WO2003028731A1

WO2003028731A1 - Chk1 kinase inhibitors

Info

Publication number: WO2003028731A1
Application number: PCT/US2002/031901
Authority: WO
Inventors: Cynthia A. Parrish; James F. Callahan; Zehong Wan; Joelle L. Burgess; Robert A. Stavenger; Dennis A. Holt
Original assignee: Smithkline Beecham Corporation
Priority date: 2001-10-04
Filing date: 2002-10-04
Publication date: 2003-04-10

Abstract

Novel compounds useful in the inhibition of damage response kinases are provided.

Description

CHK1 KINASE INHIBITORS

FIELD OF THE INVENTION

The present invention relates to damage response kinase inhibitors, especially checkpoint kinase ("chkl kinase") inhibitors, pharmaceutical compositions comprising these compounds and methods of using these compounds to treat various forms of cancer and hyperproliferative disorders.

BACKGROUND OF THE INVENTION

The cellular response to DNA damage involves cell cycle delays, increased repair, and apoptosis (Zhou and Elledge Nature 2000 408: 433-439). Although many effective cancer therapies work by causing DNA damage induced apoptosis, resistance to these agents remains a significant limitation in the treatment of cancer. One important mechanism of drug resistance is attributed to cell cycle delays (also called checkpoints) and repair activation, which provides both the opportunity and capacity for cells to repair DNA damage. It is likely that approaches abrogating these survival DNA damage responses would have significant clinical utility.

Among different DNA damage response kinases, Chkl was linked to survival responses including checkpoints. Mice lacking CHK1 die in early embryogenesis (Liu et al. Gene & Dev. 2000 14: 1448-1459; Takai et al. Gene & Dev. 2000 14: 1439-1447). ES cells expressing a conditional CHK1 gene die of p53-independent apoptosis after loss of CHKL Prior to their death, these cells become incapable of preventing mitotic entry in response to HR (Liu et al. Gene & Dev. 2000 14: 1448-1459), demonstrating that Chkl is required for the G2 DNA damage checkpoint in mammals as previously observed in other organisms.

Chkl prevents mitotic entry as follows. Arrest in G2 is regulated by the maintenance of inhibitory phosphorylation of Cdc2 (Nurse Cell 1997 91: 865-867). Cdc2 dephosphorylation and activation is catalyzed by the dual specificity phosphatase Cdc25 (Morgan Nature 1995 374: 131-134). Recent evidence indicates that part of the G2/M DNA checkpoint mechanism involves inactivation and translocation of Cdc25C into the cytoplasm. This is at least partially mediated by phosphorylation on Ser-216 in Cdc25C and its consequent binding with 14-3-3 proteins (Peng et al. Science 1997 277: 1501-1505; Dalai et al. Mol. Cell Bio. 1999 19: 4465-4479; Yang et al. EMBO J. 1999 18: 2174-2183). Chkl (Sanchez et al. Science 1997 277: 1497-1501) has been shown to phosphorylate Cdc25C at Ser-216 in vitro. This modification is thought to maintain Cdc25C phosphorylation in cells arrested at G2/M in response to DNA damage. Recently, staurosporine-like kinase inhibitors, UCN-01 and SB-218078, have been shown to be potent Chkl inhibitors (Jackson et al. Cancer Res. 2000 60: 566-572; Graves et al. J. Biol. Chan. 2000 275: 5600-5605). In vivo, they can abrogate the G2/M checkpoint induced by DNA damaging agents and enhance the cytotoxicities of the DNA damaging agents. Thus it is likely that a specific Chkl inhibitor could be used clinically in combination treatment with coventional therapies. Since Chkl is an essential kinase for regular cell cycle (Liu et al., Gene & Dev. 2000 14: 1448-1459), it is possible that Chkl inhibitors could also be used alone in cancer therapy.

Based on the foregoing, there is a need to identify a potent chkl kinase inhibitors for the treatment of the various aforementioned indications.

SUMMARY OF THE INVENTION

The present invention involves 3-ureidothiophene compounds represented by Formula (I) hereinbelow, pharmaceutical compositions comprising such compounds and methods of inhibiting chkl kinase as well as specific assays to detect inhibition of chkl kinase activity.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compounds of Formula (I), hereinbelow:

(I)

wherein: RI is selected from the group consisting of H, C,_₂ alkyl, XH, XCH₃, C_j_₂ alkyl-XH,

C ₂ alkyl-XCH₃, C(O)NH₂, C(O)NHCH₃, and C(O)-Cι_₂ alkyl, with the preferred substitution being H or CH₃;

X is selected from the group consisting of O, S, and NH;

R2 is selected from the group consisting of C(O)R⁵, CO₂R⁵, C(O)NHR⁵,

C(O)NHC(=NH)R⁵, C(O)NHC(=NH)NR⁵R⁶, C(O)NHC(O)R⁵, C(O)NHC(O)NR⁵R⁶,

SO₂R⁵, S(O)R⁵, SO₃R⁵, and PO₃R⁵R⁶;

R⁵ and R⁶ are, independently, selected from the group consisting of hydrogen, C, 1-10 alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, and C₀₆ alkylheteroaryl, or R⁵ and R⁶, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_{1 6} alkyl or (CH₂)_0.3aryl, wherein any of the foregoing may be optionally substituted by one or more of group A and on any position;

R3 is H or halogen, with the preferred substitution being H;

R4 is aryl or heteroaryl optionally substituted by one or more of group A and on any position;

A is selected from the group consisting of C_{l Q} alkyl, C_wo alkanoyl, C₂_₁₀ alkenyl, C_{2 10} alkynyl, C₃,_I0 cycloalkyl, C₀_₆ alkylaryl, C_M alkylheterocyclyl, C₀₆ alkylheteroaryl, C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R⁸, N(O)R⁷R⁸, NR⁷R⁸R⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C ₀ alkyl, C_M0 alkanoyl, C,_₁₀ alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, C₀₆ alkylheteroaryl, (CH₂)₀₆heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group D and on any position; Y is an organic or inorganic anion including, but not limited to, bisulfate, chloride, fumarate, iodide, maleate, methanesulfonate, trifluoromethanesulfonate, nitrate, or sulfate;

D is selected from the group consisting of C_j.10 alkyl, C ₀ alkanoyl, C_{2 10} alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, C_0.6 alkylheteroaryl, C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R⁸, N(O)R⁷R⁸, NR⁷R⁸R⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C_M0 alkyl, C_uo alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, ' C₀₆ alkylheterocyclyl, C₀₆ alkylheteroaryl, (CH₂)₀₆heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group E and on any position;

R⁷, R⁸, and R⁹ are, independently, selected from the group consisting of hydrogen, C,_.10 alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_M alkylaryl, ' C_M alkylheterocyclyl, and C_0.6 alkylheteroaryl, or R⁷ and R⁸, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms, optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_{I 6} alkyl or (CH₂)_0.3aryl, wherein any of the foregoing may be optionally substituted by one or more of group E and on any position;

E is selected from the group consisting of C,_.10 alkyl, C _l0 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C₃_₁₀ cycloalkyl, C₀₆ alkylaryl, C₀₆ alkylheterocyclyl, C_0.6 alkylheteroaryl, C(=NH)R¹⁰, COR¹⁰, CONR^lϋR", CON(O)R¹⁰Rⁿ, CONR¹⁰R^πR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰Rⁿ, N(O)R^I0Rⁿ, NR¹⁰R^πR¹²Y, NR¹⁰COR¹⁰, NR^IOCONR'°R^U, NR^I0CON(O)R¹⁰Rⁿ, NR¹⁰CONR¹⁰R^πR¹²Y, NR¹⁰CO₂R¹⁰, NR¹⁰C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR¹⁰SO₂NR¹⁰R", nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR¹⁰Rⁿ, SO₃R¹⁰, PO₃R¹⁰Rⁿ, and halo, wherein C,_.10 alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C₀₆ alkylaryl, C_M alkylheterocyclyl, C_M alkylheteroaryl may be optionally substituted by one or more of C(=NH)R¹⁰, COR¹⁰, CONR¹⁰R^U, CON(O)R¹⁰R", CONR¹⁰R^πR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰R^U, N(O)R¹⁰Rⁿ, NR¹⁰R^UR¹²Y, NR¹⁰COR¹⁰, NR¹⁰CONR¹⁰R^π, NR¹⁰CON(O)R¹⁰R^π, NR¹⁰CONR¹⁰R^UR¹²Y, NR¹⁰CO₂R¹⁰, NR¹⁰C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR¹⁰SO₂NR¹⁰Rⁿ, nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃,

S(O)₂CF ₃₃,, _^ SO₂₂N-^R¹¹⁰⁰RR^uU,, SSOO₃₃RR¹¹⁰⁰,, PPOO₃₃IR¹⁰R^U, or halo, and on any position; R¹⁰, R^u, and R¹²are, independently, selected from the group consisting of hydrogen, C_j_₁₀ alkyl, C_U10 alkanoyl, C_2.10 alkenyl, C₂_₁₀ alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C₀_₆ alkylheterocyclyl, and C_0.6 alkylheteroaryl; or R¹⁰ and R¹¹, taken together with the nitrogen to which they are attached, forms a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C,_.6 alkyl or (CH₂)_0.3aryl.

This invention also covers pharmaceutically acceptable inorganic or organic salts, esters, and other prodrugs of formula (I).

As used herein, "alkyl" refers to an optionally substituted hydrocarbon group joined together by single carbon-carbon bonds. The alkyl hydrocarbon group may be linear, branched or cyclic, saturated or unsaturated. Preferably, the group is saturated linear or cyclic.

The term "alkanoyl " is used herein at all occurrences to mean a C(O)alkyl group, wherein the alkyl portion is as defined below, including, but not limited to, acetyl, pivaloyl, and the like.

The term "alkenyl" is used herein at all occurrences to mean a straight or branched chain radical, wherein there is at least one double bond between two of the carbon atoms in the chain, including, but not limited to, ethenyl, 1-propenyl, 2-propenyl, 2-methyl- 1-propenyl, 1-butenyl, 2-butenyl, and the like.

The term "alkoxy" is used herein at all occurrences to mean a straight or branched chain radical bonded to an oxygen atom, including, but not limited to, methoxy, ethoxy, n- propoxy, isopropoxy, and the like. The term "alkylaryl" is used herein at all occurrences to mean an aryl group as defined below attached to an alkyl group as defined above, including, but not limited to, benzyl and phenethyl, and the like. ( The term "alkylheterocyclyl" is used herein at all occurrences to mean a heterocyclic group as defined below attached to an alkyl group as defined above, including, but not limited to, (tetrahydro-3-furanyl)methyl and 3-(4- morpholinyl)propyl, and the like.

The term "alkylheteroaryl" is used herein at all occurrences to mean a heteroaryl group as defined below attached to an alkyl group as defined above, including, but not limited to, 3-(furanyl)methyl and (2-pyridinyl)propyl, and the like.

The term "alkynyl" is used herein at all occurrences to mean a straight or branched chain radical, wherein there is at least one triple bond between two of the carbon atoms in the chain, including, but not limited to, acetylene, 1- propylene, 2- propylene, and the like.

The term "aralkyl" is used herein at all occurrences to mean an aryl moiety as defined below, which is connected to an alkyl moiety as defined above, including, but not limited to, benzyl or phenethyl, and the like.

The term "aryl" is used herein at all occurrences to mean 6-14-membered substituted or unsubstituted aromatic ring(s) or ring systems which may include bi- or tri-cyclic systems, including, but not limited to phenyl, naphthalenyl, biphenyl, phenanthryl, anthracenyl, and the like.

The term "aryloxy" is used herein at all occurrences to mean an aryl group as defined above linked via an oxy group, including, but not limited to, phenoxy, and the like.

The terms "cycloalkyl" is used herein at all occurrences to mean cyclic radicals, which may be mono- or bicyclo- fused ring systems which may additionally include unsaturation, including, but not limited to, cyclopropyl, cyclopentyl, cyclohexyl, 1,2,3,4-tetrahydronaphthalenyl, and the like.

The terms "halo" or "halogen" are used interchangeably herein at all occurrences to mean radicals derived from the elements chlorine, fluorine, iodine and bromine. The term "heteroaryl" is used herein at all occurrences to mean a 5-14- membered substituted or unsubstituted aromatic ring(s) or ring systems which may include bi- or tri-cyclic systems, which ring or ring systems contain 1 to 4 heteroatoms selected from nitrogen, which may be optionally substituted with hydrogen or Cι_galkyl, oxygen, and sulfur, including, but not limited to, indolyl, benzofuranyl, thianaphthenyl, quinolyl, isoquinolyl, pyrrolyl, furanyl, thienyl, pyridyl, and the like.

The term "heteroaryloxy" is used herein at all occurrences to mean an heteroaryl group as defined above linked via an oxy group, including, but not limited to, 2-pyridinyloxy, and the like.

The term "heterocyclic" is used herein at all occurrences to mean a saturated or wholly or partially unsaturated 5-10-membered ring system (unless the cyclic ring system is otherwise limited) in which one or more rings contain one or more heteroatoms selected from nitrogen, which may be optionally substituted with hydrogen or Cj.galkyl, oxygen, and sulfur, including, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, imidazolidine, pyrazolidine, 1,2,3,6- tetrahydropyridine, hexahydroazepine, and the like.

Preferred compounds useful in the present invention include:

4-Ureido-[2,3]bithiophenyl-5-carboxylic acid amide 5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid amide 5-(4-Fluoro-phenyl)-3-ureido-thiophene-2-carboxylic acid amide 5-Phenyl-3-ureido-thiophene-2-carboxylic acid methyl ester l-(2-Acetyl-5-phenyl-thiophen-3-yl)-3-methyl-urea 4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid amide 5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid amide 3-(3-Methyl-ureido)-5~phenyl-thiophene-2-carboxylic acid methyl ester 5-(4-Fluoro-phenyl)-3-ureido-thiophene-2-carboxylic acid methyl ester (2-Acetyl-5-phenyl-thiophen-3-yl)-urea

5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide 5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid methylamide 5-(3 ,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide 4-(3-Methyl-ureido)-[2,3 ^bithiophenyl-5-carboxylic acid methylamide -(3-Ethyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid methylamide -biuret-[2,3']bithiophenyl-5-carboxylic acid methylamide - 4-Amino-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide - 3-Hydroxymethyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid methylamide - 3-Memyl-ureido)-[2,3^bithiophenyl-5-carboxy lie acid phenylamide - 3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid benzo[l,3]dioxol-5-ylamide 3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (4-methoxy-phenyl)-amide - 3-Methyl-ureido)-[2,3^:]bithiophenyl-5-carboxy: lie acid (2-hydroxy-phenyl)-amide -^ι 3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy: lie acid ethylamide -(3-Methyl-ureido)-[2,3^bithiophenyl-5-carboxy lie acid propylamide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid isopropylamide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy: lie acid isobutyl-amide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid pyrrolidin-3-ylamide -(3-Methyl-ureido)-[2,3^bithiophenyl-5-carboxy lie acid (2-methoxy-ethyl)-amide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (2-amino-ethyl)-amide ~(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (3-dimethylamino-propyl)-amide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (3-amino-2-hydroxy-propyl)-amid -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (pyridin-4-ylmethyl)-amide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid benzylamide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxy lie acid (furan-2-ylmethyl)-amide -(3-Methyl-ureidoH2,3]bithiophenyl-5-carboxy lie acid (lH-imidazol-2-ylmethyl)-amide -(3 -Methyl-ureido)- [2 ,3 ]bithiophenyl-5 -carbox lie acid (2-morpholin-4-yl-ethyl)-amide

More preferred compounds useful in the present invention include: -Ureido-[2,3']bithiophenyl-5-carboxylic acid amide -(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid amide -(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid amide -(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid amide -(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide -(3 ,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide -(3-Methyl-ureido)-[2,3 lbithiophenyl-5-carboxylic acid methylamide -(3-Methyl-ureido)-[2,3 'Jbithiophenyl-S-carboxylic acid phenylamide -(3-Methyl-ureido)-[2,3 lbithiophenyl-5-carboxylic acid benzo[ 1 ,3]dioxol-5-ylamide -(3-Methyl-ureido)-[2,3 ^bithiophenyl-S-carboxylic acid (2-amino-ethyl)-amide The compounds of the invention can exist in unsolvated as well as solvated forms, including hydrated forms. In general, the solvated forms, with pharmaceutically acceptable solvents such as water, ethanol, and the like, are equivalent to the unsolvated forms for purposes of this invention.

The compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All of these compounds and diastereomers are contemplated to be within the scope of the present invention.

Geometric isomers and tautomers of the present compounds are also within the scope of the present invention.

The present compounds can also be formulated as pharmaceutically acceptable salts and complexes thereof. Pharmaceutically acceptable salts are non- toxic salts in the amounts and concentrations at which they are administered.

Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p- toluenesulfonate, cyclohexylsulfamate and quinate. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present.

Thiophenes of formula (I) can be prepared readily from various commercially-available 3-aminothiophenes. Treatment of the 3-aminothiophene with an isocyanate, for example methyl isocyanate or chlorosulfonyl isocyanate, under standard conditions, for example in pyridine or dichloromethane from 23-50

°C, affords the corresponding 3-ureidothiophene (Scheme I). Treatment of the 3- aminothiophene with a carbamic ester (for example, ethyl allophanate) in a solvent such as dimethylsulfoxide at elevated temperatures such as 80-100 °C can also provide 3-ureidothiophene derivatives (Scheme II). More diverse urea compounds can be prepared using standard alkylation methods, such as the use of an aldehyde (for example, formaldehyde) in pyridine, of a 3-ureidothiophene as indicated in Scheme in.

Scheme I

Scheme II

Scheme III

Alternatively, the 3-aminothiophenes can be prepared following published procedures (for example, WO200158890A1) starting from commercially-available aryl methyl ketones and mercapto derivatives (for example, mercapto acetic acid methyl ester and mercapto methyl acetamide; Scheme IN). The corresponding 3- ureidothiophenes can then be prepared as stated above.

Scheme IV

Further elaboration of the R2 position can occur starting from a suitable 3- ureidothiophene precursor, such as a 3-(3-methyl-ureido)-5-aryl-thiophene-2- carboxylic acid. Utilizing known amine coupling methods with the carboxylic acid, for example the use of BOP and an amine in the presence of a base, for example triethylamine, in a solvent, such as acetonitrile, will lead to variously-substituted 2- amido-3-ureidothiophenes (Scheme V).

Scheme V

The following experimentals are intended to illustrate the embodiments of the present invention only, but not be limiting in any way.

Example 1

Preparation of 5-phenyl-3-ureido-thiophene-2-carboxylic acid methyl ester

To a room temperature solution of 3-amino-5-phenyl-2-thiophene carboxylic acid methyl ester (0.20 mmol) in dichloromethane (1.0 mL) was added chlorosulfonyl isocyanate (0.22 mmol). The reaction mixture was stirred for 4 h, quenched with the addition of water (0.5 mL), and then stirred overnight. After concentrating in vacuo, the reaction mixture was dissolved in DMSO and purified by Gilson reverse phase HPLC to afford the title product. 1H NMR (400 MHz, CDC1₃) δ 9.56 (br s, 1H), 8.26 (s, 1H), 7.68 (d, 2H, J = 8.1 Hz), 7.40 (m, 3H), 4.81 (br s, 2H), 3.89 (s, 3H). ES S [M+H]⁺: 276.2. 3/028731

Example 2

Preparation of l-(2-acetyl-5-phenyl-thiophen-3-yl)-3-methyl-urea

To a room temperature solution of 2-acetyl-3-amino-5-phenyl-thiophene (0.23 mmol) in pyridine (1.0 mL) was added methyl isocyanate (0.42 mmol). The reaction mixture was stirred 40 h at 50 °C, cooled, and then diluted with methanol and concentrated in vacuo. The reaction mixture was dissolved in DMSO and purified by Gilson reverse phase HPLC to provide the title compound. 1H NMR (400 MHz, OMSO-dβ) δ 10.20 (br s, IH), 8.36 (s, IH), 7.72 (d, 2H, J= 6.8 Hz), 7.68 (br s, IH), 7.49 (m, 3H), 2.66 (d, 3H, J= 4.4 Hz), 2.44 (s, 3H). ESLMS [M+H]⁺: 275.0.

Example 3

Preparation of (2-acetyl-5-phenyl-thiophen-3-yl)-urea

Following the procedure described in Example 1 with 2-acetyl-3-amino-5- phenyl-thiophene provided the title compound. ESIMS [M+H]⁺: 261.2.

Example 4

Preparation of 4-(3-methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid amide

To a room temperature solution of 4-amino-[2,3 bithiophenyl-5- carboxamide (0.22 mmol) in pyridine (1.0 mL) was added methyl isocyanate (0.42 mmol). The reaction mixture was stirred overnight and then diluted with methanol and concentrated in vacuo. The reaction mixture was dissolved in DMSO and purified by Gilson reverse phase HPLC to provide the title compound. 1H NMR (400 MHz, OMSO-d₆) δ 10.16 (br s, IH), 8.11 (s, IH), 7.82 (dd, IH, J= 2.9, 1.3 Hz), 7.69 (dd, IH, J = 5.0, 2.9 Hz), 7.39 (br s, 3H), 7.36 (dd, IH, J= 5.0, 1.4 Hz), 2.63 (3, 3H, J = 4.5 Hz). ESIMS [M+H]⁺: 283.2.

Example 5 Preparation of 5-(4-methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid amide

Following the procedure described in Example 4 with 3-amino-5-(4- methoxy-phenyι)-2-thiophene carboxamide provided the title compound. 1H NMR (400 MHz, DMSO- e) δ 10.17 (br s, IH), 8.13 (s, IH), 7.56 (d, 2H, J= 8.7 Hz), 7.38 (br s, 3H), 7.03 (d, 2H, J = 8.8 Hz), 3.80 (s, 3H), 2.63 (d, 3H, J = 4.4 Hz). ES S [M+H]⁺: 306.0.

Example 6

Preparation of 3-(3-methyl-ureido)-5-phenyl-thiophene-2-carboxylic acid methyl ester

Following the procedure described in Example 2 with 3-amino-5-ρhenyl-2- thiophene carboxylic acid methyl ester provided the title compound. ESIMS [M+H]⁺: 291.0.

Example 7

Preparation of 4-ureido-r2,31bithiophenyl-5-carboxylic acid amide

Following the procedure described in Example 1 with 4-amino- [2,3]bithiophenyl-5-carboxamide provided the title compound. Η NMR (400 MHz, CD₃OD) δ 8.08 (s, IH), 7.70 (dd, IH, J = 2.9, 1.3 Hz), 7.53 (dd, IH, J = 5.1, 2.9 Hz), 7.39 (dd, IH, 7 = 5.1, 1.3 Hz).

3/028731

Example 8

Preparation of 5-(4-methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid amide

Following the procedure described in Example 1 with 3-amino-5-(4- methoxy-phenyl)-2-thiophene carboxamide provided the title compound. Η NMR (400 MHz, DMSO-d₆) δ 10.07 (s, IH), 8.12 (s, IH), 7.55 (d, 2H, J= 8.8 Hz), 7.38 (br s, 2H), 7.03 (d, 2H, J= 8.8 Hz), 6.61 (br s, 2H), 3.80 (s, 3H).

Example 9

Preparation of 5-(4-fluoro-phenyl)-3-ureido-thiophene-2-carboxylic acid amide

Following the procedure described in Example 1 with 3-amino-5-(4-fluoro- phenyl)-2-thiophene carboxamide provided the title compound. Η NMR (400 MHz, DMSO-d₆) δ 10.06 (s, IH), 8.21 (s, IH), 7.67 (m, 2H), 7.46 (br s, 2H), 7.31 (m, 2H), 6.67 (br s, 2H).

Example 10

Preparation of 5-(4-fluoro-ρhenyl)-3-ureido-thiophene-2-carboxylic acid methyl ester

Following the procedure described in Example 1 with 3-amino-5-(4-fluoro- phenyl)-thiophene-2-carboxylic acid methyl ester provided the title compound. ESIMS [M+H]⁺: 295.2.

Example 11

Preparation of 4-biuret-r2.31bithiophenyl-5-carboxylic acid methylamide

a) 4-Amino-[2,3']bithiophenyl-5-carboxylic acid methylamide

Phosphorous oxy chloride (17 mmol) was added to an ice-cooled solution of N,N-dimethylformamide (3.0 mL). After 35 min, l-thiophen-3-yl-ethanone (7.9 mmol) was added portionwise and the resulting mixture was heated briefly at 50 °C until the solution was homogeneous. Hydroxylamine hydrochloride (36 mmol) was added slowly to the reaction mixture at room temperature. The reaction mixture was stirred for 30 min, quenched with the addition of water, and stirred for an additional 30 min. The reaction mixture was poured into water and the organics were extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate, were filtered, and were concentrated by rotary evaporation. The crude oil was diluted with methanol (5.0 mL). 2-Mercapto-N-methyl-acetamide (12 mmol) and sodium methoxide (12 mmol of a 25% solution in methanol) were sequentially added and the reaction mixture was heated to reflux. After 6 h, the cooled reaction mixture was quenched with water, poured into water, and the organics were extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate, were filtered, and were concentrated. The residue was purified by flash chromatography (20-60% ethyl acetate hexanes) to provide the title compound (25%). ESIMS [M+H]⁺: 239.2.

b) 4-Biuret-[2,3^:]bithiophenyl-5-carboxylic acid methylamide

Ethyl allophanate (0.23 mmol) was added to a solution of 4-amino- [2,3']bithiophenyl-5-carboxylic acid methylamide (0.21 mmol) in dimethylsulfoxide (1.0 mL). After stirring at 80 °C for 65 h and 100 °C for 43 h, the reaction mixture was purified directly by Gilson reverse phase HPLC to afford the title product in low yield. ESIMS [M+H]⁺: 325.2.

Example 12

Preparation of 4-(3-Hydroxymethyl-ureido)-r2,3'1bithiophenyI-5-carboxylic acid methylamide

a) 4-Ureido-[2,3']bithiophenyl-5-carboxylic acid methylamide

Following the procedure of Example 1 with 4-amino-[2,3 bithiophenyl-5- carboxylic acid methylamide afforded the title compound._, ESIMS [M+H]⁺: 282.0.

b) 4-(3-Hydroxymethyl-ureido)-[2,3]bithiophenyl-5-carboxylic acid methylamide To a solution of 4-ureido-[2,3^:]bithiophenyl-5-carboxylic acid methylamide (0.060 mmol) in pyridine (0.5 mL) was added a 37% solution of formaldehyde (0.010 mL). The reaction mixture was heated at 50 °C for 18 h, then cooled, concentrated in vacuo, dissolved in DMSO, and purified by Gilson reverse phase HPLC to afford the title product in low yield. ESIMS [M+H]⁺: 312.2.

Example 13

Preparation of 5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

a) 3-Amino-5-(4-methoxy-phenyl)-thiophene-2-carboxylic acid methylamide

Using the procedure of Example 11(a) but replacing, l-thiophen-3-yl- ethanone with 4-methoxy-acetophenone provided the title compound.

b) 5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

Following the procedure of Example 4 with 3-amino-5-(4-methoxy-phenyl)- thiophene-2-carboxylic acid methylamide provided the title compound. ESIMS [M+H]⁺: 291.0.

Example 14

Preparation of 5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid methylamide

Following the procedure of Example 1 with 3-amino-5-(4-methoxy-phenyl)- thiophene-2-carboxylic acid methylamide provided the title compound. ESIMS [M+H]⁺: 306.0.

Example 15

Preparation of 5-(3,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2- carboxylic acid methylamide a) 3-Amino-5-(3,4-dimethoxy-phenyl)-thiophene-2-carboxylic acid methylamide

Using the procedure of Example 11(a) but replacing l-thiophen-3-yl- ethanone with 3,4-dimethoxy-acetophenone provided the title compound.

b) 5-(3,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

Following the procedure of Example 4 with 3-amino-5-(3,4-dimethoxy- phenyl)-thiophene-2-carboxylic acid methylamide provided the title compound. ESIMS [M+H]⁺: 350.0.

Example 16

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid methylamide

Following the procedure of Example 4 with 4-amino-[2,3']bithiophenyl-5- carboxylic acid methylamide provided the title compound. ESIMS [M+H]⁺: 296.0.

Example 17

Preparation of 4-(3-Ethyl-ureido)-r2,31bithiophenyl-5-carboxylic acid methylamide

Following the procedure of Example 16 with ethyl isocyanate provided the title compound. ESIMS [M+H]⁺: 310.2.

Example 18

Preparation of 5-(4-Amino-phenyl -3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

a) 3-Amino-5-(4-nitro-phenyl)-thiophene-2-carboxylic acid methylamide

Using the procedure described in Example 11(a) but replacing l-thiophen-3- yl-ethanone with 4-nitroacetophenone provided the title compound. b) 5-(4-Nitro-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

Following the procedure of Example 2 with 3-amino-5-(4-nitro-phenyl)- thiophene-2-carboxylic acid methylamide overnight provided the title compound. ESIMS [M+H]⁺: 334.8.

c) 5-(4-Amino-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

Palladium (22 mg of 10% Pd/C) was added to a solution of 5-(4-nitro- phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide (0.066 mmol) in ethanol (5.0 mL). The reaction mixture was degassed and backfilled with argon three times and then degassed and backfilled with hydrogen gas (balloon) three times. The reaction mixture was then stirred at room temperature under a hydrogen balloon for 2h. The reaction mixture was filtered through Celite and rinsed three times with ethanol and three times with ethyl acetate. The combined filtrate was concentrated in vacuo, dissolved in DMSO, filtered, and purified by Gilson reverse phase HPLC to afford the title product. ESIMS [M+H]⁺: 305.0.

Example 19

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid phenylamide

Following the procedure described in Example 21(d) with aniline provided the title compound. ESIMS [M+H]⁺: 358.0.

Example 20

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid benzo \ 1 , 31 dioxol-5 -y lamide

Following the procedure described in Example 21(d) with benzo[l,3]dioxol- 5-ylamine provided the title compound. ESIMS [M+H]⁺: 402.2. Example 21

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid (4-methoxy- phenyP-amide

a) 4-Amino-[2,3']bithiophenyl-5-carboxylic acid methyl ester

Using the procedure described in Example 11(a) but replacing N-methyl mercaptoacetamide with methyl mercaptoacetate provided the title compound.

b) 4-Amino-[2,3']bithiophenyl-5-carboxylic acid

4-Amino-[2,3']bithiophenyl-5-carboxylic acid methyl ester (17 mmol) was suspended in IN aqueous lithium hydroxide solution (150 mL). The reaction mixture was heated at 70 °C for 24 h. The reaction mixture was then cooled to room temperature and extracted three times with ethyl acetate. The combined extracts were dried over MgSO₄, filtered, and concentrated in vacuo to yield 1.2 g of the title compound. The aqueous layer was neutralized with 12 M HC1 (1 mL) and desired product precipitated out of solution. The white solid was filtered and dried to yield 1 g of the desired product. The remaining aqueous filtrate was extracted three times with ethyl acetate, dried over MgSO₄, filtered and concentrated in vacuo to yield another 1.1 g of the title compound. The combined products were concentrated in vacuo to yield a total of 3 g of desired product (88%) which was used directly in the next reaction without further purification. ESIMS [M+H]⁺: 227.0.

c) 4-(3-Methyl-ureido)-[2,31bithiophenyl-5-carboxylic acid

Following the procedure described in Example 2 with 4-amino- [2,31bithiophenyl-5-carboxylic acid for overnight provided the title compound.

d) 4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid (4-methoxy-phenyl)- amide

4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid (0.17 mmol) and benzotriazole-l-yl-oxy-tris-(dimethylamino)-phophoniumhexafluorophosphate (BOP, 0.17 mmol) were dissolved together in acetonitrile. This mixture was then treated with -methoxyaniline (0.51 mmol) and triethylamine (0.85 mmol). The reaction mixture was stirred for 24 h at room temperature. The crude mixture was concentrated in vacuo, dissolved in DMSO, filtered, and purified by Gilson reverse phase HPLC to afford the title product. ESIMS [M+H]⁺: 388.2.

Example 22

Preparation of 4-(3-Methyl-ureido)-r2,3Hbithiophenyl-5-carboxyric acid (2-hydroxy- phenyD-amide

Following the procedure described in Example 21(d) with 2-amino-phenol provided the title compound. ESIMS [M+H]⁺: 374.2.

Example 23

Preparation of 4-(3-Methyl-ureido)-r2.31bithiophenyl-5-carboxylic acid ethylamide

Following the procedure described in Example 21(d) with ethylamine provided the title compound. ESIMS [M+H]⁺: 310.0.

Example 24

Preparation of 4-(3-Methyl-ureido)-r2.31bithiophenyl-5-carboxylic acid propylamide

Following the procedure described in Example 21(d) with n-propylamine provided the title compound. ESIMS [M+H]⁺: 323.6.

Example 25

Preparation of 4-(3-Methyl-ureido - r2.3Hbithiophenyl-5-carboxylic acid isopropylamide Following the procedure described in Example 21(d) with isopropylamine provided the title compound. ESIMS [M+H]⁺: 324.4.

Example 26

Preparation of 4-(3-Methyl-ureido)-[2,31bithiophenyl-5-carboxylic acid isobutyl- amide

Following the procedure described in Example 21(d) with isobutylamine provided the title compound. ESIMS [M+H]⁺: 338.2.

Example 27

Preparation of 4-(3-Methyl-ureido)-[2,31bithiophenyl-5-carboxylic acid pyrrolidin- 3-ylamide

Following the procedure described in Example 21(d) with 3- aminopyrrolidine provided the title compound. ESIMS [M+H]⁺: 350.8.

Example 28

Preparation of 4-(3-Methyl-ureido)-r2₁31bithiophenyl-5-carboxylic acid (2-methoxy- ethvD-amide

Following the procedure described in Example 21(d) with 2- methoxyethylamine provided the title compound. ESIMS [M+H]⁺: 340.0.

Example 29

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid (2-amino- e hvP-amide

Following the procedure described in Example 21(d) with ethylenediamine provided the title compound. ESIMS [M+H]⁺: 324.8. Example 30

Preparation of 4-(3-Methyl-ureido)-r2,3Hbithiophenyl-5-carboxylic acid (3- dimethylamino-propyD-amide

Following the procedure described in Example 21(d) with 3- dimethylaminopropylamine provided the title compound. ESIMS [M+H]⁺: 367.0.

Example 31

Preparation of 4-(3-Methyl-ureido)-r2,3'1bithiophenyl-5-carboxylic acid (3-amino-2- hydroxy-propyD-amide

Following the procedure described in Example 21(d) with l,3-diamino-2- propanol provided the title compound. ESIMS [M+H]⁺: 355.0.

Example 32

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid (pyridin-4- ylmethyP-amide

Following the procedure described in Example 21(d) with 4- (aminomethyl)pyridine provided the title compound. ESIMS [M+H]⁺: 373.0.

Example 33

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid benzylamide

Following the procedure described in Example 21(d) with benzylamine provided the title compound. ESIMS [M+H]⁺: 372.0.

Example 34

Preparation of 4-(3-Methyl-ureido -r2,3Hbithiophenyl-5-carboxyric acid (furan-2- ylmethyD-amide Following the procedure described in Example 21(d) with furfurylamine provided the title compound. ESIMS [M+H]⁺: 362.0.

Example 35

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid (1H- imidazol-2-ylmethyl)-amide

Following the procedure described in Example 21(d) with (lΗ-imidazole-2- yl)-methylamine provided the title compound. ESIMS [M+H]⁺: 362.0.

Example 36

Preparation of 4-(3-Methyl-ureido)-r2,31bithiophenyl-5-carboxylic acid (2- morpholin-4-yl-ethyl)-amide

Following the procedure described in Example 21(d) with N-(2- aminoethyl)morpholine provided the title compound. ESIMS [M+H]⁺: 395.0.

With appropriate manipulation and protection of any chemical functionality, synthesis of the remaining compounds of Formula (I) is accomplished by methods analogous to those above.

In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the treatment of humans and other mammals, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.

Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients.

Examples of such formulations are given below:

The present ligands can be administered by different routes including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, topical, transdermal, or transmucosal administration. For systemic administration, oral administration is preferred. For oral administration, for example, the compounds can be formulated into conventional oral dosage forms such as capsules, tablets and liquid preparations such as syrups, elixirs and concentrated drops.

Alternatively, injection (parenteral administration) may be used, e.g., intramuscular, intravenous, intraperitoneal, and subcutaneous. For injection, the compounds of the invention are formulated in liquid solutions, preferably, in physiologically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution. In addition, the compounds may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms can also be produced.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, bile salts and fusidic acid derivatives. In addition, detergents may be used to facilitate permeation. Transmucosal administration, for example, may be through nasal sprays, rectal suppositories, or vaginal suppositories.

For topical administration, the compounds of the invention can be formulated into ointments, salves, gels, or creams, as is generally known in the art. The amounts of various compounds to be administered can be determined by standard procedures taking into account factors such as the compound IC₅₀, EC_J0, the biological half-life of the compound, the age, size and weight of the patient, and the disease or disorder associated with the patient. The importance of these and other factors to be considered are known to those of ordinary skill in the art.

Amounts administered also depend on the routes of administration and the degree of oral bioavailability. For example, for compounds with low oral bioavailability, relatively higher doses will have to be administered.

Preferably the composition is in unit dosage form. For oral application, for example, a tablet, or capsule may be administered, for nasal application, a metered aerosol dose may be administered, for transdermal application, a topical formulation or patch may be administered and for transmucosal delivery, a buccal patch may be administered. In each case, dosing is such that the patient may administer a single dose.

Each dosage unit for oral administration contains suitably from 0.01 to 500 mg/Kg, and preferably from 0.1 to 50 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, calculated as the free base. The daily dosage for parenteral, nasal, oral inhalation, transmucosal or transdermal routes contains suitably from 0.01 mg to 100 mg/Kg, of a compound of Formula(I). A topical formulation contains suitably 0.01 to 5.0% of a compound of Formula (I).

The active ingredient may be administered from 1 to 6 times per day, preferably once, sufficient to exhibit the desired activity, as is readily apparent to one skilled in the art.

As used herein, "treatment" of a disease includes, but is not limited to prevention, retardation and prophylaxis of the disease. As used herein, "diseases" treatable using the present compounds include, but are not limited to leukemias, solid tumor cancers and metastases, lymphomas, soft tissue cancers, brain cancer, esophageal cancer, stomach cancer, pancreatic cancer, liver cancer, lung cancer, bladder cancer, bone cancer, prostate cancer, ovarian cancer, cervical cancer, uterine cancer, testicular cancer, kidney cancer, head cancer and neck cancer, chronic inflammatory proliferative diseases such as psoriasis and rheumatoid arthritis; proliferative cardiovascular diseases such as restenosis; prolifertive ocular disorders such as diabetic retinopathy; and benign hyperproliferative diseases such as hemangiomas.

Composition of Formula (I) and their pharmaceutically acceptable salts which are active when given orally can be formulated as syrups, tablets, capsules and lozenges. A syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid carrier for example, ethanol, peanut oil. olive oil, glycerine or water with a flavoring or coloring agent. Where the composition is in the form of a tablet, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers in a hard gelatin capsule shell. Where the composition is in the form of a soft gelatin shell capsule any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils, and are incorporated in a soft gelatin capsule shell.

Typical parenteral compositions consist of a solution or suspension of a compound or salt in a sterile aqueous or non-aqueous carrier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpyrrolidone, lecithin, arachis oil or sesame oil.

Typical compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional propellant such as dichlorodifluoromethane or trichlorofluoromethane.

A typical suppository formulation comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent, for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogs.

Typical dermal and transdermal formulations comprise a conventional aqueous or non-aqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane.

Preferably the composition is in unit dosage form, for example a tablet, capsule or metered aerosol dose, so that the patient may administer a single dose.

No unacceptable toxological effects are expected when compounds of the present invention are administered in accordance with the present invention.

The biological activity of the compounds of Formula (I) are demonstrated by the tests indicated hereinbelow. Chkl Kinase Assay:

Compounds capable of inhibiting Chkl kinase can be identified with in vitro assays and cellular assays as described below. Variations of these assays would be obvious to those skilled in the art. Streptavidin coated SPA beads, ATP and ³³P-ATP were obtained from Amersham Pharmacia Biotech, Biotin labeled peptide KNSRSGLYRSPSMPENLNRK(Biotin-xx)NH₂ was obtained from Affiniti Research Products Ltd, assay buffer reagents were obtained from Sigma- Aldrich Co.Ltd. fff84 well assay plates were obtained from Corning Inc. Assay buffer: 50 mM HEPES, 50 mM KC1, 5% Glycerol, 1 mM EGTA, 0.001% Tween-20; enzyme/peptide mix: 25 nM Chkl, 2.5μM biotin peptide, 7.5 mM 2- mercaptoethanol in assay buffer; ATP mix: 20μM ATP at 650kBq/mL, 5mM MgCl₂ in assay buffer.

Inhibitors of decreasing concentration, from lOμM were incubated at room temperature for 1 hour together with 5μL enzyme/peptide mix and 5μL ATP mix. The reaction was stopped with 5μL of 0.5M EDTA followed by a further addition of 65μL of 0.2mg/mL SPA beads. Plates were spun at 2500 rpm for 10 minutes and the amount of ³³P incorporated onto the peptide was quantified by a Wallac Trilux scintillation counter at a read time of 1 minute per well. IC50's were fitted to the data using SDM Explorer version 2.5 software (©GlaxoSmithKline Pic).

Expression of GST-Chkl:

A GST-Chkl expression construct was constructed which has the glutathione-S-transf erase gene fused to the amino terminus of Chkl kinase via a linker containing a thrombin cleavage site. This construct was cloned into the Baculovirus expression vector, pFASTBAC, and this was used to make the viral stock for the subsequent infection. Spodoptera frugiperda cells (Sf9) were infected with the virus expressing the GST-Chkl and the cells were grown for 3 days, then harvested and frozen down.

Purification of GST-Chkl:

The GST-Chkl protein was purified as follows: An Sf9 cell pellet expressing GST-Chkl was resuspended on ice in lysis buffer (50mM Tris-Cl, pH 7.5, 250mM NaCl2, lmM dithiothreitol (DTT), 0.1%Brij, 5% (v/v) protease inhibitor cocktail, lmM sodium orthovanadate), cells were lysed by sonication and centrifuged at 100,000xg for 30min The supernatant was added to Glutathione Sepharose 4B, beads, equilibrated in wash buffer (20mM Tris-Cl, pH 7.0, lOmM MgCl2, lOOmM NaCl2, lmM DTT, 0.5%(v/v) protease inhibitor cocktail, lmM sodium orthovanadate). The mixture was rocked for 30min The resin with the bound GST- Chkl was spun down at 500xg for 5min and washed with 14mls of wash buffer. The beads were spun as above and resuspended in another 14mls of wash buffer. The suspension was transferred into a column and allowed to pack, then the wash buffer was allowed to flow through by gravity. The GST-Chkl was eluted from the column with lOmM Glutathione in 50mM Tris-Cl, pH 8.0 in 500ul fractions. Protein concentrations were determined on the fractions using Bio-Rad's Protein assay kit as per instructions. Fractions containing the GST-Chkl were pooled and diluted to a concentration of ~0.5mg/ml and dialyzed for 4 hours at 4^C in dialysis buffer (20mM HEPES, pH 7.0, lmM Manganese Acetate, lOOmM NaCl₂, 0.05% Brij-35, 10% glycerol, lmM DTT, 0.2% (v/v) protease inhibitor cocktail, lmM sodium orthovanadate). The protein was aliquoted and stored at -80^.

Cell Cycle Studies:

Drug studies considering cellular effects were performed in the Hela S3 adherent cell line. Cells were plated at a concentration sufficiently low such that 24 hours later they were at 10-20% confluence (typically 2x10^ cells/15cm e3). Cells were then synchronized in S phase by a repeated thymidine block. Briefly, cells were treated with 2mM thymidine for 18hours, released for 8 hours by 3 washes, and then treated again with thymidine. Following the second release from thymidine, 95% of cells were in S phase. Synchronized cells were then returned to complete media containing a DNA-damaging drug such as 50nM topotecan (a dosage we have found to be sufficient to arrest cells in early G2 phase without inducing apoptosis) alone and in combination with test compounds for up to 18 hours. Cell cycle profiles were then performed cytometrically using a procedure for propidium iodide staining of nuclei. (Vindelov et al, Cytometry Vol.3, No.5, 1983, 323-327) CHK1 inhibitors would be expected to reverse the G2 arrest caused by the DNA damaging agent. Typical concentration ranges for such activity would be 0.001 to 10 uM. Proliferation/Apoptosis Studies:

Proliferation studies were performed in a variety of adherent and non- adherent cell lines including Hela S3, HT29, and Jurkat. The proliferation assay utilized a colorimetric change resulting from reduction of the tetrazolium reagent XTT into a formazan product by metabolically active cells (Scudiero et al. Cancer Research, 48, 1981, 4827-4833) Cells were seeded in lOOul in 96 well plates to roughly 10% confluence (cell concentration varied with cell lines) and grown for 24 hours. Compounds were then added with or without sufficient vehicle- containing media to raise the cells to a 200ul final volume containing chemical reagents in 0.2% DMSO. Cells received multiple concentrations of DNA-damaging anti-proliferative drugs such as topotecan, test compounds, and combination treatment at 37°C 5% CO₂. 72 hours later, 50 uls of an XTT/ phenazine methosulfate mixture were added to each well and cells were left to incubate for 90mins. Plate was read at 450nm, and anti-proliferative effects were compared relative to vehicle treated cells. CHK1 inhibitors are expected to enhance the cytotoxicity of DNA-damaging chemotherapeutic drugs. Typical concentration ranges for such activity would be 0.001 to 10 uM. Other assays for cellular proliferation or cytotoxicity could also be used with test compounds, and these assays are known to those skilled in the art.

Ikk-β Kinase Assay

IKK-β was expressed as a GST-tagged protein, and its activity was assessed in a 96-well scintillation proximity assay (SPA). Briefly, IKK-β was diluted in assay buffer (20 mM Hepes, pH 7.7, 2 mM MgCl₂, 1 mM MnCl₂, 10 mM β- glycerophosphate, 10 mM NaF, 10 mM PNPP, 0.3 mM Na3NO4, 1 mM benzamidine, 2 μM PMSF, 10 μg/ml aprotinin, 1 ug/mL leupeptin, 1 ug/mL pepstatin, lmM DTT; 20 nM final), with various concentrations of compound or DMSO vehicle, 240 nM ATP and 200 nCi [^»-³³P]-ATP (10 mCi/mL, 2000 Ci/mmol; ΝEΝ Life Science Products, Boston, MA). The reaction was started with the addition of a biotinylated peptide comprising amino acids 15 - 46 of IκB-α (American Peptide) to a final concentration of 2.4 μM, in a total volume of 50 uL. The sample incubated for one hour a 30 °C, followed by the addition of 150 uL of stop buffer (PBS w/o Ca²+, Mg ²+ 0.1% Triton X-100 (v/v), 10 mM EDTA) containing 0.2 mg streptavidin-coated SPA PNT beads (Amersham Pharmacia Biotech, Piscataway, ΝJ). The sample was mixed, incubated for 10 min. at room temperature, centrifuged (1000 xg, 2 minutes), and measured on a Hewlett-Packard TopCount.

All publications, including but not limited to patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference as though fully set forth.

Claims

What is claimed is:

1. A method of inhibiting angiogenesis or damage response kinase activity which comprises administering to a subject in need thereof, an effective amount of a compound according to Formula (I) hereinbelow:

(I)

wherein:

RI is selected from the group consisting of H, C^ alkyl, XH, XCH₃, C_j_₂ alkyl-XH,

C,.₂ alkyl-XCH₃, C(O)NH₂, C(O)NHCH₃, and CCO)-^ alkyl;

X is selected from the group consisting of O, S, and NH;

R2 is selected from the group consisting of C(O)R⁵, CO₂R⁵, C(O)NHR⁵,

C(O)NHC(=NH)R⁵, C(O)NHC(=NH)NR⁵R⁶, C(O)NHC(O)R⁵, C(O)NHC(O)NR⁵R⁶,

SO₂R⁵, S(O)R⁵, SO₃R⁵, and PO₃R⁵R⁶;

R⁵ and R^δ are, independently, selected from the group consisting of hydrogen, C,_.10 alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.I0 cycloalkyl, C_M alkylaryl, C_0.6 alkylheterocyclyl, and C₀₆ alkylheteroaryl, or R⁵ and R⁶, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_j_₆ alkyl or (CH₂)₀₃aryl, wherein any of the foregoing may be optionally substituted by one or more of group A and on any position;

R3 is H or halogen;

A is selected from the group consisting of C_{1 I0} alkyl, C_U10 alkanoyl, C_2.10 alkenyl, C₂,₁₀ alkynyl, C_3.10 cycloalkyl, C₀_₆ alkylaryl, C₀_₆ alkylheterocyclyl, C₀₆ alkylheteroaryl,

C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R⁸, N(O)R⁷R⁸, NR⁷R⁸R⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C,_.10 alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C₃_₁₀ cycloalkyl, C₀₆ alkylaryl, C_M alkylheterocyclyl, C₀₆ alkylheteroaryl, (CH₂)_{0 6}heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group D and on any position;

Y is an organic or inorganic anion;

D is selected from the group consisting of C,_₁₀ alkyl, C ₀ alkanoyl, C_{2 10} alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C₀₆ alkylheterocyclyl, C₀₆ alkylheteroaryl, C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R⁸, N(O)R⁷R⁸, NR⁷R^SR⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C ₀ alkyl, C ₀ alkanoyl, C_2.10 alkenyl, C_{2 10} alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C₀₆ alkylheterocyclyl, C_0.6 alkylheteroaryl, (CH₂)_0.6heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group E and on any position;

R⁷, R⁸, and R⁹ are, independently, selected from the group consisting of hydrogen, C,_.I0 alkyl, C ₁₀ alkanoyl, C₂__I0 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C₀₆ alkylaryl, C_0.6 alkylheterocyclyl, and C₀₆ alkylheteroaryl, or R⁷ and R⁸, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms, optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_j.6 alkyl or (CH₂)_0.3aryl, wherein any of the foregoing may be optionally substituted by one or more of group E and on any position;

E is selected from the group consisting of C_{l 0} alkyl, C,_.10 alkanoyl, C_{2 10} alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_M alkylaryl, C_0.6 alkylheterocyclyl, C₀₆ alkylheteroaryl, C(=NH)R¹⁰, COR¹⁰, CONR'V¹, CON(O)R¹⁰R^U, CONR¹⁰RⁿR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰Rⁿ, N(O)R¹⁰R^U, NR¹⁰R^UR¹²Y, NR¹⁰COR¹⁰, NR¹⁰CONR¹⁰R^U, NR¹⁰CON(O)R¹⁰R^U, NR¹⁰CONR¹⁰R"R¹²Y, NR¹⁰CO₂R¹⁰, NR'°C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR'⁰SO₂NR¹⁰Rⁿ, nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR¹⁰R^U, SO₃R¹⁰, PO₃R¹⁰R", and halo, wherein C ₀ alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.I0 alkynyl, C_{3 10} cycloalkyl, C₀₆ alkylaryl, C_M alkylheterocyclyl, C₀₆ alkylheteroaryl may be optionally substituted by one or more of C(=NH)R¹⁰, COR¹⁰, CONR¹⁰Rⁿ, CON(O)R¹⁰Rⁿ, CONR¹⁰R^UR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰R", N(O)R¹⁰R^U, NR'⁰R"R¹²Y, NR¹⁰COR¹⁰, NR¹⁰CONR¹⁰Rⁿ, NR¹⁰CON(O)R¹⁰R¹¹, NR¹⁰CONR¹⁰RⁿR¹²Y, NR¹⁰CO₂R¹⁰, NR¹⁰C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR^SO^R^R¹¹, nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR¹⁰R^π, SO₃R¹⁰, PO₃R¹⁰R^U, or halo, and on any position; R¹⁰, R¹¹, and R¹²are, independently, selected from the group consisting of hydrogen, C_j__I0 alkyl, C ₀ alkanoyl, C₂_₁₀ alkenyl, C_{2 10} alkynyl, C_3.10 cycloalkyl, C₀₆ alkylaryl, C_M alkylheterocyclyl, and C₀₆ alkylheteroaryl, or R¹⁰ and R", taken together with the nitrogen to which they are attached, forms a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_{1 6} alkyl or (CH₂)_0.3aryl; or a pharmaceutically acceptable inorganic or organic salt, esters, or other prodrug of formula (I).

2. A method according to claim 1 wherein R3 is H.

3. A method according to claim 2 wherein RI is H or CH₃.

4. A method according to claim 2 wherein Y is selected from the group consisting of bisulfate, chloride, fumarate, iodide, maleate, methanesulfonate, nitrate, trifluoromethanesulfonate, and sulfate.

5. A method according to claim 4 wherein the compound is selected from the group consisting of: 4-Ureido-[2,3']bithiophenyl-5-carboxylic acid amide

5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid amide

5-(4-Fluoro-phenyl)-3-ureido-thiophene-2-carboxylic acid amide

5-Phenyl-3-ureido-thiophene-2-carboxylic acid methyl ester l-(2-Acetyl-5-phenyl-thiophen-3-yl)-3-methyl-urea

4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid amide

5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid amide

3-(3-Methyl-ureido)-5-phenyl-thiophene-2-carboxylic acid methyl ester

5-(4-Fluoro-phenyl)-3-ureido-thiophene-2-carboxylic acid methyl ester

(2-Acetyl-5-phenyl-thiophen-3-yl)-urea

5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid methylamide

5-(3,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide

4-(3-Methyl-ureido)-[2,3 ']bithiophenyl-5-carboxylic acid methylamide

4-(3-Ethyl-ureido)-[2,3 ']bithiophenyl-5-carboxylic acid methylamide 4-biuret-[2,3']bithiophenyl-5-carboxylic acid methylamide

5-(4-Amino-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide 4-(3-Hydroxymethyl-ureido)-[2,3 'j^'bithiophenyl-S-carboxylic acid methylamide

4-(3-Methyl-ureido> 2,3 bithiopheny -5-carboxylic acid phenylamide 4-(3-Methyl-ureido)- 2,3']bithiopheny: -5-carboxylic acid benzo[l,3]dioxol-5-ylamide 4-(3-Methyl-ureido 2,3 bithiopheny -5-carboxylic acid (4-methoxy-phenyl)-amide 4-(3-Methyl-ureido 2,3^:]bithiopheny -5-carboxylic acid (2-hydroxy-phenyl)-amide 4-(3-Methyl-ureido 2,3']bithiopheny -5-carboxylic acid ethylamide 4-(3-Methyl-ureido 2,3']bithiopheny -5-carboxylic acid propylamide 4-(3-Methyl-ureido 2,3]bithiopheny: -5-carboxylic acid isopropylamide 4-(3-Methyl-ureido)- 2,3 bithiopheny -5-carboxylic acid isobutyl-amide 4-(3-Mefhyl-ureido 2,3']bithiopheny -5-carboxylic acid pyrrolidin-3-ylamide 4-(3-Methyl-ureido 2,3']bithiopheny -5-carboxylic acid (2-methoxy-ethyl)-amide 4-(3-Methyl-ureido)- 2,3']bithiopheny -5-carboxylic acid (2-amino-ethyl)-amide 4-(3-Methyl-ureido 2,31bithiopheny -5-carboxylic acid (3-dimethylamino-propyl)-amide 4-(3-Methyl-ureido)- 2,3']bithiopheny. -5-carboxylic acid (3-amino-2-hydroxy-propyl)-amic 4-(3-Methyl-ureido)- 2,3']bithiopheny^ -5-carboxylic acid (pyridin-4-ylmethyl)-amide 4-(3-Methyl-ureido)- 2,31bithiophen -5-carboxylic acid benzylamide

4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid (furan-2-ylmethyl)-amide

4-(3-Methyl-ureido)- 2,3']bithiophenyl-5-carboxylic acid (lH-imidazol-2-ylmethyl)-amide 4-(3-Methyl-ureido)- 2,3']bithiophenyl-5-carboxylic acid (2-morpholin-4-yl-ethyl)- amide

6. A method according to claim 4 wherein the compound is selected from the group consisting of:

4-Ureido-[2,31bithiophenyl-5-carboxylic acid amide 5-(4-Methoxy-phenyl)-3-ureido-thiophene-2-carboxylic acid amide 4-(3-Methyl-ureido)-[2,3']bithiophenyl-5-carboxylic acid amide 5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid amide 5-(4-Methoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide 5-(3,4-Dimethoxy-phenyl)-3-(3-methyl-ureido)-thiophene-2-carboxylic acid methylamide 4-(3-Methyl-ureido)-[2,3 lbithiophenyl-5-carboxylic acid methylamide 4-(3-Methyl-ureido)-[2,3]bithiophenyl-5-carboxylic acid phenylamide 4-(3-Methyl-ureido)-[2,3]bithiophenyl-5-carboxylic acid benzo[l,3]dioxol-5-ylamide 4-(3-Methyl-ureido)-[2,3 ]bithiophenyl-5 -carboxylic acid (2-amino-ethyl)-amide

7. A method according to claim 1 wherein the kinase being inhibited is chk-1 kinase.

8. A method according to claim 1 wherein the present compounds are administered as IKKβ/Chkl kinase dual inhibitors for the treatment of cancer and cancer-related diseases.

9. A method according to claim 1 wherein the disease or disorder being treated is selected from the group consisting of leukemia, solid tumor cancer, metastases, lymphomas, soft tissue cancers, brain cancer, esophageal cancer, stomach cancer, pancreatic cancer, liver cancer, lung cancer, bladder cancer, bone cancer, prostate cancer, ovarian cancer, cervical cancer, uterine cancer, testicular cancer, kidney cancer, head cancer and neck cancer, chronic inflammatory proliferative diseases, proliferative cardiovascular diseases, proliferative ocular disorders and benign hyperproliferative diseases.

10. A method according to claim 9 wherein the disease or disorder treated is selected from the group consisting of psoriasis, rheumatoid arthritis, diabetic retinopathy and hemangiomas.

11. A compound according to Formula (I) hereinbelow:

(I)

wherein:

RI is selected from the group consisting of H, C,_₂ alkyl, XH, XCH₃, C^ alkyl-XH,

C_j.₂ alkyl-XCH₃, C(O)NH₂, C(O)NHCH₃, and C(O)-C₁.₂ alkyl, provided that when

RI is H, R2 is not CONH₂, or provided that when RI is C,_₂ alkyl, R2 is not

CONH₂; with the preferred substitution being H or CH₃;

X is selected from the group consisting of O, S, and NH;

R2 is selected from the group consisting of C(O)R⁵, CO '₂2R-. ⁵, C(O)NHR⁵,

C(O)NHC(O)NR⁵R⁶,

SO₂R⁵, S(O)R⁵, SO₃R⁵, and PO₃R⁵R⁶;

R⁵ and R⁶ are, independently, selected from the group consisting of hydrogen, C^ alkyl, C,_.I0 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, and C_{0 fi} alkylheteroaryl, or R⁵ and R⁶, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C_j_₆ alkyl or (CH₂)₀₃aryl, wherein any of the foregoing may be optionally substituted by one or more of group A and on any position;

R3 is H or halogen; with the preferred substitution being H;

R4 is aryl or heteroaryl optionally substituted by one or more of group A and on any position, provided that when RI is CH₃ and R2 is CO₂R⁵, R4 is not phenyl, or provided that when RI is H, R4 is not 4-pyridyl; A is selected from the group consisting of C,_.10 alkyl, C,_.10 alkanoyl, C_{2 10} alkenyl, C_{2 10} alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, C_0.6 alkylheteroaryl, C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R^S, N(O)R⁷R⁸, NR⁷R⁸R⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C,_.I0 alkyl, C,_₁₀ alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C₀_₆ alkylaryl, C₀_₆ alkylheterocyclyl, C₀₆ alkylheteroaryl, (CH₂)_0.6heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group D and on any position;

Y is an organic or inorganic anion;

D is selected from the group consisting of C_{1 10} alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C₃_₁₀ cycloalkyl, C₀_₆ alkylaryl, C₀₆ alkylheterocyclyl, C₀₆ alkylheteroaryl, C(=NH)R⁷, COR⁷, CONR⁷R⁸, CON(O)R⁷R⁸, CONR⁷R⁸R⁹Y, CO₂R⁷, C(O)SR⁷, C(S)R⁷, cyano, trifluoromethyl, NR⁷R⁸, N(O)R⁷R⁸, NR⁷R⁸R⁹Y, NR⁷COR⁷, NR⁷CONR⁷R⁸, NR⁷CON(O)R⁷R⁸, NR⁷CONR⁷R⁸R⁹Y, NR⁷CO₂R⁷, NR⁷C(O)SR⁷, NR⁷SO₂R⁷, NR⁷SO2NR⁷R⁸, nitro, OR⁷, OCF₃, aryloxy, heteroaryloxy, SR⁷, S(O)R⁷, S(O)₂R⁷, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR⁷R⁸, SO₃R⁷, PO₃R⁷R⁸, and halo, wherein C,_₁₀ alkyl, C,_₁₀ alkanoyl, C_2.10 alkenyl, C₂_₁₀ alkynyl, C_{3 10} cycloalkyl, C₀₆ alkylaryl, C₀₆ alkylheterocyclyl, C₀₆ alkylheteroaryl, (CH₂)_0.6heteroaryl, aryloxy, and heteroaryloxy may be optionally substituted by one or more of group E and on any position;

R⁷, R⁸, and R⁹ are, independently, selected from the group consisting of hydrogen, C_M0 alkyl, C ₀ alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C_M alkylaryl, C_0.6 alkylheterocyclyl, and C₀₆ alkylheteroaryl, or R⁷ and R⁸, taken together with the nitrogen to which they are attached, may optionally form a ring having 3 to 7 carbon atoms, optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C,_₆ alkyl or (CH₂)₀₃aryl, wherein any of the foregoing may be optionally substituted by one or more of group E and on any position; E is selected from the group consisting of C,_₁₀ alkyl, C,_.10 alkanoyl, C₂_₁₀ alkenyl, C_{2 10} alkynyl, C_3.10 cycloalkyl, C_0.6 alkylaryl, C_0.6 alkylheterocyclyl, C₀₆ alkylheteroaryl, C(=NH)R¹⁰, COR¹⁰, CONR¹⁰Rⁿ, CON(O)R¹⁰R^H, CONR¹⁰RⁿR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰Rⁿ, N(O)R¹⁰R^U, NR¹⁰RⁿR¹²Y, NR¹⁰COR¹⁰, NR¹⁰CONR¹⁰R", NR¹⁰CON(O)R¹⁰Rⁿ, NR¹⁰CONR¹⁰R^UR¹²Y, NR¹⁰CO₂R¹⁰, NR'°C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR^SO.NR'V, nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR¹⁰R^U, SO₃R¹⁰, PO₃R¹⁰R", and halo, wherein C,_.,₀ alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C_2.10 alkynyl, C_3.10 cycloalkyl, C₀₆ alkylaryl, C_0.6 alkylheterocyclyl, C₀_₆ alkylheteroaryl may be optionally substituted by one or more of C(=NH)R¹⁰, COR¹⁰, CONR¹⁰R^π, CON(O)R¹⁰Rⁿ, CONR¹⁰R^πR¹²Y, CO₂R¹⁰, C(O)SR¹⁰, C(S)R¹⁰, cyano, trifluoromethyl, NR¹⁰R", N(O)R'°R^u, NR¹⁰R"R¹²Y, NR¹⁰COR¹⁰, NR¹⁰CONR¹⁰Rⁿ, NR¹⁰CON(O)R¹⁰Rⁿ, NR¹⁰CONR^I0R¹¹R¹²Y, NR¹⁰CO₂R¹⁰, NR¹⁰C(O)SR¹⁰, NR¹⁰SO₂R¹⁰, NR¹⁰SO₂NR¹⁰Rⁿ, nitro, OR¹⁰, OCF₃, aryloxy, heteroaryloxy, SR¹⁰, S(O)R¹⁰, S(O)₂R¹⁰, SCF₃, S(O)CF₃, S(O)₂CF₃, SO₂NR¹⁰R^U, SO₃R¹⁰, PO₃R¹⁰R^U, or halo, and on any position; R¹⁰, R¹¹, and R^I are, independently, selected from the group consisting of hydrogen, C, ₁₀ alkyl, C,_.10 alkanoyl, C_2.10 alkenyl, C,__I0 alkynyl, C_3.10 cycloalkyl, C_M alkylaryl, C_0. ₆ alkylheterocyclyl, and C₀₆ alkylheteroaryl; or R¹⁰ and R¹¹, taken together with the nitrogen to which they are attached, forms a ring having 3 to 7 carbon atoms optionally containing 1, 2, or 3 heteroatoms selected from nitrogen, sulfur, oxygen, or nitrogen, substituted with hydrogen, C,_₆ alkyl or (CH,)₀_₃aryl; or a pharmaceutically acceptable inorganic or organic salt, esters, or other prodrug of formula (I).