WO2005000813A1 - Heteroarylamino-phenylketone derivatives and their use as kinase inhibitors - Google Patents

Abstract

The present invention relates to compounds that inhibit VEGF receptor tyrosine kinases, especially KDR, pharmaceutical compositions that contain such compounds, methods of treating VEGF receptor kinase-dependent diseases and conditions in mammals using such compounds and composition and methods for their manufacture.

Description

HETEROARYLAMINOPHENYLKETONE DERIVATIVES AND THEIR USE AS KINASE INHIBITORS

RELATED APPLICATION

₅ The present application claims benefit of U.S. Provisional Application Serial

No. 60/474,206, filed May 30, 2003, which is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION 0 The present invention relates to heteroarylaminophenylketone derivatives that inhibit kinases, pharmaceutical compositions that contain the compounds, methods for making the compounds, and methods of treating kinase-dependent diseases and conditions in mammals by administering a therapeutically effective amount of the compounds to said mammals. s BACKGROUND OF THE INVENTION

Many biological mechanisms involving regulation of cellular processes are dependent on the phosphorylation of proteins by protein tyrosine kinases. Overexpression or aberrations in the pathways coupled to these kinases can result in a number of pathological outcomes including pathogenic angiogenesis or tumor growth.o By developing appropriate inhibitors, modulators or regulators of the kinase activity of this receptor, the signaling pathway can be modulated. Such compounds are considered therapeutically relevant agents that can be used alone or in combination to treat or prevent kinase dependent diseases or conditions.

Angiogenesis is a highly complex process of developing new blood vessels₅ that involves the proliferation and migration of, and tissue infiltration by capillary endothelial cells from pre-existing blood vessels, cell assembly into tubular structures, joining of newly forming tubular assemblies to closed-circuit vascular systems, and maturation of newly formed capillary vessels.

Angiogenesis is important in normal physiological processes includingo embryonic development, follicular growth, and wound healing, as well as in pathological conditions such as tumor growth and in non-neoplastic diseases involving abnormal neovascularization, including neovascular glaucoma (Folkman, J. and Klagsbrun, M. Science 235:442-447 (1987). Other disease states include but are not limited to, neoplastic diseases, including but not limited to solid tumors, 5 atherosclerosis and other inflammatory diseases such as rheumatoid arthritis, and ophthalmological conditions such as diabetic retinopathy and age-related macular

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Express Mail No. EN 331382753 US degeneration. Conditions or diseases to which persistent or uncontrolled angiogenesis contribute have been termed angiogenic dependent or angiogenic associated diseases.

One means for controlling such diseases and pathological conditions comprises restricting the blood supply to those cells involved in mediating or causing the disease or condition, for example, by occluding blood vessels supplying portions of organs in which tumors are present. Such approaches require the site of the tumor to be identified and are generally limited to treatment to a single site, or a small number of sits. An additional disadvantage of direct mechanical restriction of a blood supply is that collateral blood vessels develop, often quite rapidly, restoring the blood supply to the tumor.

Other approaches have focused on the modulation of factors that are involved in the regulation of angiogenesis. While usually quiescent, vascular endothelial proliferation is highly regulated, even during angiogenesis. VEGF is a factor that has been implicated as a regulator of angiogenesis in vivo (Klagsbrun, M. and D'Amore, P. (1991) Annual Rev. Physiol. 53: 217-239).

An endothelial-cell specific mitogen, VEGF acts as an angiogenesis inducer by specifically promoting the proliferation of endothelial cells. It is a homodimeric glycoprotein consisting of two 23 kD subunits. Four different monomeric isoforms of NEGF resulting from alternative splicing of mRΝA have been identified. These include two membrane bound forms (VEGF206 and VEGF₁₈₉) and two soluble forms (VEGF_]65 and VEGFι₂₁). VEGF₁₆₅ is the most abundant isoform in all human tissues except placenta.

NEGF is expressed in embryonic tissues (Breier et al., Development (Camb.) 114:521 (1992)), macrophages, and proliferating epidermal keratinocytes during wound healing (Brown et al., J. Exp. Med., 176:1375 (1992)), and may be responsible for tissue edema associated with inflammation (Ferrara et al., Endocr. Rev. 13:18 (1992)). In situ hybridization studies have demonstrated high levels of VEGF expression in a number of human tumor lines including glioblastoma multiforme, hemangioblastoma, other central nervous system neoplasms and AiDS-associated Kaposi's sarcoma (Plate, K. et al. (1992) Nature 359: 845-848; Plate, K. et al. (1993) Cancer Res. 53: 5822-5827; Berkman, R. et al. (1993) J. Clin. Invest. 91 : 153-159; Nakamura, S. et al. (1992) AIDS Weekly, 13 (1)). High levels of VEGF expression has also been found in atherosclerotic lesions, plaques and in inflammatory cells.

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Express Mail No. EV 331382753 US VEGF mediates its biological effect through high affinity VEGF receptors which are selectively expressed on endothelial cells during, for example, embryogenesis (Millauer, B., et al. (1993) Cell 72: 835-846) and tumor formation, and which have been implicated in modulating angiogenesis and tumor growth. These receptors comprise a tyrosine kinase cytosolic domain that initiates the signaling pathway involved in cell growth.

VEGF receptors typically are class III receptor-type tyrosine kinases characterized by having several, typically 5 or 7, immunoglobulin-like loops in their amino-terminal extracellular receptor ligand-binding domains (Kaipainen et al., J. Exp. Med. 178:2077-2088 (1993)). The other two regions include a transmembrane region and a carboxy-terminal intracellular catalytic domain interrupted by an insertion of hydrophilic interkinase sequences of variable lengths, called the kinase insert domain (Terman et al., Oncogene 6:1677-1683 (1991)). VEGF receptors include VEGFR-1 (or flt-1), sequenced by Shibuya M. et al., Oncogene 5, 519-524 (1990); and VEGFR-2 (or flk-1/KDR). KDR, described in PCT/US92/01300, filed February 20, 1992, and in Terman et al., Oncogene 6:1677-1683 (1991), is the human homologue of flk-lk, sequenced by Matthews W. et al. Proc. Natl. Acad. Sci. USA, 88:9026-9030 (1991).

Release of VEGF by a tumor mass stimulates angiogenesis in adjacent endothelial cells. When VEGF is expressed by the tumor mass, endothelial cells adjacent to the VEGF+ tumor cells will up-regulate expression of VEGF receptor molecules, e.g., VEGFR-1 and VEGFR-2. It is generally believed that KDR/VEGFR- 2 is the main VEGF signal transducer that results in endothelial cell proliferation, migration, differentiation, tube formation, increase of vascular permeability, and maintenance of vascular integrity. VEGFR-1 possesses a much weaker kinase activity, and is unable to generate a mitogenic response when stimulated by VEGF, although it binds to VEGF with an affinity that is approximately 10- fold higher than KDR. VEGFR-1 has also been implicated in VEGF and placenta growth factor (P1GF) induced migration of monocytes and macrophages and production of tissue factor.

High levels of VEGFR-2, for example, are expressed by endothelial cells that infiltrate gliomas (Plate, K. et al., (1992) Nature 359: 845-848), and are specifically upregulated by VEGF produced by human glioblastomas (Plate, K. et al. (1993)

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Express Mail No. EV 331382753 US Cancer Res. 53: 5822-5827). The finding of high levels of VEGR-2 expression in glioblastoma associated endothelial cells (GAEC) suggests that receptor activity is induced during tumor formation, since VEGFR-2 transcripts are barely detectable in normal brain endothelial cells, indicating generation of a paracrine VEGF/VEGFR loop. This upregulation is confined to the vascular endothelial cells in close proximity to the tumor. Blocking VEGF activity with neutralizing anti-VEGF monoclonal antibodies (mAbs) results in inhibition of the growth of human tumor xenografts in nude mice (Kim, K. et al. (1993) Nature 362: 841-844), suggesting a direct role for VEGF in tumor-related angiogenesis. Accordingly, VEGFR antagonists have been developed to treat vascularized tumors and other angiogenic diseases. These have included neutralizing antibodies that block signaling by VEGF receptors expressed on vascular endothelial cells to reduce tumor growth by blocking angiogenesis through an endothelial-dependent paracrine loop. See, e.g., U.S. Patent No. 6,365,157 (Rockwell et al.), WO 00/44777 (Zhu et al.), WO 01/54723 (Kerbel); WO 01/74296 (Witte et al.), WO 01/90192 (Zhu), Bispecific Antibodies That Bind to VEGF Receptors (Zhu, International PCT application filed June 26, 2002), and Method of Treating Atherosclerosis and Other Inflammatory Diseases (Carmeliet et al.; international PCT application filed Jun. 20, 2002). VEGF receptors have also been found on some non-endothelial cells, such as tumor cells producing VEGF, wherein an endothelial-independent autocrine loop is generated to support tumor growth. For example, it has been demonstrated that a VEGF/human VEGFR-2 autocrine loop mediates leukemic cell survival and migration in vivo. Dias et al., "Autocrine stimulation of VEGFR-2 activates human leukemic cell growth and migration," J. Clin. Invest. 106:511 -521 (2000); Witte et al., "Treatment of non-solid mammalian tumors with vascular endothelial growth factor receptor antagonsits;" and WO 01/74296 (Witte et al.). Similarly, VEGF production and VEGFR expression also have been reported for some solid tumor cell lines in vitro. (See Tohoku, Sato, J. Exp. Med., 185(3): 173-84 (1998): Nippon, Sanka Fujinka Gakkai Zasshi,:47(2): 133-40 (1995); and Ferrer, FA, Urology, 54(3):567-72 (1999)). It has further been demonstrated that VEGFR-1 Mabs inhibit an autocrine VEGFR/human VEGFR-1 loop in breast carcinoma cells. Wu, et al., "Monoclonal antibody against VEGFRl inhibits fltl-positive DU4475 human breast tumor growth

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Express Mail No. EV 331382753 US by a dual mechanism involving anti-angiogenic and tumor cell growth inhibitory activities," AACR_NCI_EORTC International Conference on Molecular Targets and Cancer Therapeutics, October 29-November 2, 2001, Abstract #7; and Carmeliet et al. (international PCT application filed June 20, 2002).

There remains a need for compounds which inhibit VEGF receptor tyrosine kinase activity to treat or prevent VEGF-receptor kinase dependent diseases or conditions, by inhibiting, for example, pathogenic angiogenesis or tumor growth through inhibition of the paracrine and/or autocrine VEGF/ VEGFR loop.

SUMMARY OF THE INVENTION

In one aspect, the invention relates to heteroarylaminophenylketone derivatives, which are kinase inhibitors that have the following formula I:

or pharmaceutically acceptable salts, stereoisomers, hydrates or pro-drugs thereof, wherein Ri is selected from hydrogen, lower alkyl, lower alkenyl, lower alkynyl and aralkyl, wherein the lower alkyl, lower alkenyl, lower alkynyl and aralkyl may be unsubstituted or substituted with one or more substituents selected from R₂;

R₃ is selected from:

1) lower alkyl, optionally substituted with one or more substituents selected from R2,

2) lower alkenyl, optionally substituted with one or more substituents selected from R2,

3) aralkyl, optionally substituted with one or more substituents selected

4) lower alkynyl, optionally substituted with one or more substituents selected from R₂,

5) phenyl, optionally substituted with 1 to 5 substituents selected from R₂,

6) halogen,

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Express Mail No. EV 331382753 US 7) nitro,

8) cyano,

9) a group of the formula -CO₂R, -OR, -SR, -SO₂R, -NH₂, -NHR, -NRR, where R is independently selected from H, lower alkyl, aralkyl, aryl, and heteroaryl,

₅ 10) pyridine, optionally substituted with 1 to 4 substituents selected from

R₂,

11) pyrazine, optionally substituted with 1 to 3 substituents selected from

R₂,

12) pyrimidine, optionally substituted with 1 to 3 substituents selectedo from R2, and

13) a group selected from the following:

or tetrazole 5 wherein X is O, S, or N-R, a is 0 to 3, b is 0 to 2; c is 0 or 1 ; ₀ R-i is selected from:

1) lower alkyl, optionally substituted with one or more substituents selected from R₂,

2) lower alkenyl, optionally substituted with one or more substituents selected from R₂, 5 3) aralkyl, optionally substituted with one or more substituents selected from R₂,

4) lower alkynyl, optionally substituted with one or more substituents selected from R₂,

5) phenyl, optionally substituted with 1 to 5 substituents selected from R ,o 6) pyridine, optionally substituted with 1 to 4 substituents selected from

R2,

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Express Mail No. EV 331382753 US 7) pyrazine, optionally substituted with 1 to 3 substituents selected from

R₂,

8) pyrimidine, optionally substituted with 1 to 3 substituents selected from R₂,

₅ 9) a group selected from the following:

or tetrazole wherein X is O, S, or N-R, o a is 0 to 3,

6 is 0 to 2; c is 0 or 1 ; and

10) hydrogen. 5 each R₂ is independently selected from:

1) halogen,

2) lower alkyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy, 0 3) lower alkenyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy,

4) nitro,

5) cyano,

6) a group of the formula -CO₂R, -OR, -SR, -SO₂R, -NH₂, -NHR, -NRR,5 or R can occupy two adjacent positions to form a fused 5- or 6-membered carbocyclic or heterocychc ring, which may be substituted or unsubstituted and wherein the heterocychc ring may contain from 1 to 2 heteroatoms selected from N, O or S, o each R is independently selected from hydrogen, halogen, lower alkyl, lower alkoxy, aralkyl, aryl, and heteroaryl, and

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Express Mail No. EV 331382753 US each Y is independently selected from C-R and N.

The invention further relates to pharmaceutical compositions containing a therapeutically effective amount of the compounds of formula I.

The invention is also directed to methods of inhibiting VEGF receptor tyrosine kinases, especially KDR, and methods of treating VEGF receptor tyrosine kinase- dependent diseases and conditions in mammals using the VEGF receptor kinase inhibitors of Formula I. The diseases and conditions that may be treated or prevented by the present methods include, for example, conditions or diseases in which pathogenic angiogenesis is implicated, including neoplastic diseases such as solid or liquid tumors, atherosclerosis, age related macular degeneration, retinal vascularization, inflammatory diseases, or cell proliferative disorders, in mammals. The method includes administering to a mammal in need of such treatment a therapeutically effective amount of one or more compounds of the present invention.

The invention further relates to a method of making the compounds of formula I.

The present invention will now be described in detail for specific preferred embodiments of the invention. These embodiments are intended only as illustrative examples and the invention is not intended to be limited thereto.

DETAILED DESCRIPTION OF THE INVENTION The invention provides compounds of formula I as described above.

The term "lower alkyl" as used herein refers to a saturated hydrocarbon derived radical containing from 1 to 6 carbon atoms. The lower alkyl group may be straight, branched or cyclic. Straight or branched lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl and t-butyl, and the like. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "lower alkenyl" as used herein refers to a non-aromatic hydrocarbon radical, straight, branched or cyclic, containing from 2 to 6 carbon atoms and at least one carbon to carbon double bond. Lower alkenyl groups include ethenyl, propenyl, butenyl cyclohexenyl, and the like. The term "lower alkynyl" as used herein refers to a hydrocarbon radical that is straight, or branched, containing from 2 to 6 carbon atoms and at least one carbon to carbon triple bond. Lower alkynyl groups include ethynyl, propynyl and butynyl, and the like.

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Express Mail No. EV 331382753 US The term "aralkyl" as used herein contemplates a lower alkyl group which has as a substituent an aryl group.

The term "aryl" as used herein refers to unsubstituted or substituted 5- or 6- membered aromatic rings, such as, phenyl, substituted phenyl and the like, as well bicyclic rings such as naphthyl and heterocychc rings, such as pyridine, imidazole, oxazole, thiazole and the like. Thus, aryl denotes a group containing at least one aromatic ring having at least 5 atoms, with up to two such rings being present, containing up to 10 atoms therein. Preferred aryl groups are phenyl and naphthyl. The terms heterocycle, heteroaryl or heterocychc, as used contemplates a stable 5- to 6-membered mono- or 7- to 10-membered bicyclic heterocychc ring system, any ring of which may be saturated or unsaturated, aromatic or non-aromatic, and which includes carbon atoms and from one to three heteroatoms selected from the group consisting of N, O and S. The nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quatemized. "Heterocycle" includes the abovementioned heteroaryls, as well as dihydro and tetrahydro analogs thereof. Heterocycles include any bicyclic group in which any of the above-defined rings is fused to a benzene ring. The heterocychc ring may be attached at any heteroatom or carbon atom, which results in the creation of a stable structure. Examples of such heterocychc components include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofiiryl, tetrahydropyranyl, thiophenyl, imidazopyridinyl, tetrazolyl, triazinyl, thienyl, benzothienyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl.

The term "alkoxy" as used herein refers to a substituent with an alkyl group in either a straight-chained or branched configuration, and may include a double or a triple bond, which is attached via an oxygen atom. The alkyl portion may be substituted or unsubstituted. Substituents on the alkyl group may include for example, a phenyl ring, in which the alkoxy may be for example, a benzyloxy group.

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Express Mail No. EV 331382753 US Examples of alkoxy groups are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tertiary butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy allyloxy, propargyloxy, vinyloxy, and the like.

The term "halo" or "halogen" as used herein is intended to include the halogen atoms fluorine, chlorine, bromine and iodine.

All value ranges provided herein, for example those given for a and b, are inclusive over the entire range. Thus, a range between 0-4 would include the values 0, 1, 2, 3 and 4.

When one or more chiral centers are present in the compounds of the present invention, the individual isomers and mixtures thereof (e.g., racemates, etc.) are intended to be encompassed by the formulae depicted herein.

As used herein the terms "pharmaceutically acceptable salts" and "hydrates" refer to those salts and hydrated forms of the compound that would be apparent to those in the art, i.e., those which favorably affect the physical or pharmacokinetic properties of the compound, such as solubility, palatability, absorption, distribution, metabolism and excretion. Other factors, more practical in nature, which those skilled in the art may take into account in the selection include the cost of the raw materials, ease of crystallization, yield, stability, solubility, hygroscopicity and flowability of the resulting bulk drug. When a compound of the present invention is present as a salt or hydrate that is non-pharmaceutically acceptable, that compound can be converted in certain circumstances to a salt or hydrate form that is pharmaceutically acceptable in accordance with the present invention.

When the compound is negatively charged, it is balanced by a counterion, such as, an alkali metal cation such as sodium or potassium. Other suitable counterions include calcium, magnesium, zinc, ammonium, or alkylammonium cations, such as tetramethylammonium, tetrabutylammonium, choline, triethylhydroammonium, meglumine, triethanol-hydroammoniurn, and the like. An appropriate number of counterions are associated with the molecule to maintain overall charge neutrality. Likewise, when the compound is positively charged, e.g., protonated, an appropriate number of negatively charged counterions are present to maintain overall charge neutrality. These pharmaceutically acceptable salts are within the scope of the present invention.

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Express Mail No. EV 331382753 US Pharmaceutically acceptable salts may be prepared by the addition of an appropriate acid. Thus, the compound can be used in the form of salts derived from inorganic or organic acids. Examples include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, pamoate, pectinate, persulfate, 3-phenylpropionate, pivalate, propionate, succinate, tartrate and undecanoate.

If the compound has an acidic proton, a salt may be form by the addition of base to form a pharmaceutically acceptable base addition salt. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and so forth.

The basic nitrogen-containing groups may be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl; and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides and others.

The presence of pharmaceutically acceptable salts within the scope of the present compounds is not intended to limit the compounds of the present invention to those that are synthetically prepared. The compounds of the present invention also include compounds that are converted within the body and prodrugs. "Pro-drug" means a form of the compounds of the present invention suitable for administration to a patient without undue toxicity, irritation, allergic response, and the like, and effective for their intended use. A pro-drug is transformed in vivo to yield the parent compound of the formula I herein, for example by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems Vol. 14 of the A. C. S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987.

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Express Mail No. EV 331382753 US The compounds of the present invention may have asymmetric centers and occur as racemates, mixtures of enantiomers and as individual enantiomers with all isomeric forms being included in the present invention.

Preferably, the compounds of the invention have the following formula (II):

or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, wherein: Y' is selected from CH and N;

Y is selected from C-R and N;

X is selected from O, S, and N-R;

Ri is selected from hydrogen, lower alkyl, lower alkenyl, lower alkynyl and aralkyl, wherein the lower alkyl, lower alkenyl, lower alkynyl and aralkyl may be unsubstituted or substituted with one or more substituents selected from R ;

R₃ is selected from:

1) hydrogen

2) lower alkoxy, optionally substituted with one or more substituents selected from R₂, 3) lower alkyl, optionally substituted with one or more substituents selected from R ,

4) lower alkenyl, optionally substituted with one or more substituents selected from R₂,

5) aralkyl, optionally substituted with one or more substituents selected from R₂,

6) lower alkynyl, optionally substituted with one or more substituents selected from R₂,

7) phenyl, optionally substituted with 1 to 5 substituents selected from R₂,

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Express Mail No. EV 331382753 US 8) halogen,

9) nitro,

10) cyano,

11) a group of the formula -CO₂R, -SR, -SO₂R, -NH₂, -NHR, -NRR, where R is independently selected from H, lower alkyl, aralkyl, aryl, and heteroaryl,

12) pyridine, optionally substituted with 1 to 4 substituents selected from

R₂,

13) pyrazine, optionally substituted with 1 to 3 substituents selected from

R₂,

14) pyrimidine, optionally substituted with 1 to 3 substituents selected from R₂, and

15) a group selected from the following:

or tetrazole wherein X is O, S, or N-R, a is 0 to 3, b is 0 to 2; c is 0 or 1 ;

R₂ is independently selected from:

1) halogen,

2) lower alkyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy, 3) lower alkenyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy,

4) nitro,

5) cyano,

6) a group of the formula -CO₂R, -OR, -SR, -SO₂R, -NH₂, -NHR, -NRR,

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Express Mail No. EV 331382753 US or R₂ can occupy two adjacent positions to form a fused 5- or 6-membered carbocyclic or heterocychc ring, which may be substituted or unsubstituted with R₃ and wherein the heterocychc ring may contain from 1 to 2 heteroatoms independently selected from N, O or S; and

R is independently selected from hydrogen, halogen, lower alkyl, lower alkoxy, aralkyl, aryl and heteroaryl.

The invention further relates to a method for making the compounds of formula II.

Method A. The method entails reacting a compound of formula V with a benzyne intermediate as shown below, wherein R₃, and Y are as defined above:

VI benzyne intermediate

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Express Mail No. EV 331382753 US

VII VIII

DC

Method B. The method entails reacting an N-protected arylamine of formula X with a strong base (RLi) such as n-BuLi, LDA, etc., followed by a heterocychc acid chloride of formula XI or XII, or other acylating agents, such as carbonyl derivatives, wherein R₂, R₃, X and Y are as defined above:

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Express Mail No. EV 331382753 US

XI

xπ

Method C. The method entails reacting a orthonitrocarbonyl compound of formula IN with a bicycle of formula XIII as shown below, wherein R , R₄, X and Y are as defined above (Org. Lett. 2000, 2/10, 1485; Synth. Commun. 1997, 27, 2125):

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Express Mail No. EV 331382753 US

xrv

XV

Tables 1 and 2 provide representative embodiments of compounds of Formula

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Express Mail No. EV 331382753 US Table 1

Comp. No. R Ri R₃

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Express Mail No. EV 331382753 US

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Express Mail No. EV 331382753 US

22 H C-CH₃ H

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Express Mail No. EV 331382753 US

31 CH₃ N H

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Express Mail No. EV 331382753 US

36 H N H

..A

37 H n N H

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Express Mail No. EV 331382753 US 42 H 1 I 1 C-H 5-CH₃

"or'

Table 2

Comp. No. R Ri R₃

43 H H C-H 5-CH₃

44 H H C-H 6-CH₃

45 H H C-H 6-F

46 H H C-H 5-F

47 H H C-CH₃ H

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Express Mail No. EV 331382753 US Tyrosine kinase inhibition can be determined using well-known methods. The compounds of the present invention generally involve inhibition or regulation of phosphorylation events. Accordingly, phosphorylation assays are useful in determining antagonists useful in the context of the present invention. Tyrosine kinase inhibition may be determined by measuring the autophosphorylation level of recombinant kinase receptor, and/or phosphorylation of natural or synthetic substrates. Phosphorylation can be detected, for example, using an antibody specific for phosphotyrosine in an ELISA assay or on a Western blot. Some assays for tyrosine kinase activity are described in Panek et al., J. Pharmacol. Exp. Thera., 283: 1433.44 (1997) and Batley et al., Life Sci., 62: 143-50 (1998).

In addition, methods for detection of protein expression can be utilized, wherein the proteins being measured are regulated by tyrosine kinase activity. These methods include immunohistochemistry (IHC) for detection of protein expression, fluorescence in situ hybridization (FISH) for detection of gene amplification, competitive radioligand binding assays, solid matrix blotting techniques, such as Northern and Southern blots, reverse transcriptase polymerase chain reaction (RT- PCR) and ELISA. See, e.g., Grandis et al., Cancer, 78:1284-1292. (1996); Shimizu et al., Japan J. Cancer Res., 85:567-571 (1994); Sauter et al., Am. J. Path., 148:1047- 1053 (1996); Collins, Glia, 15:289-296 (1995); Radinsky et al., Clin. Cancer Res., 1:19-31 (1995); Petrides et al., Cancer Res., 50:3934-3939 (1990); Hoffmann et al., Anticancer Res., 17:4419-4426 (1997); Wikstrand et al., Cancer Res., 55:3140-3148 (1995).

In vivo assays can also be utilized. For example, VEGF receptor tyrosine kinase inhibition can be observed by mitogenic assays using HUVEC cells (Cornell University Medical College ) stimulated with VEGF in the presence and absence of inhibitor. Another method involves testing for inhibition of growth of VEGF- expressing tumor cells, using for example, human tumor cells injected into a mouse. (See, U.S. Patent No. 6,365,157 to Rockwell et al.) Also included within the scope of the present invention are methods of inhibiting VEGF receptor tyrosine kinases, especially KDR, and/or treating or preventing VEGF receptor kinase-dependent diseases and conditions in mammals using the VEGF receptor kinase inhibitors of Formula I. The VEGF receptor is

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Express Mail No. EV 331382753 US usually bound to a cell, such as an endothelial or tumor cell. Alternatively, the VEGF receptor may be free from the cell, preferably in soluble form.

The diseases and conditions which may be treated or prevented by the present methods include, for example, those in which pathogenic angiogenesis or tumor growth is stimulated through a VEGF/NEGFR paracrine and/or autocrine loop. For example, paracrine VEGFR stimulation of vascular endothelium is associated with angiogenic diseases and vascularization of tumors. VEGF receptors are also found on non-endothelial cells, such as tumor cells, indicating the presence of an autocrine and/or paracrine loop in these cells. Thus, the method is also useful for neutralizing VEGF receptors on such cells, thereby inhibiting autocrine and/or paracrine stimulation and inhibiting tumor growth.

The method is effective for treating subjects with tumors and neoplasms, particularly where disease development involves angiogenesis. Tumors and neoplasms include, for example, malignant tumors and neoplasms, such as blastemas, carcinomas or sarcomas, and highly vascular-dependent tumors and neoplasms. Cancers that may be treated by the methods of the present invention include, for example, cancers of the brain, genitourinary tract, lymphatic system, stomach, renal, colon, larynx and lung and bone. Νon-limiting examples further include epidermoid tumors, squamous tumors, such as head and neck tumors, colorectal tumors, prostate tumors, breast tumors, lung tumors, including lung adenocarcinoma and small cell and non-small cell lung tumors, pancreatic tumors, thyroid tumors, ovarian tumors, and liver tumors. The method is also used for treatment of vascularized skin cancers, including squamous cell carcinoma, basal cell carcinoma, and skin cancers that can be treated by suppressing the growth of malignant keratinocytes, such as human malignant keratinocytes. Other cancers that can be treated include Kaposi's sarcoma, CΝS neoplasms (neuroblastomas, capillary hemangioblastomas, meningiomas and cerebral metastases), melanoma, gastrointestinal and renal carcinomas and sarcomas, rhabdomyosarcoma, glioblastoma, including glioblastoma multiforme, histiocytic lymphoma, and leiomyosarcoma. A further aspect of the present invention includes methods of treating or preventing angiogenic diseases, inflammatory diseases or other diseases characterized by paracrine stimulation through VEGF receptors, by administering to a mammal in need of such treatment a therapeutically effective amount of one or more of the

SJO 51087-1 25

Express Mail No. EV 331382753 US compounds set forth herein. Examples of non-neoplastic angiogenic diseases include diseases characterized by retinal vascularization, such as neovascular glaucoma, proliferative retinopathy, including diabetic retinopathy, retinopathy of prematurity (retrolental fibroplastic), macular degeneration, and corneal graft rejection. Examples of inflammatory diseases that may be treated include, but are not limited to, atherosclerosis, rheumatoid arthritis (RA), insulin-dependent diabetes mellitus, multiple sclerosis, myasthenia gravis, Chron's disease, autoimmune nephritis, primary biliary cirrhosis, psoriasis, acute pancreatitis, allograph rejection, allergic inflammation, contact dermatitis and delayed hypersensitivity reactions, inflammatory bowel disease, septic shock, osteoporosis, osteoarthritis, and cognition defects induced by neuronal inflammation. Other non-limiting examples of angiogenic diseases are hemangiomas, angiofibromas, Osier- Weber syndrome, restinosis, and fungal, parasitic and viral infections, including cytomegaloviral infections.

The present invention also includes methods for treating tumors that express VEGF receptors, especially KDR, for example through inhibition of an autocrine VEGF/NEGFR loop, wherein one or more of the Formula I compounds are adminstered in an amount effective to reduce tumor growth or size. In an aspect of the invention, the method is effective for treating a solid or liquid tumor that is not vascularized, or is not yet substantially vascularized. Examples of solid tumors which may be accordingly treated include breast carcinoma, lung carcinoma, colorectal carcinoma, pancreatic carcinoma, glioma and lymphoma. Examples of liquid tumors include leukemia, multiple myeloma and lymphoma. Some examples of leukemias include acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), erythrocytic leukemia or monocytic leukemia. Some examples of lymphomas include Hodgkin's and non-Hodgkin's lymphoma.

Moreover, the compounds of formula I may be used for in vivo and in vitro investigative, diagnostic, or prophylactic methods, which are well known in the art. In the methods of the present invention, a therapeutically effective amount of one or more of the Formula I compounds is administered to a mammal in need. The term "administering" as used herein means delivering the compounds of the present invention to a mammal by any method that may achieve the result sought. They may be administered, for example, orally, parenterally (intravenously or intramuscularly),

SJO 51087-1 26

Express Mail No. EV 331382753 US topically, transdermally or by inhalation. The term "mammal" as used herein is intended to include, but is not limited to, humans, laboratory animals, domestic pets and farm animals. "Therapeutically effective amount" means an amount of compound of the present invention that when administered to a mammal is effective in producing the desired therapeutic effect, such as inhibiting kinase activity. Another aspect of the present invention relates to pharmaceutical compositions, which include at least one compound of the present invention as described herein (that is, a compound of Formula I) or a pharmaceutically acceptable salt, hydrate or pro-drug thereof, in combination with a pharmaceutically acceptable carrier.

The compounds of the present invention may be employed in solid or liquid form including for example, powder or crystalline form, in solution or in suspension. They may be administered in numerous different ways, such as orally, parenterally (intravenously or intramuscularly), topically, transdermally or by inhalation. The choice of carrier and the content of active compound in the carrier are generally determined in accordance with the solubility and chemical properties of the desired product, the particular mode of administration and the provisions to be observed in pharmaceutical practice. Thus, the carrier employed may be, for example, either a solid or liquid. One method of administering a solid dosage form is to form solid compositions for rectal administration, which include suppositories formulated in accordance with known methods and containing at least one compound of the present invention. Examples of solid carriers include lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Examples of liquid carriers include syrup, peanut oil, olive oil, water and the like. For parenteral administration, emulsions, suspensions or solutions of the compounds according to the invention in vegetable oil, for example sesame oil, groundnut oil or olive oil, or aqueous-organic solutions such as water and propylene glycol, injectable organic esters such as ethyl oleate, as well as sterile aqueous solutions of the pharmaceutically acceptable salts, are used. Injectable forms must be fluid to the extent they can be easily syringed, and proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

SJO 51087-1 27

Express Mail No. EV 331382753 US Prolonged absorption of the injectable compositions can be brought about by use of agents delaying absorption, for example, aluminum monostearate and gelatin. The solutions of the salts of the products according to the invention are especially useful for administration by intramuscular or subcutaneous injection. Solutions of the active compound as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxypropyl- cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. The aqueous solutions, also including solutions of the salts in pure distilled water, may be used for intravenous administration with the proviso that their pH is suitably adjusted, that they are judiciously buffered and rendered isotonic with a sufficient quantity of glucose or sodium chloride and that they are sterilized by heating, irradiation, microfiltration, and/or by various antibacterial and antifungal agents, for example, Xparabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. Examples of injectable dosage forms include sterile injectable liquids, e.g., solutions, emulsions and suspensions. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation may include vacuum drying and a freeze-dry technique that yields a powder of the active ingredient plus any additional desired ingredient from previously sterile-filtered solution thereof.

Examples of injectable solids include powders that are reconstituted, dissolved or suspended in a liquid prior to injection. In injectable compositions, the carrier typically includes sterile water, saline or another injectable liquid, e.g., peanut oil for intramuscular injections. Also, various buffering agents, preservatives and the like can be included within the compositions of the present invention.

For oral administration, the active compound may be administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or

SJO 51087-1 28

Express Mail No. EV 331382753 US it may be incorporated directly with the food of the diet, or may be incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Examples of oral solid dosage forms include tablets, capsules, troches, lozenges and the like. Examples of oral liquid

₅ dosage forms include solutions, suspensions, syrups, emulsions, soft gelatin capsules and the like. Carriers for oral use (solid or liquid) may include time delay materials known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax. To prepare a capsule, it may be advantageous to use lactose and liquid carrier, such as high molecular weight polyethylene glycols. 0 Topical administration, in the form of gels (water or alcohol based), creams or ointments, for example, containing compounds of the invention may be used. Topical applications may be formulated in carriers such as hydrophobic or hydrophilic bases to form ointments, creams, lotions, in aqueous, oleaginous or alcoholic liquids to form paints or in dry diluents to form powders. Such topical formulations can be used fors example, to treat ocular diseases as well as inflammatory diseases such as rheumatoid arthritis, psoriasis, contact dermatitis, delayed hypersensitivity reactions and the like. Compounds of the invention may be also incorporated in a gel or matrix base for application in a patch, which would allow a controlled release of compound through transdermal barrier. o For administration by inhalation, compounds of the invention may be dissolved or suspended in a suitable carrier for use in a nebulizer or a suspension or solution aerosol, or may be absorbed or adsorbed onto a suitable solid carrier for use in a dry powder inhaler.

Compositions according to the invention may also be formulated in a manner₅ that resists rapid clearance from the vascular (arterial or venous) wall by convection and/or diffusion, thereby increasing the residence time of the viral particles at the desired site of action. A periadventitial depot comprising a compound according to the invention may be used for sustained release. One such useful depot for administering a compound according to the invention may be a copolymer matrix,o such as ethylene-vinyl acetate, or a polyvinyl alcohol gel surrounded by a Silastic shell. Alternatively, a compound according to the invention may be delivered locally from a silicone polymer implanted in the adventitia.

SJO 51087-1 29

Express Mail No. EV 331382753 US An alternative approach for minimizing washout of a compound according to the invention during percutaneous, transvascular delivery comprises the use of nondiffusible, drug-eluting microparticles. The microparticles may be included a variety of synthetic polymers, such as polylactide for example, or natural substances, including proteins or polysaccharides. Such microparticles enable strategic manipulation of variables including total dose of drug and kinetics of its release. Microparticles can be injected efficiently into the arterial or venous wall through a porous balloon catheter or a balloon over stent, and are retained in the vascular wall and the periadventitial tissue for at least about two weeks. Formulations and methodologies for local, intravascular site-specific delivery of therapeutic agents are discussed in Reissen et al. (J. Am. Coll. Cardiol. 1994; 23: 1234-1244).

A composition according to the invention may also comprise a hydrogel which is prepared from any biocompatible or non-cytotoxic (homo or hetero) polymer, such as a hydrophilic polyacrylic acid polymer that can act as a drug absorbing sponge. Such polymers have been described, for example, in application WO93/08845.

Certain of them, such as, in particular, those obtained from ethylene and/or propylene oxide are commercially available.

Another embodiment of the invention provides for a compound according to the invention to be administered by means of perfusion balloons. These perfusion balloons, which make it possible to maintain a blood flow and thus to decrease the risks of ischaemia of the myocardium, on inflation of the balloon, also enable the compound to be delivered locally at normal pressure for a relatively long time, more than twenty minutes, which may be necessary for its optimal action.

Alternatively, a channeled balloon catheter (such as "channelled balloon angioplasty catheter", Mansfield Medical, Boston Scientific Corp., Watertown, Mass.) may be used. This catheter includes a conventional balloon covered with a layer of 24 perforated channels that are perfused via an independent lumen through an additional infusion orifice. Various types of balloon catheters, such as double balloon, porous balloon, microporous balloon, channel balloon, balloon over stent and hydrogel catheters, all of which may be used to practice the invention, are disclosed in Reissen et al. (1994).

Another aspect of the present invention relates to a pharmaceutical composition including a compound according to the invention and poloxamer, such as

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Express Mail No. EV 331382753 US Poloxamer 407, which is a non-toxic, biocompatible polyol, commercially available (e.g., from BASF, Parsippany, N.J.). A poloxamer impregnated with a compound according to the invention may be deposited for example, directly on the surface of the tissue to be treated, for example during a surgical intervention. Poloxamer possesses essentially the same advantages as hydrogel while having a lower viscosity. The use of a channel balloon catheter with a poloxamer impregnated with a compound according to the invention may be advantageous in that it may keep the balloon inflated for a longer period of time, while retaining the properties of facilitated sliding, and of site-specificity of the poloxamer. The composition may also be administered to a patient via a stent device. In this embodiment, the composition is a polymeric material in which the compound of the invention is incorporated, which composition is applied to at least one surface of the stent device.

Polymeric materials suitable for incorporating the compound of the invention include polymers having relatively low processing temperatures such as polycaprolactone, poly(efhylene-co-vinyl acetate) or poly(vinyl acetate or silicone gum rubber and polymers having similar relatively low processing temperatures. Other suitable polymers include non-degradable polymers capable of carrying and delivering therapeutic drugs such as latexes, urethanes, polysiloxanes, styrene- ethylene/butylene-styrene block copolymers (SEBS) and biodegradable, bioabsorbable polymers capable of carrying and delivering therapeutic drugs, such as poly-DL-lactic acid (DL-PLA), and poly-L-lactic acid (L-PLA), polyorthoesters, polyiminocarbonates, aliphatic polycarbonates, and polyphosphazenes.

In addition to the active compound and the pharmaceutically acceptable carrier, the compositions of the present invention optionally contain one or more excipients that are conventional in the art. For example, excipients such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and disintegrating agents such as starch, alginic acids and certain complex silica gels combined with lubricants such as magnesium stearate, sodium lauryl sulfate and talc may be used for preparing tablets, troches, pills, capsules and the like.

Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. When aqueous suspensions are used they may

SJO 51087-1 31

Express Mail No. EV 331382753 US contain emulsifying agents or agents which facilitate suspension. Diluents such as sucrose, ethanol, polyols such as polyethylene glycol, propylene glycol and glycerol, and chloroform or mixtures thereof may also be used. In addition, the active compound may be incorporated into sustained-release preparations and formulations. The percentage of active ingredient in the compositions of the invention may be varied. Several unit dosage forms may be administered at about the same time. A suitable dose employed may be determined by a physician or qualified medical professional, and depends upon various factors including the desired therapeutic effect, the nature of the illness being treated, the route of administration, the duration of the treatment, and the condition of the patient, such as age, weight, general state of health and other characteristics, which can influence the efficacy of the compound according to the invention. In adults, doses are generally from about 0.001 to about 50, preferably about 0.001 to about 5, mg/kg body weight per day by inhalation; from about 0.01 to about 100, preferably 0.1 to 70, more preferably 0.5 to 10, mg/kg body weight per day by oral administration; from about 0.1 to about 150 mg applied externally; and from about 0.001 to about 10, preferably 0.01 to 10, mg/kg body weight per day by intravenous or intramuscular administration.

The compounds and compositions according to the invention may be administered as frequently as necessary as determined by a skilled practitioner in order to obtain the desired therapeutic effect. Some patients may respond rapidly to a higher or lower dose and may find much weaker maintenance doses adequate. For other patients, it may be necessary to have long-term treatments at the rate of 1 to 4 doses per day, in accordance with the physiological requirements of each particular patient. Generally, the active product may be administered orally 1 to 4 times per day. For other patients, it may be necessary to prescribe not more than one or two doses per day.

The compounds of the present invention may also be formulated for use in conjunction with other therapeutically active compounds or in connection with the application of therapeutic techniques to address pharmacological conditions, which may be ameliorated through the application of a compound according to the present invention.

Detailed descriptions of conventional assays, such as those employed in phosphorylation assays, can be obtained from numerous publication, including

SJO 51087-1 32

Express Mail No. EV 331382753 US Sambrook, J. et al., (1989) Molecular Cloning: A Laboratory Manual, 2^nd ed., Cold Spring Harbor Laboratory Press. All references mentioned herein are incorporated in their entirety.

s EXAMPLES

EXAMPLE 1,

(lH-Indazol-3-vD- {2-r(pyridin-4-ylmethylVaminol-pheny -methanone (1) o 2-Diazo-l-(2-nitro-phenyl)-ethanone (49). A solution of 2-nitrobenzoyl chloride (20 g, 108 mmol) in anhydrous ether (100 ml) was added dropwise to 2.0 M solution of trimethylsilyl diazomenthane in hexane (54 ml, 108 mmol) and triethylamine (15 ml) at 0 C. The solution was allowed to warm up to room temperature and was stirred for one hour. The reaction mixture was then filtered and the filtrate was concentrated ins vacuo. The crude material was purified on silica gel column (eluting with a gradient of 75%- 100% dichloromethane/hexane) to give 7.0 g (34%) of 2-nitrodiazophenone as a pale yellow solid. TLC 25%hexane/dichloromethane, UV, Rf=0.28. 1H NMR (300 MHz, CDC1₃): δ 5.53 (s, 1H), 7.50 (d, 1H), 7.58 (m, 2H), 7.93 (t, 1H), 8.02 (t, 1H) ppm. 0

(lH-Indazol-3-yl)-(2-nitro-phenyI)-methanone (50). To a gently refluxed solution of 49 (7 g, 36.8 mmol) and isoamylnitrite (5.45 ml, 40.3 mmol) in dichloromethane (180 ml) was added dropwise a solution of anthranilic acid (5.50 g,₅ 38.6 mmol) in acetone (30 ml). The resulting solution was refluxed for 30 minutes, cooled to room temperature, and the solvent was evaporated under high vacuum. The crude product was triturated with dichloromethane to give 5.87 g (60%) of the desired compound as a pink solid. TLC dichloromethane, UV, Rf=0.34. 1H NMR (300 MHz, DMSO-d₆): δ 7.42 (t, 1H), 7.53 (t, 1H), 7.71 (d, 1H), 7.80 (m,₀ 2H), 7.92 (d, 1H), 8.22 (d, 1H), 8.29 (d, 1H) ppm.

3-(2-Nitro-benzoyl)-indazole-l-carboxyIic acid tert-butyl ester (51). To a solution of 50 (0.5 g, 1.87 mmol) in acetonitrile (20 ml) was added Boc anhydride

SJO 51087-1 33

Express Mail No. EV 331382753 US (0.408 g, 1.87 mmol) followed by catalytic amount (20 mg) of DMAP. After stirring at room temperature for 30 minutes, no compound 50 was detected on TLC. Solvent was evaporated and EtOAc (30 ml) was added followed by water (30 ml). Organic layer was separated, washed with 0.5N HCl and brine, dried (Na₂SO₄) and concentrated to give 748 g of crude product that was used for the next step without purification.

3-(2-Amino-benzoyl)-indazole-l-carboxylic acid tert-butyl ester (52). To ao solution of 51 (0.73 g, 1.94mmol) in ethanol (30 ml), was added Pd/C (10%) (0.204 g) and the mixture was allowed to stir under H₂ (1 atm) until no starting material was left (about 5 hours). The reaction mixture was filtered over celite and concentrated to give 0.336 g of pure 52 (99% yield). 5

(lH-IndazoI-3-yl)-{2-[(pyridin-4-ylmethyl)-amino]-phenyl}-methanone (l)

A solution of 52 (leq) and 4-pyridinecarboxaldehyde (2.4eq) in benzene (30 ml) was refluxed in the presence of 3 A molecular sieve or acetic acid (2% volume) for 16hrs. The sieves were filtered off and the solvent was evaporated to give corresponding₀ imine that was further dissolved in methanol. NaBH₃CN (4.4 eq) was added and the mixture was stirred for 48 hrs (The pH was adjusted to 6 with acetic acid). Solvent was evaporated, dichloromethane was added and washed with saturated sodium bicarbonate solution. The organic layer was dried (Na₂SO₄) and concentrated to give crude product. This material was purified on silica gel to give pure desired Boc₅ protected product, which was treated with a solution of HCl (4N) in dioxane for 16 hours or TFA / DCM (1/1) for 4 hours. Solvent was removed, EtOAc and saturated sodium bicarbonate were added and organic layer was separated. It was washed with brine and concentrated to give desired final compound that were purified on silica gel (TLC Bakerflex silica gel IB2-F, 2.5x7.5 cm, 5% MeOH/CH₂Cl₂, UV, Rf=0.32) too give pure materials (1).

Η NMR (300 MHz, DMSO-d₆): δ 4.89 (d, 2H, CH₂), 6.60 (d, 1H), 6.71 (t, 1H), 7.35 (m, 2H), 7.50 (t, 1H), 7.70 (d, 1H), 7.96 (d, 2H), 8.23 (d, 1H), 8,47 (d, 1H),

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Express Mail No. EV 331382753 US 8.84 (d, 2H), 9.00 (br. S, IH) ppm; APCI MS: (M+1) 327; UV (MeOH) λ_max 205 nm, 237 nm, 309 nm, 400 nm.

EXAMPLE 2,

(l-Methyl-lH-indazol-3-vπ-{2-r(pyridin-4-ylmethyl)-aminol-phenyl|-methanone (5)

(l-Methyl-l/y-indazol-3yl)-(2-nitro-phenyl)-methaιιone (53). To a solution of 50 (0.8g, 2.99 mmol) in DMF (10 ml) was added methyl iodide (205 μl, 3.29 mmol) and potassium carbonate (0.454 g). After 16 hrs stirring at room temperature, water (70 ml) was added, and extracted with EtOAc (2x30ml). Organic layer was washed with water (2x30ml), brine (20 ml) and dried (Na₂SO₄). 0.83 g of the desired compound was obtained and was used without purification for the next step.

(2-Amino-phenyl)-(l-methyl-ljH-indazol-3-yl)-methanone (54). This compound was prepared according to a procedure analogous to the procedure for the synthesis of compound 52.

(l-MethyI-lH-indazol-3-yl)-{2-[(pyridin-4-ylιτιethyl)-amino]-phenyl}- methanone (5) A solution of 52 (leq) and pyridine-4-carboxaldehyde (2.4eq) in benzene (30 ml) was refluxed in the presence of 3 A molecular sieve or acetic acid (2% volume) for 16 hrs. The sieves were filtered off and the solvent was evaporated to give corresponding imine that was further dissolved in methanol. NaBH₃CN (4.4 eq) was added and the mixture was stirred for 48 hrs (The pH was adjusted to 6 with acetic acid). Solvent was evaporated, dichloromethane was added and washed with saturated sodium bicarbonate solution. The organic layer was dried (Na₂SO₄) and concentrated to give crude product. This material was purified on silica gel to give pure desired product (5). m.p.193-195 °C (decomp.). 1H NMR (400 MHz, DMSO-d₆): δ 4.17 (s, 3H, CH₃), 4.82 (s, 2H, CH₂), 6.58 (d, IH), 6.67 (t, IH), 7.36 (m, 2H), 7.49 (t, IH), 7.79 (d, IH), 7.85 (d, 2H), 8.21 (d, IH), 8,40 (d, IH), 8J8 (d, 2H), 8.90 (br. S, IH) ppm; ESI MS:

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Express Mail No. EV 331382753 US (M+1) 343. Elemental analysis, calcd. for C₂ιHι₈N₄OHCl: C, 66.57; H, 4.76; N, 14.17. Found: C, 66.15; H, 5.05; N, 14.63.

PREPARATION OF ALDEHYDES

lH-Pyrazole-4-carbaldehyde (55). To a solution of l-(4-Methoxy-benzyl)-lH- pyrazole (0.21 mol) (Synth. Commun. 1990, 20/18; 2849) in dry DMF (235 ml) was added POCl₃ (1.5 eq.) dropwise at room temperature. The mixture was then heated at 95°C for 16 hrs. After cooling to room temperature, the reaction mixture was neutralized by adding ice and saturated sodium carbonate solution. EtOAc (800 ml) was added and the organic layer was separated and washed several times with water. After drying (Na₂SO₄), the solvent was removed and the crude product was purified over silica gel to give 16.4 g (35% yield) desired aldehyde l-(4-Methoxy-benzyl)-lH- pyrazole-4-carbaldehyde. It was heated with 150 ml TFA for 2 hrs. TFA was evaporated and the basified crude product was purified on silica gel to give 5.3 g of 55.

4-FormyI-pyrazoIe-l-carboxylic acid tert-butyl ester (56). To a solution of

55 (0.31 g, 3.26 mmol) in acetonitrile (30 ml) was added BOC anhydride (0.71 g, 3.26 mmol) followed by DMAP (0.02 g). After 30 minutes of stirring at room temperature, the solvent was evaporated, water and EtOAc were added. Organic layer was separated, washed with 0.5 N HCl (10 ml) and brine (15 ml), dried (Na₂SO₄) and concentrated to give 0.41 g (76%yield) of the desired product which was used in the next step to prepare di-Boc-protected 2 without purification.

(lH-Indazol-3-yl)-{2-[(lH-pyrazol-4-ylmethyl)-amino]-phenyl}-methanone (2).

Η NMR (400 MHz, DMSO-d₆): δ 4.36 (s, 2H, CH₂), 6.65 (t, IH), 6.92 (d IH), 7.27 (t, IH), 7.42 (m, 2H), 7.54 (br. s, IH), 7.68 (d, IH), 7.78 (br. s, IH), 8.13 (d, IH), 8.43 (d, IH), 8.69 (t, IH), 12.75 (br. S, IH), 13.79 (br. S, IH) ppm; API-ES MS: (M+1) 318.

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Express Mail No. EV 331382753 US lH-IndazoIe-5-carbaldehyde (57) To a solution of commercially available 5- bromoindazole (0.5 g, 2.53 mmol) in dry THF (10 ml) was added 2.5 M solution of n-BuLi in hexane (6.42 mmol) at -40 C and under argon atmosphere. After 15 minute ₅ of stirring at this temperature, DMF (0.4 ml, 5.06 mmol) was added and the mixture stiπed for additional 30 minutes at -40 C. The cold bath was then removed and the mixture was allowed to stir for one hrs at room temeperature. Water (30 ml) was added and the aqueous layer was extracted with EtOAc (2x30 ml). The organic layer was washed with brine, dried (Na₂SO₄) and concentrated to give 0.54 g of crude 57o that was purified on silica gel to give 0.21 g (57% yield) aldehyde 57.

1H NMR (400 MHz, CDC1₃): δ 7.62 (dd, IH), 7.98 (d, IH), 8.35 (m, 2H), 10.11 (s, IH), 10.59 (br. S, IH) ppm.

s 5-Formyl-indazole-l-carboxylic acid tert-butyl ester (58) To a solution of

57 (0.21 g, 1.43 mmol) in acetonitrile (10 ml) was added BOC anhydride (0.314 g, 1.43 mol) followed by catalytic amounts of DMAP (0.03 g). After 30 minutes of stirring at room temperature, solvent was evaporated, EtOAc (30 ml) was added and the organic layer was washed with 0.5 N HCl solution (10 ml) and brine. It was then₀ dried (Na₂SO₄) and concentrated to give 0.35 g (99% yield) desired product that was used to prepare di-Boc-protected 3 without purification.

6-Formyl-indazole-l-carboxylic acid tert-butyl ester (59). This compound₅ was prepared according to a procedure analogous to the procedure for the synthesis of compound 58 and was used to prepare di-Boc-protected 4 without purification.

(lH-Indazol-3-yI)-{2-[(lH-indazol-5-ylmethyl)-amino]-phenyl}-o methanone (3).

This compound was prepared according to a procedure analogous to the procedure for the synthesis of compound 1.

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Express Mail No. EV 331382753 US m.p.250-252 °C ; 1H NMR (400 MHz, DMSO-d₆): δ 4.60 (s, 2H, CH₂), 6.61 (t, IH), 6.85 (d IH), 7.25-7.58 (m, 4H), 7.67 (d, IH), 7.77 (s, IH), 8.03 (s, IH), 8.18 (d, IH), 8.45 (d, IH), 8.13 (d, IH), 9.01 (t, IH), 13.06 (br. S, 2H) ppm; ESI MS: (M+1) 368.

(lH-Indazol-3-yl)-{2-[(lH-indazol-6-ylmethyl)-amino]-phenyl}-methanone

(4).

This compound was prepared according to a procedure analogous to the procedure for the synthesis of compound 1. ,o 1H NMR (400 MHz, DMSO-d₆): δ 4.66 (s, 2H, CH₂), 6.62 (t, IH), 6.78 (d, IH), 7.16 (d, IH), 7.33 (m, 2H), 7.48 (m, 2H), 7.71 (m, 2H), 7.01 (s, IH), 8.19 (d, IH), 8.45 (d, IH), 9.09 (t, IH), 12.90 (br. S, IH), 13.83 (br. S, IH) ppm; ESI MS: (M+1) 368.

,₅ EXAMPLE 3,

(lH-Indol-3-yl)-{2-r(pyridin-4-ylmethyl)-amino]-phenylj-methanone (6)

(lH-IndoI-3-yl)-(2-nitro-phenyl)-methanone (60). To a suspension of indole (2.9 g, 0.025 mol), and ZnC12 (6.8 g, 0.050 mol) in 100 ml of CH2C12 was added

₂₀ propylmagnesium chloride (11.25 ml of 2.0 M solution, 0.025 mol) at room temperature. This mixture was stirred for one hour at room temperature and aluminum chloride (4.00 g, 0.03 mol) and 2-nitrobenzoyl chloride (5.0 g, 0.027 mol) were added and the mixture stirred for an additional hour. The reaction was quenched with ammonium chloride solution and the aqueous layer was extracted with EtOAc (2x30

₂₅ ml). The organic layer was washed with brine, dried (Na₂SO₄) and concentrated to give of crude 60 that was purified on silica gel to give 0.55 g (9%) of 60 as yellow crystalline solid.

Η NMR (400 MHz, DMSO-d₆): δ 7.25 (m, 2H), 7.56 (t, IH), 7.78 (m, 4H), 8.16 (m, 2H), 10.33 (br. s, IH) ppm.

30

(2-Amino-phenyl)-(lH-indol-3-yl)-methanone (61) The nitro compound 60 (0.55 g, 2.1 mmol) was reduced with 10% Pd/C (0.05 g) in ethanol (50 ml) using Rαrr hydrogenator to obtain 0.39 g of amine 61 (80 %) after column chromatography.

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Express Mail No. EV 331382753 US 1H NMR (400 MHz, CDC1₃): δ 5.48 (br. s, 2H, NH₂), 6.71 (m, 2H), 7.30 (m, 5H), 8.28 (m, IH), 9.13 (br. s, IH) ppm; ESI MS: (M+1) 236.

₅ (lH-Indol-3-yl)-{2-[(pyridin-4-ylmethyl)-amino]-phenyl}-methanone (6) A solution of the amino compound 61 (0.33 g, 1.4 mmol) in methanol (10 ml) and pyridine-4-carboxaldhyde (0.19 g, 1.8 mmol) was refluxed for 3 hours. Solvent was removed in vacuo and the residue was subjected to hydrogenation in ethanol (50 ml) in the presence of 10% Pd/C (0.05 g). The crude product was purified by careful flasho chromatography to obtain 105 mg of 6 (21%) as bright yellow solid.

1H NMR (400 MHz, CDC1₃): δ 4.39 (s, 2H, CH₂), 6.44 (d, IH), 6.62 (t, IH), 7.21 (m, 5H), 7.34 (m, IH), 7.56 (d, IH), 7.69 (d, IH), 8.14 (br. S, IH), 8.22 (m, IH), 8.46 (d, 2H), 9.32 (br. s, IH) ppm; ESI MS: (M+1) 328. s EXAMPLE 4, f5-Methyl-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl -amino -phenyl|- methanone hydrochloride (13)

(5-Methyl-l H-indol-3-yl)-(2-nitro-phenyl)-methanone (62) 0 To a solution of 5-methylindole (5g, 38.1 mmol) in THF (95mL) at -78 °C was added 3.0 M Ethyl magnesium bromide (14.0 mL, 41.9 mmol) dropwise. The pale white solution turns a pale yellow. After about 5 mins, the solution was allowed to warm up by removing from the dry ice bath for about 3-5 mins. Then cooled it down again to -78 °C and added the acid chloride (90%Technical grade) (6.1 mL,₅ 41.9 mmol) dissolved in THF (10 mL). The reaction mixture was stiπed for 1 hr at - 78 °C and then allowed to warm upto room temperature. Stiπed for another 3 hrs and then quenched with saturated NH₄C1 solution. Allowed to stir for 10 mins and then took up the solution in a separatory funnel and worked up with water followed by sat. NaCl solution. Dried the organic layer with NaSO₄, then filtered and concentrated theo solution to get a pale yellow oil that was purified via flash chromatography. The diacylated product was taken up in THF (35 mL) and 0.5M LiOH solution in methanol (152.4 mL, 72.6 mmol) was added to it. After stirring for 2 hours the reaction mixture was concentrated to one-third of the volume, neutralized with 10%

SJO 51087-1 39

Express Mail No. EV 331382753 US HCl and then filtered off. Washed the solids with copious amounts of water and dried in a vacuum oven overnight to yield 4.5 g (42%) of 62.

1H-NMR (300 MHz, DMSO-d₆) 12.05 (br s, IH), 8.17 (d, J = 7.8Hz, IH), 7.98 (br s, IH), 7.88 (t, J = 7.2Hz, IH), 7.78 (t, J = 7.5Hz, IH), 7.70 (br d, J = 5.4Hz, 2H), 5 7.40 (d, J = 8.1Hz, IH), 7.11 (d, J = 8.4Hz, IH), 2.44 (s, 3H) ppm. ¹³C-NMR (300 MHz, DMSO-d₆) 186.5, 147.0, 136.6, 135.9, 135.3, 133.9, 131.2, 130.5, 129.2, 125.8, 124.8, 124.4, 120.9, 115.3, 112.1, 21.2 ppm.

(2-Amino-phenyl)-(5-methyl-l/-r-indol-3-yl)-methanone (63) ιo The nitro compound 62 (1.04 g, 3.71mmol) was dissolved in a 1 : 1 mixture of ethyl acetate and methanol (30 mL) with heating and subjected to reduction in a hydrogen atmosphere with 10% Pd/C (0.371 g). The reaction mixture was filtered through a pad of celite and silica gel and cconcentrated to yield 0.913g (98%) of 63 after purification via flash chromatography.

,₅ 1H-NMR (300 MHz, DMSO-d₆) 11.89 (br s, IH), 8.04 (s, IH), 7.80 (br s, IH),

7.61 (d, J = 7.8Hz, IH), 7.44 (d, J = 8.1Hz, IH), 7.28 (t, J = 8.4Hz, IH), 7.11 (d, J = 8.4Hz, IH), 6.85 (d, J = 8.4Hz, IH), 6.65 (t, J = 7.2Hz, IH), 6.31 (br s, 2H), 2.47 (s, 3H) ppm. ¹³C-NMR (300 MHz, DMSO-d₆) 191.7, 149.3, 134.9, 134.2, 131.8, 131.3, 130.2, 126.7, 124.2, 121.4, 121.1, 116.2, 115.8, 114.6, 111.7, 21.3 ppm.

₂o

(5-Methyl-lH-indol-3-yI)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone hydrochloride (13)

To a stiπed solution of 63 (0.128g, 0.51 mmol) and 6-formyl quinoline ₂₅ (0.104g, 0.66mmol) in benzene (5.1 mL) was added glacial acetic acid (0.10 mL) and the solution was refluxed at 100 °C overnight in an oil bath. After 16 hrs, the reaction vessel was removed from the oil bath and sodium triacetoxyborohydride (0.216g, 1.02 mmol) was added slowly. The solution turns a bright red color which eventually changes to bright yellow. After stirring for 2-3 hours the reaction mixture is diluted ₃o with ethyl acetate and quenched with a few drops of sat. NaHCO3 solution. Allowed to stir for 10-15 mins and then took up the solution in a separatory funnel and worked up with NH₄C1 solution and water followed by sat. NaCl solution. Dried over sodium

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Express Mail No. EV 331382753 US sulfate, concentrated and chromatographed via flash chromatography to give 0.128 g (64%) of a yellow solid as product.

1H NMR (300 MHz, DMSO-d₆) S 11.69 (br s, IH), 8.64 (br s, IH), 8.09 (d, J = 7.8 Hz, IH), 7.79 (s, IH), 7.76-7.69 (m, 3H), 7.57 (br s, 2H), 7.44 (d, J = 7.5Hz, IH), ₅ 7.28 (dd, J = 8.4, 4.2Hz, IH), 7.17 (d, J = 8.1Hz, IH), 7.05 (t, J = 7.8Hz, IH), 6.85 (d, J = 8.1Hz, IH), 6.53 (d, J = 8.4Hz, IH), 6.43 (t, J = 7.2Hz, IH), 4.42 (d, J = 6.0Hz, 2H), 2.19 (s, 3H) ppm.

The following compounds were prepared according to procedures described ino the examples above.

{2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(lH-indoI-3-yl)-methanone

(7)

1H NMR (300 MHz, CD₃OD) δ 8.21 (d, J = 9.0 Hz, IH), 7.95 (br s, IH), 7.68-₅ 7.63 (m, 3H), 7.51 (br s, IH), 7.47 (d, J = 6.3Hz, IH), 7.27-7.20 (m, 3H), 7.13 (d, J = 8.4Hz, IH), 6.72 (d, J = 8.4Hz, IH), 6.62 (t, J = 7.5Hz, IH), 4.51 (s, 2H) ppm.

{2-[(lH-Benzoimidazol-5-ylmethyl)-amino]-phenyl}-(lH-indazol-3-yl)-o methanone (8)

1H NMR (300 MHz, CD₃OD) δ 8.29 (d, J = 8.1 Hz, IH), 8.07-8.04 (m, 2H), 7.58-7.52 (m, 3H), 7.38-7.15 (m, 4H), 6.76 (d, J = 8.1 Hz, IH), 6.54 (t, J = 7.2 Hz, IH), 4.59 (s, 2H) ppm. 5

[2-(4-Imidazol-l-yl-benzyIamino)-phenyl]-(lH-indazol-3-yl)-methanone

(9)

Η NMR (300 MHz, DMSO-d₆) δ 12.06 (br s, IH), 9.21 (s, IH), 9.09 (br t, IH), 8.51 (d, J = 8.1 Hz, IH), 8.27 (d, J = 8.1 Hz, IH), 8.15 (br s, IH), 7.83-7.74 (m,0 3H), 7.66 (d, J = 8.4Hz, 2H), 7.56-7.51 (m, 2H), 7.43-7.35 (m, 2H), 6.84 (d, J = 8.7Hz, IH), 6.72 (t, J = 7.8Hz, IH), 4.69 (d, J = 6.0Hz, 2H) ppm.

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Express Mail No. EV 331382753 US (lH-Indazol-3-yl)-{2-[(3-methyl-lH-indazol-5-ylmethyl)-amino]-phenyl}- methanone (10)

1H NMR (300 MHz, DMSO-d₆) δ 13.72 (br s, IH), 12.49 (s, IH), 8.86 (br t, IH), 8.35 (br d, J = 7.8Hz, IH), 8.04 (d, J = 8.1Hz, IH), 7.61 (s, IH), 7.56 (d, J = 8.4Hz, IH), 7.36-7.24 (m, 4H), 7.17 (t, J = 7.2Hz, IH), 6.78 (d, J = 8.4Hz, IH), 6.52 (t, J = 7.2Hz, IH), 4.47 (d, J = 5.7Hz, 2H), 2.34 (s, 3H) ppm.

{2-[(Benzo[l,3]dioxol-5-yImethyl)-amino]-phenyl}-(lH-indazol-3-yl)- methanone (12)

1H NMR (300 MHz, CD₃OD) δ 8.18 (d, J = 8.1 Hz, IH), 7.96 (d, J = 8.1 Hz, IH), 7.44 (d, J = 8.4 Hz, IH), 7.27 (t, J = 6.9Hz, IH), 7.18 (t, J = 8.7Hz, IH), 7.09 (t, J = 8.1Hz, IH), 6.73-6.70 (m, 2H), 6.64-6.59 (m, 2H), 6.45 (t, J = 8.1Hz, IH), 5.74 (s, 2H), 4.26 (s, 2H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(5-methyl-lH-indol-3-yl)- methanone (14)

Η-NMR (300 MHz, DMSO-d₆) 12.95 (br s, IH), 11.90 (br s, IH), 8.02 (br s, 2H), 7.92 (t, J = 6.0Hz, IH), 7.80 (br s, IH), 7.72 (d, J = 8.1Hz, IH), 7.68 (d, J = 7.8Hz, IH), 7.50 (br s, IH), 7.40 (d, J = 8.1Hz, IH), 7.29 (d, J = 7.8Hz, IH), 7.14 (d, J = 8.4Hz, IH), 7.08 (d, J = 8.1Hz, IH), 6.74 (d, J = 8.4Hz, IH), 6.65 (t, J = 7.2Hz, IH), 4.57 (d, J = 5.7Hz, 2H), 2.43 (s, 3H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(5-methyl-lH-indol-3-yl)- methanone (15)

1H-NMR (300 MHz, DMSO-d₆) 11.91 (br s, IH), 8.86 (dd, J = 4.2, 1.5Hz, IH), 8.35-8.26 (br d, IH), 7.99 (d, J = 8.7Hz, IH), 7.92-7.73 (m, 5H), 7.65 (dd, J = 7.5, 1.2Hz, IH), 7.50 (dd, J = 8.1, 4.2Hz, IH), 7.41 (d, J = 8.7Hz, IH), 7.27 (t, J = 7.2Hz, IH), 4.57 (dd, J = 8.7, 2.4Hz, IH), 6.73 (d, J = 8.1Hz, IH), 6.53 (t, J = 7.2Hz, IH), 4.65 (d, J = 6.0Hz, 2H), 3.78 (s, 3H) ppm.

(1 H-Indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}-methanone (16)

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Express Mail No. EV 331382753 US 1H NMR (300 MHz, DMSO-d₆) δ 12.06 (br s, IH), 8.91 (dd, J = 4.2, 1.8Hz, IH), 8.36 (d, J = 7.5Hz, IH), 8.23 (d, J = 6.9Hz, IH), 8.06-7.97 (m, 3H), 7.90 (s, IH), 7.82 (dd, J = 8.7, 1.8Hz, IH), 7.73 (dd, J = 7.8, 1.5Hz, IH), 7.58-7.53 (m, 2H), 7.36- 7.23 (m, 3H), 6.79 (d, J = 8.1Hz, IH), 6.70 (t, J = 6.9Hz, IH), 4.70 (d, J = 6.0Hz, 2H) ppm.

(lH-IndazoI-3-yl)-{2-[(pyridin-3-ylmethyl)-amino]-pheιιyI}-methanoιιe (17) 1H NMR (300 MHz, DMSO-d₆) δ 14.01 (br s, IH), 9.11 (t, J = 6.0Hz, IH),

8.78 (d, J = 1.5Hz, IH), 8.64-8.58 (m, IH), 8.35 (d, J = 8.1Hz, IH), 7.94 (d, J = 7.8Hz, IH), 7.84 (d, J = 8.48Hz, IH), 7.62 (t, J = 7.2Hz, IH), 7.55-7.44 (m, 2H), 6.95 (d, J = 8.75Hz, IH), 6.81 (t, J = 7.2Hz, IH), 4.74 (d, J = 6.0Hz, 2H) ppm.

(5-Fluoro-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (18)

1H NMR (300 MHz, DMSO-d₆) δ 12.19 (br s, IH), 8.91 (br s, IH), 8.37 (d, J = 7.8Hz, IH), 8.08-7.91 (m, 5H), 7.83 (dd, J = 8.7, 1.8Hz, IH), 7.75 (dd, J = 7.8, 1.5Hz, IH), 7.58 (td, J = 9.0, 4.5Hz, 2H), 7.35 (t, J = 7.5Hz, IH), 7.17 (dt, J = 9.3, 2.7Hz, IH), 6.81 (d, J = 8.4Hz, IH), 6.72 (t, J = 7.5Hz, IH), 4.71 (d, J = 6.0Hz, 2H) ppm.

(5-Fluoro-lH-indoI-3-yl)-{2-[(lH-indazol-6-ylmethyl)-amino]-phenyl}- methanone (19)

1H NMR (300 MHz, DMSO-d₆) δ 12.88 (br s, IH), 12.06 (br s, IH), 7.96-7.88 (m, 3H), 7.81 (dd, J = 10.2, 2.4Hz, IH), 7.65 (t, J = 8.4Hz, 2H), 7.50-7.43 (m, 2H), 7.23 (t, J = 7.2Hz, IH), 7.09-7.03 (m, 2H), 6.68 (d, J - 8.1Hz, IH), 6.60 (t, J = 7.2Hz, IH), 4.52 (d, J = 5.7Hz, 2H) ppm.

(5-Chloro-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (20)

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Express Mail No. EV 331382753 US 1H NMR (300 MHz, DMSO-d₆) δ 12.22 (br s, IH), 8.89 (dd, J = 4.2, 1.5Hz, IH), 8.34 (d, J = 8.4Hz, IH), 8.21 (d, J = 2.1Hz, IH), 8.05-7.95 (m, 4H), 7.80 (dd, J = 8.7, 2.1Hz, IH), 7.73 (dd, J = 7.8, 1.2Hz, IH), 7.59-7.51 (m, 2H), 7.35-7.29 (m, 2H), 6.79 (d, J = 8.4Hz, IH), 6.69 (t, J = 7.5Hz, IH), 4.68 (d, J = 5.4Hz, 2H) ppm.

(2-Methyl-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (21)

Η-NMR (300 MHz, DMSO-d₆) 12.30 (br s, IH), 8.93 (br s, IH), 8.46 (d, J = 7.5Hz, IH), 8.05 (br s, IH), 7.88 (d, J = 8.7Hz, IH), 7.70-7.33 (m, 8H), 7.10 (t, J = 6.9Hz, IH), 6.95 (d, J = 7.8Hz, IH), 6.78 (s, 2H), 6.55 (t, J - 6.9Hz, IH), 2.56 (s, 3H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(2-methyl-lH-indol-3-yl)- methanone (22)

1H NMR (300 MHz, DMSO-d₆) δ 13.16 (br s, IH), 12.21 (br s, IH), 8.12 (s, IH), 7.82 (d, J = 8.4Hz, IH), 7.64-7.52 (m, 2H), 7.43-7.24 (m, 6H), 7.10 (t, J = 7.2Hz, IH), 6.95 (d, J = 8.1Hz, IH), 6.74 (br s, 2H), 6.56 (t, J = 7.2Hz, IH), 3.39 (s, 3H) ppm.

(5-Chloro-lH-indol-3-yl)-{2-[(lH-indazol-6-ylmethyl)-ammo]-phenyl}- methanone (23) 1H NMR (300 MHz, DMSO-d₆) δ 12.87 (br s, IH), 12.12 (br s, IH), 8.25 (s,

IH), 8.12 (d, J = 1.8Hz, IH), 7.95-7.89 (m, 3H), 7.67-7.62 (m, 2H), 7.48 (d, J = 8.4Hz, IH), 7.43 (s, IH), 7.26-7.19 (m, 2H), 7.07 (dd, J = 8.4, 1.2Hz, IH), 6.68 (d, J 8.1Hz, IH), 6.59 (t, J = 7.2Hz, IH), 4.52 (d, J = 5.7Hz, IH) ppm.

(lH-Indol-3-yl)-{2-[(isoquinolin-6-ylmethyl)-amino]-phenyl}-methanone

(24)

Η NMR (300 MHz, DMSO-d₆) δ 11.89 (br s, IH), 9.13 (s, IH), 8.33 (d, J = 5.7Hz, IH), 8.06 (d, J = 7.2Hz, IH), 7.96 (d, J = 8.4Hz, IH), 7.85 (t, J = 6.0Hz, IH),

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Express Mail No. EV 331382753 US 7.75 (d, J = 9.0Hz, 2H), 7.63 (d, J = 5.7Hz, IH), 7.58-7.54 (m, 2H), 7.39 (d, J = 8.1Hz, IH), 7.16-7.08 (m, 3H), 6.59-6.50 (m, 2H), 5.7 (d, J = 5.7Hz, 2H) ppm.

(6-Methoxy-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (25)

1H NMR (300 MHz, DMSO-d₆) δ 11.69 (br s, IH), 8.73 (br s, IH), 8.18 (d, J = 7.8Hz, IH), 7.93-7.79 (m, 4H), 7.66-7.53 (m, 3H), 7.39-7.35 (m, IH), 7.14 (t, J =0 7.8Hz, IH), 6.87 (s, IH), 6.73 (d, J = 8.7Hz, IH), 6.60 (d, J = 8.4Hz, IH), 6.52 (t, J = 7.5Hz, IH), 4.51 (d, J = 5.4Hz, 2H), 3.68 (s, 3H) ppm.

(6-Fluoro-l H-indoI-3-yl)-{2- [(quiπolin-6-ylmethyl)-amino] -phenyl}-5 methanone (26)

1H NMR (300 MHz, DMSO-d₆) δ 11.86 (br s, IH), 8.67 (br s, IH), 8.12 (br d,

IH), 7.97 (dd, J = 8.7,5.7Hz, IH), 7.82-7.79 (m, 2H), 7.73 (br s, IH), 7.68 (br s, IH), 7.58 (d, J = 8.4Hz, IH), 7.49 (d, J = 6.3Hz, IH), 7.31 (dd, J = 8.4, 4.2Hz, IH), 7.13- 7.07 (m, 2H), 6.88 (br t, J = 9.0Hz, IH), 6.55 (d, J = 8.4Hz, IH), 6.60 (t, J = 7.5Hz,o IH), 4.45 (d, J = 5.7Hz, 2H) ppm.

(6-Fluoro-lH-indol-3-yl)-{2-[(lH-indazol-6-ylmethyl)-amino]-phenyl}- methanone (27) 5 1H NMR (300 MHz, DMSO-d₆) δ 12.89 (br s, IH), 11.99 (br s, IH), 8.12 (dd,

J = 9.0,5.7Hz, IH), 7.97 (s, IH), 7.94 (t, J = 5.7Hz, IH), 7.82 (s, IH), 7.66 (t, J = 8.1Hz, 2H), 7.44 (s, IH), 7.28-7.22 (m, 2H), 7.10-7.00 (m, 2H), 6.69 (d, J = 8.4Hz, IH), 6.60 (t, J = 7.5Hz, IH), 4.53 (d, J = 5.7Hz, 2H) ppm. 0

(7-Methyl-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (28)

Η-NMR (300 MHz, DMSO-d₆) 12.10 (br s, IH), 8.92 (br s, IH), 8.38 (d, J = 8.4Hz, IH), 8.08-7.82 (m, 6H), 7.73 (d, J = 7.8Hz, IH), 7.57 (dd, J = 8.1, 4.2Hz, IH),

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Express Mail No. EV 331382753 US 7.32 (t, J = 8.7Hz, IH), 7.20-7.10 (m, 2H), 6.81 (d, J = 8.7Hz, IH), 6.72 (t, J = 7.5Hz, IH), 4.71 (d, J = 5.7Hz, 2H), 2.59 (s, 3H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-amino]-plιenyl}-(7-methyl-lH-indol-3-yl)- methanone (29)

1H-NMR (300 MHz, DMSO-d₆) 13.03 (br s, IH), 12.11 (br s, IH), 8.10-8.04 (m, 3H), 7.89-7.75 (m, 3H), 7.58 (s, IH), 7.37 (t, J - 8.1Hz, IH), 7.23-7.12 (m, 3H), 6.82 (d, J = 8.1Hz, IH), 6.74 (t, J = 7.2Hz, IH), 4.66 (d, J = 6.0Hz, 2H), 2.61 (s, 3H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(4-methoxy-lH-indol-3-yl)- methanone (30)

1H-NMR (300 MHz, DMSO-d₆) 12.97 (br s, IH), 11.71 (br s, IH), 8.95 (t, J = 6.0Hz, IH), 8.03 (br s, IH), 7.74 (d, J = 8.4Hz, IH), 7.55-7.50 (m, 3H), 7.27 (t, J = 7.2Hz, IH), 7.17-7.07 (m, 3H), 6.75 (d, J = 8.4Hz, IH), 6.59 (d, J = 7.2Hz, IH), 6.49 (t, J = 7.8Hz, IH), 4.65 (d, J = 6.0Hz, 2H), 3.60 (s, 3H) ppm.

{2-[(lH-Indazol-5-ylmethyl)-amino]-phenyl}-(l-methyl-lH-indazol-3-yl)- methanone (31)

1H NMR (300 MHz, DMSO-d₆) δ 13.04 (br s, IH), 9.00 (t, J = 5.4Hz, IH), 8.46 (d, J - 8.1Hz, IH), 8.18 (d, J = 8.1Hz, IH), 8.04 (s, IH), 7.82-7.64 (m, 2H), 7.55-7.31 (m, 5H), 6.86 (d, J = 8.1Hz, IH), 6.64 (t, J = 7.8Hz, IH), 4.61 (d, J = 5.4Hz, 2H), 4.19 (s, 3H) ppm.

{2-[(lH-Indazol-5-ylmethyl)-amino]-phenyl}-[l-(2-methoxy-ethyl)-lH- indazol-3-yl]-methanone (32)

1H NMR (300 MHz, DMSO-d₆) δ 13.13 (br s, IH), 9.13 (t, J = 5.4Hz, IH), 8.53 (d, J = 8.1Hz, IH), 8.26 (d, J = 8.4Hz, IH), 8.13 (s, IH), 7.93 (d, J = 8.4Hz, IH), 7.86 (s, IH), 7.65-7.39 (m, 5H), 6.96 (d, J = 8.7Hz, IH), 6.73 (t, J = 7.2Hz, IH), 4.82 (t, J = 5.1Hz, 2H), 4.71 (d, J = 5.7Hz, 2H), 3.92 (t, J = 5.4Hz, 2H), 3.31 (s, 3H) ppm.

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Express Mail No. EV 331382753 US {2-[(lH-Indazol-6-ylmethyl)-amino]-phenyl}-(l-πιethyl-lH-indazoI-3-yl)- methanone (33)

1H NMR (300 MHz, CDC1₃) δ 9.86 (br s, IH), 9.18 (t, J = 6.3Hz, IH), 8.42 (d, J = 8.4Hz, IH), 8.22 (d, J = 8.1Hz, IH), 7.97 (s, IH), 7.66 (d, J = 8.7Hz, IH), 7.46- 7.40 (m, 3H), 7.29-7.12 (m, 3H), 6.65-6.59 (m, 2H), 4.63 (d, J = 6.0Hz, 2H), 4.14 (s, 3H) ppm.

(lH-Indazol-3-yl)-{2-[(qumoIm-6-ylmethyl)-amino]-plιenyl}-methanone (34)

Η NMR (300 MHz, DMSO-d₆) δ 13.80 (br s, IH), 9.07 (t, J = 5.7Hz, IH), 8.81 (dd, J = 1.8, 4.2Hz, IH), 8.43 (d, J = 8.1Hz, IH), 8.29 (d, J = 8.4Hz, IH), 8.16 (d, J = 8.4Hz, IH), 7.97 (d, J = 8.7Hz, IH), 7.90 (s, IH), 7.75 (dd, J = 2.1, 8.7Hz, IH), 7.61 (d, J = 8.4Hz, IH), 7.48-7.39 (m, 2H), 7.32-7.23 (m, 2H), 6.76 (d, J = 8.4Hz, IH), 6.60 (t, J = 7.5Hz, IH), 4.70 (d, J = 6.0Hz, 2H) ppm.

(lH-Indazol-3-yl)-{2-[(isoquinolin-6-ylmethyl)-amino]-phenyl}- methanone (35)

Η NMR (300 MHz, DMSO-d₆) δ 13.80 (br s, IH), 9.21 (s, IH), 9.07 (t, J -

6.0Hz, IH), 8.43-8.39 (m, 2H), 8.16 (d, J = 8.1Hz, IH), 8.06 (d, J = 8.4Hz, IH), 7.87

(br s, IH), 7.73 (d, J = 5.7Hz, IH), 7.68-7.62 (m, 2H), 7.41 (d, J = 7.8Hz, IH), 7.30- 7.23 (m, 2H), 6.70 (d, J = 8.1Hz, IH), 6.59 (t, J = 7.8Hz, IH), 4.72 (d, J = 6.0Hz, 2H) ppm.

N-(4-{[2-(lH-IndazoIe-3-carbonyl)-phenylamino]-methyl}-phenyl)- acetamide (36)

1H NMR (300 MHz, DMSO-d₆) δ 13.84 (br s, IH), 9.93 (s, IH), 8.94 (t, J = 5.4Hz, IH), 8.47 (d, J = 8.1Hz, IH), 8.18 (d, J = 8.1Hz, IH), 7.69 (d, J = 8.1Hz, IH),

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Express Mail No. EV 331382753 US 7.55 (d, J = 8.4Hz, 2H), 7.48-7.27 (m, 5H), 6.80 (d, J = 8.4Hz, IH), 6.64 (t, J = 6.9Hz, IH), 4.46 (d, J = 5.7Hz, 2H), 2.1 (s, 3H) ppm.

4-{[2-(lH-Indazole-3-carbonyl)-phenyIamino]-methyl}-N-isopropyl- benzamide (37)

1H NMR (300 MHz, DMSO-d₆) δ 13.86 (br s, IH), 9.03 (t, J = 5.7Hz, IH), 8.48 (d, J = 8.1Hz, IH), 8.23-8.15 (m, 2H), 7.82 (d, J = 8.4Hz, 2H), 7.71 (d, J = 8.4Hz, IH), 7.51-7.30 (m, 5H), 6.74 (d, J = 8.7Hz, IH), 6.66 (t, J = 6.9Hz, IH), 4.61 (d, J = 6.0Hz, 2H), 4.13-4.03 (m, IH), 1.16 (d, J = 6.6 Hz, 6H) ppm.

(6-Methyl-lH-indol-3-yl)-{2-[(quinolin-6-ylmethyl)-amino]-phenyl}- methanone (38) 1H NMR (300 MHz, DMSO-d₆) δ 11.95 (s, IH), 8.93 (m, IH), 8.38 (d, J =

8.0Hz, IH), 8.13-7.99 (m, 4H), 7.86-7.82 (m, 2H), 7.73 (d, J = 8.0Hz, IH), 7.57 (dd, J = 8.4, 4.2 Hz, IH), 7.37 (s, IH), 7.34 (t, J = 9.0 Hz, IH), 7.11 (d, J = 8.4Hz, IH), 6.80 (d, J = 8.4Hz, IH), 6.72 (t, J = 7.2 Hz, IH), 4.71 (d, J = 6.0 Hz, 2H), 2.49 (s, 3H) ppm.

{2-[(lH-Indazol-6-ylmethyl)-ammo]-phenyl}-(6-methyl-lH-indol-3-yl)- methanone (39)

1H NMR (300 MHz, DMSO-d₆) δ 12.94 (s, IH), 11.87 (s, IH), 8.06-8.01 (m, 2H), 7.93 (t, J = 5.7 Hz, IH), 7.77-7.66 (m, 3H), 7.49 (s, IH), 7.30 (s, IH), 7.28 (t, J = 7.2Hz, IH), 7.13 (d, J = 8.4 Hz, IH), 7.04 (d, J = 8.1 Hz, IH), 6.72 (d, J = 8.4 Hz, IH), 6.41 (t, J = 7.2Hz, IH), 4.57 (d, J = 5.7Hz, 2H), 2.43 (s, 3H) ppm.

EXAMPLE 5 In-vitro assay for VEGFR-2 (KDR) Kinase Inhibition

VEGFR tyrosine kinase inhibition is determined by measuring the phosphorylation level of poly-Glu-Ala-Tyr-biotin (pGAT-biotin) peptide in a Homogeneous Time-Resolved Fluorescence (HTRF) assay. Into a black 96-well

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Express Mail No. EV 331382753 US Costar plate is added 2 μl/well of 25x compound in 100% DMSO (final compound concentration in the 50-μl kinase reaction is typically 1 nM to 10 μM). Next, 38 μl of reaction buffer (25 mM Hepes pH 7.5, 5 mM MgC12, 5 mM MnC12, 2 mM DTT, 1 mg/ml BSA) containing 0.5 pmoles polyGAT-biotin and 3-4 ng KDR enzyme is added to each well. After 5-10 min preincubation, the kinase reaction is initiated by the addition of 10 μl of 10 μM ATP in reaction buffer, after which the plate is incubated at room temperature for 45 min. The reaction is stopped by the addition of 50 μl of KF buffer (50 mM Hepes pH 7.5, 0.5 M KF, 1 mg/ml BSA) containing 100 mM EDTA and 0.36 μg/ml PY20K (Eu-cryptate labeled anti-phosphotyrosine antibody, CIS bio international). After 30 min, 100 μl of 5 nM SV-XL (modified- APC-labeled Streptavidin, CIS bio international) in KF buffer is added, and after an addition 2-hr incubation at room temperature, the plate is read in a RUBYstar HTRF Reader.

Representative compounds of the invention were tested for their inhibition of VEGFR (KDR) tyrosine kinase. The results are reported in Table 3.

Table 3

Comp. No. IC50 (μM)

1 24

2 58

3 0.7

4 0.54

5 2.9

6 8.6

7 0.14

9 >40

10 4.3

1 1 0.61

13 2.0

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Express Mail No. EV 331382753 US 14 9.1

15 1.9

18 5.9

20 3.1

21 >40

31 0.4

32 0.5

33 0.43

34 0.17

35 10.5

36 >40

37 >40

38 0.88

EXAMPLE 6

Cell-based assay for VEGFR-2 (KDR) Kinase Inhibition Transfection of 293 cells with DNA expressing FGFR1/KDR chimera

DNA for transfection is diluted to a final concentration of 5 μg/ml DNA in IXBBS, 125 mM CaCk and incubated at room temperature for 30 min. 293 cells are seeded in 15 cm tissue culture plates using 2 x 10⁷ cells per plate and incubated for 4 hrs, followed by dropwise addition of 3 ml of DNA solution. The plates are incubated overnight.

The next morning, cells are trypsinized, seeded at 4 x 10⁵ cell/ml into wells of 48 well tissue culture plates (1 ml/well) and incubated overnight. Compounds of the invention are added to individual wells to a final concentration of 10-30 μM and incubated for 2 hours. Generally, 10 mM stock solution are diluted 1/300-1/1000, yielding a final DMSO concentration of 0.1-0.3%. Cells in each well were lysed by resuspension in 100 μL Lysis buffer (150 mM NaCl, 50 mM Hepes pH 7.5, 0.5% Trition X-100, 10 mM NaPPi, 50 mM NaF, 1 mM Na3VO4) and rocked for lh at 4°C. ELISA for detection of tyrosine-phosphorylated chimeric receptor

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Express Mail No. EV 331382753 US 96 well ELISA plates are coated using 100 μl/well of 10 μg/ml αFGFRl antibody, and incubated overnight at 4°C. αFGFRl is prepared in a buffer made with 16 ml 0.2M Na₂CO₃ and 34 ml 0.2M NaHCO₃ and the pH adjusted to 9.6. Concuπent with lysis of the transfected cells, αFGFRl coated ELISA plates are washed three times with PBS+0.1 % Tween-20 and blocked by addition of 200 μl/well of 3% BSA in PBS for lhr upon removal of blocking solution. 80 μl of lysate is then transfeπed to the coated and blocked wells and incubated for lh at 4°C. The plates are washed three times with PBS+0.1% Tween-20.

To detect bound phosphorylated chimeric receptor, 100 μl of anti- phosphotyrosine antibodies (RC20:HRP, Transduction Laboratories) was added per well (final concentration 0.5 μg/ml in PBS) and incubated for lh. The plates are washed six times with PBS+0.1% Tween-20. Enzymatic activity of HRP is detected by adding 50 μl/well of equal amounts of the Kirkegaard & Perry Laboratories (KPL) Substrate A and Substrate B. (KPL cat. #54-61-0). The reaction is stopped by the addition of 50 μl/well 0.1N H₂SO₄ and absorbance is detected at 450nm

Representative compounds of the invention were tested using the ELISA Cell- based assay for KDR inhibition. The results are reported in Table 4

Table 4

Comp. No. IC50 (μM)

1 0.85

2 16

3 135

4 0.075

5 0.8

6 2.5

7 0.16

8 0.26

1 1 0.38

12 2.3

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Express Mail No. EV 331382753 US 13 0.14

14 0.077

15 0.3

16 3.4

17 >40

18 >40

19 0.27

20 0.53

21 >40

31 12

32 1.8

33 0.3

34 0.13

35 21

36 15.1

37 35.7

38 0.35

39 0.071

40 0.33

The above examples are intended to be illustrative only. In particular, the invention is not intended to be limited to the methods, protocols, conditions and the like specifically recited herein, insofar as those skilled in the art would be able to substitute other conditions, methods, amounts, materials, etc. based on the present disclosure to arrive at compounds within the scope of this disclosure. While the present invention is described with respect to particular examples and prefeπed embodiments, the present invention is not hmited to these examples and embodiments. In particular, the compounds of the present invention are not limited to the exemplary species' recited herein. Moreover, the methods of the present invention are not limited to treating only the exemplified diseases and conditions, but

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Express Mail No. EV 331382753 US rather any disease or condition that may be treated by regulation of kinases. Additionally, the methods of synthesis of the present invention are not limited to the methods exemplified in the example. The methods of the present invention include methods of making any of the compounds set forth in the present invention that those skilled would be able to make in view of the present disclosure, and are not limited to the exemplified method. For example, methods encompassed by the present invention may involve the use of a different starting material depending on the desired final compound, different amounts of various ingredients, or substitution of different ingredients such as other reactants or catalysts that would be suitable depending on the starting material and result to be achieved.

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Express Mail No. EV 331382753 US

Claims

What is claimed is:

1. A compound having the following formula (II):

or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, wherein:

Y' is selected from CH and N;

Y is selected from C-R and N;

X is selected from O, S, and N-R; Ri is selected from hydrogen, lower alkyl, lower alkenyl, lower alkynyl and aralkyl, wherein the lower alkyl, lower alkenyl, lower alkynyl and aralkyl may be unsubstituted or substituted with one or more substituents selected from R₂;

R₃ is selected from:

1) hydrogen 2) lower alkoxy, optionally substituted with one or more substituents selected from R₂,

3) lower alkyl, optionally substituted with one or more substituents selected from R₂,

4) lower alkenyl, optionally substituted with one or more substituents selected from R₂,

5) aralkyl, optionally substituted with one or more substituents selected

6) lower alkynyl, optionally substituted with one or more substituents selected from R₂, 7) phenyl, optionally substituted with 1 to 5 substituents selected from R₂,

8) halogen,

9) nitro,

10) cyano,

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Express Mail No. EV 331382753 US 11) a group of the formula -CO₂R, -SR, -SO₂R, -NH₂, -NHR, -NRR, where R is independently selected from H, lower alkyl, aralkyl, aryl, and heteroaryl,

12) pyridine, optionally substituted with 1 to 4 substituents selected from

R₂,

13) pyrazine, optionally substituted with 1 to 3 substituents selected from

R₂,

14) pyrimidine, optionally substituted with 1 to 3 substituents selected from R2, and

15) a group selected from the following:

or tetrazole wherein X is O, S, or N-R, a is 0 to 3, b is 0 to 2; c is 0 or 1 ; R₂ is independently selected from: 1) halogen, 2) lower alkyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy,

3) lower alkenyl, which may be optionally substituted with one or more halogen, hydroxy, lower alkoxy,

4) nitro, 5) cyano,

6) a group of the formula -CO₂R, -OR, -SR, -SO₂R, -NH₂, -NHR, -NRR,

or R₂ can occupy two adjacent positions to form a fused 5- or 6-membered carbocyclic or heterocychc ring, which may be substituted or unsubstituted with R₃ and wherein the heterocychc ring may contain from 1 to 2 heteroatoms independently selected from N, O or S; and

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Express Mail No. EV 331382753 US R is independently selected from hydrogen, halogen, lower alkyl, lower alkoxy, aralkyl, aryl and heteroaryl. 2. A compound of claim 1 , wherein X is N-R. 3. A compound of claim 2, wherein Y' is CH.

4. A compound of claim 2, wherein Y' is N.

5. A compound of claim 3, wherein: Ri is aralkyl;

R is selected from hydrogen, lower alkyl and lower alkoxy; R₃ is selected from hydrogen, lower alkyl, lower alkoxy, and halogen.

6. A compound of claim 4, wherein: Ri is aralkyl;

R is selected from hydrogen, lower alkyl and lower alkoxy;

R₃ is selected from hydrogen, lower alkyl, lower alkoxy, and halogen.

7. A method to treating or preventing a VEGF receptor kinase-dependent disease or condition comprising administering to a mammal a therapeutically effective amount of a compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof.

8. The method according to claim 7, wherein the kinase-dependent disease is a neoplastic disease.

9. The method according to claim 7, wherein the kinase-dependent disease or condition is selected from the group consisting of anigogenic disease, solid or liquid tumors, atherosclerosis, age related macular degeneration, retinal vascularization, inflammatory diseases, or cell proliferative disorders.

10. A method of inhibiting a VEGF receptor tyrosine kinase in a mammal comprising administering to said mammal a tyrosine kinase inhibiting amount of a compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof.

11. The method of claim 10, wherein the VEGF receptor tyrosine kinase is KDR.

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Express Mail No. EV 331382753 US