MXPA01010099A - Pyrazole compositions useful as inhibitors of erk - Google Patents

Abstract

Described herein are compounds that are useful as protein kinase inhibitors having formula (I) where R<1>, R<2>, T and Ht are described in the specification. The compounds are useful for treating disease states inmammals that are alleviated by a protein kinase inhibitor, particularly diseases such as cancer, inflammatory disorders, restenosis, and cardiovascular disease.

Description

PIRAZOL COMPOSITIONS USEFUL AS ERK INHIBITORS This application claims the benefits of United States Provisional Application serial number 60 / 180,506 filed on February 5, 2000; Provisional Application of the United States serial number 60 / 191,956 filed on March 24, 2000; and United States Provisional Application serial number 60 / 255,309 filed on December 13, 2000.

FIELD OF THE INVENTION The present invention pertains to the field of medical chemistry and relates to pyrazole compounds that are inhibitors of protein kinase, especially inhibitors of ERK, compositions containing said compounds and methods of use. The compounds are useful for treating cancer and other diseases that are alleviated by protein kinase inhibitors.

BACKGROUND OF THE INVENTION Mammalian mitogen-activated protein kinases (MAP) 1 are serine / serotonin kinases that mediate intracellular signal transduction pathways (Cobb and Goldsmith, 1995, J. Biol. Chem., 270, 14843; Davis, 1995 Mol. Reprod. Dev. 42, 459). Members of the MAP kinase family share sequential similarity and conserved structural domains and include the kinase ERK (kinase regulated by extracellular signal (extracellular signal regulated kinase), JNK (Jun N terminal kinase) and p38.The kinase JNK and p38 are activated as response to the proinflammatory cytokines TNF-alpha and interleukin 1 and by cellular stress, for example, thermal shock, hyperosmolarity, ultraviolet radiation, lipopolysaccharides and inhibitors of protein synthesis Derijard et al., 1994, Cell 76, 1025; Han et al. al., 1994, Science 265, 808; Raingeaud et al., 1995, J. Biol. Chem. 270, 7420; Shapiro and Dinarello, 1995, Proc. Nati, Acad. Sci. USA 92, 12230). In contrast, ERKs are activated through mitogens and growth factors (Bokemeyer et al., 1996, Kidney Int. 49, 1187). ERK2 is a widely distributed protein kinase that achieves its maximum activity when both the thr83 and Tyrl85 are phosphorylated by the MAP kinase kinase towards the 5 'end, MEK 1 (Anderson et al., 1990, Nature 343, 651; et al., 1992, Science 258, 478). Upon activation, ERK2 phosphorylates many regulatory proteins that include Rsk90 protein kinases (Bjorbaek et al., 1995, J. Biol. Chem. 270, 18848) and MAPKAP2 (Rouse et al., 1994, Cell 78, 1027) and transcription factors such as ATF2 (Raingeaud et al., 1996 Mol.Cell Biol. 16, 1247), Elk-1 (Raingeaud et al., 1996), c- Fos (Chen et al., 1993 Proc. Nati, Acad. Sci. USA 90, 10952) and c-Myc (Oliver et al., 1995, Proc. Soc. Exp. Biol. Med. 210, 162). ERK2 is also a target in the 3 'direction of the Ras / Raf dependent pathways (Moodie et al., 1993, Science 260, 1658) and can help diffuse the signals from those potentially oncogenic proteins. ERK2 has been shown to play a role in the control of the negative growth of breast cancer cells (Frey and Mulder, 1997, Cancer Res. 57, 628) and the overexpression of ERK2 in human breast cancer has been reported (Sivaraman et al., 1997, J \ Clin. Invest. 99, 1478). Activated ERK2 has also been implicated in the proliferation of airway smooth muscle cells stimulated by endothelin, which suggests that this kinase has a role in asthma (Elchel et al., 1997, Am. J. Respir. Cell. Mol. Biol. 16, 589). The JNK family of kinases (MAP) 1 have been implicated for having a role in mediating a cellular response to a variety of disorders including cancer [Oncogene 13: 135-42 (1996)], hepatic disorders [Hepatology 28: 1022 -30 (1998)], cardiovascular disorder [Circ. Res. 83: 167-78 (1998)]; Circulation 97: 1731-7 (1998); J. Biol. Chem. 272: 28050-6 (1997); Circ. Res. 79: 162-73 (1996); Circ. Res. 78: 947-53 (1996): J. Clin.

Invest. 97: 508-14 (1996)] and immunological disorders [J. Im unol. 162: 3176-87 (1999); Eur. J. Immunol. 28: 3867-77 (1998); J. Exp. Med. 186: 941-53 (1997); Eur. J. Immunol. 26: 989-94 (1996)], among others. Aurora2 is a serine / threonine protein kinase that has been linked to human cancer, for example, colon, breast and other solid tumors. It is believed that this kinase is involved in phosphorylation events of proteins that regulate the cell cycle. Specifically, aurora2 may play a role in the control of exact segregation of chromosomes during mitosis. The erroneous regulation of the cell cycle can cause cell proliferation and other abnormalities. In colon cancer tissue, it has been found that the aurora2 protein is overexpressed. See Bischoff et al., EMBO J., 1998, 17, 3052-3065; Schumacher et al., J. "Cell Biol., 1998, 143, 1635-1646; Kimura et al., J". Biol. Chem., 1997, 272, 13766-13771. Glycogen synthase kinase-3 (GSK-3) is a serine / threonine protein kinase composed of the isoforms a and ß which are each encoded by different genes [Coghlan et al., Chemistry & Biology, 7, 793-803 (2000); Kim and Kimmel, Curr. Opinion Genetics Dev., 10, 508-514 (2000)]. GSK-3 is implicated in several diseases including diabetes, Alzheimer's disease, CNS (central nervous system) disorders for example, manic depressive disorder and neurodegenerative diseases and hypertrophy of cardiomyocetes [WO 99/65897; WO 00/38675; and Haq et al., J. Cell. Biol. (2000) 151, 117]. These diseases can be triggered or lead to the abnormal functioning of certain cell signaling pathways in which the GSK-3 plays a role. KDR is a tyrosine kinase receptor that also binds to VEGF (vascular endothelial growth factor) (Neufeld et al., 1999, FASEB J., 13, 9). The binding of VEGF to the KDR receptor results in angiogenesis, which is the rapid growth of capillary vessels from preexisting blood vessels. In several types of cancer are high levels of VEGF that cause tumor angiogenesis and that allow the rapid growth of cancer cells. Therefore, suppressing VEGF activity is a way to inhibit tumor growth and it has been shown that this can be achieved by inhibiting the KDR tyrosine kinase receptor. AKT, also known as protein kinase B, is a serine / tyrosine kinase that plays a central role in promoting the survival of a wide range of cell types [Khwaja, A., Nature, 33-34 (1990)]. Zang et al., Have shown that ovarian cancer cells Pl372 show elevated levels of AKT-1 and AKT-2. The inhibition of AKT induces apoptosis in these human ovarian cancer cells, demonstrating that AKT may be a target or important target in the treatment of ovarian cancer [Zang, QY, et al., Oncogene, 19 (2000)] and other proliferative disorders. The AKT pathway is also involved in mononeural survival and nerve regeneration [Kazuhiko, N. et al., The Journal of Neuroscience, 20 (2000)]. There is a great unmet medical need to develop protein kinase inhibitors, especially ERK inhibitors, that are useful for treating various conditions associated with ERK activation, especially if relatively inadequate treatment options are considered for most of these conditions, that are available today. Accordingly, there is still a great need to develop potent protein kinase inhibitors, including ERK inhibitors, that are useful for treating various conditions associated with the activation of the protein kinase.

DESCRIPTION OF THE INVENTION It has now been found that the compounds of this invention and the pharmaceutical compositions thereof, are effective as protein kinase inhibitors, especially as inhibitors of ERK. These compounds have the general formula I: or a pharmaceutically acceptable derivative or prodrug thereof, wherein: Ht is a heterocyclic ring selected from pyrazol-3-yl, [1, 2, 4] triazol-3-yl, [1, 2, 3] triazole-4 -yl or tetrazol-5-yl, pyrazol-3-yl has the substituents R3 and QR4 and [1,2,4] triazol-3-yl or [1, 2, 3] triazol-4-yl are substituted either by R3 or QR4; R1 is selected from R, F, Cl, N (R8) 2 / OR, NRCOR, NRCON (R8) 2, CON (R8) 2, S02R, NRS02R or S02N ((R8) 2; T is selected from a valence bond or a linking group, each R independently is selected from a hydrogen or an optionally substituted aliphatic group having from one to six carbons; R2 is selected from hydrogen, CN, halogen or an optionally substituted group selected from aryl, aralkyl, heteroaryl, heterocyclyl, an acyclic aliphatic chain group having from one to six carbons or a cyclic aliphatic group having from three to ten carbons; R3 is selected from R, OH, OR, N (R8) 2, F, Cl or CN; Q is a valence bond, J or an optionally substituted C 1 -C 6 alkylidene chain, wherein up to two non-adjacent carbons of the alkylidene chain are optionally and independently replaced by J; J is selected from -C (= 0) -, -C02-, -C (0) C (0) -, -NRCONR8-, -N (R) N (R8) -, -C (= 0) NR8- , -NRC (= 0) -, -0-, -S-, -SO-, -S02-, - N (R) 0, -0N (R8) -, -OC- (= 0) N (R8) -, -N (R) C00-, -S02N (R8) -, - N (R) S02- ON (R8) -; R4 is selected from -R8, -R5, -NH2, -NHR5, -N (R5) 2 or -NR5 (CH2) and N (R5) 2; each R5 independently is selected from R6, R7, (CH2) and CH (R6) (R7), - (CH2) and R6, - (CH2) and CH (R6) 2, - (CH2) and CH (R7) 2 or - (CH2) and R7; and is from 0 to 6; each R6 is an optionally substituted group which independently is selected from an aliphatic, aryl, aralkyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy, heterocyclyl, heterocyclylalkyl or heterocyclylalkoxy group; each R7 independently is selected from an optionally substituted hydroxyalkyl, alkoxyalkyl, aryloxyalkyl or alkoxycarbonyl group; each R8 independently is selected from R or two R8 in the same nitrogen which together with the nitrogen optionally form a four to eight membered heterocyclic ring, saturated or unsaturated having from one to three heteroatoms; and each nitrogen of the substitutable ring is independently substituted by R, NR2, COR, C02 / (optionally substituted C? -C6 alkyl), S02 (optionally substituted C? -C6 alkyl), CONR or S02NR2. In the sense used herein, the following definitions will apply unless otherwise indicated. Also, combinations of substituents or variables are allowed, provided that these combinations give rise to stable compounds. The term "aliphatic" in the sense that is used herein, refers to straight chain, branched or cyclic C? -C? 2 hydrocarbons that are completely saturated or that contain one or more saturation units. For example, suitable aliphatic groups include linear, branched or cyclic substituted or unsubstituted linear, branched or cyclic alkyl, alkenyl or alkynyl groups and hybrids thereof, for example, alkyl (cycloalkyl), alkyl (cycloalkenyl) or alkenyl (cycloalkyl). The term "alkyl" and "alkoxy" employed alone or as part of a larger entity, is Pl37 refer to both straight and branched chains containing from one to twelve carbon atoms. The terms "alkenyl" and "alkynyl" employed alone or as part of a larger entity, will include both straight and branched chains containing from two to twelve carbon atoms. The terms "haloalkyl", "haloalkenyl" and "haloalkoxy" refer to alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. The term "halogen" refers to F, Cl, Br or I. The term "heteroatom" refers to N, 0 or S and will include any oxidized form of nitrogen and sulfur and the quaternized form of any basic nitrogen. The term "aryl" employed alone or as part of a larger entity as in "aralkyl", refers to aromatic ring groups having from five to fourteen members, for example, phenyl, benzyl, 1-naphthyl, 2-naphthyl , 1-anthracyl and 2-anthracyl and heterocyclic aromatic groups or heteroaryl groups, for example, 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl , 5-isoxazolyl, 2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, pyrimidyl, 5-pyrimidyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 5-tetrazolyl, 2-triazolyl, 5- Pl372 triazolyl, 2-thienyl or 3-thienyl. The term "aryl ring" also refers to rings that are optionally substituted. Aryl groups also include polycyclic aromatic ring systems in which a carbocyclic aromatic ring is fused to one or more rings. Examples include tetrahydronaphthyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzothiazolyl, benzooxazolyl, benzimidazolyl, isoquinolinyl, isoindolyl, acridinyl, benzoisoxazolyl and the like. Within the scope of the term "aryl" in the sense that is used herein, a group is also included in which one or more carbocyclic aromatic rings and / or heteroaryl rings are fused to a cycloalkyl or non-aromatic heterocyclic ring., for example, indanyl or tetrahydrobenzopyranyl. The non-aromatic heterocyclic rings are non-aromatic carbocyclic rings in which one or more carbons of the ring are replaced by a heteroatom such as nitrogen, oxygen or sulfur in the ring. The ring may be five, six, seven or eight members and / or be fused with another ring, for example, a cycloalkyl or an aromatic ring. Examples include 3-lH-benzimidazol-2-one, 3- (1-alkyl) benzimidazol-2-one, 2- Pl372 tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl , 1-piperazinyl, 2-piperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl, diazolonyl, N-substituted diazolonyl, 1-phthalimidinyl, benzoxane, benzotriazol-1-yl, benzopyrrolidine, benzopiperidine, benzoxolane, benzothiolane and benzothiane. The term "heterocyclic ring", whether saturated or unsaturated, also refers to rings that are optionally substituted. An aryl group (carbocyclic or heterocyclic) or an aralkyl group, for example, benzyl or phenethyl, may contain one or more substituents. Examples of suitable substituents on the unsaturated carbon atom of an aryl group include halogen, -R, -OR, -SR, protected OH (eg, acyloxy), phenyl (Ph), substituted Ph, -OPh substituted, -N02, -CN, -N (R) 2, -NRN (R) 2, -NRCOR, -NRC02 (aliphatic), -C02R, -COR, -C (0) C (0) R, -CON (R) 2, -CONRN (R) 2, -S (0) 2R, -SON (R) 2, -S (0) (aliphatic), -S02N (R) 2 or NRS (0) 2R, wherein each R independently is selected of hydrogen, an aliphatic group or a substituted aliphatic group. An aliphatic group or a heterocyclic ring does not Pl372 aromatic may contain one or more substituents. Examples of suitable substituents, on the saturated carbon of an aliphatic group or a non-aromatic heterocyclic ring, include those stated above for the unsaturated carbon, as well as the following: = 0, = S, = NNHR, = NNR2, = N -, OR, = NNHC0R, = NNHC02 (aliphatic), = NNHS02 aliphatic) or = NR, wherein each R independently is selected from hydrogen, an aliphatic group or a substituted aliphatic group. The term "alkylidene chain" refers to a straight or branched, optionally substituted carbon chain that may be fully saturated or have one or more units of unsaturation. Optional substituents of the C? -C6 alkylidene chain of Q, include those described above for an aliphatic group. A substitutable nitrogen in an aromatic or non-aromatic heterocyclic ring may be optionally substituted. Suitable substituents for nitrogen include, R, COR, N (R) 2, C0N (R) 2 / C0NRN (R) 2, S (0) 2R, and C02R, wherein R independently is selected from hydrogen, a group optionally substituted or aliphatic aryl. The term "linking group" or "linker" refers to an organic fraction that connects two parts of a compound. Liaison groups are usually Pl37 constituted by an oxygen or sulfur atom, a unit of -NH- or -CH2- or a chain of atoms, for example, an alkylidene chain. The molecular mass of a linking group is generally in the range of about 14 to 200. Examples of linking groups include saturated and unsaturated C 1 -C 6 alkylidene chains which as an option may be substituted and wherein up to two saturated carbons of the chain are optionally replaced by -C (= 0) -, -CONH-, CONHNH-, -C02-, -NHC02-, -O-, -NHCONH-, -OC (= 0) -, -0C (= 0) NH-, -NHNH-, -NHCO-, -0-, -S-, -SO-, -S02-, -NH-, -S02NH- or -NHS02-. It will be apparent to one skilled in the art that certain compounds of this invention can exist as tautomeric forms, that tautomeric forms of the compounds are within the scope of the invention. Unless stated otherwise, it is assumed that the structures represented here include all the stereochemical forms of the structure; that is, the R and S configurations for each asymmetric center. Therefore, the stereochemical isomers alone, as well as the enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention. Unless stated otherwise, the structures represented here are assumed to also include compounds that are distinguished only by the presence of one or Pl372 plus isotopically enriched atoms. For example, compounds having the structures present except for the substitution of a hydrogen with deuterium or tritium or the substitution of a carbon with a carbon enriched in 13C- or 14C-, are within the scope of this invention. Below are embodiments of this invention for the Ht ring when it is pyrazol-3-yl (II-A), [1,2,4] triazol-3-yl (II-B), [1, 2, 3] triazol-4-yl (II-C and II-D) and tetrazol-5-yl (II-E): II-A ll-D ll-E wherein R1"4, T and Q are as defined above The preferred compounds of the formulas II-A, Pl372 II-B, II-C, II-D and II-E, include those having one or more and more preferably all of the following features: (a) Q is -CO-, -C02- or -CONH-; (b) T is a valence bond; (c) R1 is hydrogen or NHR; (d) R2 is an optionally substituted aryl ring, more preferably an optionally substituted phenyl ring; (e) R3 is hydrogen; (f) R4 is selected from R5, -NHR5, N (R5) 2, -NR5R6, -NHCHR5R6 or -NHCH2R5; and / or (g) R5 is an optionally substituted group selected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl groups. heterocyclylalkyl, (CH2) and Re, (CH2) and R7, or (CH2) and CH (R6) (R7). Examples of substitutions in the phenyl group R2 include halo, nitro, alkoxy and amino. When R 4 is R 5, examples of preferred R 5 groups include pyrrolidin-1-yl, morpholin-1-yl, piperidin-1-yl and piperazin-1-yl, wherein each group is optionally substituted. When R4 is -NHR5 or -N (R5) 2, preferred R5 groups further include (CH2) and R6, (CH2) and R7 and (CH2) and CH (R6) (R7). Examples of preferred R6 and R7 include pyridin-3-yl, pyridin-4-yl, imidazolyl, furan-2-yl, tetrahydrofuran-2-yl, cyclohexyl, phenyl, -CH20H, (CH2) 2OH and isopropyl, wherein optionally each group is replaced. Preferred embodiments of this invention are represented by formulas III-A, III-B, III-C, III-D and Pl372 III-E: wherein R2, R4 and T are as defined above. Exemplary compounds of the formulas II-A, II-B, II-C and II-E are presented in Table 1 below. Table 1. Compounds II-A, II-B, II-C and II-E Pl372 Pl372 Pl37 Other embodiments of this invention relate to compounds of formula IV-A to IV-E: Pl372 IV-D IV-E wherein T, R, R and R4 are as defined above. The preferred compounds of formulas IV-A, IV-B, IV-C, IV-D and IV-E, include those having one or more and more preferably all of the following features: (a) T is a valence bond; (b) R3 is hydrogen; and / or (c) R2 is an optionally substituted aryl ring, more preferably an optionally substituted phenyl ring. The exemplifying compounds of formula IV-A, IV-B, IV-C, IV-D and IV-E, are presented below in Table 2. Table 2. Compounds IV-A Pl372 Pl37 Pl372 The present compounds can be prepared in general by methods known to those skilled in the art for analogous compounds. The compounds of formula II-A can be prepared according to a modified method of Finar, I.L., J. Chem. Soc., (1995), p.1205, such as Pl37 shows below in Scheme 1, for the preparation of compound II-A6. Scheme 1 II-A6 The compounds of formula II-B can be prepared according to the methods of Clitherow, J.W. , et al., Bioorg. Med. Chem. Lett., (1996) p.833-838, as shown below in Scheme 2, for the preparation of compound II-B3. Scheme 2 II-B3 The compounds of formula II-C can be prepared according to the methods of Beck, G. et al., Chem. Ver., (1973), p.106, as shown below in Scheme 3, for Pl37 the preparation of compound II -C4 Scheme 3 II-C4 The compounds of formula II-E can be prepared according to the methods of Kaltenbronn, J.S., et al., Eur. J. Med. Chem., (1997) p.425-431-838 and Norman, M.H. , et al., (1995) p. 4670-4678, as shown below in Scheme 4, for the preparation of compound II-E3. Scheme 4 II-E3 According to another embodiment, the invention provides a method for inhibiting the activity of the kinase in a biological sample. This method comprises the step of contacting the biological sample with a compound of this invention.

P1372 The term "biological sample", in the sense used herein, includes cell cultures or extracts thereof; material extracted by biopsy or extracts thereof, from a mammal; and blood, saliva, urine, feces, semen, tears or other body fluid or extracts thereof. The term "biological sample" also includes living organisms, in which case "contacting a compound of this invention with a biological sample, is synonymous with the term" administering the compound (or composition comprising the compound) to an animal. " aspect of this invention relates to a method for treating a disease state in a mammal, which is alleviated by treatment with a protein kinase inhibitor, this method comprises administering to a mammal in need thereof, a therapeutically effective amount of a compound of Formula I: or a pharmaceutically acceptable derivative or prodrug thereof, wherein: P1372 Ht is a heterocyclic ring selected from pyrazol-3-yl, [1, 2, 4] triazol-3-yl, [1, 2, 3] triazol-4-yl or tetrazol-5-yl, pyrazole-3 -yl has the substituents R3 and QR4 and [1, 2,4] triazol-3-yl or [1, 2, 3] triazol-4-yl is substituted with either R3 or QR4; R1 is selected from R, F, Cl, N (R8) 2, OR, NRCOR, NRCON (R8) 2, -CON (R8) 2, S02R, NRS02R or S02N ((R8) 2; T is selected from a valence bond or a linking group, each R independently is selected from a hydrogen or an aliphatic group optionally substituted having one to six carbons; R2 is selected from hydrogen, CN, halogen or an optionally substituted group selected from aryl, aralkyl, heteroaryl, heterocyclyl, an acyclic aliphatic chain group having from one to six carbons or a group cyclic aliphatic having three to ten carbons; R3 is selected from R, OH, OR, N (R8) 2, F, Cl or CN; Q is a valence bond, J, or an optionally substituted Ci-C6 alkylidene chain; , wherein up to two non-adjacent carbons of the alkylidene chain are optionally and independently replaced by J; J is selected from -C (= 0) -, -C02-, -C (0) C (0) -, -NRCONR8- , -N (R) N (R8) -, -C (= 0) NR8-, -NRC (= 0) -, -O-, -S-, -SO-, -S02-, -N (R) 0, -ON (R8) -, -OC- (= 0) N (R8) -, -N (R) COO-, -S02N (R8) -, - Pl372 N (R) S02- OR N (R8) -; R4 is selected from -R8, -R5, -NH2, -NHR5, -N (R5) 2 or -N R5 (CH2) and N (R5) 2; each R5 independently is selected from R6, R7, (CH2) and CH (R6) (R7), - (CH2) and R6, - (CH2) and CH (R6) 2, - (CH2) and CH (R7) 2 or - (CH2) ) and R7; and is from 0 to 6; each R6 is an optionally substituted group which independently is selected from an aliphatic, aryl, aralkyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy, heterocyclyl, heterocyclylalkyl or heterocyclylalkoxy group; each R7 independently is selected from an aliphatic hydroxyalkyl, alkoxyalkyl, aryloxyalkyl or optionally substituted alkoxycarbonyl group; each R8 independently is selected from R or two R8 in the same nitrogen which together with the nitrogen optionally form a four to eight membered heterocyclic ring, saturated or unsaturated having from one to three heteroatoms; and each substitutable nitrogen ring is independently substituted by R, NR2 / COR, C02; (optionally substituted C? -C6 alkyl), S02 (optionally substituted C? -C6 alkyl), C0NR2 or S02NR2. One embodiment comprises administering a compound of formula II-A, II-B, II-C, II-D or II-E. One modality Preferred compound comprises administering a compound of formula II-A or II-B, more preferably a compound of formula II-A and preferably superlatively a compound included in Table 1. Another preferred embodiment comprises administering a compound of formula III-A or III-B, preferably a superlative compound of formula III -A or a compound included in Table 2. The pharmaceutical compositions useful in those methods are described below. The present method in particular is useful for treating a disease state that is alleviated by the use of an ERK inhibitor, JAK, JNK, Aurora, GSK, KDR or AKT. In the sense used herein, unless otherwise indicated, the terms "ERK", "JAK" "JNK", "Aurora", "KDR" and "GSK" refer to all known isoforms of the respective enzymes, which include non-exclusively, ERK1, ERK2, ERK3, ERK4, ERK5, ERK6, ERK7, JAK1, JAK2, JAK3, JAK4, JNK1, JNK2, JNK3, Auroral, Aurora2, KDR, GSK3-alpha and GSK3-beta. The activity of the compounds as protein kinase inhibitors, for example, ERK inhibitors, can be analyzed in vi tro, in vivo or in a cell line. Using ERK as an example, in vi tro trials include assays that determine the inhibition of either the activity of the kinase or the activity of the Atpase of the ERK.

Pl372 activated. Alternate in vitro assays quantify the ability of the inhibitor to bind to ERK and can be measured by either radioactively labeling the inhibitor before it binds, isolating the inhibitor / ERK complex and determining the amount of radiolabeled binding or by performing a competition experiment in which new inhibitors are incubated with ERK bound to known radioligands. Any type or isoform of ERK can be used, depending on which type or isoform of ERK is going to be inhibited. The protein kinase inhibitors of this invention or the pharmaceutical salts thereof can be formulated in pharmaceutical compositions for administration in animals or in humans. These pharmaceutical compositions effective to treat or prevent a protein kinase-mediated condition, comprising the protein kinase inhibitor in an amount sufficient to inhibit the activity of the protein kinase and a pharmaceutically acceptable carrier in a form capable of being detected, are another embodiment of the invention. present invention. The term "inhibiting in a susceptible form", in the sense that is used herein, refers to a change in activity capable of being measured, between a sample containing the inhibitor and a sample containing only the protein kinase. The term "condition mediated by ERK", in the P1372 meaning that is used herein, refers to any disease or other harmful condition of which ERK is known to play a role. These conditions include, but are not limited to, cancer, stroke, diabetes, hepatomegaly, cardiovascular disease including cardiomegaly, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune diseases, atherosclerosis, restenosis, psoriasis, allergic disorders including asthma, inflammation, neurological disorders and diseases related to hormones. The term "cancer" includes, but is not limited to, the following types of cancer: breast, ovarian, cervical, prostate, testes, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, carcinoma epidermoid, long-cell carcinoma, small cell carcinoma, pulmonary adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, gallbladder carcinoma, liver carcinoma and bile ducts, kidney carcinoma, myeloid disorders, lymphoid disorders, hairy cells of Hodgkin, oral cavity and pharynx (oral), lips, tongue, mouth, pharynx, small intestine, colon-rectum, intestine Pl372 thick, rectum, brain and central nervous system and leukemia. The compounds of the present invention are also useful as inhibitors of related kinases. The term "related kinases" refers to protein kinases with residues that are similar to the residues that are aligned with the ERK binding site. Without attempting to be limited to the theory, the applicants assume that this inhibitory activity is due to the close structural similarity between the active sites of the ERK and the related kinases. Alignment of the ERK sequence with other kinases can be derived from common software programs, for example, the most suitable program available from Genetics Computer Group. This program uses the local homology algorithm described by Smith and Waterman in Advances in Applied Mathematics 2; 282 (1981). Related kinases that are inhibited by the compounds of this invention would contain residues, identified by the aforementioned protein sequence alignment software, corresponding to the ERK residues: 131, E33, G34, A35, Y36, G37, M38, V39, A52, K54, R67, T68, E71, L75, 184, 186, 1103, Q105, D106, L107, M108, E109, DII, K114, D149, K151, S153, N154, L156, C166 and D167, with a degree of similarity of 80% or greater. The similarity score can be determined Pl372 using a standard table of amino acid substitution, for example, those described by Dayhoff (Dayhoff, MO, et al., Atlas of Protein Sequence and Structure, 1979) and Blosom-Henikoff (Blosum-Henikoff, S and Henikoff, JG, PNAS, 1992, 89: 10915-10919). The term "related kinases" also includes those that contain residues with a degree of similarity of 80% or greater than the following ERK residues: 131, G37, A52, 1103, E109 and N154. The compounds of the present invention are also useful as inhibitors of kinases of the JAK family. Without intending to be limited by theory, the applicants assume that this inhibitory activity is due to the close structural similarity between the active sites of ERK and JAK, as determined by the standard methods mentioned above. It has been found, from experiments, developed at home, of X-ray crystal structure with inhibitors attached to ERK, that three amino acid residues in the active site of ERK generate key hydrogen bonding interactions with these types of inhibitors. These three amino acid residues are M108, D106 and Q105. This amino acid numbering corresponds to the entry to the Swiss-Prot database for access # P28482. The Swiss-Prot database is an international database of protein sequences, distributed by the European P1372 Bioinformatics Institute (EBI) in Geneva, Switzerland. The database can be found at www.ebi.ac.uk/swissprot. The atoms of the main chain of M108 and D106 and the associated interactions are common to all kinases. M108 provides both a hydrogen bond donor and an acceptor and D106 provides a hydrogen bond acceptor through its CO chain. It is expected that an inhibitor that could form a hydrogen bond with one or more of these groups that bind to hydrogen within the active site will bind to the enzyme and thus exhibit inhibition. The Q105 residue of glutamine is comprised in a subset of kinases including ERK and JAK as determined by examining the alignment data obtained with the aforementioned software programs. Q105 provides a side chain CO accepting hydrogen bonding key. Modeling experiments reveal that for both ERK and JARK, the hydrogen bond donor of the Ht ring is within the hydrogen bond distance to the Q105 residue. Because of these interactions at the active site, the ERK inhibitors of the present invention will also inhibit JAK. Accordingly, these compounds are expected to be useful for treating conditions mediated by JAK. The term "JAK-mediated condition," in the sense used herein, refers to any disease or other deleterious condition in which JAK is known to play a role. These conditions include non-exclusively, allergic disorders such as asthma and atopic dermatitis, autoimmune diseases for example, SLE lupus and psoriasis and conditions associated with organ transplantation. The compounds of this invention will also inhibit kinases of the JAK family, useful for treating conditions mediated by JAK. The term "JAK-mediated condition", as used herein, refers to any disease or other deleterious condition in which JAK is known to play a role.These conditions include, but are not limited to, degenerative induced by apoptosis, for example, Alzheimer's disease, parkinson's disease, ALS (amyotrophic lateral sclerosis), epilepsy and attacks, Huntington's disease, traumatic brain injuries as well as hemorrhagic and ischemic stroke, heart disease, immunodeficiency disorders, inflammatory diseases, allergic disorders, autoimmune diseases, destructive bone disorders, for example, osteoporosis, proliferative disorders, infectious diseases, viral diseases, disorders related to cell death and hyperplasia, among P1372 including reperfusion / ischemia in stroke, cardiac infarctions and organ hypoxia, platelet aggregation induced by thrombin, chronic myelogenous leukemia (CML), rheumatoid arthritis, asthma, osteoarthritis, ischemia, cancer, liver disease including hepatic ischemia, heart disease such as myocardial infarction and congestive heart failure, immunopathological conditions that involve the activation of T cells and neurodegenerative disorders. The compounds of this invention will also inhibit Aurora, which is useful for treating conditions mediated by Aurora. The term "Aurora-mediated condition," in the sense used herein, refers to any disease or other deleterious condition in which Aurora is known to play a role. These conditions include, but are not limited to, various types of cancer, for example, colon and ovarian cancer. The compounds of this invention will also inhibit kinases of the KDR family, which is useful for treating conditions mediated by KDR. The term "KDR-mediated condition," in the sense used herein, refers to any disease or other harmful condition in which KDR is known to play a role. These conditions include, but are not limited to, cancers, for example, brain cancer, genitourinary tract, lymphatic system, stomach, larynx, lung, pancreatic, breast, Kaposi's sarcoma, and leukemia; endometriosis, benign prostatic hyperplasia; vascular diseases such as restenosis and atherosclerosis; autoimmune diseases such as rheumatoid arthritis and psoriasis; ocular conditions, for example, proliferative or angiogenic retinopathy and macular degeneration; and inflammatory diseases, for example, contact dermatitis, asthma and delayed hypersensitivity reactions. The compounds of this invention will also inhibit kinases of the GKS family, which is useful for treating conditions mediated by GKS. The term "GKS mediated condition", in the sense used herein, refers to any disease or other deleterious condition in which the GKS is known to play a role. These conditions include, but are not limited to, bipolar disorder, mania, Alzheimer's disease, diabetes and leukopenia. In addition to the compounds of this invention, pharmaceutically acceptable derivatives or prodrugs thereof may also be employed in compositions for treating or preventing the above-mentioned disorders.

Pl372 A "pharmaceutically acceptable derivative or prodrug" refers to any salt, ester, salt of an ester or other pharmaceutically acceptable derivative of a compound of this invention, which, when administered to the recipient, is capable of providing either directly or indirectly indirectly, a compound of this invention or a metabolite or residue thereof with inhibitory activity. In particular, derivatives or prodrugs that increase the bioavailability of the compounds of this invention when administered to a mammal are preferred (e.g., by allowing a compound that is orally administered to be more readily absorbed into the blood) or that it reinforces the supply of the precursor compound to a biological compartment (for example, the brain or the lymphatic system) in relation to the precursor species. The pharmaceutically acceptable salts of the compounds of this invention include those derived from bases and from pharmaceutically acceptable organic and inorganic acids. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorrate, camphorsulfonate, cyclopentanpropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride. , hydrobromide, yodhydrate, 2- P1372 hydroxyethane sulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, for example oxalic, can also be used, although they are not themselves pharmaceutically acceptable, in the preparation of salts that are used as intermediates to obtain the compounds of the invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal salts (e.g., sodium and potassium salts), alkaline earth metal salts (e.g., magnesium), ammonium salts and N + (C 1 -C 4) alkyl salts. This invention also contemplates the quaternization of any of the nitrogen-containing groups of the compounds set forth herein. By means of said quaternization, soluble or dispersible compounds can be obtained in water or in oil. Pharmaceutically acceptable carriers that can be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer-like substances, eg, phosphates, glycine, acid Sorbic P1372, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes, for example, protamine sulphate, disodium hydrogenated phosphate, hydrogenated potassium phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and wool wax. The compositions of the present invention can be administered orally, parenterally, by atomization for inhalation, topically, rectally, nasally, buccally, vaginally or by implanted reservoir. The term "parenteral" in the sense that is used herein, includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection. Preferably, the compositions are administered orally, intraperitoneally or intravenously. The sterile injectable forms of the compositions of this invention may be an aqueous or oily suspension. These suspensions can be formulated according to techniques known in the technical field, using dispersing or wetting and suspending agents.

P1372 suitable. The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oily suspension. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic, parenterally-acceptable diluent or solvent, for example, a solution in 1,3-butane diol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, fixed oils are used conventionally as a solvent or suspension medium. For this purpose, any soft fixed oil can be used, including synthetic mono or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, since they are natural pharmaceutically acceptable oils, for example, olive oil or castor oil, especially their polyoxyethylated forms. These oily solutions or suspensions may also contain a long chain alcohol diluent or dispersant, for example, carboxymethylcellulose or similar dispersing agents which are generally used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. You can also P1372 use other commonly used surfactants such as Tween, Span and other emulsifying agents or bioavailability enhancers, which are commonly used in the manufacture of solid, liquid or other dosage forms. The pharmaceutical compositions of this invention can be administered orally in any acceptable oral dosage form, including non-exclusively, capsules, tablets, suspensions or aqueous solutions. In the case of tablets for oral use, the vehicles that are usually used include lactose and corn starch. It is also common to add lubricating agents such as magnesium stearate. For administration in the form of capsules, the diluents that are used include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying or suspending agents. If desired, some sweetening, flavoring or coloring agents may also be added. Alternatively, the pharmaceutical compositions of the invention can be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the agent with a suitable non-irritating excipient, which is solid Pl372 at room temperature but liquid at the rectal temperature and therefore it melts in the rectum and releases the active components. These materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols. The pharmaceutical compositions of this invention can also be administered topically, especially when the target or target of the treatment involves areas or organs that are easily accessible by topical application, including diseases of the eyes, skin or lower intestinal tract. . With ease, suitable topical formulations are prepared for each of these areas or organs. Topical application in the lower intestinal tract can be carried out with a rectal suppository formulation (see above) or in an adequate enema formulation. Topical transdermal patches can also be used.

For topical applications, the pharmaceutical compositions can be formulated in a suitable ointment containing the active component suspended or dissolved in one or more vehicles. The vehicles for topical administration of the compounds of this invention include, in non-exclusive form, mineral oil, liquid petrolatum, Pl372 white petrolatum, propylene glycol, polyoxyethylene and polyoxypropylene compounds, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with an appropriate lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. For ophthalmic use, the pharmaceutical compositions can be formulated as micronized suspensions in saline, with regulated, isotonic pH or preferably as sterile saline solution, with regulated, isotonic pH, either with or without preservative of the benzalkonium chloride type. Alternatively, for ophthalmic uses, the pharmaceutical compositions can be formulated into an ointment such as petrolatum. The pharmaceutical compositions of this invention can also be administered by aerosol or nasal inhalation. These compositions are prepared according to techniques well known in the field of pharmaceutical formulation and can be prepared as saline solutions, employing benzyl alcohol or other suitable preservatives, absorption promoters that increase bioavailability, Pl372 fluorocarbons and / or other conventional solubilizing or dispersing agents. The amount of ERK inhibitor that can be combined with the carriers to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. Preferably, the compositions should be formulated in such a way that a dose of approximately between 0.01 and 100 mg / kg of body weight / day of the inhibitor can be administered to a patient receiving these compositions. It should be understood that a specific dosing and treatment regimen for any particular patient will depend on a variety of factors, including, activity of the specific compound being used, age, body weight, general health status. , sex, diet, time of administration, speed of excretion, combination of medications, the criteria of the doctor who attends and the severity of the particular disease that is treated. The amount of inhibitor will also depend on the particular compound in the composition. The kinase inhibitors of this invention or pharmaceutical compositions thereof can also be incorporated into compositions for coating a device Pl372 implantable doctor, for example, prostheses, artificial valves, vascular grafts, stents and catheters. For example, vascular stents have been used to overcome restenosis (by re-narrowing the vessel wall after the injury). Nevertheless, patients who use stents or other implantable devices run the risk of clot formation or platelet activation. These undesirable effects can be prevented or mitigated by applying a prior coating with a composition comprising a kinase inhibitor to the device. Compositions comprising a kinase inhibitor of this invention and a suitable carrier or coating are another embodiment of the present invention. Suitable coatings and general preparation of the coated implantable devices are described in U.S. Patent Nos. 6,099,562; 5,886,026; and 5,304,121. The coatings are generally biocompatible polymeric materials, for example, hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene-vinyl acetate and mixtures thereof. The coatings in optional form can be further covered by a suitable layer of fluorosilicone, polysaccharides, polyethylene glycol, phospholipids or combinations thereof to impart characteristics P137 controlled release in the composition. Implantable devices coated with a kinase inhibitor of this invention are another embodiment of the present invention. According to another embodiment, the invention provides methods for treating or preventing a condition mediated by ERK, JAK, JNK, Aurora, KDR or GSK or disease state, comprising the step of administering to a patient one of the pharmaceutical compositions described above. The term "patient", in the sense that is used herein, refers to an animal, preferably a mammal and preferably a human superlative. Preferably, this method is used to treat or prevent a condition selected from several types of cancer, for example, breast, colon, prostate, skin, pancreatic, brain, genitourinary tract, lymphatic system, of stomach, larynx and lung, including pulmonary adenocarcinoma and small cell lung cancer, stroke, diabetes, hepatomegaly, cardiomegaly, Alzheimer's disease, cystic fibrosis and viral disease or any specific disease or disorder described above. Depending on the particular condition or the condition of the disease to be treated or prevented, it can be administered together with the inhibitors of this P1372 invention, additional therapeutic agents that are usually administered to treat or prevent that condition. For example, chemotherapeutic agents or other antiproliferative agents can be combined with the inhibitors of this invention to treat proliferative diseases and cancer. Examples of known chemotherapeutic agents include, in non-exclusive form, adriamycin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, taxol, interferons and platinum derivatives. Other examples of agents with which the inhibitors of this invention may be combined, include non-exclusively, anti-inflammatory agents, such as coricosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide and sulfasalazine; immunomodulatory and immunosuppressive agents, for example, cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil, interferons, corticosteroids, cyclophosphamide, azathioprine and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole and anti-Parkinson's agents; agents for treating cardiovascular diseases, for example, beta blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers and statins; agents P1372 for treating liver diseases, for example, corticosteroids, cholestyramine, interferons and antiviral agents; agents for treating blood disorders, for example, corticosteroids and anti-leukemia agents and growth factors; agents for treating diabetes, for example, insulin, insulin analogues, alpha glucosidase inhibitors, biguanides and insulin sensitizers; and agents for treating immunodeficiency disorders, for example, gamma globulin. These additional agents can be administered separately, as part of a multiple dosage regimen, from the composition containing the inhibitor. Alternatively, these agents can be part of a single dosage form, mixed together with the inhibitor in a single composition. In order that the invention described herein be better understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are in no way to be construed as limiting this invention. EXAMPLES Example 1 ERK inhibition assay For the inhibition of ERK2 the compounds can be analyzed by a spectrophotometric assay P1372 coupled to enzyme (Fox et al (1998) Protein Sci. 7, 2249). In this assay, a certain concentration of activated ERK2 (10 nM) is incubated with various concentrations of the compound in DMSO (2.5%) for 10 minutes at 30 ° C in 0.1 M HEPES buffer, pH 7.5, coning 10 mM MgCl 2, 2.5 mM phosphoenolpyruvate, 200 μM NADH, 150 μg / mL pyruvate kinase, 50 μg / mL lactate dehydrogenase and 200 μM erktido peptide. The reaction is initiated by the addition of 65 μM ATM. The rate of decrease in absorbance at 340 nm is monitored. The IC50 is evaluated from the velocity data as a function of the concentration of the inhibitor.

Example 2: Cell proliferation assay by inhibition of ERK The compounds can be analyzed to assess the inhibition of ERK2 by a cell proliferation assay. In this assay, a complete medium is prepared by the addition of 10% fetal bovine serum and penicillin / streptomycin solution to RPMI 1640 medium (JHR Biosciences), Colon cancer cells are added (HT-29 cell line) to each of 84 wells of a 96-well plate at a seeding density of 10,000 cells / well / 150 μL. The cells are allowed to adhere to the plate when incubated at 37 ° C for 2 hours. One prepares Pl372 solution of the test compound in complete medium, by serial dilution, to obtain the following concentrations: 20 μM, 6.7 μM, 2.2 μM, 0.74 μM, 0.25 μM and 0.08 μM. The solution of the test compound (50 μL) is added to each well of 72 cells. To the remaining 12 wells containing cells, only complete medium (200 μL) is added to form a control group in order to measure maximum proliferation. To the remaining 12 empty wells, complete medium is added to form a control group with vehicle in order to measure the initial conditions. The plates are incubated at 37 ° C for 3 days. A stock solution of 3H-thymidine (1 mCi / mL, New England Nuclear, Boston MA) is diluted to 20 μCi / mL in RPMI medium and then 20 μL of this solution is added to each well. The plates are then incubated at 37 ° C for 8 hours, then harvested and analyzed for 3 H-thymidine consumption by means of a liquid scintillation counter.

Example 3 JAK inhibition assay JAK inhibition can be evaluated by the method described by G.R. Brown, et al., Bioorg. Med. Chem.

Lett. 2000, vol. 10, p.575-579 in the following manner. In Maxisorb plates that have been previously coated at 4 ° C with Pl372 Poly (Glu, Ala, Tyr) 6: 3: 1 and then washed with saline buffered with 0.05% phosphate and Tween (PBST), add 2 μM ATP, 5 mM MgCl2 and a solution of the compound in DMSO. The reaction is started with JAK enzyme and the plates are incubated for 60 minutes at 30 ° C. Then, the plates are washed with PBST, 100 μL HRP-Conjugate 4G10 antibody is added and the plate is incubated for 90 minutes at 30 ° C. The plate is washed again with PBST, 100 μL of TMB solution is added and then the plates are incubated for another 30 minutes at 30 ° C. Sulfuric acid (100 μL μM) is added to stop the reaction and the plate is read at 450 nM to obtain the optical densities for analysis and determination of the IC 50 values.

Example 4 JNK inhibition assay: The compounds can be screened in the following manner to check their ability to inhibit JNK by an enzyme-coupled spectrophotometric assay. To a buffer buffer for assay containing 0.1 M HEPES buffer (pH 7.5), 10 mM MgCl2, 2.5 mM phosphoenolpyruvate, 200 μM NADH, 150 μg / mL pyruvate kinase, 50 μg / mL lactate dehydrogenase and receptor peptide of EGF 200 μM (with sequence KRELVEPLTPSGEAPNQALLR), several additions are added P137 concentrations of the compound in DMSO and a fixed concentration of activated JNK (10 nM). The resulting mixture is incubated at 30 ° C for 10 minutes, then the reaction is initiated by the addition of 10 μM ATP. The decrease in absorbance at 340 nM at 30 ° C is monitored as a function of time and the resulting data are adjusted to a kinetic model of competitive inhibition to determine Ki.

Example 5 Aurora inhibition assay The compounds can be screened in the following manner to check their ability to inhibit Aurora by an enzyme coupled standard assay. To a buffer buffer for assay containing 0.1 M HEPES, pH 7.5, 10 mM MgCl 2, 25 mM NaCl, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 30 μg / mL pyruvate kinase, 10 μg / mL lactate dehydrogenase, ATP 40 μM and 800 μM of peptide (LRRASLG, American Peptide, Sunnyvale, CA)), a solution of 30 μM of the compound in DMSO is added and the resulting mixture is incubated at 30 ° C for 10 minutes. The reaction is initiated by the addition of 10 μL of 70 nM Aurora and 1 mM DTT. The reaction rates are obtained by monitoring the absorbance at 340 nM for a reading time of 5 minutes at 30 ° C using a BioRad Ultramark plate reader (Hercules, CA). The IC50 is P1372 determines from the velocity data, as a function of inhibitor concentration.

EXAMPLE 6 GSK-3 Inhibition Assay Compounds can be screened as follows to test their ability to inhibit glycogen kinase 3 (GSK-3) by an enzyme-coupled standard assay (Fox et al. (1998) Protein). Sci. 7, 2249). To a buffer buffer for assay containing 0.1 M HEPES, pH 7.5, 10 mM MgCl 2, 25 mM NaCl, 2.5 mM phosphoenolpyruvate, 300 μM NADH, 1 mM DTT, 30 μg / mL pyruvate kinase, 10 μg / mL lactate dehydrogenase, 300 μM peptide (HSSPHQp-SEDEEE, American Peptide, Sunnyvale, Calif.)), a solution of 30 μM of the compound in DMSO is added and the resulting mixture is incubated at 30 ° C for 5 minutes. The reaction is initiated by the addition of 10 μM ATP. Reaction rates are obtained by monitoring the absorbance at 340 nM for a 5 minute reading time at 30 ° C using a Molecular Devices plate reader (Sunnyvale, CA). The IC50 is determined from the velocity data, as a function of the inhibitor concentration.

P1372 Example 7 KDR Inhibition Assay The compounds can be screened as follows to test their ability to inhibit KDR by standard enzyme-linked assay (Fox et al (1998) Protein Sci. 1, 2249). The assays were carried out in a mixture of 200 M HEPES, pH 7.5, 10 mM MgCl 2, 25 mM NaCl, 1 mM DTT and 1.5% DMSO. The final concentrations of the substrate in the assay were 300 μM ATP (Sigma Chemicals) and 10 μM E4Y poly (Sigma). The tests were carried out at 37 ° C and 30 nM KDR. The final concentrations of the components of the coupled enzyme system were 2.5 nM phosphoenolpyruvate, 200 μM NADH, 30 μg / mL pyruvate kinase and 10 μg / mL lactate dehydrogenase. A buffer buffer for assay was prepared, containing all the reagents listed above, with the exception of the ATP and the test compound of interest. 177 μl of the stock solution was placed in a 96-well plate and then 3 μl of 2 mM DMSO stock containing the test compound was added. (final concentration of the compound 30 μM). The plate was preincubated for approximately 10 minutes at 37 ° C and the reaction was initiated by the addition of 20 μl of ATP (final concentration 300 μM). Reaction rates were obtained using a Molecular plate reader P1372 Devices (Sunnyvale, CA) for a reading time of 5 minutes at 37 ° C. Compounds that showed greater than 50% inhibition with respect to the standard wells containing the assay mixture and DMSO without the test compound, were titrated to determine the IC50 values. While various embodiments of this invention have been described, it is evident that our basic examples can be modified to provide other embodiments using the compounds and methods of this invention. Therefore, it will be appreciated that the scope of this invention is defined by the attached modalities rather than by the specific modalities that have been presented as an example.

P1372

Claims (27)

1. CLAIMS; A compound of formula I or a pharmaceutically acceptable derivative or prodrug thereof, wherein: Ht is a heterocyclic ring selected from pyrazol-3-yl, [1, 2, 4] triazol-3-yl, [1, 2, 3] triazole-4 -yl or tetrazol-5-yl, pyrazol-3-yl has the substituents R3 and QR4 and [1, 2, 4] triazol-3-yl or [1, 2, 3] triazol-4-yl are substituted either by R3 or QR4; R1 is selected from R, F, Cl, N (R8) 2 / OR, NRCOR, NRCON (R8) 2, CON (R8) 2, S02R, NRS02R or S02N ((R8) 2; T is selected from a valence bond or a linking group, each R independently is selected from a hydrogen or an optionally substituted aliphatic group having from one to six carbons; R2 is selected from hydrogen, CN, halogen or an optionally substituted group selected from aryl, aralkyl, heteroaryl, heterocyclyl, a chain group P1372 acyclic aliphatic having from one to six carbons or a cyclic aliphatic group having from three to ten carbons; R3 is selected from R, OH, OR, N (R8) 2, F, Cl or CN; Q is a valence bond, J or an optionally substituted C 1 -C 6 alkylidene chain, wherein up to two non-adjacent carbons of the alkylidene chain are optionally and independently replaced by J; J is selected from -C (= 0) -, -C02-, -C (0) C (0) -, -NRCONR8-, - N (R) N (R8) -, -C (= 0) NR8- , -NRC (= 0) -, -O-, -S-, -SO-, -S02-, -N (R) 0, -ON (R8) -, -OC- (= 0) N (R8) -, -N (R) COO-, -S02N (R8) -, - N (R) S02- or N (R8) -; R4 is selected from -R8, -R5, -NH2, -NHR5, -N (R5) 2 or -N R5 (CH2) and N (R5) 2; each R5 independently is selected from R6, R7, (CH2) and CH (R6) (R7), - (CH2) and R6, - (CH2) and CH (R6) 2, - (CH2) and CH (R7) 2 or - (CH2) ) and R7; and is from 0 to 6; each R6 is an optionally substituted group which independently is selected from an aliphatic, aryl, aralkyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy, heterocyclyl, heterocyclylalkyl or heterocyclylalkoxy group; each R7 independently is selected from an optionally substituted hydroxyalkyl, alkoxyalkyl, aryloxyalkyl or alkoxycarbonyl group; P1372 each R8 independently is selected from R or two R8 in the same nitrogen which together with the nitrogen optionally form a four to eight membered heterocyclic ring, saturated or unsaturated having from one to three heteroatoms; and each substitutable nitrogen of the ring, independently is substituted by R, NR2, COR, C02, (optionally substituted C? -C6 alkyl), S02 (optionally substituted C? -C6 alkyl), CONR2 or S02NR2; provided that: (a) TR2 and QR4 are not equal; (b) TR2 and R3 are not equal; (c) when Ht is tetrazol-5-yl and R1 is methyl, then TR2 is other than hydrogen; (d) when Ht is [1, 2, 3] triazol-4-yl and both R 1 and R 2 are methyl, then TR 2 is other than hydrogen; and (e) when Ht is pyrazol-3-yl and both R1 and R3 are hydrogen, then TR2 is different from methyl when QR4 is phenyl at the 4-position.
2. 2. The compound according to claim 1, the compound is selected from of the following: P1372 ll-A ll-C ll-D ll-E or a pharmaceutically acceptable derivative or prodrug thereof.
3. 3. The compound according to claim 2 having the formula ll-A or a pharmaceutically acceptable derivative or prodrug thereof. P1372
4. 4. The compound according to any of claims 1, 2 or 3, which also one or more of the following features: (a) Q is -CO-, -C02- or -CONH-; (b) T is a valence bond; (c) R1 is hydrogen or NHR; (d) R2 is an optionally substituted aryl ring; (e) R3 is hydrogen; (f) R4 is selected from R5, -NHR5, -N (R5) 2, -NR5R6, -NHCHR5R6 or -NHCH2R5; or (g) R5 is an optionally substituted group selected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl groups. heterocyclylalkyl, (CH2) and R6, (CH2) and R7, or (CH2) and CH (R6) (R7).
5. 5. The compound according to claim 4, which has the formula III-A or a pharmaceutically acceptable derivative or prodrug thereof.
6. 6. The compound according to claim 5, having the following features: (a) T is a valence bond; (b) R2 is an optionally substituted aryl ring; (c) R4 is selected from R5, -NHR5, -N (R5) 2, -NR5R6, -NHCHR5R6 or -NHCH2R5; and (d) R5 is a group Optionally substituted p1372 selected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl groups. heterocyclylalkyl, (CH2) and R6 [CH2) and R, (CH2) and CH (R6) (R7).
7. 7. The compound according to claim 1, wherein the compound is selected from those included in Table 1.
8. 8. The compound according to claim 1, the compound is selected from the following: IV-D IV-E or a pharmaceutically acceptable derivative or prodrug thereof. Pl372
9. 9. The compound according to claim 8, which also one or more of the following features: (a) Q is -CO-, -C02- or -CONH-; (b) T is a valence bond; (c) R2 is an optionally substituted aryl ring; (d) R3 is hydrogen; (e) R4 is selected from R5, -NHR5, -N (R5) 2, -NR5R6, -NHCHR5R6 or -NHCH2R5; or (f) R5 is an optionally substituted group selected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl groups. heterocyclylalkyl, (CH2) and R6, (CH2) and R7, or (CH2) and CH (R6) (R7).
10. 10. The compound according to claim 9 having the formula IV-A or a pharmaceutically acceptable derivative or prodrug thereof.
11. 11. The compound according to claim 10, having the following features: (a) T is a valence bond; (b) R2 is an optionally substituted aryl ring; (c) R4 is selected from R5, -NHR5, -N (R5) 2, -NR5R6, -NHCHR5R6 or -NHCH2R5; and (d) R5 is a group Optionally substituted Pl37 selected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl groups. heterocyclylalkyl, - (CH2) and R6, - (CH2) and R7, or (CH2) and CH (R6) (R7). The compound according to claim 1, wherein the compound is selected from those included in Table 2. 13. A composition comprising a compound according to any of claims 1 to 12, in an amount sufficient to inhibit in a form capable of detecting the activity of the protein kinase, the protein kinase is selected from one or more of ERK, JAK, JNK, Aurora, GSK, KDR, AKT or a protein kinase related thereto; and a pharmaceutically acceptable vehicle. The composition according to claim 13, wherein the compound is formulated in a pharmaceutically acceptable manner for administration to a patient. 15. A composition according to claim 13, further comprising a therapeutic agent, either as part of a multiple dose form together with the compound or as a separate dose. 16. A method for inhibiting the activity of the protein kinase in a biological sample, wherein the Pl37 protein kinase is selected from among ERK, JAK, JNK, Aurora, GSK, KDR, AKT or a protein kinase related thereto, which comprises the step of contacting the sample with a compound according to any of claims 1 to
12. 12. 17. A method for treating in a patient a disease state mediated by a protein kinase, wherein the protein kinase is selected from among ERK, JAK, JNK, Aurora, KDR, AKT or a protein kinase related thereto, comprising the step of administering to the patient a composition according to claim
13. 13. 18. The method according to claim 17, comprising the additional step of administering to the patient a therapeutic agent either as part of a multiple dose form together with the compound or in the form of separate dose. 19. A method for treating a disease state in a patient, wherein the disease state is selected from cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, Alzheimer's disease, cystic fibrosis, viral disease, autoimmune diseases, atherosclerosis, restenosis, psoriasis, allergic disorders, inflammation, neurological disorders, diseases related to hormones, conditions associated with the transplant of P1372 organs, immunodeficiency disorders, bone destructive disorders, proliferative disorders, infectious diseases, conditions associated with cell death, platelet aggregation induced by thrombin, chronic myelogenous leukemia (CML), liver disease, immunopathological conditions involving the activation of T cells or CNS disorders, comprising the step of administering a composition to the patient according to claim 13. 20. The method according to claim 19, wherein the disease is cancer. The method according to claim 20, wherein the disease state is a cancer selected from breast, ovarian, cervical, prostate, testicular, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, squamous cell carcinoma, long cell carcinoma, small cell carcinoma, pulmonary adenocarcinoma, bone cancer, colon adenoma, pancreas adenoma, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma , gallbladder carcinoma, liver and bile duct carcinoma, kidney carcinoma, myeloid disorders, lymphoid disorders, Hodgkin hair cells, oral cavity and pharynx (oral), Pl372 lips, tongue, mouth, pharynx, small intestine, colon-rectum, large intestine, rectum, brain and central nervous system or leukemia. 22. The method according to claim 20, comprising the additional step of administering to the patient a chemotherapeutic agent either as part of a multiple dose form together with the compound or as a separate dose form. 23. The method according to claim 19, wherein the disease state is a cardiovascular disease. The method according to claim 23, wherein the disease state is a cardiovascular disease, selected from restenosis, cardiomegaly, atherosclerosis, myocardial infarction or congestive heart failure. 25. The met5d according to claim 23, comprising the additional step of administering to the patient a therapeutic agent for treating cardiovascular disease, either as part of a multiple dose form together with the compound or as a separate dose. 26. A composition for coating an implantable device, comprising a compound according to claim 1 and a suitable vehicle for coating the P137 implantable device. 27. An implantable device coated with a composition according to claim 26. P1372