a day for 18 days or p.o. (methotrexate) at day 0 and then 3 times a week until day 18. Paw swelling was measured by the increased volume of the legs using a plethysmometer. Table 14 shows the change in the swelling expressed as percent swelling relative to the volume of legs on day 0. Table 14. Effect of Compounds on the Collagen-induced Arthritis Model
Figure imgf000092_0001
Significant difference from control group: *P<0.05, **P<0.01 (Least Significant Difference Method).
Example 24: Collagen-induced Arthritis Model in Rat
Collagen-induced arthritis was induced in female Lewis rats as follows. Rats were injected subcutaneously with the emulsion of bovine collagen type II (3.3mg/kg at day 0 and 1.67mg/kg at day 7) and complete Freund's adjuvant (CFA) at the base of the tail at day 0 and immunized additionally at day 7. The test compounds were administered p.o. at day 0 and then twice a day for 21 days or 3 times a week until day 21 was also administered p.o. from day 13 to day 21 in another group. Paw swelling was measured by the increased volume of the legs using a plethysmometer. Table 15 shows the change in the swelling expressed as percent swelling relative to the volume of legs on day 0.
Table 15. Effect of Compounds on the Collagen-induced Arthritis Model
Figure imgf000092_0002
Figure imgf000093_0001
Significant difference from control group: *P<0.05, **P<0.01 (Least Significant Difference Method). Methotrexate is used as a positive control.
Example 25: Restenosis Model in Rat Effects of MCP-1 antagonists on rat carotid artery responses to the balloon catheter injury.
Male, δweeks old Sprague-Dawley rats were anaesthetized with sodium pentobarbital (50 mg/kg, i.p.) The endothelium of the left common carotid artery was denuded by 3 passages of an inflated 2F Fogarty embolectomy catheter. Test compounds were administered as food admixture to 7 days prior to balloon injury and continued for 14 days after injury. Two weeks after injury, rats were sacrificed under ether anesthesia. The carotid arteries were removed and fixed with 10% neutral buffered formalin for 24 hours. To determine cross-sectional lesion areas, the carotid arteries were embedded in paraffin and sectioned 4-6 μm. Four cross sections of each artery were stained with elastic van Gieson. Intimal and medial areas were measured by a computer-based image analyzing system (IPAP-WIN, Sumica techos). Serum MCP-1 volume was determined with rat MCP-1 ELISA kit (Cosmo bio).
Table 16 Intimal and Medial areas of rat carotid arteries 14 days after balloon injury.
Figure imgf000093_0002
Data are shown as mean±SE. *, ***: Significantly different from the control at p<0.05 and p<0.001 , respectively. Example 26: Anti-Thy-1 Antibody Induced Nephritis Model
Anti-Thy-1 nephritis was induced by intravenous injection of anti-Thy-1 -antibody to male Wistar rats. The test compound was per os administered 2 h before, immediately after, and 5 h after the anti-Thy-1 antibody treatment, and then twice a day for the following 2 days. Seven days after the anti-Thy-1 antibody treatment, the rats were sacrificed. The amount of urinary protein was determined with the DC protein assay kit (Bio-Rad, Hercules, CA). The effect of a representative test compound on the level of urinary protein excretion is shown in Table 17.
Table 17.a Effect of Compound on Anti-Thy-1 Antibody Induced Nephritis
Figure imgf000094_0001
Example 27: Oral pharmaceutical composition - Solid dosage formulation
A pharmaceutical composition for oral administration may be prepared by combining the following:
% w/w
Compound of this invention 10.0
Magnesium stearate 0.5
Starch 2.0
HPM cellulose 1.0
Microcrystalline cellulose 86.5
The mixture may be compressed to tablets, or filled into hard gelatin capsules. The tablet may be coated by applying a suspension of film former
(e.g., HPM cellulose), pigment (e.g., titanium dioxide) and plasticiser (e.g., diethyl phthalate) and drying the film by evaporation of the solvent. The film coat can comprise 2.0% to 6.0% of the tablet weight, preferably about 3.0%.
Example 28: Oral pharmaceutical composition preparation - Capsule
A pharmaceutical composition of a compound of the invention suitable for oral administration may also be prepared by combining the following:
% w/w Compound of this invention 20
Polyethylene glycol 400 80
The medicinal compound is dispersed or dissolved in the liquid carrier, with a thickening agent added, if required. The formulation is then enclosed in a soft gelatin capsule by suitable technology. Example 29: Pharmaceutical composition for parenteral administration
A pharmaceutical composition for parenteral administration may be prepared by combining the following:
Preferred Level (%) Compound of this invention 1.0 Saline 99.0
The solution is sterilized and sealed in sterile containers.
All references cited above are herein incorporated in their entirety by reference. Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as disclosed should not be unduly limited to such specific embodiments. Various modifications of the described modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of this invention.

Claims

We claim:
1 . A compound of formula I or formula II:
Figure imgf000097_0001
(0 (ii) wherein
Y is O, S or N-R7,
Z is N or C-R8,
R1, R2, R3, and R8 are independently, hydrogen, or optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl (lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR9, -SR9, -NR9R10, -NR9(carboxy(lower alkyl)), -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -OC(=O)R9, -SO2R9, -OSO2R9, -SO2NR9R10, -NR9SO2R10 or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(d-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl(lower alkyl), aryl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group,
R7 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -SO2R9, or -SO2NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl(lower alkyl), aryl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2) .6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or
N-(optionally substituted Cι-2 alkyl) group, R4 and R5 are independently, hydrogen, lower alkyl optionally substituted lower alkyl, optionally substituted aryl, or optionally substituted aryl(lower alkyl), or, together, are -(CH2)2. -, R6 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), -C(=O)R11, -C(=O)OR11, -C(=O)NR11R12, -SO2R11, or -SO2NR11R12, wherein R11 and R12 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R11 and R12 together are -(CH2)4.6-, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
2. The compound of claim 1 , wherein said compound is a compound of Formula I or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
3. The compound of claim 1 , wherein said compound is a compound of Formula II or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
4. The compound of claim 1 , wherein Y is O or N-R7. 5. The compound of claim 1 , wherein R1 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), aryl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl (lower alkyl).
6. The compound of claim 5, wherein R1 is optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι.2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), aryl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl). 7. The compound of claim 1 , wherein R2 is hydrogen, optionally substituted lower alkyl, cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9(carboxy(lower alkyl)), -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2) -6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
8. The compound of claim 7, wherein R2 is optionally substituted lower alkyl, cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halogen, -OR9, -NR9(carboxy(lower alkyl)), -C(=O)OR9, -C(=O)NR9R10, -SO2NR9R10, or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(d-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl (lower alkyl), or R9and R10 together are -(CH2)4-6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
9. The compound of claim 1, wherein R3 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, or -C(=O)NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9and R10 together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
10. The compound of claim 9, wherein R3 is optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), halogen, -OR9, -NR9R10, -C(=O)OR9, or -C(=O)NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C-|.2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι.2 alkyl) group.
11. The compound of claim 1 , wherein Y is N-R7, and R7 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -SO2R9, or -SO2NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, heteroaryl, or heteroaryl(lower alkyl).
12. The compound of claim 1 , wherein Z is C-R8, and R8 is hydrogen, optionally substituted lower alkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl(lower alkyl), halo(lower alkyl), -CF3, halogen, -OR9, -NR9R10, -C(=O)R9, -C(=O)OR9, -C(=O)NR9R10, -OC(=O)R9, -SO2R9, -SO2NR9R10, -NR9SO2R10 or -NR9C(=O)R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(C1.2 alkyl)2, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
13. The compound of claim 1 , wherein R4 and R5 are independently, hydrogen or lower alkyl.
14. The compound of claim 1 , wherein R6 is hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), -C(=O)R11, -C(=O)OR11, -C(=O)NR11R12, -SO2R11, or -SO2NR11R12, wherein R11 and R12 are independently, hydrogen, optionally substituted lower alkyl, cycloalkyl, cycloalkyl(lower alkyl), aryl, heteroaryl, heteroaryl(lower alkyl), or R11 and R12 together are -(CH2)4-6-.
5. The compound of claim 1 that is a compound of formula la or formula lla:
Figure imgf000102_0001
(la)
(lla) wherein:
Y is O, S or N-R7,
Z is N or C-R8,
R1, R2, R3, R4, R5, R7 and R8 are as defined in claim 1 ,
R13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, -NR15SO2R16 or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), or, together, are -(CH2) -6- optionally interrupted by one O, S, NH or N-(C-|.2 alkyl) group, each R14 is independently selected from optionally substituted lower alkyl, optionally substituted aryl, optionally substituted heteroaryl, hydroxy, halogen, -CF3, -OR17, -NR17R18, -C(=O)R18, -C(=O)OR18, -C(=O)NR17R18, wherein R17 and R18 are independently, hydrogen, lower alkyl, alkenyl, alkynyl, -CF3, optionally substituted heterocycloalkyl, cycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted aryloxy, heteroaryl, heteroaryl(lower alkyl), or, together, are -(CH2) .6-, optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group, and where n is an integer of 0 to 4, or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers.
16. The compound of claim 15, wherein said compound is a compound of Formula la or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers.
17. The compound of claim 15, wherein said compound is a compound of Formula lla or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers.
18. The compound of claim 15, wherein R13 is -OR15, and R15 is hydrogen, lower alkyl optionally substituted with -C(=O)OR19, wherein R19 is hydrogen or lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heteroaryl(lower alkyl).
19. The compound of claim 15, wherein R13 is hydrogen, optionally substituted lower alkyl, alkenyl, heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, or -NR15C(=O)R16, wherein R 5 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl) or, together, are -(CH2)4.6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group.
20. The compound of claim 19, wherein R13 is optionally substituted lower alkyl, alkenyl, heterocycloalkyl, optionally substituted aryl, optionally substituted aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl), halo(lower alkyl), -CF3, halogen, nitro, -CN, -OR15, -SR15, -NR15R16, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaryl (lower alkyl) or, together, are -(CH2)4-6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group.
21. The compound of claim 15, wherein R14 is independently selected from optionally substituted lower alkyl, optionally substituted aryl, optionally substituted heteroaryl, hydroxy, halogen, -CF3, -OR17 -NR17R18, -C(=O)R18, -C(=O)OR18, -C(=O)NR17R18, wherein R17 and R18 are, independently, hydrogen, lower alkyl, alkenyl, or optionally substituted aryl.
22. The compound of claim 20, wherein n is an integer of 1 to 2.
23. The compound of claim 22, wherein n is 1.
24. The compound of claim 15, wherein Y is O, and R1 is lower alkyl. 25. The compound of claim 15, wherein Y is N-R7, R7 is hydrogen or lower alkyl, and R1 is lower alkyl.
26. The compound of claim 15, wherein R1 is methyl, Y is N-R7, and R7 is methyl.
27. The compound of claim 15, wherein Z is N. 28. The compound of claim 15, wherein Z is C-R8, and R8 is hydrogen.
29. The compound of claim 15, wherein R2 and R3 are independently selected from hydrogen, lower alkyl, halogen, OR9, -NR9R10, where R9 and R10 are independently lower alkyl, substituted lower alkyl, or substituted aryl, or R9 and R10 together are -(CH2)4.6- optionally interrupted by one O, S, NH, N-(aryl), N- (aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι-2 alkyl) group.
30. The compound of claim 20, wherein R13 is independently selected from halogen, optionally substituted aryl, -CF3, -CH3, -CN, -OR15, -C(=O)R15, -C(=O)OR15, -C(=O)NR15R18 , or -CO2H.
31. The compound of claim 15, wherein R14 is independently selected from halogen, optionally substituted lower alkyl, -CF3, -OR17, aryl, heteroaryl, -NR17R18, -C(=O)R17, -C(=O)OR17, -C(=O)NR17R18, or -CO2H, where R17 and R18 are, independently, lower alkyl, substituted lower alkyl, or substituted aryl, or, together, are -(CH2) .6- optionally interrupted by one O, S, NH or N-(Cι-2 alkyl) group.
32. The compound of claim 15, wherein Z is N, R2 is 4-methylpiperazinyl, R 3 is 3-CF3, and R14 is 4-F.
33. The compound of claim 14, wherein Y is N-R7 and Z is N, R1 is lower alkyl, and R4 and R5 are hydrogen.
34. The compound of claim 33, wherein R2 is -NR9R10.
35. The compound of claim 15, wherein Y is N-R7 and Z is N,
R1 is lower alkyl, R2 is -NR9R10, wherein R9 and R10 are independently, hydrogen, optionally substituted lower alkyl, lower alkyl-N(Cι-2 alkyl)2, lower alkyl(optionally substituted heterocycloalkyl), alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkyl(lower alkyl), benzyl, optionally substituted aryl, heteroaryl, heteroaryl(lower alkyl), or R9 and R10 together are -(CH2)4.6- optionally interrupted by one O, S,
NH, N-(aryl), N-(aryl(lower alkyl)), N-(carboxy(lower alkyl)) or N-(optionally substituted Cι.2 alkyl) group, and
R13 is hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl(lower alkyl), heterocycloalkyl, optionally substituted aryl, aryl(lower alkyl), optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl), halo(lower alkyl),
-CF3, halogen, nitro, -CN, -OR 1150, -SR ,1153, -NR )1η5°DR116, -C(=O)R >15 -C(=O)OR15, -C(=O)NR15R16, -OC(=O)R15, -SO2R15, -SO2NR15R16, -NR15SO2R16 or -NR15C(=O)R16, wherein R15 and R16 are independently, hydrogen, optionally substituted lower alkyl, alkenyl, alkynyl, -CF3, cycloalkyl, optionally substituted heterocycloalkyl, cycloalkyl(lower alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heteroaryl(lower alkyl) or, together, are -(CH2)4-6- optionally interrupted by one O, S, NH or N-(Ci.2 alkyl) group.
36. The compound of claim 35, wherein R4 and R5 are hydrogen. 37. The compound of claim 1 that is:
Figure imgf000106_0001
or a pharmaceutically acceptable salt thereof.
38 The compound of claim 1 that is:
Figure imgf000106_0002
, or a pharmaceutically acceptable salt thereof.
39. A compound of claim 1 that is:
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(4-fluorophenyl)amino]carbonyl}- carboxamide;
Λ/-{[(3,4-Dichlorophenyl)amino]carbonyl}(1 ,3-dimethylpyrazolo[5,4-b]pyridin- 5-yl)carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -Λ/-{[(3-chlorophenyl)amino]carbonyl} -carboxamide;
Λ/-{[(2,4-Dichlorophenyl)amino]carbonyl (1 ,3-dimethylpyrazolo[5,4-b]pyridin- 5-yl)carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -Λ/-{[(4-chlorophenyl)amino]carbonyl}
-carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl >-N-{[ 4-iodophenyl)amino]carbonyl}- carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -N-{[ 3-cyanophenyl)amino]carbonyl} -carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -Λ/-{[ 3-bromophenyl)amino]carbonyl}- carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl ■N-{[ 3-chloro-4-hydroxyphenyl)amino]- carbonyl}carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl ■N-({\ 3,5-bis(trifluoromethyl)phenyl]amino}
-carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl ■N-({\ 4-fluoro-3-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl >-N-({ 3-fluoro-4-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -Λ/-({[4-nitro-3-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
Λ/-{[(2,3-Dichlorophenyl)amino]carbonyl (1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- carboxamide; Λ/-{[(2,6-Dichlorophenyl)amino]carbonyl (1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- carboxamide;
Λ/-{[(2,5-Dichlorophenyl)amino]carbonyl (1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- carboxamide;
Λ/-{[(3,5-Dichlorophenyl)amino]carbonyl (1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl -Λ/-({[2-chloro-5-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[4-chloro-2-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[2-chloro-4-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[4-chloro-3-(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[2,5-bis(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[3,4-bis(trifluoromethyl)phenyl]- amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[3-iodophenyl)amino]carbonyl}- carboxamide;
2-[4-({[(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)carbonylamino]carbonyl}- amino)-2-chlorophenoxy] acetic acid; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[3-(trifluoromethyl)phenyl]amino}
-carbonyl )carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[4-hydroxy-3-(trifluoromethyl)- phenyl]amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[5-hydroxy-3-(trifluoromethyl)- phenyl]amino}carbonyl)carboxamide;
3-({[(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)carbonylamino)carbonyl}amino)- 5-(trifluoromethyl)benzoic acid;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[3-(1- methylethoxy)phenyl]amino}-carbonyl)carboxamide; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(3-phenoxyphenyl)amino]- carbonyljcarboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(3-phenylphenyl)amino]carbonyl }-carboxamide;
4-({[(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)carbonylamino]carbonyl}amino)- 2-(1-methylethoxy)benzoic acid;
Ethyl2-[4-({[(1 ,3-dimethypyrazolo[5,4-b]pyridin-5-yl)carbonylamino]carbonyl}- amino)-2-(trifluoromethyl)phenoxy]acetate; 2-[4-({[(1 ,3-Dimethypyrazolo[5,4-b]pyridin-5-yl)carbonylamino]carbonyl}amino )-2-(trifluoromethyl)phenoxy]acetic acid;
(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(3-chlorophenyl)amino] -carbonyljcarboxamide; Λ/-{[(3,4-Dichlorophenyl)amino]carbonyl}(4-chloro-1 ,3-dimethyl- pyrazolo[5,4-b]pyridin-5-yl)carboxamide;
(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-({[4-(trifluoromethyl)- phenyl]amino}carbonyl)carboxamide;
(4-Chloro-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)-Λ/-{[(4-chlorophenyl)amino]- carbonyljcarboxamide;
2-[4-({[(1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)carbonylamino]- carbonyl}amino)-2-chlorophenoxy]acetic acid;
4-({[(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)carbonylamino]carbonyl}amino)- benzoic acid; (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[3-
(trifluoromethyl)phenyl]amino}-carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)(4-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
[4-(Dimethylamino)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(4-{[2-(Dimethylamino)ethyl]amino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(4-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4- b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(4-{[3-(Dimethylamino)propyl]amino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl)- N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{4-[bis(2-Hydroxyethyl)amino]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; {1 ,3-Dimethyl-4-[(2-morpholin-4-ylethyl)amino]pyrazolo[5,4-b]pyridin-5-yl}-N-
({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; {1 ,3-Dimethyl-4-[methyl(1-methylpyrrolidin-3-yl)amino]pyrazolo[5,4-b]pyridin- 5-yl}-N-({[4.f|Uoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{4-[(4-Aminocyclohexyl)amino]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; {1 ,3-Dimethyl-4-[(perhydropyrrolizin-7a-ylmethyl)amino]pyrazolo[5,4- b]pyridin-5-yl}-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{1 ,3-Dimethyl-4-[(2-perhydropyrrolizin-7a-ylethyl)amino]pyrazolo[5,4- b]pyridin-5-yl}-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{1 ,3-Dimethyl-4-[benzylamino]pyrazolo[5,4-b]pyridin-5-yl}-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{4-[3-(Dimethylamino)pyrrolidinyl]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; (1 ,3-Dimethyl-4-piperidylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3-
(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(1 ,3-Dimethyl-4-morpholin-4-ylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(1 ,3-Dimethyl-4-piperazinylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3- chlorophenyl)amino]carbonyl}carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3- bromophenyl)amino]carbonyl}carboxamide; [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(4- chlorophenyl)amino]carbonyl}carboxamide;
N-{[(3,4-Dichlorophenyl)amino]carbonyl}[1 ,3-dimethyl-4-(4- methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- (methylethyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3-(1- methylethoxy)phenyl]amino}carbonyl)carboxamide;
4-[({[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5- yl]carbonylamino}carbonyl)amino]-2-(1-methylethoxy)benzoic acid; [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3-
(trifluoromethoxy)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3-(1 ,3- thiazol-2-yl)phenyl)amino]carbonyl}carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3- phenylphenyl)amino]carbonyl}carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-{[(3- phenoxyphenyl)amino]carbonyl}carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4- methyl-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-
3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3-fluoro- 4-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4- hydroxy-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- hydroxy-5-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3,5- bis(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; Methylethyl-5-[({[1 ,3-dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5- yl]carbonylamino}carbonyl)amino]-2-chlorobenzoate;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- (phenylcarbonyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[3- chloro-4-(morpholin-4-ylcarbonyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4- chloro-3-(morpholin-4-ylcarbonyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-[({4- chloro-3-[(4-methylpiperazinyl)carbonyl]phenyl}amino)carbonyl]carboxamide;
[1 ,3-Dimethyl-4-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-[({3- chloro-4-[(4-methylpiperazinyl)carbonyl]phenyl}amino)carbonyl]carboxamide; [4-(4-methylpiperazinyl)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-
3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[4-(4ι4-Dimethylpiperazinyl)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[4-(4-Ethylpiperazinyl)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl){4-[4-(2- hydroxyethyl)piperazinyl]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}carboxamide;
[1 ,3-Dimethyl-4-(4-phenylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro- 3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; {1 ,3-Dimethyl-4-[4-benzylpiperazinyl]pyrazolo[5,4-b]pyridin-5-yl}-N-({[4-fluoro-
3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide
4-Amino-1-{5-[N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl) carbamoyl]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-4-yl}pyridine;
[1 ,3-Dimethyl-4-(4-methyl(1 ,4-diazaperhydroepinyl))pyrazolo[5,4-b]pyridin-5- yl]-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(6-hydroxy-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
[1 ,3-Dimethyl-6-(1-methylethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-6-(phenylmethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3-
(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(6-methoxy-1 ,3- dimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
(1 ,3-Dimethyl-6-morpholin-4-ylpyrazolo[5,4-b]pyridin-5-yl)-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[6-(Dimethylamino)-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-y|]-N-({[4-fluoro-3- (trifluoromethyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-6-(4-methylpiperazinyl)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro- 3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(6-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin- 5-yl)-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; {6-[2-(Dimethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{6-[2-(Dimethylamino)-isopropoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{6-[3-(Dimethylamino)propoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N- ({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
{6-[2-(Diethylamino)ethoxy]-1 ,3-dimethylpyrazolo[5,4-b]pyridin-5-yl}-N-({[4- fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
[1 ,3-Dimethyl-6-(2-pyrrolidinylethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro- 3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide; [1 ,3-Dimethyl-6-(2-piperidylethoxy)pyrazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3-
(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(1-methyl-3- phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(1- methylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(3-methyl-1- phenylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-[4-(4- methylpiperazinyl)-1 ,3-diphenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide; N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-[1-methyl-4-(4- methylpiperazinyl)-3-phenylpyrazolo[5,4-b]pyridin-5-yl]carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(1 ,3,6- trimethylpyrazolo[5,4-b]pyridin-5-yl)carboxamide;
Ethyl 1 -{1 ,3-dimethyl-5-[N-({[3-(1 -methylethoxy)phenyl]amino}carbonyl) carbamoyl]-pyrazolo[5,4-b]pyridin-4-yl}piperidine-4-carboxylate; tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1-methylethoxy)phenyl]amino}carbonyl)- carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}amino)acetate carboxamide; tert-Butyl 2-({1 ,3-dimethyl-5-[N-({[3-(1-methylethoxy)phenyl]amino}carbonyl)- carbamoyl]pyrazolo[5,4-b]pyridin-4-yl}methylamino)acetate;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-[3-methyl-4-(4- methylpiperazinyl)isoxazolo[5,4-b]pyridin-5-yl]carboxamide; [4-(Dimethylamino)-3-methylisoxazolo[5,4-b]pyridin-5-yl]-N-({[4-fluoro-3-
(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
(4-{[2-(Dimethylamino)ethyl]methylamino}-3-methylisoxazolo[5,4-b]pyridin-5- yl)-N-({[4-fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)carboxamide;
N-({[4-Fluoro-3-(trifluoromethyl)phenyl]amino}carbonyl)-(3-methyl-4- morpholin-4-ylisoxazolo[5,4-b]pyridin-5-yl)carboxamide;
(6-{[2-(Dimethylamino)ethyl]methylamino}-1 ,3-dimethylpyrazolo[5,4-b]pyridin- 5-yl)-N-({[3-(1-methylethoxy)phenyl]amino}carbonyl)carboxamide;
(1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-({[3-chloro-4-(morpholin-4- ylcarbonyl)phenyl]amino}carbonyl)carboxamide; and (1 ,3-Dimethylpyrazolo[5,4-b]pyridin-5-yl)-N-[({3-chloro-4-[(4- methylpiperazinyl)carbonyl]phenyl}amino)carbonyl]carboxamide; or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
40. A pharmaceutical composition comprising: (a) a therapeutically effective amount of a compound of any one of claims 1 to 39; and
(b) a pharmaceutically acceptable excipient.
41. The pharmaceutical composition of claim 40, further comprising an anti- inflammatory drug, cytokine, or immunomodulator. 42. The pharmaceutical composition of claim 40 for treating an allergic, inflammatory, or autoimmune disorder or disease.
43. The pharmaceutical composition of any one of claims 40 or 42, wherein the allergic, inflammatory, or autoimmune disorder or disease is selected from the group consisting of asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, and transplant rejection. 44. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is asthma.
45. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is atherosclerosis.
46. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is glomerulonephritis.
47. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is pancreatitis.
48. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is restenosis.
49. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is rheumatoid arthritis.
50. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is diabetic nephropathy.
51. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is pulmonary fibrosis.
52. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is inflammatory bowel disease.
53. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is Crohn's disease.
54. The pharmaceutical composition of claim 43, wherein the allergic, inflammatory, or autoimmune disorder or disease is transplant rejection.
55. A method of treating an allergic, inflammatory, or autoimmune disorder or disease, comprising administering a therapeutically effective dose of at least one compound of any one of claims 1 to 39 to a mammal in need of such treatment.
56. The method of claim 55, wherein the compound is administered in combination with an anti-inflammatory drug, cytokine, or immunomodulator.
57. The method of claim 55, wherein the allergic, inflammatory, or autoimmune disorder or disease is selected from the group consisting of asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, and transplant rejection.
58. The method of claim 55, wherein the allergic, inflammatory, or autoimmune disorder or disease is associated with lymphocyte and/or monocyte accumulation.
59. A method of inhibiting leukocyte migration, comprising administering a therapeutically effective dose of at least one compound of any one of claims 1 to 39 to a mammal in need of such treatment.
60. The use of a compound of any one of claims 1 to 39 in the preparation of a medicament for the treatment of an allergic, inflammatory, or autoimmune disorder or disease.
61. The use of claim 60, wherein the allergic, inflammatory, or autoimmune disorder or disease is selected from the group consisting of asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, and transplant rejection. 62. The use of claim 60, wherein the allergic, inflammatory, or autoimmune disorder or disease is associated with lymphocyte and/or monocyte accumulation.
63. A process for preparing a compound of formula I or formula II
Figure imgf000117_0001
(I) OO
wherein Y, Z, and R1 - R6 are as defined in claim 1 , said process comprising:
(a) contacting a compound of formula lb or lib
Figure imgf000117_0002
(lb) (lib) with a compound of the formula
R° — N=C=O
under conditions sufficient to produce a compounds of formula I or formula II, wherei nn RR44 aanndd RR55 aarree bbootthh HH;; oorr (b) contacting a compound of formula lc or formula lie
Figure imgf000117_0003
(lc) (lie)
wherein X is halogen, nitro, -CN, or -OR9, with a compound of the formula
R -H, under conditions sufficient to produce a compound of formula I or formula
I; or
(c) contacting a compound of formula lb or formula Mb
Figure imgf000118_0001
(lb) (Mb) with a haloformylation reagent and a compound of the formula
Figure imgf000118_0002
under conditions sufficient to produce a compound of formula I or formula II, wherein R4 is H; or (d) elaborating substituents of a compound of formula I or formula II in a manner known per se; or
(e) reacting the free base of a compound of formula I or formula II with an acid to give a pharmaceutically acceptable addition salt; or
(f) reacting an acid addition salt of a compound of formula I or formula II with a base to form the corresponding free base; or
(g) converting a salt of a compound of formula I or formula II to another pharmaceutically acceptable salt of a compound of Formula I or II; or
(h) resolving a racemic mixture of any proportions of a compound of formula I or formula II to yield a stereoisomer thereof. 64. The process of claim 63(a) wherein step (a) is carried out in the presence of an organic solvent or a mixture of solvents at elevated temperatures.
65. The process of claim 64 where the organic solvent is toluene and the reaction is carried out under refluxing conditions.
66. The process of claim 63(b), wherein step (b) is carried out using the salt of the compound of the formula R2-H in an organic solvent. 67. The process of claim 63(b) wherein the salt of the compound of the formula R2-H is the lithium, sodium, or potassium salt.
68. The process of claim 63(c) wherein step (c) is carried out in an organic solvent or a mixture of solvents at elevated temperatures.
69. The process of claim 63(c) where the haloformylation reagent is oxalyl chloride, the organic solvent is THF, and the reaction is heated above 50 °C.
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Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005014600A1 (en) * 2001-01-31 2005-02-17 Telik, Inc. Antagonists of mcp-1 function and methods of use thereof
WO2005058892A1 (en) * 2003-12-19 2005-06-30 Glaxo Group Limited Pyrazolo [3,4-b] pyridine compounds, and their use as phosphodiesterase inhibitors
US6992086B2 (en) 2001-01-31 2006-01-31 Telik, Inc. Antagonists of MCP-1 function and methods of use thereof
JP2007529486A (en) * 2004-03-16 2007-10-25 グラクソ　グループ　リミテッド Pyrazolo [3,4-b] pyridine compounds and their use as PDE4 inhibitors
WO2008055945A1 (en) 2006-11-09 2008-05-15 Probiodrug Ag 3-hydr0xy-1,5-dihydr0-pyrr0l-2-one derivatives as inhibitors of glutaminyl cyclase for the treatment of ulcer, cancer and other diseases
WO2008065141A1 (en) 2006-11-30 2008-06-05 Probiodrug Ag Novel inhibitors of glutaminyl cyclase
WO2008104580A1 (en) 2007-03-01 2008-09-04 Probiodrug Ag New use of glutaminyl cyclase inhibitors
US7528148B2 (en) 2002-12-23 2009-05-05 Glaxo Group Limited Pyrazolo[3,4-B]pyridine compounds, and their use as phosphodiesterase inhibitors
WO2009100167A1 (en) * 2008-02-06 2009-08-13 Glaxo Group Limited Dual pharmacophores-pde4-muscarinic antagonistics
WO2009100169A1 (en) * 2008-02-06 2009-08-13 Glaxo Group Limited Dual pharmacophores - pde4-muscarinic antagonistics
WO2009100170A1 (en) * 2008-02-06 2009-08-13 Glaxo Group Limited Dual pharmacophores - pde4-muscarinic antagonistics
WO2009100168A1 (en) * 2008-02-06 2009-08-13 Glaxo Group Limited Dual pharmacophores - pde4-muscarinic antagonistics
US7645752B2 (en) 2006-01-13 2010-01-12 Wyeth Llc Sulfonyl substituted 1H-indoles as ligands for the 5-hydroxytryptamine receptors
US7732162B2 (en) 2003-05-05 2010-06-08 Probiodrug Ag Inhibitors of glutaminyl cyclase for treating neurodegenerative diseases
US20100210644A1 (en) * 2007-06-25 2010-08-19 Boehringer Ingelheim International Gmbh Chemical compounds
US7838541B2 (en) 2002-02-11 2010-11-23 Bayer Healthcare, Llc Aryl ureas with angiogenesis inhibiting activity
US7846928B2 (en) * 2006-06-13 2010-12-07 Bayer Schering Pharma Ag Substituted pyrazolopyridines and salts thereof, pharmaceutical compositions comprising same, methods of preparing same and uses of same
US7897623B2 (en) 1999-01-13 2011-03-01 Bayer Healthcare Llc ω-carboxyl aryl substituted diphenyl ureas as p38 kinase inhibitors
WO2011029920A1 (en) 2009-09-11 2011-03-17 Probiodrug Ag Heterocylcic derivatives as inhibitors of glutaminyl cyclase
US7915286B2 (en) 2005-09-16 2011-03-29 Ranbaxy Laboratories Limited Substituted pyrazolo [3,4-b] pyridines as phosphodiesterase inhibitors
CN102153539A (en) * 2002-11-27 2011-08-17 因塞特公司 3-aminopyrrolidine derivatives as modulators of chemokine receptors
WO2011107530A2 (en) 2010-03-03 2011-09-09 Probiodrug Ag Novel inhibitors
WO2011110613A1 (en) 2010-03-10 2011-09-15 Probiodrug Ag Heterocyclic inhibitors of glutaminyl cyclase (qc, ec 2.3.2.5)
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US8796250B2 (en) 2003-05-20 2014-08-05 Bayer Healthcare Llc Diaryl ureas for diseases mediated by PDGFR
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EP3461819A1 (en) 2017-09-29 2019-04-03 Probiodrug AG Inhibitors of glutaminyl cyclase

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10153259A1 (en) 2001-10-31 2003-05-22 Osram Opto Semiconductors Gmbh Optoelectronic component
EP1692506A4 (en) * 2003-11-17 2008-01-09 Janssen Pharmaceutica Nv Modeling of systemic inflammatory response to infection
EP2041568A4 (en) * 2006-06-15 2009-08-12 Ccr2 antagonists for chronic organ transplantation rejection
US20080015465A1 (en) * 2006-06-15 2008-01-17 Scuderi Gaetano J Methods for diagnosing and treating pain in the spinal cord
US20080076120A1 (en) * 2006-09-14 2008-03-27 Millennium Pharmaceuticals, Inc. Methods for the identification, evaluation and treatment of patients having CC-Chemokine receptor 2 (CCR-2) mediated disorders
CN101616689A (en) * 2006-10-05 2009-12-30 森托科尔奥索生物科技公司 Be used for the treatment of Fibrotic CCR2 antagonist
US7709215B2 (en) 2007-06-01 2010-05-04 Cytonics Corporation Method for diagnosing and treating acute joint injury
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ES2968371T3 (en) 2013-10-10 2024-05-09 Eastern Virginia Medical School 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as 12-lipoxygenase inhibitors
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WO2024108386A1 (en) * 2022-11-22 2024-05-30 中国科学院深圳先进技术研究院 Use of anti-mcp1 neutralizing antibody in preparing medicament for treating systemic inflammation caused by neurodegenerative diseases

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998002151A2 (en) * 1996-07-12 1998-01-22 Leukosite, Inc. Chemokine receptor antagonists and methods of use therefor
WO1998006703A1 (en) * 1996-08-14 1998-02-19 Warner-Lambert Company 2-phenyl benzimidazole derivatives as mcp-1 antagonists
WO2000046198A1 (en) * 1999-02-05 2000-08-10 Astrazeneca Ab Anti-inflammatory indole derivatives

Family Cites Families (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IE32700B1 (en) 1968-02-03 1973-10-31 Beecham Group Ltd Penicillins
GB1250611A (en) 1968-02-03 1971-10-20
DE2131034A1 (en) 1971-06-23 1973-01-11 Hoechst Ag ACYLIC URNANE AND METHOD FOR MANUFACTURING IT
DE2652004C3 (en) 1976-11-15 1979-09-13 Basf Ag, 6700 Ludwigshafen l-Acyl-2-arylamino-2-imidazolines, process for their preparation and their use
FR2672888B1 (en) 1991-02-14 1994-02-04 Fabre Medicament Pierre NEW UREAS AND THIOUREAS, THEIR PREPARATION AND THEIR APPLICATION IN THERAPEUTICS.
JP3964478B2 (en) 1995-06-30 2007-08-22 エーザイ・アール・アンド・ディー・マネジメント株式会社 Heterocycle-containing carboxylic acid derivative and pharmaceutical containing the same
WO1997024325A1 (en) 1995-12-28 1997-07-10 Takeda Chemical Industries, Ltd. DIPHENYLMETHANE DERIVATIVES AS MIP-1α/RANTES RECEPTOR ANTAGONISTS
JPH09255572A (en) 1996-03-26 1997-09-30 Takeda Chem Ind Ltd Medicine for antagonizing chemokine receptor
WO1997044329A1 (en) 1996-05-20 1997-11-27 Teijin Limited Diarylalkyl cyclic diamine derivatives as chemokine receptor antagonists
AU3633997A (en) 1996-07-29 1998-02-20 Banyu Pharmaceutical Co., Ltd. Chemokine receptor antagonists
US6228869B1 (en) * 1996-10-16 2001-05-08 American Cyanamid Company Ortho-sulfonamido bicyclic hydroxamic acids as matrix metalloproteinase and TACE inhibitors
WO1998027815A1 (en) 1996-12-20 1998-07-02 Merck & Co., Inc. Substituted aminoquinolines as modulators of chemokine receptor activity
GB9716656D0 (en) 1997-08-07 1997-10-15 Zeneca Ltd Chemical compounds
JP2002502844A (en) 1998-02-03 2002-01-29 ベーリンガー インゲルハイム ファルマ コマンディトゲゼルシャフト 5-membered heterocyclic fused benzo derivatives, their preparation and their use as pharmaceuticals
AU753360B2 (en) 1998-07-31 2002-10-17 Nippon Soda Co., Ltd. Phenylazole compounds, process for producing the same and drugs for hyperlipemia
US6326379B1 (en) * 1998-09-16 2001-12-04 Bristol-Myers Squibb Co. Fused pyridine inhibitors of cGMP phosphodiesterase
US6316449B1 (en) 1999-07-08 2001-11-13 Millennium Pharmaceuticals, Inc. Chemokine receptor antagonists
DE60036726T2 (en) 1999-11-16 2008-02-07 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield UREA DERIVATIVES AS ANTI-INFLAMMATORY AGENTS
EP1255743A1 (en) 2000-02-01 2002-11-13 Millenium Pharmaceuticals, Inc. 3,4-DIHYDRO-2H-BENZO[1,4]OXAZINE INHIBITORS OF FACTOR Xa
AU2001230537A1 (en) 2000-02-01 2001-08-14 Daiichi Pharmaceutical Co., Ltd. Pyridoxazine derivatives
WO2001057021A2 (en) 2000-02-01 2001-08-09 Cor Therapeutics, Inc. 2-[1H]-QUINOLONE AND 2-[1H]-QUINOXALONE INHIBITORS OF FACTOR Xa
US6670364B2 (en) * 2001-01-31 2003-12-30 Telik, Inc. Antagonists of MCP-1 function and methods of use thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998002151A2 (en) * 1996-07-12 1998-01-22 Leukosite, Inc. Chemokine receptor antagonists and methods of use therefor
WO1998006703A1 (en) * 1996-08-14 1998-02-19 Warner-Lambert Company 2-phenyl benzimidazole derivatives as mcp-1 antagonists
WO2000046198A1 (en) * 1999-02-05 2000-08-10 Astrazeneca Ab Anti-inflammatory indole derivatives

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 100, no. 21, 1984 Columbus, Ohio, US; abstract no. 174751, page 626; column 1; XP002206320 & OREMEK, G. ET AL.: "Ring-opening reactions of potentially biologically active benzothienooxazines" ARCH. PHARM., vol. 317, no. 2, 1984, pages 117-120, Weinheim *
CHEMICAL ABSTRACTS, vol. 103, no. 17, 1985 Columbus, Ohio, US; abstract no. 141873, page 714; column 2; XP002206319 & KUCZINSKI, L. ET AL.: "Synthesis of new 4 and 5 disubstiuted isothiazoles" POL. J. PHARMACOL. PHARM., vol. 36, no. 5, 1984, pages 485-491, *

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US7528148B2 (en) 2002-12-23 2009-05-05 Glaxo Group Limited Pyrazolo[3,4-B]pyridine compounds, and their use as phosphodiesterase inhibitors
US7732162B2 (en) 2003-05-05 2010-06-08 Probiodrug Ag Inhibitors of glutaminyl cyclase for treating neurodegenerative diseases
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