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WO2004026302A1 - Methods of inhibiting tgf beta with substituted pyrazoles - Google Patents

Methods of inhibiting tgf beta with substituted pyrazoles Download PDF

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Publication number
WO2004026302A1
WO2004026302A1 PCT/US2003/026296 US0326296W WO2004026302A1 WO 2004026302 A1 WO2004026302 A1 WO 2004026302A1 US 0326296 W US0326296 W US 0326296W WO 2004026302 A1 WO2004026302 A1 WO 2004026302A1
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Prior art keywords
quinoline
pyrazol
pyridin
methylpyridin
quinolyl
Prior art date
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Ceased
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PCT/US2003/026296
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French (fr)
Inventor
Jason Scott Sawyer
Beverly Ann Teicher
Jonathan Michael Yingling
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Eli Lilly and Co
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Eli Lilly and Co
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Priority to AU2003268155A priority Critical patent/AU2003268155A1/en
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Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • TGF-Beta transforming growth factor-beta
  • TGF- ⁇ The transforming growth factor-beta (TGF-Beta)("TGF- ⁇ ") polypeptides influence growth, differentiation, and gene expression in many cell types.
  • the first polypeptide of this family that was characterized, TGF- ⁇ l has two identical 112 amino acid subunits that are covalently linked.
  • TGF- ⁇ l is a highly conserved protein with only a single amino acid difference distinguishing humans from mice.
  • TGF- ⁇ 2 is 71% homologous to TGF- ⁇ l (de Martin, et al. (1987) EMBO J.
  • TGF- ⁇ 3 is 80% homologous to TGF- ⁇ l (Derynck, et al. (1988) EMBO J 7:3737-3743).
  • the structural characteristics of TGF- ⁇ l as determined by nuclear magnetic resonance agree with the crystal structure of TGF- ⁇ 2 (Daopin, et al. (1992) Science 257:369-374; Schlunegger and Grutter (1992) Nature 358:430-434).
  • TGF- ⁇ receptors Type I, II and III that are involved in the biological functions of TGF- ⁇ l, - ⁇ 2 and - ⁇ 3 (For reviews, see Derynck (1994) TIBS 19:548-553 and Massague (1990) Ann. Rev. Cell Biol. 6:597-641).
  • the type I and type II receptors are transmembrane serine/threonine kinases which in the presence of TGF- ⁇ form a heteromeric signaling complex (Wrana, et al (1992) Cell 71 : 1003-1014). The mechanism of activation of the heteromeric signaling complex at the cell surface has been elucidated (Wrana, et al.
  • TGF- ⁇ first binds the type II receptor which is a constitutively active transmembrane serine/threonine kinase.
  • the type I receptor is subsequently recruited into the complex, phoshorylated at the GS domain and activated to phosphorylate downstream signaling components (e.g. Smad proteins) to initiate the intracellular signaling cascade.
  • a constitutively active type I receptor (T204D mutant) has been shown to effectively transduce TGF- ⁇ responses, thus bypassing the requirement for TGF- ⁇ and the type II receptor (Wieser, et al. (1995) EMBO J 14: 2199-2208).
  • TGF- ⁇ 2's affinity for the type II receptor makes it essentially equipotent with TGF- ⁇ l and TGF- ⁇ 3 (Lopez-Casillas, et al. (1993) Cell 73: 1435-1444).
  • Vascular endothelial cells lack the Type III receptor. Instead endothelial cells express a structurally related protein called endoglin (Cheifetz, et al. (1992) J. Biol. Chem. 267:19027-19030), which only binds TGF- ⁇ l and TGF- ⁇ 3 with high affinity. Thus, the relative potency of the TGF- ⁇ 's reflect the type of receptors expressed in a cell and organ system. In addition to the regulation of the components in the multi-factorial signaling pathway, the distribution of the synthesis of TGF- ⁇ polypeptides also affects physiological function. The distribution of TGF- ⁇ 2 and TGF- ⁇ 3 is more limited (Derynck, et al.
  • TGF- ⁇ 3 is limited to tissues of mesenchymal origin, whereas TGF- ⁇ l is present in both tissues of mesenchymal and epithelial origin.
  • TGF- ⁇ l is a multifunctional cytokine critical for tissue repair. High concentrations of TGF- ⁇ l are delivered to the site of injury by platelet granules (Assoian and Sporn (1986) J. Cell Biol. 102:1217-1223). TGF- ⁇ l initiates a series of events that promote healing including chemotaxis of cells such as leukocytes, monocytes and fibroblasts, and regulation of growth factors and cytokines involved in angiogenesis, cell division associated with tissue repair and inflammatory responses. TGF- ⁇ l also stimulates the synthesis of extracellular matrix components (Roberts, et al. (1986) Proc. Natl. Acad. Sci.
  • TGF- ⁇ l autoregulates its own synthesis (Kim, et al. (1989) J. Biol. Chem. 264:7041-7045).
  • TGF- ⁇ l over production A number of diseases have been associated with TGF- ⁇ l over production. Fibrotic diseases associated with TGF- ⁇ l overproduction can be divided into chronic conditions such as fibrosis of the kidney, lung and liver and more acute conditions such as dermal scarring and restenosis. Synthesis and secretion of TGF- ⁇ l by tumor cells can also lead to immune suppression such as seen in patients with aggressive brain or breast tumors (Arteaga, et al. (1993) J. Clin. Invest. 92:2569-2576). The course of Leishmanial infection in mice is drastically altered by TGF- ⁇ l (Barral-Netto, et al. (1992) Science 257:545-547). TGF- ⁇ l exacerbated the disease, whereas TGF- ⁇ l antibodies halted the progression of the disease in genetically susceptible mice. Genetically resistant mice became susceptible to Leishmanial infection upon administration of TGF- ⁇ l.
  • TGF- ⁇ l levels are elevated in human diabetic glomerulosclerosis (advanced neuropathy) (Yamamoto, et al. (1993) Proc. Natl. Acad. Sci. 90:1814-1818).
  • TGF- ⁇ l is an important mediator in the genesis of renal fibrosis in a number of animal models (Phan, et al.
  • TGF- ⁇ l leads to dermal scar-tissue formation.
  • Neutralizing TGF- ⁇ l antibodies injected into the margins of healing wounds in rats have been shown to inhibit scarring without interfering with the rate of wound healing or the tensile strength of the wound (Shah, et al. (1992) Lancet 339:213-214).
  • angiogenesis reduced number of macrophages and monocytes in the wound, and a reduced amount of disorganized collagen fiber deposition in the scar tissue.
  • TGF- ⁇ l may be a factor in the progressive thickening of the arterial wall which results from the proliferation of smooth muscle cells and deposition of extracellular matrix in the artery after balloon angioplasty.
  • the diameter of the restenosed artery may be reduced 90% by this thickening, and since most of the reduction in diameter is due to extracellular matrix rather than smooth muscle cell bodies, it may be possible to open these vessels to 50% simply by reducing extensive extracellular matrix deposition.
  • TGF- ⁇ l gene expression was associated with both extracellular matrix synthesis and hyperplasia (Nabel, et al. (1993) Proc. Natl. Acad. Sci. USA 90:10759-10763).
  • TGF- ⁇ l induced hyperplasia was not as extensive as that induced with PDGF-BB, but the extracellular matrix was more extensive with TGF- ⁇ l transfectants. No extracellular matrix deposition was associated with FGF-1 (a secreted form of FGF) induced hyperplasia in this gene transfer pig model (Nabel (1993) Nature 362:844-846).
  • TGF- ⁇ l produced by the tumor may be deleterious.
  • MATLyLu rat cancer cells Steiner and Barrack (1992) Mol. Endocrinol 6:15-25
  • MCF-7 human breast cancer cells Arteaga, et al. (1993) Cell Growth and Differ. 4:193-201 became more tumorigenic and metastatic after transfection with a vector expressing the mouse TGF- ⁇ 1.
  • poor prognosis is associated with elevated TGF- ⁇ (Dickson, et al. (1987) Proc. Natl. Acad. Sci. USA 84:837-841; Kasid, et al. (1987) Cancer Res. 47:5733-5738; Daly, et al.
  • TGF- ⁇ l antibodies inhibit the growth of MDA-231 human breast cancer cells in athymic mice (Arteaga, et al. (1993) J. Clin. Invest. 92:2569-2576), a treatment which is correlated with an increase in spleen natural killer cell activity.
  • CHO cells transfected with latent TGF- ⁇ l also showed decreased NK activity and increased tumor growth in nude mice (Wallick, et al. (1990) J. Exp. Med. 172:1777-1784).
  • TGF- ⁇ secreted by breast tumors may cause an endocrine immune suppression.
  • High plasma concentrations of TGF- ⁇ l have been shown to indicate poor prognosis for advanced breast cancer patients (Anscher, et al.
  • TGF- ⁇ transforming growth factor- ⁇
  • TGF- ⁇ transforming growth factor- ⁇
  • TGF-bl can act as a potent tumor suppressor and may mediate the actions of some chemopreventive agents.
  • tumor cells appear to escape from TGF- ⁇ -dependent growth inhibition in parallel with the appearance of bioactive TGF- ⁇ in the microenvironment.
  • the production of TGF- ⁇ l by malignant cells in primary tumors appears to increase with advancing stages of tumor progression.
  • Studies in many of the major epithelial cancers suggest that the increased production of TGF- ⁇ by human cancers occurs as a relatively late event during tumor progression. Further, this tumor-associated TGF- ⁇ provides the tumor cells with a selective advantage and promotes tumor progression.
  • TGF- ⁇ may allow tumor cells to escape from immune surveillance since it is a potent inhibitor of the clonal expansion of activated lymphocytes.
  • TGF- ⁇ has also been shown to inhibit the production of angiostatin.
  • Cancer therapeutic modalities such as radiation therapy and chemotherapy induce the production of activated TGF- ⁇ in the tumor, thereby selecting outgrowth of malignant cells that are resistant to TGF- ⁇ growth inhibitory effects.
  • these anticancer treatments increase the risk and hasten the development of tumors with enhanced growth and invasiveness.
  • agents targeting TGF- ⁇ l itself might be a very effective therapeutic strategy.
  • TGF- ⁇ 1 neutralizing agent The resistance of tumor cells to TGF- ⁇ has been shown to negate much of the cytotoxic effects of radiation therapy and chemotherapy and the treatment-dependent activation of TGF- ⁇ in the stroma may even be detrimental as it can make the microenvironment more conducive to tumor progression and contributes to tissue damage leading to f ⁇ brosis.
  • the development of a TGF- ⁇ 1 neutralizing agent is likely to benefit the treatment of progressed cancer alone and in combination with other therapies.
  • WO 98/52940 (PCT/US98/10436) as p38 kinase inhibitors.
  • the compounds and examples disclosed in this application were all made by the applicants, with the exception of Example 41, which is commercially available, and were not specifically disclosed or claimed WO 98/52940. Nor did this application make any reference to the fact that the compounds disclosed herein would be useful as TGF-beta inhibitors. We believe that all the compounds disclosed in the WO 98/52940 application, which we incorporate by reference, would therefore be useful as TGF-beta inhibitors as well.
  • the invention herein disclosed is directed to the treatment of cancer and other disease states influenced by TGF beta by inhibiting TGF- ⁇ in a patient in need thereof by administering to said patient a compound of from the group consisting essentially of the following: (1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
  • (22) 1 -(4-phenylbutyl)-3 -(2-pyridyl)-4-(4-quinolyl)pyrazole.
  • (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol.
  • 6-Propylpyridine-2-carboxylic Acid Methyl Ester Dissolve dimethyl oxalate (5.90 g, 50 mmol, Aldrich) and KOH (2.80 g, 50 mmol) in 9/1 - MeOH/water (25 mL) and stir at RT 4 h. Evaporate the oxalate salt to a residue and re-dissolve in water (40 mL).
  • MV1LU P3TP-LUX/PURO CLONE CI ASSAY A stable MvlLu clone (CI) containing the p3TP-Lux reporter was created by standard transfection and puromycin selection protocols. This stable clone was used to screen the example compounds for their ability to inhibit TGF- ⁇ dependent luciferase production as briefly described below:
  • the 6X-HIS tagged cytoplasmic kinase domain of the receptor was expressed and purified from Sf9 insect cell lysates as briefly described below:
  • lysis buffer 50 mM Tris pH 7.5, 150 mM NaCl, 50 mM NaF, 0.5% NP40 with freshly added 20 mM ⁇ - mercaptoethanol, 10 mM imidazole, 1 mM PMSF, IX EDTA-free Complete Protease Inl ⁇ ibitor(Boehringer Mannheim).
  • reactions were incubated at 30 °C for 1 lir. Reactions were stopped and quantitated using standard TCA/BSA precipitation onto Millipore FB glass fiber filter plates and by liquid scintillation counting on a MicroBeta JET.
  • compositions of the present invention are therapeutically effective amounts of the TGF- ⁇ antagonists, noted above.
  • the composition may be formulated with common excipients, diluents or carriers, and compressed into tablets, or formulated elixirs or solutions for convenient oral administration or administered by intramuscular intravenous routes.
  • the compounds can be administered transdermally and maybe formulated as sustained relief dosage forms and the like.
  • the method of treating a human patient according to the present invention includes administration of the TGF- ⁇ antagonists.
  • the TGF- ⁇ antagonists are formulated into formulations which may be administered by the oral and rectal routes, topically, parenterally, e.g., by injection and by continuous or discontinuous intra-arterial infusion, in the form of, for example, tablets, lozenges, sublingual tablets, sachets, cachets, elixirs, gels, suspensions, aerosols, ointments, for example, containing from 1 to 10% by weight of the active compound in a suitable base, soft and hard gelatin capsules, suppositories, injectable solutions and suspensions in physiologically acceptable media, and sterile packaged powders adsorbed onto a support material for making injectable solutions.
  • compositions may be provided in dosage unit form, preferably each dosage unit containing from about 5 to about 500 mg (from about 5 to 50 mg in the case of parenteral or inhalation administration, and from about 25 to 500 mg in the case of oral or rectal administration) the compounds.
  • Dosages from about 0.5 to about 300 mg/kg per day, preferably 0.5 to 20 mg/kg, of active ingredient may be administered although it will, of course, readily be understood that the amount of the compound actually to be administered will be determined by a physician, in the light of all the relevant circumstances including the condition to be treated, the choice of compound to be administered and the choice of route of administration and therefore the above preferred dosage range is not intended to limit the scope of the present invention in any way.
  • the formulations useful for separate administration of the TGF- ⁇ antagonists will normally consist of at least one compound selected from the compounds specified herein mixed with a carrier, or diluted by a carrier, or enclosed or encapsulated by an ingestible carrier in the form of a capsule, sachet, cachet, paper or other container or by a disposable container such as an ampoule.
  • a carrier or diluent may be a solid, semi-solid or liquid material which serves as a vehicle, excipient or medium for the active therapeutic substance.
  • diluents or carrier which may be employed in the pharmaceutical compositions of the present invention are lactose, dextrose, sucrose, sorbitol, mannitol, propylene glycol, liquid paraffin, white soft paraffin, kaolin, fumed silicon dioxide, microcrystalline cellulose, calcium silicate, silica, polyvinylpyrrolidone, cetostearyl alcohol, starch, modified starches, gum acacia, calcium phosphate, cocoa butter, ethoxylated esters, oil of theobroma, arachis oil, alginates, tragacanth, gelatin, syrup, methyl cellulose, polyoxyethylene sorbitan monolaurate, ethyl lactate, methyl and propyl hydroxybenzoate, sorbitan trioleate, sorbitan sesquioleate and oleyl alcohol and propellants such as trichloromonofluoromethane, dichlorodifluoromethane and dich
  • a lubricant may be incorporated to prevent sticking and binding of the powdered ingredients in the dies and on the punch of the tableting machine.
  • a lubricant may be employed for instance aluminum, magnesium or calcium stearates, talc or mineral oil.
  • Preferred pharmaceutical forms of the present invention are capsules, tablets, suppositories, injectable solutions, creams and ointments.
  • formulations for inhalation application such as an aerosol, for injection, and for oral ingestion.

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Abstract

Substituted pyrazoles are disclosed useful in the treatment of cancer and other disease states influenced by TGF beta by inhibiting TGF-β in a patient in need thereof.

Description

METHODS OF INHIBITING TGF BETA WITH SUBSTITUTED PYRAZOLES
BACKGROUND OF INVENTION The transforming growth factor-beta (TGF-Beta)("TGF-β") polypeptides influence growth, differentiation, and gene expression in many cell types. The first polypeptide of this family that was characterized, TGF-βl, has two identical 112 amino acid subunits that are covalently linked. TGF-βl is a highly conserved protein with only a single amino acid difference distinguishing humans from mice. There are two other members of the TGF-β gene family that are expressed in mammals. TGF-β2 is 71% homologous to TGF-βl (de Martin, et al. (1987) EMBO J. 6:3673-3677), whereas TGF- β3 is 80% homologous to TGF-βl (Derynck, et al. (1988) EMBO J 7:3737-3743). The structural characteristics of TGF-βl as determined by nuclear magnetic resonance (Archer, et al. (1993) Biochemistry 32:1164-1171) agree with the crystal structure of TGF-β2 (Daopin, et al. (1992) Science 257:369-374; Schlunegger and Grutter (1992) Nature 358:430-434).
There are at least three different extracellular TGF-β receptors, Type I, II and III that are involved in the biological functions of TGF-βl, -β2 and -β3 (For reviews, see Derynck (1994) TIBS 19:548-553 and Massague (1990) Ann. Rev. Cell Biol. 6:597-641). The type I and type II receptors are transmembrane serine/threonine kinases which in the presence of TGF-β form a heteromeric signaling complex (Wrana, et al (1992) Cell 71 : 1003-1014). The mechanism of activation of the heteromeric signaling complex at the cell surface has been elucidated (Wrana, et al. (1994) Nature 370: 341-347). TGF-β first binds the type II receptor which is a constitutively active transmembrane serine/threonine kinase. The type I receptor is subsequently recruited into the complex, phoshorylated at the GS domain and activated to phosphorylate downstream signaling components (e.g. Smad proteins) to initiate the intracellular signaling cascade. A constitutively active type I receptor (T204D mutant) has been shown to effectively transduce TGF-β responses, thus bypassing the requirement for TGF-β and the type II receptor (Wieser, et al. (1995) EMBO J 14: 2199-2208). Although no signaling function has been discovered for the type III receptor, it does increase TGF-β2's affinity for the type II receptor making it essentially equipotent with TGF-βl and TGF-β3 (Lopez-Casillas, et al. (1993) Cell 73: 1435-1444).
Vascular endothelial cells lack the Type III receptor. Instead endothelial cells express a structurally related protein called endoglin (Cheifetz, et al. (1992) J. Biol. Chem. 267:19027-19030), which only binds TGF-βl and TGF-β3 with high affinity. Thus, the relative potency of the TGF-β's reflect the type of receptors expressed in a cell and organ system. In addition to the regulation of the components in the multi-factorial signaling pathway, the distribution of the synthesis of TGF-β polypeptides also affects physiological function. The distribution of TGF-β2 and TGF-β3 is more limited (Derynck, et al. (1988) EMBO J 7:3737-3743) than TGF-βl, e.g., TGF-β3 is limited to tissues of mesenchymal origin, whereas TGF-βl is present in both tissues of mesenchymal and epithelial origin.
TGF-βl is a multifunctional cytokine critical for tissue repair. High concentrations of TGF-βl are delivered to the site of injury by platelet granules (Assoian and Sporn (1986) J. Cell Biol. 102:1217-1223). TGF-βl initiates a series of events that promote healing including chemotaxis of cells such as leukocytes, monocytes and fibroblasts, and regulation of growth factors and cytokines involved in angiogenesis, cell division associated with tissue repair and inflammatory responses. TGF-βl also stimulates the synthesis of extracellular matrix components (Roberts, et al. (1986) Proc. Natl. Acad. Sci. USA 83:4167-4171; Sporn, et al. (1983) Science 219:1329-1330; Massague (1987) Cell 49:437-438) and most importantly for understanding the pathophysiology of TGF-βl, TGF-βl autoregulates its own synthesis (Kim, et al. (1989) J. Biol. Chem. 264:7041-7045).
A number of diseases have been associated with TGF-βl over production. Fibrotic diseases associated with TGF-βl overproduction can be divided into chronic conditions such as fibrosis of the kidney, lung and liver and more acute conditions such as dermal scarring and restenosis. Synthesis and secretion of TGF-βl by tumor cells can also lead to immune suppression such as seen in patients with aggressive brain or breast tumors (Arteaga, et al. (1993) J. Clin. Invest. 92:2569-2576). The course of Leishmanial infection in mice is drastically altered by TGF-βl (Barral-Netto, et al. (1992) Science 257:545-547). TGF-βl exacerbated the disease, whereas TGF-βl antibodies halted the progression of the disease in genetically susceptible mice. Genetically resistant mice became susceptible to Leishmanial infection upon administration of TGF-βl.
The profound effects of TGF-βl on extracellular matrix deposition have been reviewed (Rocco and Ziyadeh (1991) in Contemporary Issues in Nephrology v.23, Hormones, autocoids and the kidney, ed. Jay Stein, Churchill Livingston, New York pp.391-410; Roberts, et al. (1988) Rec. Prog. Hormone Res. 44:157-197) and include the stimulation of the synthesis and the inhibition of degradation of extracellular matrix components. Since the structure and filtration properties of the glomerulus are largely determined by the extracellular matrix composition of the mesangium and glomerular membrane, it is not surprising that TGF-βl has profound effects on the kidney. The accumulation of mesangial matrix in proliferate glomerulonephritis (Border, et al. (1990) Kidney Int. 37:689-695) and diabetic nephropathy (Mauer, et al. (1984) J. Clin. Invest. 74:1143-1155) are clear and dominant pathological features of the diseases. TGF-βl levels are elevated in human diabetic glomerulosclerosis (advanced neuropathy) (Yamamoto, et al. (1993) Proc. Natl. Acad. Sci. 90:1814-1818). TGF-βl is an important mediator in the genesis of renal fibrosis in a number of animal models (Phan, et al. (1990) Kidney Int. 37:426; Okuda, et al. (1990) J. Clin. Invest. 86:453). Suppression of experimentally induced glomerulonephritis in rats has been demonstrated by antiserum against TGF-βl (Border, et al. (1990) Nature 346:371) and by an extracellular matrix protein, decorin, which can bind TGF-βl (Border, et al. (1992) Nature 360:361-363).
Too much TGF-βl leads to dermal scar-tissue formation. Neutralizing TGF-βl antibodies injected into the margins of healing wounds in rats have been shown to inhibit scarring without interfering with the rate of wound healing or the tensile strength of the wound (Shah, et al. (1992) Lancet 339:213-214). At the same time there was reduced angiogenesis, reduced number of macrophages and monocytes in the wound, and a reduced amount of disorganized collagen fiber deposition in the scar tissue.
TGF-βl may be a factor in the progressive thickening of the arterial wall which results from the proliferation of smooth muscle cells and deposition of extracellular matrix in the artery after balloon angioplasty. The diameter of the restenosed artery may be reduced 90% by this thickening, and since most of the reduction in diameter is due to extracellular matrix rather than smooth muscle cell bodies, it may be possible to open these vessels to 50% simply by reducing extensive extracellular matrix deposition. In uninjured pig arteries transfected in vivo with a TGF-βl gene, TGF-βl gene expression was associated with both extracellular matrix synthesis and hyperplasia (Nabel, et al. (1993) Proc. Natl. Acad. Sci. USA 90:10759-10763). The TGF-βl induced hyperplasia was not as extensive as that induced with PDGF-BB, but the extracellular matrix was more extensive with TGF-βl transfectants. No extracellular matrix deposition was associated with FGF-1 (a secreted form of FGF) induced hyperplasia in this gene transfer pig model (Nabel (1993) Nature 362:844-846).
There are several types of cancer where TGF-βl produced by the tumor may be deleterious. MATLyLu rat cancer cells (Steiner and Barrack (1992) Mol. Endocrinol 6:15-25) and MCF-7 human breast cancer cells (Arteaga, et al. (1993) Cell Growth and Differ. 4:193-201) became more tumorigenic and metastatic after transfection with a vector expressing the mouse TGF-β 1. In breast cancer, poor prognosis is associated with elevated TGF-β (Dickson, et al. (1987) Proc. Natl. Acad. Sci. USA 84:837-841; Kasid, et al. (1987) Cancer Res. 47:5733-5738; Daly, et al. (1990) J. Cell Biochem. 43:199-211; Barrett-Lee, et al. (1990) Br. J Cancer 61:612-617; King, et al. (1989) J. Steroid Biochem. 34:133-138; Welch, et al. (1990) Proc. Natl. Acad. Sci. USA 87:7678-7682; Walker, et al. (1992) Eur. J. Cancer 238:641-644) and induction of TGF-βl by tamoxifen treatment (Butta, et al. (1992) Cancer Res. 52:4261- 4264) has been associated with failure of tamoxifen treatment for breast cancer (Thompson, et al. (1991) Br. J. Cancer 63:609-
614). Anti TGF-βl antibodies inhibit the growth of MDA-231 human breast cancer cells in athymic mice (Arteaga, et al. (1993) J. Clin. Invest. 92:2569-2576), a treatment which is correlated with an increase in spleen natural killer cell activity. CHO cells transfected with latent TGF-βl also showed decreased NK activity and increased tumor growth in nude mice (Wallick, et al. (1990) J. Exp. Med. 172:1777-1784). Thus, TGF-β secreted by breast tumors may cause an endocrine immune suppression. High plasma concentrations of TGF-βl have been shown to indicate poor prognosis for advanced breast cancer patients (Anscher, et al. (1993) N. Engl. J. Med. 328:1592-1598). Patients with high circulating TGF-β before high dose chemotherapy and autologous bone marrow transplantation are at high risk for hepatic veno-occlusive disease (15-50% of all patients with a mortality rate up to 50%) and idiopathic interstitial pneumonitis (40-60% of all patients). The implication of these findings is 1) that elevated plasma levels of TGF-βl can be used to identify at risk patients and 2) that reduction of TGF-βl could decrease the morbidity and mortality of these common treatments for breast cancer patients.
Many malignant cells secrete transforming growth factor-β (TGF-β), a potent immunosuppressant, suggesting that TGF-β production may represent a significant tumor escape mechanism from host immunosurveillance. Establishment of a leukocyte sub- population with disrupted TGF-β signaling in the tumor-bearing host offers a potential means for immunotherapy of cancer. Down regulation of TGF-β secretion in tumor cells results in restoration of immunogenicity in the host, while T-cell insensitivity to TGF-β results in accelerated differentiation and autoimmunity, elements of which may be required in order to combat self-antigen-expressing tumors in a tolerized host. The rationale, approaches, and potential pitfalls of this strategy will be discussed.
Cancer: During the earliest stages of carcino genesis, TGF-bl can act as a potent tumor suppressor and may mediate the actions of some chemopreventive agents. However, at some point during the development and progression of malignant neoplasms, tumor cells appear to escape from TGF-β-dependent growth inhibition in parallel with the appearance of bioactive TGF-β in the microenvironment. Thus, the production of TGF- βl by malignant cells in primary tumors appears to increase with advancing stages of tumor progression. Studies in many of the major epithelial cancers suggest that the increased production of TGF-β by human cancers occurs as a relatively late event during tumor progression. Further, this tumor-associated TGF-β provides the tumor cells with a selective advantage and promotes tumor progression. The effects of TGF-β on cell/cell and cell/stroma interactions result in a greater propensity for invasion and metastasis. Tumor-associated TGF-β may allow tumor cells to escape from immune surveillance since it is a potent inhibitor of the clonal expansion of activated lymphocytes. TGF-β has also been shown to inhibit the production of angiostatin. Cancer therapeutic modalities such as radiation therapy and chemotherapy induce the production of activated TGF-β in the tumor, thereby selecting outgrowth of malignant cells that are resistant to TGF-β growth inhibitory effects. Thus, these anticancer treatments increase the risk and hasten the development of tumors with enhanced growth and invasiveness. In this situation, agents targeting TGF-βl itself might be a very effective therapeutic strategy. The resistance of tumor cells to TGF-β has been shown to negate much of the cytotoxic effects of radiation therapy and chemotherapy and the treatment-dependent activation of TGF-β in the stroma may even be detrimental as it can make the microenvironment more conducive to tumor progression and contributes to tissue damage leading to fϊbrosis. The development of a TGF-β 1 neutralizing agent is likely to benefit the treatment of progressed cancer alone and in combination with other therapies.
We now go on to show that certain compounds exhibit both TGF beta inhibition as well as activity against cancer cells in vitro.
The compounds and examples disclosed in this application were generically disclosed in PCT application
WO 98/52940 (PCT/US98/10436) as p38 kinase inhibitors. The compounds and examples disclosed in this application were all made by the applicants, with the exception of Example 41, which is commercially available, and were not specifically disclosed or claimed WO 98/52940. Nor did this application make any reference to the fact that the compounds disclosed herein would be useful as TGF-beta inhibitors. We believe that all the compounds disclosed in the WO 98/52940 application, which we incorporate by reference, would therefore be useful as TGF-beta inhibitors as well.
SUMMARY OF THE INVENTION
The invention herein disclosed is directed to the treatment of cancer and other disease states influenced by TGF beta by inhibiting TGF-β in a patient in need thereof by administering to said patient a compound of from the group consisting essentially of the following: (1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(5) 4-[3-(6-[n-butylamino)pyridin-2-yl]-lH-pyrazol-4-yl]-quinoline. (6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(11) 6-trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline.
(12) 4-[3-(3 -chlorophenyl)- 1 H-pyrazol-4-yl] -quinoline . (13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
(14) 6-sec-butyl-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
(15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)- 1 H-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (18) 4-[3-(6-propylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole.
(20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) 1 -benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3 -(2-pyridyl)-4-(4-quinolyl)pyrazole. (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol.
(25) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate.
(26) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]morpholine.
(27) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine. (28) 4-(4-Pyridin-2-yl- 1 H-pyrazol-3 -yl)-quinoline. (29) 4-(3-Pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
DETAILED DESCRIPTION OF THE P VENTION The compounds disclosed herein can be made according to the following examples. The examples should in not be understood to be limiting in any way as to how the compounds may be made. EXAMPLE 1 6-Methylpyridine-2-carboxylic acid Methyl Ester
Figure imgf000009_0001
To a solution of 6-methylpyridine-2-carboxylic acid (TCI America, 5.48 g, 40 mmol) in 80 ml MeOH and 280 ml acetonitrile add slowly (trimethylsilyl)diazomethane (Aldrich, 2.0M in hexanes, 25 ml, 50 mmol). Stir at RTfor 1 hour and check the reaction for completeness (tic; 9/1 CH2Cl2/MeOH). Add AcOH(3 drops) to the reaction mixture which dissipates the gold color. Stir 30 minutes, remove the solvent in vacuo (rotary evaporator). Dissolve the resulting residue in EtOAc and wash with NaHCO solution.
Purify by silica gel chromatography to afford the title compound (3.2 g, 53%) as a colorless oil.
]H NMR (CDC13): δ 7.83(d, 1H,J=7.7 Hz), 7.61 (t, 1H,J=7.8 Hz), 7.33 (d, 1H,J=7.7 Hz), 3.9 (s, 3H), 2.55 (s, 3H).
MS (APCI)(M+H) 152.
EXAMPLE 2
2-(4-Fluorophenyl)- 1 -(6-methylpyridin-2-yl)- 1 -ethanone
Figure imgf000009_0002
Combine a solution of methyl-4-fluorophenylacetate (1M THF, 15 ml), potassium t-butoxide (1M THF, 45 ml), and 6-methylpyridine-2-carboxylic acid methyl ester (1M THF, 15 ml). Heat the mixture (oil bath) to 65 °C /48hrs. Evaporate the THF to give a brownish paste (ion spray MS m/z m+1 : 288). Add cautiously to the paste cone. HC1 (10 ml, 120 mmol) and heat to 100 °C for 12 hours. Add 6N sodium hydroxide to neutralize (pH = 8-9). Extract with EtOAc, dry (MgSO_ι), and filter to give the crude product.
Chromatography (silica gel, Hexanes/EtOAc gradient) provided the pure product (1.12 g, 4.1mmol, 27% yield).
MS (APCI)(M+1) 231.
EXAMPLE 3 3-(6-Methylpyridin-2-yl)-4-(4-fluorophenyl)- 1 H-pyrazole
Figure imgf000010_0001
Dissolve 2-(4-fluorophenyl)-l-(6-methylpyridin-2-yl)-l-ethanone (1,12 g, 4.88 mmol) in 20 ml THF. Add dimethyformamide dimethylacetal (5.82g, 6.49 ml, 48.8 mmol)and stir 40 hours. Evaporate off solvent in vacuo to give a dark residue. Add 10 ml EtOH and hydrazine monohydrate (5.2 ml, 7.43 g, 148 mmol) and stir overnight. Evaporate the solvent and partition the residue between EtOAc and water. Additionally wash the aqueous with EtOAc. Combine the organic extracts and evaporate to give the crude product residue. Chromatography (silica gel, Hx/EtOAc-EtOAc/MeOH gradient provided the title product (65 mg, 5% yield).
1H NMR (CDC13): δ 7.58 (s, 1H), 7.4 (t, 1H), 7.33-7.37 (m, 2H), 7.02-7.1 (m, 4H), 2.51 (s, 3H).
MS (APCI)(M+1) 254. EXAMPLE 4 4-Methyl-7-ethoxyquinoline
Figure imgf000011_0001
Make a solution A of: 135.0 g FeCl3-6H2O, 5.0 g ZnCl , 25 ml cone. HC1, and add EtOH to bring volume to 375 ml.
Add 3-ethoxyaniline (m-phenetidine; 1.72 g, 12.5 mmol), solution A (15 ml), and 1,3,3-trimethoxybutane (1.6 ml, 10 mmol) into a large screw cap vial. Place into a heating block/shaker with heat set to 63 °C. Heat for 6 hours and let cool to RT overnight. Pour the reaction mixture into 100 ml of 1M sodium citrate and extract with EtOAc. Wash the EtOAc with 1M sodium citrate and then IN sodium hydroxide. Dry with hydromatrix and silica gel and shake well. Filter through fluted filter paper. Add 4 g DIEA-Resin (Argonaut Technologies, approximate loading 3.3 mmol/g), and 5g polystyrene carbonyl chloride resins, (approximate loading 2.0 mmol/g). Shake for several hours and filter. Silica gel chromatography (hexane/EtOAc or CH2Cl2/EtOAc) affords the title compound (0.9 g, 4.8 mmol, 38% yield).
TLC: hexane/EtOAc (1 :1, Rf 0.4) (Note: the 6-isomer is also obtained; Rj-: 0.5).
1H NMR: δ 8.64(d, 1H,J=4.4 Hz), 7.83(d, 1H,J=9.2 Hz), 7.36(d, 1H,J=2.7 Hz), 7.15(dd, 1H,J=9.2, 2.7 Hz), 7.04 (dd, 1H,J=4.4,0.9 Hz), 4.15(q, 2H), 2.6(s, 3H), 1.45(t, 3H).
MS (APCI)(M+1) 188. EXAMPLE 5 4-(3-(6-Methylpyridin-2-yl)- 1 H-pyrazol-4-yl)-7-ethoxyquinoline
Figure imgf000012_0001
Dissolve 4-methyl-7-ethoxyquinoline (0.45 g, 2.41 mmol) in THF (10 ml). Add lithium bis(trimethylsilyl)amide (IM THF, 7.22 ml, 7.22 mmol)and stir 10 minutes. Add 6-methylpyridine-2-carboxylic acid methyl ester (364 mg, 2.41 mmol) and stir overnight at RT. Evaporate the solvent and partition the residue between NH4CI solution and EtOAc. The EtOAc is washed with brine twice. Dry (hydromatrix and silica gel). Filter to give a residue which is purified via silica gel chromatography (gradient: 100% EtOAc - 9:1 EtOAc/MeOH). Combine column fractions containing the product l-(6-methylpyridin-2-yl)-2-(7-ethoxyquinolin-4-yl)-l-ethanone and use without further purification in the next reaction. Dissolve the l-(6-methylpyridin-2-yl)-2-(7-ethoxyquinolin-4-yl)-l-ethanone in
20 ml THF. Add dimethylformamide dimethyl acetal (DMF-DMA)(2.86g, 3.2 ml, 24 mmol) and shake overnight at RT. Evaporate the solvent to give a residue. Add EtOH (20 ml) and hydrazine monohydrate (2.52 ml, 3.6 g, 72 mmol) to the residue and shake overnight. Evaporate the EtOH and hydrazine and partition the residue between ammonium chloride solution and EtOAc. Evaporate the EtOAc to provide the crude product. Purification with silica gel chromatography provides the desired pyrazole product (56 mg, 0.17 mmol, 7% yield from the quinoline).
Η NMR: δ 8.84(d, 1H), 8.57(d, 1H), 7.68(s, 1H), 7.63(d, 1H), 7.46(d, 1H), 7.32(m, 1H), 7.25(d, 1H), 7.12(m, 1H), 7.06(dd, 1H), 6.78(d, 1H), 4.2(q, 2H), 1.48(t, 3H). MS (APCI)(M+1) 331. EXAMPLE 6 4-(3-Pyridin-2-yl- 1 H-pyrazol-4-yl)-7-ethoxyquinoline
Dissolve 4-methyl-7-ethoxyquinoline (0.45g, 2.41 mmol) in 10 ml THF. Add lithium hexamethyldisilylazide (IM THF, 7.22 ml, 7.22 mmol). Add 6-methylpyridine-2- carboxylic acid methyl ester (330 mg, 2.41 mmol)and stir 18 hours. Evaporate the solvent to give a residue which is partitioned between EtOAc and NH4CI solution. Wash the EtOAc layer twice with brine and dry with hydromatrix and a small amount of silica gel. Filter to give crude l-(pyridin-2-yl)-2-(7-ethoxyquinolin-4-yl)-l-ethanone, MS(APCI)(M+1) 293.
Dissolve l-(pyridin-2-yl)-2-(7-ethoxyquinolin-4-yl)-l-ethanone in 20 ml THF and add 10 molar equivalents of DMF-DMA (24 mmol, 2.86 g) and stir overnight. Evaporate the solvent to give a dark residue. Add EtOH (20 ml) to dissolve the residue and add hydrazine monohydrate (72 mmol, 3.6 g) and stir 18 hours. Evaporate the solvent and partition the residue between NH4CI solution and EtOAc. Evaporation of the EtOAc gives a dark residue. Chromatography (silica gel, CH2θ2/EtOAC gradient) provides the title compound (110 mg, 24% yield).
1H NMR (CDC13): δ 8.84(d, 1H), 8.58(d, 1H), 7.68(s, 1H), 7.64(d, 1H), 7.47(d, 1H), 7.30-7.36(m, 1H), 7.26(d, 1H), 7.1-7.14(m, 1H), 7.05(dd, 1H), 6.78(d, 1H), 4.18(q, 2H), 1.46(t, 3H).
MS(APCI)(M+1) 307.
EXAMPLE 7
7-Fluoro-4-methylquinoline
Using a method similar to EXAMPLE 4, 3-fluoroaniline was reacted with 1,3,3- trimethoxybutane in the same manner to produce 1.5 g 7-fluoro-4-methyl-quinoline. MS (APCI)(M+1) 162. EXAMPLE 8 7-Fluoro-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Figure imgf000014_0001
Dissolve 4-methyl-7-fluoroquinoline (1.50g, 9.32mmol, HL5-A3B-87-10B) in
THF (50mL) in a round bottomed flask. Cool solution to 0 °C on an ice bath. Add lithium bis(trimethylsilyl)amide (20mL, 21.2mmol, 1.06M in THF, Lancaster) via syringe. Let stir 15 minutes, then add 6-methylpyridine-2-carboxylic acid methyl ester (1.697g, 11.2mmol) dissolved in THF (15mL) slowly over 15 minutes via syringe. Let warm to RT with stirring overnight.
Remove THF in vacuo and dissolve remaining residue in EtOAc (lOOmL). Wash with saturated ammonium chloride solution (2 X lOOmL), and brine (1 x lOOmL). Dry over anhydrous MgSO4, filter and concentrate in vacuo to give 2-(7-fluoroquinolin-4-yl)- l-(6-methylpyridin-2-yl)ethanone as a dark red oil.
Combine a solution of 2-(7-fluoroquinolin-4-yl)-l-(6-methylpyridin-2-yl)ethanone (4.0g, 14.2mmol) and N,N-dimethylformamide dimethyl acetal (9.0g, 79.6mmol, Aldrich) in THF (lOOmL). Let stir at RT. Remove THF in vacuo to yield a crude brown residue. Dissolve the residue (9.0g, 32.0mmol) in absolute EtOH (50mL) and add hydrazine (1.50g, 30mmol, Aldrich) via syringe. Let stir at RT overnight.
Remove EtOH in vacuo and dissolve resulting residue in EtOAc (lOOmL). Wash with water (2 x lOOmL), dry over anhydrous MgSO4, filter and concentrate in vacuo.
Chromatograph the residue on silica gel (elute with 98% CH2C12/MeOH) to afford the title compound (0.478g, 1.57mmol, 16.8%) as a yellow solid. MS (APCI) (M+l) 305.327. EXAMPLE 9 4-[3-(6-Bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Figure imgf000015_0001
Dissolve lepidine (5.01g, 35.1mmol, Aldrich) in THF (150mL) in a 3-necked round bottomed flask equipped with addition funnel under N . Cool solution to 0 °C on an ice bath. Add lithium bis(trimethylsilyl)amide (70mL, 74.2mmol, 1.06M in THF, Lancaster) via syringe. Let stir 15 minutes, then add 6-bromopyridine-2-carboxylic acid methyl ester (9.67g, 44.8mmol) dissolved in THF (30mL) dropwise over 30 minutes via the addition funnel. Let warm to RT with stirring.
Remove THF in vacuo and dissolve remaining residue in EtOAc (200mL). Wash with saturated ammonium chloride solution (2 X 150mL), and brine (1 x 150mL). Dry over anhydrous MgSO4, filter and concentrate in vacuo. Chromatograph on silica gel to give 2-(quinolin-4-yl)-l -(6-bromopyridin-2-yl)ethanone as a yellow oil.
Combine a solution of 2-(quinolin-4-yl)-l-(6-bromopyridin-2-yl)ethanone (7.59g, 23.2mmol) and N,N-dimethylformamide dimethyl acetal (30.5g, 256.3mmol, Aldrich) in THF (200mL). Let stir at RT. Removed THF in vacuo to yield a crude brown residue.
Dissolve the residue (8.83g, 23.2mmol) in absolute EtOH (150mL) and add hydrazine (1.50g, 30mmol, Aldrich) in one portion via syringe. Let stir at RT overnight. Remove EtOH in vacuo and dissolve resulting residue in EtOAc (lOOmL). Wash with water (2 x 150mL), dry over anhydrous MgSO4, filter and concentrate in vacuo.
Chromatograph the residue on silica gel (elute with 98% CH2Cl2/MeOH) to afford the title compound (2.1 lg, δ.Ommol, 17.1%) as a pink solid. MS (APCI) (M+l) m/z 351.3. EXAMPLE 10 4-[3-(6-[n-Butylamino)pyridin-2-yl]-lH-pyrazol-4-yl]-quinoline
Figure imgf000016_0001
Combine 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline (0.1 lOg, 0.313mmol) and n-butylamine (27.8g, 379.8mmol, Aldrich) in a sealed tube with a Teflon screw cap and heat to 150 °C on an oil bath for 48 hours. Remove excess n-butylamine in vacuo. Dissolve residue in EtOAc (25mL) and wash with water (2 x 25mL). Dry over anhydrous MgSO4, filter and concentrate in vacuo. Chromatograph residue on silica gel
(elute with 95% CH2C12/MeOH) to afford the title compound (0.05 lg, 0.149mmol, 41.6%) as a brown gum.
MS (APCI) (M+l) m/z 344.3.
EXAMPLE 11 4-[3-(6-Methylpyridin-2-yl)- 1 H-pyrazol-4-yl] -quinoline
Figure imgf000016_0002
To a solution of lepidine (1.43 g, 10 mmol, Aldrich) in THF (20 mL) add lithium bis(trimethylsilyl)amide (30 mL, 30 mmol, 1.0 M THF). Stir for 10 min and add a solution of 6-methylpyridine-2-carboxylic acid methyl ester (1.66 g, 11 mmol) in THF
(10 mL). Stir the reaction mixture overnight. Remove THF from the reaction mixture in vacuo and quench the mixture with satd. NH C1. Extract the reaction mixture in EtOAc and separate the layers. Wash the organic layer with brine, dry over hydromatrix. Treat the organic layer with Siθ2 for 30 min and filter through celite. Remove EtOAc in vacuo to afford crude 2-(quinolin-4-yl)-l-(6-methylpyridin-2-yl)ethanone.
Dissolve 2-(quinolin-4-yl)-l-(6-methylpyridin-2-yl)ethanone in THF (20 mL) and add N,N-dimethylformamide-dimethyl acetal (10.6 mL, 80 mmol). Stir the reaction mixture for two days and then remove THF in vacuo. Dissolve the residue in reagent alcohol (20 mL) and to this solution add hydrazine monohydrate (11.63 mL, 240 mmol). Stir the reaction mixture overnight. Concentrate the reaction mixture in vacuo and chromatograph the residue on silica gel (50 % EtOAc/ 49 % hexanes/ 1 % TEA) to give the title compound (572 mg, 20 %).
]H NMR (CDC13): δ8.94 (d, IH, J=4.4 Hz), 8.20 (d, IH, J=8 Hz), 7.79 (d, IH, J=8 Hz), 7.72 (m, 2H), 7.43 ( , 2H), 7.21 (t, IH, J=8 Hz), 6.97 (d, IH, J=7.7 Hz), 6.58 (d, IH, J=7.7 Hz), 2.53 (s, 3H).
MS (APCI) (M+l) calc 287.1; found 287.6.
EXAMPLE 12 6-Chloro-4-methylquinoline
Using a method similar to EXAMPLE 4, 4-chloroaniline was reacted with 1,3,3- trimethoxybutane in the same manner to produce 70 mg 6-chloro-4-methylquinoline. MS (APCI)(M+1) 179.
EXAMPLE 13
6-Chloro-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Using a procedure similar to EXAMPLE 11 , prepare the title compound starting from 6-chloro-4-methylquinoline in an overall of yield of 5%.
1H NMR (CDCI3): δ8.92 (d, IH, J=4.4 Hz), 8.29 (d, IH, J=8 Hz), 7.80 (d, IH, J=2.2 Hz), 7.73 (s, IH), 7.71 (dd, IH, J=2.2 Hz), 7.50 (d, IH, J-4.4 Hz), 7.32 (t, IH, J=8 Hz), 7.03 (d, IH, 1=1 Hz), 6.71 (d, IH, 1=1 Hz), 2.51 (s, 3H). MS (APCI) (M+l) calc 321.1; found 321.4. EXAMPLE 14 6-Trifluoromethyl-4-methylquinoline
Using a method similar to EXAMPLE 4, 4-trifluoromethylaniline was reacted with 1,3,3-trimethoxybutane in the same manner to produce 233 mg 6-trifluorornethyl-4- methyl-quinoline .
MS (APCI)(M+1) 212.
EXAMPLE 15 6-Trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl3-quinoline
Figure imgf000018_0001
Using a procedure similar to EXAMPLE 11 , prepare the title compound starting from 6-trifluoro-4-methyl quinoline in an overall of yield of 3%.
1H NMR (CDC13): δ 9.04 (d, IH, J=4.4 Hz), 8.32 (d, IH, J=8.8 Hz), 8.09 (s, IH), 7.88 (d, IH, J=8.8 Hz), 7.75 (s, IH), 7.50 (d, IH, J=4.8 Hz), 7.28 (t, IH, J=8 Hz), 7.01 (d, IH, 1=1 Hz), 6.67 (d, IH, J=7.5 Hz), 2.50 (s, 3H). MS (APCI) (M+l) calc 355.1; found 355.3.
EXAMPLE 16 7-Methyl-4-methylquinoline
Using a method similar to EXAMPLE 4, 3-methylaniline was reacted with 1,3,3- trimethoxybutane in the same manner to produce 743 mg 7-methyl-4-methyl-quinoline. MS (APCI)(M+1) 158. EX AMPLE 17 7-Methyl-4-[3-(6-methyl-pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Figure imgf000019_0001
Dissolve 4,7-dimethylquinoline (350mg, 2.23mmol) in 10ml THF. Add IM THF solution of LHMDS (5.35mmol, 2.4eq) to the solution at RT and stir for 30 mins. Add 6- methyl-pyridine-2-carboxylic acid methyl ester (337mg, 2.23mmol) to the reaction mixture. Stir overnight. Add NH C1 saturated aqueous solution and extracted with EtOAc. Wash the EtOAc extract with brine, dry (Na2SO4), filter and concentrate the filtrate to give a residue.
Dissolve the residue in 10 ml THF and add 3.4 ml of dimethylformamide dimethyl acetal. Stir for 48 hrs. Remove the solvent. Add EtOH (10ml) and 3.4 ml of hydrazine. Stir overnight. Concentrate the mixture in vacuo. Chromatograph the residue on silica gel (elute with CH2C12, 95/5 CH2Cl2:MeOH) to afford the product (52 mg, 8%).
1H NMR (CDC13): δ 8.88 (d, IH), 7.94 (s, IH), 7.68 (s, IH), 7.64 (d, IH), 7.32(d, IH), 7.24 (d, IH), 7.2 (t, IH), 6.97 (d, IH), 6.56 (d, IH), 2.57 (s, 6H). MS (ACPI) calcd. 300.4; MS (M+l) 301.5.
EXAMPLE 18
6-Methoxy-4-methylquinoline
Using a method similar to EXAMPLE 4, 4-methoxyaniline was reacted with 1,3,3-trimethoxybutane in the same manner to produce 355 mg 6-methoxy-4-methyl- quinoline.
MS (APCI)(M+1) 174. EXAMPLE 19 6-Methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Using a method similar to EXAMPLE 17, with the exception of using 4-methyl-6- methoxy-quinoline, affords the product (2% yield).
1H NMR (CDC13): δ8.79(d, IH), 8.06(d, IH), 7.70 (s, IH), 7.36 (m, 2H), 7.22 (t, IH), 6.98 (m, 2H), 6.60 (d, IH), 3.60 (s, 3H), 2.50 (s, 3H). MS (ACPI) calcd. 316.4; MS (M+l) 317.2.
EXAMPLE 20
6-Trifluoromethoxy-4-methylquinoline
Using a method similar to EXAMPLE 4, 4-trifluoromethoxyaniline was reacted with 1,3,3-trimethoxybutane in the same manner to produce 326 mg 6-trifluoromethoxy- 4-methyl-quinoline.
MS (APCI)(M+1) 228.
EXAMPLE 21
6-Trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline
Figure imgf000020_0001
Using a method similar to EXAMPLE 17, with the exception of using 4-methyl-6- trifluoromethoxyquinoline, affords the product (9% yield). 1H NMR (CDCI3): δ8.96(d, IH), 8.20 (d, IH), 7.69 (s. IH), 7.56 (m, 2H), 7.46(d,
IH), 7.22 (t, IH), 6.98 (d, IH), 6.58 (d, IH), 2.50 (s, 3H). MS (ACPI) calcd. 370.4. MS (M+l) 371.3. EXAMPLE 22 4-[3-(3-Chlorophenyl)-lH-pyrazol-4-yl]-quinoline
Figure imgf000021_0001
Using a method similar to EXAMPLE 5, with the exception of using lepidine and methyl 3-chlorobenzoate as starting materials, affords the product (54% yield). MS (APCI)(M+1) 306.
EXAMPLE 23
7-Butoxy-4-methylquinoline
Using a method similar to EXAMPLE 4, with the exception of using 4- butoxyaniline as starting material, affords the product (30% yield). MS (APCI)(M+1) 216.8.
EXAMPLE 24 6-Butoxy-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline
Using a method similar to EXAMPLE 5, with the exception of using 7-butoxy-4- methylquinoline and pyridine-2- carboxylic acid methyl ester as starting materials, affords the product (16% yield). 1H NMR (CDC13): δ 8.82 (d, IH, J=4.6 Hz), 8.62 (d, IH, J=4.6 Hz), 8.10 (d, IH,
J=9.2 Hz), 7.76 (s, IH), 7.41-7.37 (m, 3H), 7.17 (dd, IH, J=7.3, 5 Hz), 6.99 (d, IH, J=2.8 Hz), 6.85 (d, IH, J=7.8 Hz), 3.73 (t, 2H, J=6 Hz), 1.66 (quint, 2H, J=8.2 Hz), 1.41 (sext, 2H, J=7.5 Hz), 0.91 (t, 3H, J=7.3 Hz). MS (APCI)(M+1) 345.2.
EXAMPLE 25 7-sec-Butyl-4-methylquinoline
Figure imgf000022_0001
Using a method similar to EXAMPLE 4, with the exception of using 4-sec- butylaniline as starting material, affords the product (36% yield). MS (APCI)(M+1) 200.5.
EXAMPLE 26 6-sec-Butyl-4-(3-pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline
Figure imgf000022_0002
Using a method similar to EXAMPLE 5, with the exception of using 7-sec-butyl- 4-methylquinoline and pyridine-2-carboxylic acid methyl ester as starting materials, affords the product (11% yield). 1H NMR (CDC13): δ 8.93 (d, IH, J=4.1 Hz), 8.61 (d, IH, J=4.1 Hz), 8.14 (d, IH,
J=8.3 Hz), 7.76 (s, IH), 7.60 (dd, IH, J=8.7, 1.8 Hz), 7.51 (s, IH), 7.44 (d, IH, J=4.6 Hz), 7.34 (t, IH, J=7.8 Hz), 7.15 (dd, IH, J=4.6, 2.3 Hz), 6.81 (d, IH, J=7.8 Hz), 2.60 (sext, IH, J=7 Hz), 1.49 (m, 2H), 1.12 (d, 3H, J=6.9 Hz), 0.63 (t, 3H, J=7.3 Hz). MS (APCI)(M+1) 329.4. EXAMPLE 27 4-(4-Methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole
Figure imgf000023_0001
Using a method similar to EXAMPLE 27, with the exception of using (4- methoxyphenyl)acetone (Aldrich) as starting material, affords the product (3% yield).
1H NMR (CDC13): δ 7.37 (t, IH, J=7.8 Hz), 7.25 (d, 2H, J=8.3 Hz), 7.01 (d, IH, J=8.3 Hz), 6.99 (d, 2H, J=8.7 Hz), 6.94 (d, IH, J=7.8 Hz), 3.88 (s, 3H), 2.58 (s, 3H), 2.23 (s, 3H).
MS (APCI)(M+1) 281.
EXAMPLE 28 (Quinolin-4-yl)acetone
Figure imgf000023_0002
Using a method similar to EXAMPLE 5, with the exception of using lepidine and mEtOAc as starting materials, affords the product in crude form, which proved adequate for further use ( 100% yield).
MS (APCI)(M+1) 186.2. EXAMPLE 29 4-[5-Methyl-3-(6-methylpyridin-2-yl)-lH-ρyrazol-4-yl]-quinoline
Figure imgf000024_0001
Using a method similar to EXAMPLE 27, with the exception of using (quinolin-4- yl)acetone as starting material, affords the product (4% yield).
1H NMR (CDC13): δ 9.01 (d, IH, J=4.1), 8.23 (d, IH, J=8.3), 7.75 (t, IH, J=7.8), 7.70 (d, IH, J=8.3), 7.46 (t, IH, J=7.5), 7.41 (d, IH, J=4.1), 7.19 (t, IH, J=7.8), 6.97 (d, IH, J=7.8), 6.47 (d, IH, J=7.8), 2.55 (s, 3H), 2.12 (s, 3H).
MS (APCI)(M+1) 302.
EXAMPLE 30
Figure imgf000024_0002
6-Propylpyridine-2-carboxylic Acid Methyl Ester Dissolve dimethyl oxalate (5.90 g, 50 mmol, Aldrich) and KOH (2.80 g, 50 mmol) in 9/1 - MeOH/water (25 mL) and stir at RT 4 h. Evaporate the oxalate salt to a residue and re-dissolve in water (40 mL).
Dissolve sodium persulfate (2.38 g, 10 mmol, Aldrich), silver nitrate (85 mg, 0.5 mmol, Aldrich), and H SO (0.47 mL, 8.5 mmol) in water (10 mL). Add CH2C12 (20 mL) and 2-propylpyridine (0.664 mL, 5 mmol, Lancaster), and heat the mixture to reflux with rapid stirring. Add the above oxalate salt solution (10 mL portion) dropwise rapidly, and continue heating 1.5 h.
Cool the mixture and adjust to basic pH with satd aq Na2CO3. Extract with
CH C1 (15 mL), dry the organics over hydromatrix, filter and evaporate to obtain the title product as an oil suitable for further use, 209 mg (23% yield). 1H NMR (CDC13): δ 7.97 (d, IH, J=7 Hz), 7.76 (t, IH, J=7.7 Hz), 7.37 (d, IH, J=7 Hz), 4.00 (s, 3H), 2.90 (t, 2H, J=7.7 Hz), 1.77 (m, 2H), 0.98 (t, 3H, J=7.3 Hz). MS (APCI)(M+1) 180.0.
EXAMPLE 31 4-[3-(6-Propylpyridin-2-yl)- 1 H-pyrazol-4-yl]-quinoline
Figure imgf000025_0001
Using a method similar to EXAMPLE 5, with the exception of using lepidine and 6-propylpyridine-2-carboxylic acid methyl ester as starting materials, affords the product (10% yield).
1H NMR (CDC13): δ 8.98 (d, IH, J=4.6), 8.23 (d, IH, J=8.3), 7.83 (d, IH, J=8.3), 7.75 (t, IH, J=7.8), 7.72 (s, IH), 7.46 (m, 2H), 7.26 (m, IH), 7.00 (d, IH, J=7.8), 6.65 (d, IH, J=7.8), 2.77 (t, 2H, J=7.8 Hz), 1.76 (sext, 2H, J=7.3 Hz), 0.99 (t, 3H, J=7.3 Hz). MS (APCI)(M+1) 315.
EXAMPLE 32 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole
Figure imgf000025_0002
Heat a solution of 2-quinolin-4-yl-l-pyridin-2-yl ethanone (0.50 g, 2.01 mmol), cyclopropane-carboxylic acid hydrazide (0.20 g, 2.01 mmol) and HC1 (cone. 3 drops) in
THF (10 mL) under nitrogen at 40 °C for 18 h. Collect the precipitate formed in the reaction by filtration. The solid (hydrazone) is dried in vacuo, then heated in an oil bath at 185 °C for 15 min. Partition the dark residue between CHC^/aq NaHCO3. Combine the organic extracts, dry (Na2SO4), filter, and concentrate in vacuo. Chromatograph the residue on silica gel (elute with 2% MeOH/CHCl3) to afford the title compound as an off- white solid.
1H NMR (CDC13): δ 9.01 (d, IH, J=4Hz), 8.55 (d, IH, J=4Hz), 8.24 (d, IH, J=8Hz), 7.79 (t, IH, J=8Hz), 7.75 (td, IH, J=8,2Hz), 7.48 (m, 2H), 7.30 (td, IH, J=8,2H), 7.10 (ddd, IH, J=8,7,2Hz), 6.64 (m, IH), 1.45 (m, IH), 0.94 (m, 2H), 0.75 (m, 2H). MS ES (M+l) 313.
EXAMPLE 33
3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole
Figure imgf000026_0001
Combine a mixture of 2-quinolin-4-yl-l-(3-trifluoromethylphenyl)-ethanone (1.00 g, 3.17 mmol) and N,N-dimethylformamide dimethyl acetal and heat at reflux for 4 h.
Concentrate in vacuo to afford a colored oil. Dissolve 0.59 g of this dark residue in ethanol (10 mL) and treat with hydrazine monohydrate (0.25 mL, 7.95 mmol). Stir at RT overnight. Partition between CHC^/aq NaHCO3. Combine the organic extracts, dry
(Na2SO4), filter, and concentrate in vacuo. Chromatograph the residue on silica gel (elute with 2% MeOH/CH2θ2) to afford the title compound as an off-white solid.
1H NMR (CDCI3): δ 9.14 (d, IH, J=4Hz), 8.36-8.32 (m, 2H), 8.05-7.95 (m, 2H), 7.84 (t, IH, J=8Hz), 7.75-7.62 (m, 3Hz), 7.50-7.41 (m, 2H). MS ES (M+l) 340. EXAMPLE 34 l-benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. 2 HC1
Figure imgf000027_0001
Add 2.10 mL (12.06 mmol) diisopropylethylamine to a suspension of 1.18 g (6.03 mmol) benzylhydrazine.2H2θ in 10 ml ethanol. Add a 5 mL ethanol soln (approximately
2.01 mmol) of ketone reagent and heat 2 h at 60 °C. Cool and stir at rt 16 h. Evacuate and extract product between 100 ml CHCI3 and 100 ml sat'd NaHCO3 soln.. Dry (Na2SO4) and evacuate. Purify the product over silica (0 to 2.2 % MeOH in CH2CI2 with 3 drops cone. NH4OH per 150 mL) to obtain title compound as free base. Dissolve the product in
10 mL MeOH, add excess 2 M HC1 in MeOH, and evacuate to obtain 20 mg of title compound.
1H NMR (DMSO-de) δ: 5.51 (1, 3H), 7.29 (m, IH), 7.33-7.49 (m, 5H), 7.77 (t, IH), 7.88-7.93 (m, 3H), 8.03 (d, IH), 8.10 (t, IH), 8.13 (d, IH), 8.45 (d, IH), 8.55 (s, IH), 9.21 (d, IH).
ES MS 363 (M+l for free base).
EXAMPLE 35 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. 2HC1
Figure imgf000028_0001
Prepare title compound (50 mg, 18 % Yield), using 1 -chloro-4-phenylbutane as the reagent, similar as in EXAMPLE 35 (on same scale).
1H NMR (DMSO-d6) δ: 1.66 (quintet, 2H), 1.97 (quintet, 2H), 2.67 (t, 2H), 4.36 (t, 2H), 7.15 - 7.30 (m, 6H), 7.72 (t, IH), 7.85 -7.93 (m, 3H), 7.97 (d, IH), 8.10 (m, 2H), 8.38 (d, IH), 8.39 (s, IH), 9.17 (d, IH); ES MS 405 (M+l).
EXAMPLE 36 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol. HCl
Figure imgf000028_0002
Dissolve 1.0 g (4.03 mmol) ketone in 15 mL ethanol. Add 0.30 mL (4.43 mmol) hydroxyethylhydrazine and 6 drops cone HCl. Heat at reflux for 1.75 h and evacuate solvent. Dissolved residue in 10 mL N,N-dimethyformamide dimethyl acetal, heat at reflux 6 h, and stir at ambient temperature 16 h. Evacuate solvent and extract product between 100 mL CHCI3 and 100 mL dilute aqueous NaHCO3. Dry organic layer (Na2SO4), concentrate, and purify over silica (0 to 4 % MeOH in CHCI3 with 3 drops cone NH4OH per 150 mL solvent) to obtain 0.67 g (53 % Yield) of the title compound as the free base. Obtain some HCl salt (title compound) by treating a fraction of the free base in MeOH with excess 2M methanolic HCl and then evacuating solvent. ]H NMR (DMSO-d6) δ: 3.77 (t, 2H), 4.45 (t, 2H), 7.25 (d, IH), 7.38 (m, IH), 7.51 (d, IH), 7.65 - 7.73 (m, 2H), 7.97 (m, 2H), 8.03 (s, IH), 8.27 (d, IH), 8.68 (d, IH), 9.06 (d, IH); ES MS 317 (M+l).
EXAMPLE 37
2-(3 -(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate
Dissolve 0.30 g (0.95 mmol) alcohol free base prepared in EXAMPLE 37 in 4 mL pyridine. Cool soln. in salt-ice bath and slowly add 0.10 mL (1.05 mmol) methanesulfonyl chloride. Stir reaction mixture 18 h at °C, add 4 mL ice cold water, and extract between 50 mL methylene chloride and 50 mL dilute aquous NaHCO3. Extract aquous layer with 50 mL methylene chloride, combine organic layers, dry (Na2SO4), and evacuate to give an oil, to be used without further purification. ES MS 395 (M+l). EXAMPLE 38
4-[2-(3-(2-pyridyl)-3-(4-quinolyl)pyrazolyl)ethyl]morpholine
Figure imgf000029_0001
Dissolve 0.12 g (0.30 mmol) methanesulfonate intermediate EXAMPLE 38 in lmL methylene chloride. Add excess morpholene (0.2 mL) and stir at 75 °C 4 h. Extract the product between 50 mL CHCI3 and 50 mL sat'd aquous NaHCO3- Dry (Na2SO4) and concentrate organic layer. Purify product over silica (0 to 4 % MeOH in methylene chloride with 3 drops NH4OH per 150 mL solvent) and precipitate fractions collected from ether-hexane to obtain 50 mg (43 % yield) of title compound. 1H NMR (DMSO-d6) δ: 2.50 (t. 4H), 2.86 (t, 2H), 3.57 (t, 4H), 4.41 (t, 2H), 7.16 (t, IH), 7.32 (d, IH), 7.402 (t, IH), 7.70 (t, IH), 7.74 - 7.83 (m, 3H), 8.03 (d, IH), 8.22 (d, IH), 8.27 (s, IH), 8.83 (d, IH); ES MS 386 (M+l).
EXAMPLE 39 Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethyl]amine
Figure imgf000030_0001
Prepare title compound as in EXAMPLE 39 (same scale), using aniline, to obtain 80 mg (68 % yield) of precipitate material from ether-hexane.
]H NMR (DMSO-d6) δ: 3.58 (quintet, 2H), 4.94 (t, 2H), 5.88 (t, IH), 6.54 (t, IH), 6.63 (d, 2H), 7.070 (d, IH), 7.10 (d, IH), 7.17 (t, IH), 7.28 (d, IH), 7.38 (t, IH), 7.68 (d, IH), 7.69 (t, IH), 7.78 (t, IH), 7.62 (t, IH), 8.03 (d, IH), 8.07 (s, IH), 8.13 (d, IH), 8.82 (d, IH); ES MS 392 (M+l). EXAMPLE 40 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl -quinoline
Figure imgf000031_0001
Combine 2.5 g (14.4 mmol) of quinoline-4-carboxylic acid, 30 mL thionyl chloride and 2 drops of DMF. Reflux for 16 h then concentrate to dryness under vacuum. Mix residue with 1,2-dichloroethane and reconcentrate to dryness. Mix with 50 mL CH2CI2 and add 2.8 g (28.9 mmol) of N-O-dimethylhydroxylamine. Add 7.5 g (57.8 mmol) of diisoproplyethylamine and stir for 2 hours. Concentrate to dryness. Mix with 30 mL CHCI3 and chromatograph on silica using 1 % MeOH in CHCI3 to recover 2.96 g of an oil.
In a dry flask, mix 25 mL dry THF and 2-methylpyridine. Cool to -78 C and add 7 mL of IM sodium hexamethyldisilazide in THF dropwise. Warm to 0 C and stir for 1 hour. Recool to -78 C and transfer via cannula into a separate flask containing a mixture of the crude amide from the above procedure (0.54 g, 5.8 mmol) and 25 mL dry THF at - 78 C. Stirred at RT for 17 hours. Quench with 5 drops of saturated aqueous NH4CI, concentrate to dryness under vacuum and partition the residue between 50 mL brine and 50 mL EtOAc. Separate layers and extract the aqueous layer twice with 50 mL EtOAC. Combine the organic layers and dry with MgSO4- Concentrate to dryness under vacuum to recover 0.9 g of an orange oil. Purify by cliromatrography using Hexanes and EtOAc to recover 353 mg orange oil (1.4 mmol, 31%).
Combine the ketone (340 mg, 1.4 mmol), 2 mL DMF dimethylacetal and 3 drops of Et3N. Reflux for 1 hour. Concentrate under vacuum to dryness and mix the residue with 10 mL EtOH and .2 mL hydrazine hydrate. Reflux for 1 hour then concentrate to dryness. Purify by chromatography using silica and CHC^/MeOH to recover 200 mg tan solid.
Calculated for C17H12N4 C 74.98, H 4.44, N 20.57. Found C 74.71, H 4.38, N 20.40. MS calcd. 272; MS (M+l) 273.
EXAMPLE 41 4-(3-Pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline
Figure imgf000032_0001
Commercial source: Peakdale Inc. 109 East Scotland Drive
Bear, DE 19701-1756
USA Catalog #: PFC-0655 The compounds disclosed herein were tested by the following protocols for TGF- β inhibition and for anticancer activity by the following assays, as described below in the protocol description. The data collected thereby is shown in Table 1.
MV1LU P3TP-LUX/PURO CLONE CI ASSAY A stable MvlLu clone (CI) containing the p3TP-Lux reporter was created by standard transfection and puromycin selection protocols. This stable clone was used to screen the example compounds for their ability to inhibit TGF-β dependent luciferase production as briefly described below:
1. Plated MvlLu CI cells in Wallac™ Black Isoplates 2. Allowed cells to adhere overnight.
3. Removed media and replaced with 0.5% FBS DMEM media '4. Added the compound dilution series in 0.5% FBS/DMEM containing 1% DMSO such that the final compound concentration ranged from 20 uM to 0.1 nM and the final DMSO concentration was 0.2%. 5. Incubated at 37°C/5% CO2 for 2 hrs. 6. Added 0.5% FBS/DMEM as control or TGF-βl diluted in 0.5% FBS/DMEM (final concentration of 10 pM) to the -/+ TGF-β wells respectively
7. Incubated for 16-20 hrs. at 37°C/5% CO2 8. Removed media and rinsed IX with PBS.
9. Removed PBS and lysed the cells with IX Passive Lysis Buffer (Promega) at room temperature.
10. Counted relative luciferase activity on the MicroBeta JET by injecting Luciferase Assay Reagent II (PROMEGA). The use of the above assay in measuring TGF-β responsive activity is described in
Wrana, et al. Cell 71 : 1003-101 (1992).
TGF-β RECEPTOR I PURIFICATION AND P VITRO KINASE REACTIONS
For TGF-β Type I CRIT204D Receptors:
The 6X-HIS tagged cytoplasmic kinase domain of the receptor was expressed and purified from Sf9 insect cell lysates as briefly described below:
Cell pellets after 48-72 hrs of infection were lysed in lysis buffer (LB: 50 mM Tris pH 7.5, 150 mM NaCl, 50 mM NaF, 0.5% NP40 with freshly added 20 mM β- mercaptoethanol, 10 mM imidazole, 1 mM PMSF, IX EDTA-free Complete Protease Inlιibitor(Boehringer Mannheim).
Cell lysates were clarified by centrifugation and 0.45 uM filtered prior to purification by Ni/NTA affinity chromatography (Qiagen).
Chromatography Protocol:
Equilibrate with 10 CV of LB, load sample, wash with 10 CV RIPA buffer (50 mM Tris pH 7.5, 150 mM NaCl, 1% NP40, ImM EDTA, 0.25% sodium deoxycholate, added fresh 20 mM β-mercaptoethanol, 1 mM PMSF), wash with 10 CV LB, wash with 10 CV IX KB (50 mM Tris pH 7.5, 150 mM NaCl, 4 mM MgCl2, 1 mM NaF, 2 mM β- mercaptoethanol), elute with a linear gradient of IX KB containing 200 mM Imidazole. The enzyme was approximately 90% pure and had autophosphorylation activity.
Reactions: 170-200 nM enzyme in IX KB, compound dilution series in IX KB/16% DMSO (20 uM to 1 nM final concentration with 4% DMSO final concentration), reactions started by adding ATP mix (4 uM ATP/1 uCi 33P-γ-ATP final concentrations) in IX KB.
Reactions were incubated at 30 °C for 1 lir. Reactions were stopped and quantitated using standard TCA/BSA precipitation onto Millipore FB glass fiber filter plates and by liquid scintillation counting on a MicroBeta JET.
TABLE 1
Figure imgf000034_0001
PHARMACEUTICAL COMPOSITIONS
The compositions of the present invention are therapeutically effective amounts of the TGF-β antagonists, noted above. The composition may be formulated with common excipients, diluents or carriers, and compressed into tablets, or formulated elixirs or solutions for convenient oral administration or administered by intramuscular intravenous routes. The compounds can be administered transdermally and maybe formulated as sustained relief dosage forms and the like.
The method of treating a human patient according to the present invention includes administration of the TGF-β antagonists. The TGF-β antagonists are formulated into formulations which may be administered by the oral and rectal routes, topically, parenterally, e.g., by injection and by continuous or discontinuous intra-arterial infusion, in the form of, for example, tablets, lozenges, sublingual tablets, sachets, cachets, elixirs, gels, suspensions, aerosols, ointments, for example, containing from 1 to 10% by weight of the active compound in a suitable base, soft and hard gelatin capsules, suppositories, injectable solutions and suspensions in physiologically acceptable media, and sterile packaged powders adsorbed onto a support material for making injectable solutions. Advantageously for this purpose, compositions may be provided in dosage unit form, preferably each dosage unit containing from about 5 to about 500 mg (from about 5 to 50 mg in the case of parenteral or inhalation administration, and from about 25 to 500 mg in the case of oral or rectal administration) the compounds. Dosages from about 0.5 to about 300 mg/kg per day, preferably 0.5 to 20 mg/kg, of active ingredient may be administered although it will, of course, readily be understood that the amount of the compound actually to be administered will be determined by a physician, in the light of all the relevant circumstances including the condition to be treated, the choice of compound to be administered and the choice of route of administration and therefore the above preferred dosage range is not intended to limit the scope of the present invention in any way.
The formulations useful for separate administration of the TGF-β antagonists will normally consist of at least one compound selected from the compounds specified herein mixed with a carrier, or diluted by a carrier, or enclosed or encapsulated by an ingestible carrier in the form of a capsule, sachet, cachet, paper or other container or by a disposable container such as an ampoule. A carrier or diluent may be a solid, semi-solid or liquid material which serves as a vehicle, excipient or medium for the active therapeutic substance. Some examples of the diluents or carrier which may be employed in the pharmaceutical compositions of the present invention are lactose, dextrose, sucrose, sorbitol, mannitol, propylene glycol, liquid paraffin, white soft paraffin, kaolin, fumed silicon dioxide, microcrystalline cellulose, calcium silicate, silica, polyvinylpyrrolidone, cetostearyl alcohol, starch, modified starches, gum acacia, calcium phosphate, cocoa butter, ethoxylated esters, oil of theobroma, arachis oil, alginates, tragacanth, gelatin, syrup, methyl cellulose, polyoxyethylene sorbitan monolaurate, ethyl lactate, methyl and propyl hydroxybenzoate, sorbitan trioleate, sorbitan sesquioleate and oleyl alcohol and propellants such as trichloromonofluoromethane, dichlorodifluoromethane and dichlorotetrafluoroethane. In the case of tablets, a lubricant may be incorporated to prevent sticking and binding of the powdered ingredients in the dies and on the punch of the tableting machine. For such purpose there may be employed for instance aluminum, magnesium or calcium stearates, talc or mineral oil.
Preferred pharmaceutical forms of the present invention are capsules, tablets, suppositories, injectable solutions, creams and ointments. Especially preferred are formulations for inhalation application, such as an aerosol, for injection, and for oral ingestion.

Claims

WE CLAIM:
1. A method of inhibiting TGF-β for the treatment of cancer in a patient in need thereof comprising administration to said patient a therapeutically effective amount of a compound selected from the group consisting essentially of:
(1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)- 1 H-pyrazol-4-yl]-quinoline. (5) 4-[3-(6-[n-butylamino)pyridin-2-yl]- 1 H-pyrazol-4-yl]-quinoline.
(6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(11) 6-trifluoro methoxy-4- [3 -(6-methylpyridin-2-yl)- 1 H-pyrazol-4-yl] - quinoline.
(12) 4-[3-(3-chlorophenyl)-lH-pyrazol-4-yl]-quinoline.
(13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline. (14) 6-sec-butyl-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
(15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)- 1 H-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(18) 4-[3-(6-propylpvridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole.
(20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) 1 -benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol. (24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate.
(25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]moφholine.
(26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine. (27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3 -Pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline.
2. A method of inhibiting TGF-β in a patient in need thereof comprising administering to said patient a therapeutically effective amount of a compound selected from the group consisting essentially of:
( 1 ) 4-(3-(6-methyl pyridin-2-yl)- 1 H-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (4) 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(5) 4-[3-(6-[n-butylamino)pyridin-2-yl]-lH-pyrazol-4-yl]-quinoline.
(6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-l H-pyrazol-4-yl] -quinoline.
(11) 6-trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline.
(12) 4-[3-(3-chlorophenyl)-lH-pyrazol-4-yl]-quinoline. (13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
(14) 6-sec-butyl-4-(3 -pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline.
(15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)-lH-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (18) 4-[3-(6-propylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole.
(20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) l-benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol.
(24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate.
(25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]morpholine. (26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine.
(27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3-Pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline.
3. A method of treating cancer in a patient in need thereof comprising administration to said patient a therapeutically effective amount of a compound selected from the group consisting essentially of:
(1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline. (3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl3-quinoline.
(5) 4-[3-(6-[n-butylamino)pyridin-2-yl]-lH-pyrazol-4-yl]-quinoline.
(6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(11) 6-trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline. (12) 4-[3-(3-chlorophenyl)-lH-pyrazol-4-yl]-quinoline.
(13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
(14) 6-sec-butyl-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
(15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)-lH-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-l H-pyrazole. (17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(18) 4-[3-(6-propylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole.
(20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) 1 -benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. (22) l-(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan-l-ol.
(24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate. (25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]morpholine.
(26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine.
(27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3-Pyridin-2-yl-lH-pyrazol-4-yl)-quinoline and the therapeutically acceptable salts, esters and prodrugs thereof.
4. Use in the manufacture of a medicament for inhibiting TGF-β for the treatment of cancer in a patient in need thereof of a compound selected from the group consisting essentially of: (1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(5) 4-[3-(6- [n-butylamino)pyridin-2-yl] - 1 H-pyrazol-4-yl] -quinoline . (6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (11) 6-trifluoro methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline.
(12) 4- [3 -(3 -chlorophenyl)- 1 H-pyrazol-4-yl] -quinoline.
(13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
( 14) 6-sec-butyl-4-(3 -pyridin-2-yl- 1 H-pyrazol-4-yl)-quinoline. (15) 5 -mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)-l H-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(18) 4- [3 -(6-propylpyridin-2-yl)-lH-pyrazol-4-yl] -quinoline.
(19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole. (20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) 1 -benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan- 1 -ol.
(24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl ethylsulfonate.
(25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]moφholine.
(26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine. (27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3-Pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
5. Use in the manufacture of a medicament for inhibiting TGF-β in a patient in need thereof of a compound selected from the group consisting essentially of: (1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(5) 4-[3-(6- [n-butylamino)pyridin-2-yl] - 1 H-pyrazol-4-yl] -quinoline . (6) 4- [3 -(6-methylpyridin-2-yl)-lH-pyrazol-4-yl] -quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (11) 6-trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline.
(12) 4-[3-(3-chlorophenyl)-lH-pyrazol-4-yl]-quinoline.
(13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
(14) 6-sec-butyl-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline. (15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)-lH-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazo]e.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(18) 4-[3-(6-propylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
( 19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole. (20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) 1 -benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan- 1 -ol.
(24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate.
(25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl3morpholine.
(26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine. (27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3-Pyridin-2-yl-lH-pyrazol-4-yl)-quinoline.
6. Use in the manufacture of a medicament for treating cancer in a patient in need thereof of a compound selected from the group consisting essentially of: (1) 4-(3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(2) 4-(3-pyridin-2yl-lH-pyrazol-4-yl)-7-ethoxy quinoline.
(3) 7-fluoro-4-[3-(6-methyl pyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(4) 4-[3-(6-bromopyridin-2-yl)- 1 H-pyrazol-4-yl]-quinoline.
(5) 4-[3-(6-[n-butylamino)pyridin-2-yl]- 1 H-pyrazol-4-yl]-quinoline. (6) 4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(7) 6-chloro-4-[3-(6-mehylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(8) 6-trifluoromethyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(9) 7-methyl-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(10) 6-methoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline. (11) 6-trifluoromethoxy-4-[3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]- quinoline.
(12) 4-[3-(3-chlorophenyl)-lH-pyrazol-4-yl]-quinoline.
(13) 6-butoxy-4-(3-pyridin-2yl-lH-pyrazol-4-yl)-quinoline.
(14) 6-sec-butyl-4-(3-pyridin-2-yl-lH-pyrazol-4-yl)-quinoline. (15) 5-mehtyl-3-(6-methylpyridin-2-yl)-4-(-4-fluorophenyl)-lH-pyrazole.
(16) 4-(4-methoxyphenyl)-5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazole.
(17) 4-[5-methyl-3-(6-methylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(18) 4-[3-(6-propylpyridin-2-yl)-lH-pyrazol-4-yl]-quinoline.
(19) 3-Cyclopropyl-5-pyridin-2-yl-4-quinolin-4-yl-pyrazole. (20) 3-(3-Trifluoromethylphenyl)-4-quinolin-4-yl-pyrazole.
(21) l-benzyl-3-(2-pyridyl)-4-(4-quinolyl)pyrazole.
(22) 1 -(4-phenylbutyl)-3-(2-pyridyl)-4-(4-quinolyl)pyrazole. (23) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazolyl)ethan- 1 -ol.
(24) 2-(3-(2-pyridyl)-4-(4-quinolyl)pyrazoly)ethyl methylsulfonate.
(25) 4-[2-(3-(2-pyridyl)-3-(4-quinolyl)-pyrazolyl)ethyl]morpholine.
(26) Phenyl[2-(3-(2-pyridyl)-4-(4-quinolyl)-pyrazolyl)ethyl]amine.
(27) 4-(4-Pyridin-2-yl-lH-pyrazol-3-yl)-quinoline.
(28) 4-(3-Pyridin-2-yl-lH-pyrazol-4-yl)-quinoline and the therapeutically acceptable salts, esters and prodrugs thereof.
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