Substituted pyrrolo-pyridines, composition containing them, method for their producing and use thereof

The present invention relates to novel chemical compounds, in particular substituted pyrrolopyridines, to compositions containing them and to their use as medicaments.

More specifically, according to a first aspect, the invention relates to novel specifically substituted pyrrolopyridine with anticancer activity, via modulation of protein activity, in particular kinases.

To date, most of the commercial compounds used in chemotherapy cause major problems in terms of side effects and patient tolerance. These effects can be limited if the drugs used act selectively on cancer cells, excluding healthy cells. One solution to limit the harmful effects of chemotherapy may consist in the use of drugs that act on metabolic pathways or components of these pathways, primarily expressed in cancer cells, which are moderately expressed or not expressed in healthy cells.

Protein kinases are a family of enzymes that catalyze the phosphorylation of hydroxyl groups of specific protein residues such as tyrosine, serine or threonine residues. This phosphorylation can greatly modify protein function; thus, protein kinases play an important role in the regulation of various cellular processes, especially including metabolism, cell proliferation, cell differentiation, cell migration, or cell survival. Of the various cellular functions in which protein kinase is involved, certain processes are attractive targets for the treatment of gout as well as other diseases.

Accordingly, one of the objects of the invention are compositions with anticancer activity, which act specifically on kinases. Of the kinases for which modulation of activity is desired, KDR and Tie2 are preferred.

51136Β [0006] The formula (I) below corresponds to these products:

<img file="RS51136B_D0001.tif" />

Formula (I) where:

1) A and Ar are selected from the group of phenyl substituents;

2) L is NH-CO-NH;

3) W is C (R6); one of Y and Z is selected from N and N0, and the other of Y and Z is C (R5), and

4) R1, R5, iR6suH,

5) RajeH.

Acceptable substituent combinations include those wherein R1, R5 and R6 are II and one of Y and Z is selected from N and NO.

A is preferably substituted with a first substituent selected from the group consisting of: (C1-C12) alkyl, (C1-C12) alkyl halogen, (S2-C12) cycloalkyl, (C2-C12) alkylene, (C2-C12 ) alkynyl, (C8-C14) aryl, (C1-C13) heteroaryl, O- (C1-C3) alkyl, O- (C6-C14) Aryl, O- (C1-C13) heteroaryl, S- (C1-C3) ) alkyl, S- (C6-C14) Aryl, S- (CI-C13) heteroaryl, each optionally substituted with a substituent selected from: (C1-C3) alkyl, halogen, O (C1-C3) alkyl, N ) (R9); wherein R 8 and R 9 are independently selected from: H, (C 1 -C 3) alkyl, (C 1 -C 3) alkylOH, (C 1 -C 3) alkyl halogen, (C 1 -C 3) alkylNH₂, (C1-C3) alkylCOOM, (C1-C3) alkylSO₃M, in which, in the case where R8 and R9 are at the same time substituents other than H, they can be bonded to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from Ο, N and S, in which M is H or an alkali metal cation selected from Li, Na and K.

Additionally, A is suitably substituted with another substituent selected from the group consisting of: F, Cl, Br, I, OH, SH, SO₃M COOM, CN, NO₂, CON (R8) (R9), N (R8) CO (R9), (C1-C3) alkyl-OH, (C1-C3) alkyl-N (R8) (R9), (C1-C3) alkyl- R 10), (C 1 -C 3) alkyl-COOH, N (R 8) (R 9); wherein R8 and R9 are independently selected from H, (C1-C3) alkyl, (C1-C3) alkylOH, (C1-C3) alkyl halogen, (C1-C3) alkylNH₂, (C1-C3) alkylCOOM, (C1-C3) alkylSO₃M; in which R8 and R9 are simultaneously a substituent other than H, they may be linked to form 5- to 7-membered

A 51136Β ring containing from 0 to 3 heteroatoms selected from Ο, N and S, wherein M is H or an alkali metal cation selected from Li, Na and K; and wherein R 10 is H or an optionally substituted non-aromatic heterocycle of 2 to 7 carbon atoms, and 1 to 3 heteroatoms selected from N, O and S.

When A is disubstituted, the two substituents may be linked to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from N, O and S.

According to one preferred embodiment, A is phenyl substituted with at least one group selected from: halogen, (C 1 -C 4) alkyl, (C 1 -C 3) alkyl halogen, O- (C 1 -C 4) alkyl, S (C 1 -C 4) alkyl, O- (C1-C4) alkyl halogen, S- (C1-C4) alkyl halogen, and in which, in the case of A, the two substituents may be linked to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from N, O and C.

The products according to the invention can be;

1) in non-chiral form,

2) in racemic form

3) enriched with a single steroid isomer, or

4) enriched one enantiomeric form;

(C1-C3) alkylCOOM, (C1-C3) alkylSO₃M; in which R8 and R9 are simultaneously a substituent other than H, they may be attached to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from Ο, N and S, in which M is H or an alkaline cation metals selected from Li, Na and K; and wherein R 10 is H or an optionally substituted non-aromatic heterocycle of 2 to 7 carbon atoms, and 1 to 3 heteroatoms selected from Li, Na and K.

Additionally, A is suitably substituted with another substituent selected from the group consisting of: F, Cl, Br, 1, OH, SH, SO₃M COOM, CN, NO₂, CON (R8) (R9), N (R8) CO (R9), (C1-C3) alkyl-OH, (C1-C3) alkyl-N (R8) (R9), (C1-C3) alkyl- R10), (C1-C3) alkyl-COOH, N (R8) (R9); wherein R8 and R9 are independently selected from dans, (C1-C3) alkyl, (C1-C3) alkylOH, (C1-C3) alkyl halogen, (C1-C3) alkylNH₂, (C1-C3) alkylCOOM, (C1-C3) alkylSO₃M: in which R8 and R9 are simultaneously a substituent other than H, they may be linked to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from Ο, N and S, in which M is H or an alkali metal cation selected from Li, Na and K; and wherein R 10 is H or an optionally substituted non-aromatic heterocycle of 2 to 7 carbon atoms, and 1 to 3 heteroatoms selected from N, N and S.

51136 When A is disubstituted, the two substituents can be linked to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from N, O and C.

According to one preferred embodiment, A is phenyl substituted with at least one group selected from: halogen, (C 1 -C 4) alkyl, (C 1 -C 3) alkyl halogen, O- (C 1 -C 4) alkyl, S (C 1 -C 4) alkyl, O- (C1-C4) alkyl halogen, S- (C1-C4) alkyl halogen, and in which, in the case of A, the two substituents may be linked to form a 5- to 7-membered ring containing from 0 to 3 heteroatoms selected from N, O and C.

The products according to the invention can be:

1) in non-chiral form,

2) in racemic form

3) enriched with a single steroid isomer, or

4) enriched one enantiomeric form;

And the choice can be in the form of salt.

The product according to the invention can be used for the production of a drug which is useful for the treatment of pathological conditions, in particular cancer.

The present invention also relates to medicaments containing a product according to the invention and to therapeutic compositions comprising a product according to the invention in combination with a pharmaceutically acceptable excipient according to the chosen route of administration. The pharmaceutical composition may be in solid or liquid form or in the form of a liposome.

Solid compositions which may be mentioned are powders, gel capsules and tablets. We can also mention the following oral forms or solid forms that are protected from the acidic medium (medium) of the stomach (stomach). The substrates used for the solid forms in particular are mineral substrates, for example phosphates or or carbonate, or organic substrates, for example lactose, cellulose, chromium or polymers. Liquid forms are: solutions, suspensions or dispersions. They contain as a dispersive medium either water or an organic solvent (ethanol, NMP or the like) or mixtures thereof or mixtures of surfactants and solvents, or complexing agents and solvents.

The liquid forms would probably preferably be in a form suitable for injection and as a result, will have a formulation acceptable for this use.

51136 00 Methods of administration acceptable for injection are intravenous, intraperitoneal, intramuscular and subcutaneous, with intravenous being preferred.

The dose of the compound of the present invention to be administered will be adjusted to that determined by the physician, as a function of the mode of administration to the patient and depending on the condition of the patient.

The compounds of the present invention may be administered alone or in combination with another anticancer agent. Of the possible combinations we can mention the following:

• alkylating agents and in particular cyclophosphamide, melphaian, ifosfamide, chlorambucil, busuifan, thiotepa, prednimustine, carmustine, lomustine, semustine, sieptozotocin, decarbazine, temozolomide, procarbazine and hexamethylmeplatinam oplatinum or derivatives • antibiotics such as, in particular, bleomycin, mitomycin, dactinomycin • antimicrotubular agents such as, in particular, vinblastine, vincristine, vindesine, vinorelbine, taxoids (paclitaxel and docetaxel) • anthracyclines such as, in particular, doxorubicin, daunorubicin, idarubicin, epirubicins, mitoxantrone, losoxantrone • group I and II topoisomerase inhibitors such as: etoposide, teniposide, amsacnn, irinotecan, topotecan and tomudex • fluoropyrimidines such as 5-fluorouracil, UFT, floxuridine • cytidine analogues such as 5-azacitidine, cytarabine, gemcitabine, 6mercaptomurine, 6-thioguostinine cytarabine or fludarabine phosphate • methotrexate and folic acid • various enzymes and compounds such as la L-asparaginase, hydroxyurea, transretinoic acid, suramines, dexrazoxane, amifostine, herceptin as well as estrogenic and androgenic hormones • antivasculami agents such as combretastatin derivatives, e.g. CA4P, chalcone derivatives or colchicine, e.g. ZD6126, and their prodrugs.

It is also possible to combine the compounds of the present invention with radiation treatment (radiation, radiation). These treatments can be administered

51136 Βimultaneously, separately or sequentially. The treatment will be adjusted by the doctor depending on the condition of the patient being treated.

The products of the invention are useful as inhibitors of the kinase catalyzed reaction, in particular FAK, KDR, Tie2, Aurora A, Aurora B and CDK2. FAK, KDR and Tie2 are kinases for which the manufacturers of the present invention are particularly useful as inhibitors.

The reasons why these kinases were selected are given in the following text:

FAK FAK is a cytoplasmic tyrosine kinase that plays an important role in the transmission of signals transmitted by tr integrins, a family of heterodyme cell adhesion receptors. FAK and integrins are located together in periimembranous structures known as adhesion packs. It has been shown in different cell types that the activation of FAKs and their phosphorylation on tyrosine residues and especially their autophosphorylation on tyrosine 397 is dependent on the binding of integrins to their extracellular ligands and thus indicated during cell adhesion [Komberg L, et al. J. Biol. Chem. 267 (33): 23439-442. (1992)]. Autophosphorylation at FAS tyrosine 397 represents a binding site for other tyrosine kinases, Src, via its SH2 domain [Schalleri al. Mol. Cell. Biol. 14: 1680-1688. 1994; Xingi al. Mol. Cell. Biol. 5: 413421. 1994]. The heart is able to phosphorylate FAK on tyrosine 925, thus recruiting the Grb2 protein adapter and inducing cell activation of dredged races and the MAP Kinase pathway involved in the control of cell proliferation [Schlaepfer et al. Nature; 372: 786-791.1994; Schlaepfer et al. Prog. Biophy. Mol. Biol. 71: 435-478. 1999; Schlaepfer and Hunter, J. Biol. Chem. 272: 13189-13195. 1997]. Activation of FAK can also induce jun NH2-terminal kinase (JNK) signaling pathway and result in cell progression toward the G1 cell cycle phase [Oktay et al., J. Cell. Biol.145: 1461-1469. 1999]. Phosphatidylinositol-3-OH kinase (Ρ13-kinase) also binds to FAK on tyrosine 397 and this interaction may be necessary for Ρ13 kinase activation [Chen and Guan, Proc. Nat. Acad. Sci. USA. 91: 10148-10152. 1994; Ling et al. J. Cell. Biochem. 73: 533- 544. 1999], the FAK / Src complex phosphorylates various substrates, for example, paxillins and pl30CAS in fibroblasts [Vuori et al. Mol. Cell. Biol. 16: 2606-2613.1996], [0029] The results of a number of studies support the hypothesis that FAK inhibitors may be useful in the treatment of cancer. Tests have indicated that FAK can play

51136 Β an important role in cell proliferation and / or cell survival in vitro. For example, in CHO cells, some authors have shown that overexpression of p125FAK leads to an acceleration of the G1 to S transition, suggesting that pl25FAK favors cell proliferation [Zhao J.-H et al. J. Cell Biol. 143: 1997-2008. 1998], Other authors have shown that tumor cells treated with FAK antisense oligonucleotides lose their adhesion and enter apoptosis (Xu et al., Cell Growth Differ. 4: 413-4.18.1996). FAK has also been shown to promote cell migration in vitro. Thus, fibroblasts that are incomplete for FAK expression (FAK "knockoul" mouse) show round morphology and lack of cell migration in response to hemostatic signals and these deficiencies are eliminated by re-expression (re-expression) of FAK [DJ. Sieg et al., J. Cell Science. 112: 2677-91.1999]. Overexpression of the C-terminal domain of FAK (FRNA) blocks the stretching of adrenal cells and reduces cell migration in vitro [Richardson A. and Parsons JT Nature. 380: 538-540. 1996]. Excessive expression of FAK in CHO, COS cells or in human astrocytoma cells promotes cell play. The participation of FAK in promoting cell proliferation and migration in numerous cell types in vitro indicates the important role of FAK in neoplastic processes. Recent studies have effectively shown an increase in tumor cell proliferation in vivo following induction of FAK expression in human astrocytoma human cells [Sagu LA et al. J. Cell Sci. 109: 1787-94. 1996; WangD et al. J. Cell Sci. 113: 4221-4230. 2000]. In addition, immunohistochemical studies of hyman biopsies showed that FAK was overexpressed in prostate cancer, breast cancer, thioride cancer, colon cancer, melanoma, brain cancer, and lung cancer, and FAK expression levels were directly associated with tumors of the most aggressive phenotype [WeinerTM, and al. Lancet. 342 (8878): 10241025. 1993; Owens et al. Cancer Research. 55: 2752- 2755. 1995; Maung K. et al. Oncogene. 18: 6824-6828. 1999; Wang D et al. J. Cell Sci. 113: 4221-4230. 2000],

KDR | 0030] KDR (Kinase insert Domain Receptor) also known as VEGF-R2 (Vascular Endothelial Growth Factor Receptor 2), is expressed exclusively in endothelial cells. This receptor binds to angiogenic growth factor VEGF, and accordingly serves as a mediastinum transduction signal through activation of its og kinase domain. Direct inhibition of VEGF-R2 kinase activity allows to reduce the phenomenon of angiogenesis in the presence of oxogenic VEGF (Vascular Endothelial Growth

51136 Β

Factor) (Strawn et al., Cancer Research, 1996, vol. 56, p.3540-3545). This process has been specifically shown using VEGF-R2 mutants (Millauer et al., Cancer Research, 1996, vol. 56, p.1615-1620). The VEGF-R2 receptor appears to have no other function in adults than that associated with the angiogenic activity of VEGF. Thus, a selective inhibitor of VEGF-R2 kinase activity should show only low toxicity.

In addition to its central role in the dynamics of angiogenic processes, recent results suggest that VEGF expression contributes to the survival of tumor cells in chemotherapy or radiotherapy, emphasizing the potential synergy of KDR inhibitors with other agents (Lee et al. Kancer Research, 2000, vol. 60). p.5565-5570).

Tie2 Tie-2 (TEK) is a member of the endothelial cell-specific receptor tyrosine kinase family. Tie2 is the first receptor with tyrosine kinase activity for which an agonist (angiopoietin 1 or Angl) is known to stimulate phosphorylation of receptors and signaling cells [S. Davis et al (1996) Cell 87,1161-1169] and an antagonist (angiopoietin 2 or Ang2) [PC Maisonpierre et al. (1997) Science 277, 55-60], Angiopoietin 1, can be synergized with VEGF end-stage neo-angiogenesis [AsaharaT. Circ. Res. (1998) 233- 240]. Knock-out experiments and transgenic manipulation of Tie2 or Angl expression lead to animals presenting vascularization defects [DJ Dumont et al (1994) Gencs Dev. 8,1897-1909 and C. Suri (1996) Cell 87, 1171-1180], Binding of Angl to its receptors leads to autophosphorylation of kinases of the Tie2 domain which is essential for neovascularization and also for engagement and interaction of blood vessels with peridites and smooth muscle cells; this phenomenon contributes to the maturation and stability of newly formed blood vessels [PC Maisonpierre et al (1997) Science 277, 55-60], Lin et al (1997) J. Clin. Invest. 100, 8: 2072- 2078 and Lynn P. (1998) PNAS 95, 8829-8834, showed inhibition of tumor growth and vascularization and also reduction of lung metastases, during adenoviral infection or injection of the extracellular domain of Tie-2 (Tek) in melanoma and breast cancer xenograft models.

Tie2 inhibitors can be used in situations where neovascularization (i.e., diabetic retinopathy, chronic neovascularization due to macular degeneration, psoriasis, chromic inflammation, Kaposi's sarcoma, rheumatoid arthritis, and infarct arthritis) is inappropriate.

51136 00 Cell cycle progression is driven by cyclin-dependent kinases (CDKs) that are activated by the interaction of proteins belonging to the cyclin family, this activation leads to substrate dophorylation and ultimately to cell division, In addition, endogenous CDK inhibitors that are activated (INK4 family) and KIP / CIP) negatively regulate CDK activity. The growth of normal cells is a psoeldica of balance between CDK activators (cyclins) and endogenous CDK inhibitors. In several types of cancer, uncontrolled expression or activity of several cell cycle regulators has been described.

Cyclin E activates Cdk2 kinase which then acts on the phosphorylation of pRb protein (retinoblastoma protein) leading to irreversible binding in cell division and S phase transition (PL Toogood, Medicinal Research Reviews (2001), 21 (6); 487-498 The CDK2 kinase and possibly CDK3 are necessary for progression to G1 and entry into the S phase.During the formation of complexes with cyclin E, they maintain pRb hyperphosphorylation to aid continuation from the G1 phase to the S phase. In complexes with cyclin A, CDK2 plays a role in E2F inactivation and is necessary for the realization of the S phase (TD. Davies et al. (2001) Structure 9,389-3).

The CDK1 / cyclin B complex regulates cell cycle progression between the G2 phase and the M phase. Negative regulation of the CDK / Cyclin B complex prevents normal cells from entering the S phase before the G2 phase is properly and completely completed. (KK Roy and EA Sausville Current Pharmaceutical Design, 2001, 7, 1669-1687).

There is a level of regulation of CDK activity. Cyclin-dependent kinase activators (CAKs) have a positive effect in the regulation of CDK. CAK phosphorylates CDK on threonine islets to make the target enzyme fully active.

The presence of defects in the meolcules involved in the cell cycle leads to CDK activation and cycle progression, it is normal to wish to inhibit CDK enzyme activity and thus block the growth of cancer cells.

Numerous proteins involved in chromosome segregation and having a spindle arrangement have been identified in yeast and Drosophila. Disorganization of these proteins leads to non-segregation of chromosomes and monopolar or disorganized spindles. Of these proteins, certain kinases, including Aurora and Ipll, derived from Drosophila and S. cerevisiae, are necessary for chromosome segregation by centrosome separation. The human analogue Ipll from yeast was recently cloned and characterized in various laboratories. These kinases, known as Aurora2, STK15 or BTAK belong to the serine / threonine kinase family, Bischoff et al. showed that jc Aurora2 oncogene, and multiplies

51136Β in colon cancer in humans (EMBO J, 1998,17,3052- 3065). This has also been described in cancers involving epithelial tumors such as breast cancer.

Definitions The term "halogen" refers to an element selected from F, Cl, Vg, and I.

The term "alkyl" refers to a linear or branched saturated hydrocarbon substituent containing from 1 to 12 carbon atoms. Substituents methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2 , 2-Dimethylpropyl, 1-ethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 1-ethylbutyl, 2-ethylbutyl, 3,3-dimethylbutyl, heptyl, 1-ethylpentyl, octyl, nonyl, decyl, undecyl, and dodecyl are examples of alkyl substituents.

The term "alkylene" refers to a linear or branched saturated hydrocarbon substituent containing from 2 to 12 carbon atoms. Substituents ethylene, 1-methylethylene, prop-1-enyl, prop-2-enyl, Z1-methylprop-1-enyl, El-methylprop-1-enyl, Z1,2-Dimethyl-prop-1-enyl, E-1,2 - Dimethylprop-1-enyl, but-1,3-dienyl, 1-methylidenediyl-2-enyl, Z-2-methylbut-1,3-dienyl, E-2-methylbut-1,3-dienyl, 2-methyl -1-Methylideneprop-2-enyl, undec-1-enyl and undec-10-enyl are examples of alkylene substituents. The term "alkynyl" refers to a linear or branched saturated hydrocarbon substituent containing at least two unsaturated bonds on a pair of carbon atoms, and containing from 2 to 12 carbon atoms. Examples of these alkynyl substituents are: ethynyl; prop-1-ynyl; prop-2-ynyl; and but-1-ynyl.

The term "aryl" refers to monocyclic or polycyclic atomic substituents containing from 6 to 14 carbon atoms. Examples of aryl substituents are: phenyl, naphth-1-yl; naphth-2-yl; anthracen-9-yl; 1,2,3,4-tetrahydronaphth-5-yl; yl, 2,3,4-tetrahydronaphth-6-yl.

The term "heteroaryl" refers to monocyclic or polycyclic heteroaromatic substituents containing from 1 to 13 carbon atoms and from 1 to 4 heteroatoms. Examples of the heteroaryl substituent are: pyrrol-1-yl; pyrrol-2-yl; pyrrol-3-yl; furyl; thienyl; imidazolyl; oxazolyl; thiazoles 1; isoxazolyl; isothiazolyl; 1,2,4-triazolyl; oxadiazolyl; thiadiazolyl; tetrazolyl; pyridyl; pyrimidyl; pyrazinyl; 1,3,5-triazinyl; indolyl; benzo [b] furyl; benzo [b] thienyl; indazolyl; benzimidazolyl; azaindolyl; quinolyl; isoquinole; carbazolyl; and acridyl.

51136 00 The term "heteroatom" in this context refers to at least a divalent non-carbon atom. Examples of these heteroatoms are: N; 0; S; and Se.

The term "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent containing from 3 to 12 carbon atoms. Examples of these cycloalkyl substituents are: cyclopropyl; cyclobutyl; cyclopentyl; cyclopentenyl; cyclopentadienyl; cyclohexyl; cyclohexenyl; cycloheptyl; bicyclo [2.2.1] heptyl; cyclooctyl; bicyclo [2.2.2] octyl; adamantil; and perhydronaphthyl.

The term "heterocyclyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent containing from 1 to 13 carbon atoms and from 1 to 4 heteroatoms. Preferably, the saturated or partially unsaturated cyclic hydrocarbon substituent is monocyclic and has 4 or 5 carbon atoms and 1 to 3 heteroatoms.

The term "substituted" refers to one or more substituents other than H, for example halogen; alkyl; aryl; heteroaryl, cycloalkyl; heterocyclyl; alkylene; alkynyl; OH; O-alkyl; O-alkylene; O-aryl; O-heteroaryl; NH₂; NH-alkyl; NH-aryl; NH-heteroaryl; '; Nalkyl-alkyl SH; S-alkyl; S-aryl; S (O₂) H; S (O₂) -alkyl; S (O₂) -aryl; SO₃H; SO₃. alkyl; SO 2 -aryl; CHO; C (O) -alkyl; C (O) -aryl; C (O) OH; C (O) O-alkyl; C (O) O-aryl; OC (O) alkyl; OC (O) -aryl; C (O) NH₂; C (O) NH-alkyl; C (O) NH-aryl; NHCHO; NHC (O) -alkyl; NHC (O) -aryl; NH-cycloalkyl; NH-heterocyclyl.

The products of the present invention may be prepared according to methods customary in organic chemistry. Scheme 1 below illustrates the process by which the compound of Example 1 was obtained, and relates to substituted b-aza-indoles. In this regard, it cannot constitute a limitation on the scope of the present invention for processes for the preparation of the compounds for which protection is claimed by the present invention. Preparation of 6-aza-indole-2-carboxamide derivatives substituted at position 3:

51136 Β

<img file="RS51136B_D0002.tif" />

Scheme 1

<img file="RS51136B_D0003.tif" />

H_a/ 2h rdasgaon

<img file="RS51136B_D0004.tif" />

NH, / MeOH nh₄oh

<img file="RS51136B_D0005.tif" />

<img file="RS51136B_D0006.tif" />

Scheme 2 below illustrates the methods used to prepare the examples relating to the substituted 7-aza-indoles, in particular the compound of Example 7. In this regard, it cannot constitute a limitation on the scope of the invention with respect to the methods for preparing the invention. whose protection is sought. Preparation of 7-aza-indole 2-carboxamide derivatives substituted at position 3:

51136Β

Scheme 2

<img file="RS51136B_D0007.tif" />

The products of general formula (I) wherein Ra is a second substituent of H can be obtained according to conventional methods known to those of ordinary skill in the art, for example, by replacing ammonia in aminolysis with an appropriate primary alkylamine.

La The subject of the invention is also a process for the preparation of a product of formula (I) as defined above, characterized in that the product of general formula (V) is hereinafter:

<img file="RS51136B_D0008.tif" />

Has undergone the following steps:

a) by halogenation in position 3, and then

51136 Β

b) Suzuki coupling in position 3, whereby the product of general (III) is obtained, ie:

<img file="RS51136B_D0009.tif" />

followed by

c) an ester amidation reaction in plate 2 to give the product of general formula (II) or:

<img file="RS51136B_D0010.tif" />

izatim

d) acylation of the amino-phenyl group in position 3.

The subject of the invention are also, as intermediates, compounds of general nature

<img file="RS51136B_D0011.tif" />

Where Z, Y, W are as previously defined, to give products of general formula (I).

LC / MS analyzes were performed on a Micromass LCT model connected to the HP 1100. Most manufacturers measured on an HP G1315A detector with a diode detector at a wavelength in the range of 200-600 nm and a Sedex 65 light scatter detector. Mass spectra were recorded in the range of 180 to 800. Data were analyzed using Micromass MassLinx software. Dispensing was performed on a Hypcrsil BDS C18, 3 pm (50 x 4.6 mm) column, eluting with a gradient of 5 to 90% acetonitrile containing 0.05% (v / v) trifluoroacetic acid (TFA) in water containing

51136 Β

0.05% (ν / ν) TFA over 3.5 min at a flow rate of 1 mL / mn. The total analysis time including column re-equilibration was 7 min.

Mass spectra of MS were recorded in electrospray (ES⁺) mode on the Platform II instrument (Micromass). The observed main (basic) ions are described.

Melting points were measured capillary, on a Mettler FP62 instrument, in the range of 30 ° C to 300 ° C, with a temperature increase of 2 ° C per minute.

Purification by LC / MS:

The products can be purified by LC / MS on a Waters FractionsLinx system consisting of a Waters model 600 gradient pump, a Waters model 515 regeneration pump, a Waters Reagent Manager dilution pump, a Waters model 2700 auto-injector. Rheodine model LabPro with two valves, Waters model 996 diode detector, mass spectrometer Waters model ZMD and collector fraction Gilson model 204. The system is controlled by Waters FractionLinx. Separation was performed alternately on two Waters Symmetry columns (C18, 5μΜ, 19x50 mm, catalog reference 186000210), one column is in water / acetonitrile 95/5 (v / v) regeneration mode with 0.07% ( c / c) a solution of trifluoroacetic acid in water, while the second column is used for separation. The columns were eluted using a gradient of 5 to 95% acetonitrile with a 0.07% (v / v) solution of trifluoroacetic acid in water at a flow rate of 10 mL / mn. After leaving the separation column, one thousandth of the effluent was separated using an LC Packing Accurate instrument, diluted with methanol at a flow rate of 0.5 mL / mn and directed to the detectors at a ratio of 75% to the diode detector, and the remaining 25% to the mass spectrometer. The rest of the effluent (999/1000) is directed to the fraction collector, where it is discarded if the mass of the expected product is not detected using FractionLinx software. The molecular formulas of the expected products are entered into the FractionLinx software which starts the product collection process when a mass signal corresponding to [M + H] is detected.⁺ion and / or [M + Na]⁺ ion. In certain cases, depending on the analytical results of LC / MS, when a pronounced [M + 2HJ⁺⁺ion, a value corresponding to half of the calculated molecular mass (MW / 2) was also entered into the FractionLinx software. Under these conditions, collection is also initiated when the [M + 2H] signal is detected.⁴⁴”And / or [M + Na + H]⁺⁺ion. The products were collected in a measured glass tube. After collection, the solvents were evaporated, in Savant AES 2000 or Genevac HT8

51136 Β centrifugal evaporator and product weights were determined by measuring the weight of the wash after evaporation of the solvent.

The following object of the invention relates to the products of the examples given below, which illustrate the present invention in a non-limiting manner.

Prinier 1:

3- [4- [3- (2-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine -

2-carboxamide

<img file="RS51136B_D0012.tif" />

A solution of 90 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in mL of tetrahydrofuran was added, dropwise, 50 μL of 2-fluoro-5 (trifluoromethyl) phenylisocyanate. The reaction mixture was stirred at ambient temperature under argon for 16 h and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. The suspended solid was filtered off and dried. After drying in vacuo at 40 ° C, 115 mg of 3- {4 [3- (2-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide are obtained. , with the following characteristics:

IR (KBr): 3455; 1661; 1602; 1542; 1444; 1341; 1312; 1127; 1070 and 819 cm '¹1 N NMR: 6.98 (s broad, 1H); 7.39 (m broad, 1H); from 7.42 to 7.56 (m, 4H); 7.60 (broad d, J 8.0 Hz, 2H); 7.74 (s broad, 1H); 8.17 (d, J = 6.0 Hz, 1 H); 8.65 (d broad, J =

7.5 Hz, 1H); 8.82 (s, 1 H); 8.94 (s broad, 1H); 9.31 (s, 1 H); 12.15 (s broad, 1H). Mass spectrum (ES⁺): m / z = 458 [M + H]⁺

Melting point: 286 ° C (Kofler).

51136 Β

3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide:

To a solution of 600 mg of ethyl 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxylate in 62 ml. An 11N solution of ammonia in methanol was added to 11 mL of 22%. aqueous ammonia solution The reaction mixture was stirred for 20 h in an autoclave at 80 ° C (12 bar), and then concentrated under reduced pressure. The resulting residue was diluted in 100 μL of methanol, treated with activated carbon and refluxed for 30 minutes. The venison was filtered warm through Celite and then washed with 2 x 10 mL of methanol. The filtrate was concentrated under reduced pressure to give 490 mg of 3- (4-aminophenyl) -1Hpyrrolo [2,3-c] pyridine-2-carboxamide as a foam, with the following characteristics: Mass spectrum (EI) m / z = 252 [M] + °, m / z = 235 [M-NH₃] + ° ethyl 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxylate:

To a solution of ethyl 1-3-bromo-1H-pyrrolo [2,3-c] pyridine-2-carboxylate in 100 mL of dioxane, 773 mg of 4-aminophenyl-boronic acid hydrochloride, 1.1 g of potassium fluoride in 9 mL of water were added. . The reaction mixture was stirred under argon for 15 minutes. 425 mg of tetrakis (triphenylphosphine) palladium (0) and 630 [mu] L of triethylamine were added. The reaction mixture was stirred at reflux for 17 h. After treatment with activated charcoal and then filtration through Ceiite®, the le filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica (60; 35-70 μΜ), eluting with a mixture of dichloromethane, methanol and acetonitrile (90/5/5 by volume). 600 mg of ethyl 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxylate are obtained, with the following characteristics: Mass spectrum (EI) m / z = 281 [M]⁺ , m / z = 235 [M-OEt]⁺ Ethyl 3-Bromo-1H-pyrrolo [2,3-c] pyridine-2-carboxylate:

To a solution of 2.24 g of ethyl 1H-pyrrolo [2,3-c] pyridine-2-carboxylate in 150 mL of pyridine, a solution of 3.53 g of pyridine tribromide in 30 mL of pyridine was added dropwise. 5 ° C. The reaction mixture was then stirred for 16 h at temperatures of about 20 ° C and then washed with 500 mL of ice-cold water. The suspension was filtered. The resulting solid was washed with water and then dried in a vacuum funnel at 40 ° C. 1.97 g of ethyl 3-bromo-1H-pyrrolo [2,3-c] pyridine-2-carboxylate are obtained, with the following characteristics; Mass spectrum (EI) m / z = 269 [M]⁺ , m / z = 189 [M-Br]⁺, m / z = 144 [M-OEt]⁺

51136 Ethyl 1H-Pyrrolo [2,3-c] pyridine-2-carboxylate:

1.8 g of palladium-on-carbon 10% were added to the atuclav, after which the atmosphere was changed to inert with argon. A solution of 6 g of ethyl 3- (3-nitropyridin-4-yl) -2-oxo-propionate in 72 mL of absolute ethanol was added. The reaction medium is then stirred for 3 h at 20 ° C under a hydrogen pressure of 2 bar. The mixture was then filtered through Celite®. The filtrate was concentrated under reduced pressure, and dried in an oven at 40 ° C to give 4 g of ethyl 1H-pyrrolo [2,3-c] pyridine-2-carboxylate, with the following characteristics:

Mass spectrum (EI) m / z = 190 [M⁺ ], m / z = 144 [M-OEt]⁺ Ethyl 3- (3-Nitropyridin-4-yl) -2-oxo-propionate:

To a solution of 930 mg of sodium in 50 mL of absolute ethanol was rapidly added 26 mL of diethyl oxalate. The reaction medium is stirred for 15 minutes at 20 ° C. A solution of 3.8 g of 4-methyl-3-nitropyridine in 50 mL of absolute ethanol was then added dropwise over 1 h. The reaction mixture was stirred at about 20 ° C for 4 h, and then concentrated under reduced pressure. To the residue was added 100 mL of ethyl ether and then filtered.

The solid residue was mixed with 40 mL of 5N hydrochloric acid solution, washed with water and dried under vacuum at 40 ° C to give 6.2 g of ethyl 3- (3-nitropyridin-4-yl) -2-oxo-propionate, followed by characteristics:

Mass spectrum (EI) m / z = 238 [M⁺ ].

Example 2: 3- [4- [3- (2-Methoxy-5-methyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-

c] pyridine-2-carboxamide

51136Β [0067]

<img file="RS51136B_D0013.tif" />

To a solution of 100 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in 5 mL of tetrahydrofuran was added dropwise 54.4 μL of 2-methoxy-5-methylphenylisocyanate. . The reaction mixture was stirred at ambient temperature under argon for 16 h and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. The suspended solid residue was filtered, washed with water and suction. After drying in vacuo at 40 ° C, 40 mg of 3- (4 [3- (2-methoxy-5-Methyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-carboxamide are obtained, with the following characteristics:

IR (KBr), 3458; 3331; 1664; 1595; 1537; 1315; 1285; 1213; 1135; 1033 cnf¹1 H NMR: 2.24 (s, ZN); 3.86 (s, ZN); 6.75 (broad d, J 8.5 Hz, 1H); from 6.85 to 6.95 (m, 2H); 7.43 (broad d, J = 8.5 Hz, 2H); 7.46 (d, J = 5.5 Hz, 1H); 7.58 (d wide, J -

8.5 Hz, 2H); 7.73 (s broad, 1H); 8.02 (s broad, 1H); 8.16 (d, J = 5.5 Hz, 1H); 8.22 (s. 1H); 8.82 (s, 1 H); 9.44 (s broad, 1H); 12.1 (s broad, 1H).

Mass spectrum (EI): m / z = 415 [M ^]

Melting point: 227 ° C

Example 3: 3- [4- [3- (3-Chloro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate

<img file="RS51136B_D0014.tif" />

51136 Rast To a solution of 100 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in 5 mL of tetrahydrofuran was added dropwise 45.2 μL of 3-chlorophenylisocyanate. The reaction mixture was stirred for 16 h at ambient temperature under argon and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. The suspended solid residue was filtered, washed with water and dried. Purification was performed by preparative LC / MS to give, after drying in vacuo at 40 ° C, 70 mg of 3- [4- [3- (3-chloro-phenyl) -ureido] -phenyl) -1H-pyrrolo [2,3-c] pyridine -2-carboxamide in the form of the trifluoroacetate salt and having the following characteristics: IR (KBr): 3390; 1672; 1592; 1537; 1483; 1203; 1138; 836; 722 cm '¹1 N NMR: 7.03 (m, 1H); from 7.26 to 7.34 (m, 2H); from 7.42 to 7.52 (m, ZN); 7.63 (broad d, J = 8.5 Hz, 2H); 7.74 (s broad, 1H); 7.97 (d, J = 6.0 Hz, 1 H); 8.06 (s broad, 1H); 8.31 (d, J = 6.0 Hz, 1 H); 9.03 (s broad, 2H); 9.13 (s. 1H); 13.35 (m broad, 1H). Mass spectrum (ES⁺): m / z = 406 [MH⁺]

Melting point: 221 ° C

Example 4: 3- [4- [3- (3-Chloro-4-fluoro-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine -

2-carboxamide trifluoroacetate [0071]

<img file="RS51136B_D0015.tif" />

To a solution of 100 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in 5 mL of tetrahydrofuran was added dropwise 46.2 μL of 3-chloro-4- fluorophenylisocyanate. The reaction mixture was stirred for 16 h at ambient temperature under argon and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. The suspended solid residue was filtered, washed with water and dried. Purification was performed by preparative LC / MS to give a vacuum drying operation at 40 ° C, 105 mg 3- {4- [3- (3-chloro-4-fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in the form of the trifluoroacetate salt, having the following characteristics: IR (KBr): 3452; 1673; 1601; 1544; 1500; 1208; 1143; 836; 803; 722 cm '¹

51136 Β ^!Η NMR: from 7.32 to 7.38 (tn, 2H); from 7.44 to 7.54 (m, ZN); 7.64 (đ broad, J = 8.5 Ηζ, 2H); 7.84 (broad d, J = 7.5 Ηζ, 1H); 8.01 (d, J = 6.0 Hz, 1 H); 8.09 (s broad, 1H); 8.32 (d, J = 6.0 Ηζ, 1H); 9.10 (s broad, 2H); 9.16 (s, 1 H); 13.4 (m wide, W). Mass spectrum (ES⁺): m / z = 424 [MH]⁺]

Melting point: 214 ° C

Example 5: 3- {4- [3- (2-Fluoro-5-Methyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-pyridine-2-carboxamide trifluoroacetate

<img file="RS51136B_D0016.tif" />

To a solution of 100 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in 5 [mu] L of tetrahydrofuran was added dropwise 48.3 [mu] L of 2-fluoro-5-methylphenyl isocyanate. The reaction mixture was stirred for 16 h at ambient temperature under argon and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. The suspended solid residue was filtered, washed with water and dried. Final purification was performed by preparative LC / MS to give, after drying in vacuo at 40 ° C, 36 mg 3- {4- [3- (2-fluoro-5-methyl-phenyl) ureido] -phenyl} -1H-pyrrolo [2, 3-Cypyridine-2-carboxamide in the form of a trifluoroacetate salt, with the following characteristics:

IR (KBr): 3452; 1675; 1603; 1544; 1314; 1202; 1144; 836; 805; 722 black 1 H NMR: 2.29 (s, ZN); 6.82 (m, 1H); 7.12 (dd, J = 8.5 and 11.5 Ηζ, 1H); from 7.46 to 7.51 (m, ZN); 7.62 (broad d, J = 8.5 Hz, 2H); from 7.97 to 8.03 (m, 2H); 8.08 (s broad, 1H); 8.32 (d, J = 6.5 Ηζ, 1H); 8.54 (broad d, J = 2.5 Ηζ, 1H); 9.15 (s. 1H); 9.25 (s. 1H); 13.4 (m wide, 1H)

Mass spectrum (ES⁺): m / z = 404 [MH⁺] Melting point; 222 ° C

Example 6: 3- [4- (3-m-tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate

51136 Β

<img file="RS51136B_D0017.tif" />

To a solution of 100 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide in 5 mL of tetrahydrofuran was added dropwise 47.8 μL of m-tolyl isocyanate. The reaction mixture was stirred at ambient temperature under argon and then concentrated under reduced pressure. The resulting residue was stirred for 30 minutes in 2 mL of dichloromethane. Suspended The solid residue was filtered off, washed with water and dried. Final purification was performed by preparative LC / MS to give, after drying in vacuo at 40 ° C, 40 mg of 3- [4- (3-m-tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide. in the form of the trifluoroacetate salt, having the following characteristics: IR (KBr): 3408; 1699; 1595; 1526; 1203; 1138; 834; 797; 724 cm '¹

NMR. * H: 2.29 (s, ZN); 6.81 (broad d, J = 7.5 Ηζ, 1H); 7.17 (t, J = 7.5 Ηζ, 1H); 7.25 (broad d, J = 7.5 Ηζ, 1H); 7.32 (s broad, 1H); 7.44 (s broad, 1H); 7.47 (d broad, J =

8.5 Ηζ, 2H); 7.62 (broad d, J = 8.5 Ηζ, 2H); 7.96 (m broad, 1H); 8.06 (s broad, 1H); 8.30 (d, J = 6.0 Ηζ, 1H); 8.67 (s. 1H); 8.86 (s, 1 H); 9.12 (s, 1 H); 13.3 (m broad, 1H).

Mass spectrum (ES⁺): m / z = 386 [MH &amp; lt; + &amp; gt ;.⁺]

Example 7: 3- [4- [3- (2-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0077]

<img file="RS51136B_D0018.tif" />

51136Β To a solution of 130 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide in 5 mL of tetrahydrofuran was added dropwise 85 [mu] L of 2-fluoro-5 (trifluoromethyl) phenylisocyanate. . The reaction mixture was stirred for 16 h at ambient temperature under argon and then concentrated under reduced pressure. The residue obtained is chromatographed on a column of islicium dioxide (eluent dichloromethane-methanol 9-1 by volume). Fractions containing the expected compound were concentrated in vacuo. 237 mg of 3- {4- [3- (2-fluoro-5-trifluoromethyl-phenyl) ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide are obtained in the form of a white solid, with the following characteristics:

IR (KBr): 1659; 1623; 1542; 1443; 1339; 1316; 1119 cm '¹ 1 N NMR: 7.08 (m broad, 1H); 7.14 (dd, J = 5.0 and 8.0 Hz, 1H); 7.40 (m, 1 H); 7.46 (broad d, J = 8.5 Hz, 2H); 7.51 (m. 1H); 7.57 (broad d, J 8.5 Hz, 2H); from 7.55 to 7.60 (m masque, 1H); 7.92 (broad d, J = 8.0 Hz, 1H); 8.38 (broad d, J = 5.0 Hz, 1H); 8.64 (broad d, J 7.5 Hz 1H); 9.01 (s broad, 1H); 9.36 (s broad, 1H); 12.1 (s broad, 1H). Mass spectrum (ES⁺): m / z = 458 [M + H] +⁺]

Melting point: 232 ° C (Kofler).

3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide:

To a solution of 260 mg of methyl 3- (4-aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate in 30 mL of a 7N solution of ammonia in methanol was added 5 mL of a 22% aqueous solution of ammonia. The reaction mixture was stirred for 20 h in an autoclave to 80 ° C (12.6 bar), and then concentrated under reduced pressure. The residue obtained is chromatographed on a silica column (eluent dichloromethane-methanol 9-1 by volume). Fractions containing the expected compound were concentrated under reduced pressure. 140 mg of 3- (4-aminophenyl) -1H-pyrrolo [2,3-b] pyridine 2-carboxamide are obtained in the form of a pale yellow solid, with the following characteristics: Melting point: 139 ° C methyl 3- (4-Aminophenyl) - 1H-pyrrolo [2,3-b] pyridine-2-carboxylate:

To a solution of 0.64 g of methyl 3-bromo-1H-pyrrolo [2,3-b] pyridine-2-carboxylate in 50 mL of toluene and 50 ml of methanol, 1.08 g of 4-aminophenyl-boronic acid hydrochloride and 0.9 ml of triethylamine. The reaction mixture was stirred under argon for 15 minutes. Then added in order: 144 mg tetrakis (triphenylphosphine) - palladium (0), 0.3

51136Β g of lithium chloride, 0.66g of sodium carbonate, and 7.5 mL of distilled water. The reaction mixture was stirred at reflux for 8 h. After filtration through Celite®, the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica eluting with a mixture of ethyl acetate and cyclohexane (7-3 volume ratio). 400 mg of methyl 3- (4-aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxylate are obtained in the form of a yellow solid, with the following characteristics:

Melting point: 236 ° C

Methyl 3-Bromo-1H-pyrrolo [2,3-b] pyridine-2-carboxylate:

To a solution of 3.2 g of methyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate hydrochloride in

165 mL · pyridine, a solution of 5.04 g of pyridinium tribromide in 35 mL of pyridine was added dropwise at 0 ° C under an argon atmosphere. The reaction mixture was then stirred at 0 ° C, the probe was poured into a mixture of 250 g of crushed ice and 750 ml of distilled water. The suspension was filtered, the solid residue was washed twice with 25 mL of distilled water, and then air dried. 0.87 gmethyl 3-bromo-1H "pyrrolo [2,3-b] pyridine-2-carboxylate was obtained in the form of a beige solid, with the following characteristics: Mass spectrum (ES⁺): m / z = 256 [M + H]⁺]

Methyl 1H-pyrrolo [2,3-b] pyridine-2-carboxylate hydrochloride:

To a solution of 4 g of 1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid hydrochloride in 100 mL of methanol was added dropwise 6 mL of thionyl chloride at ambient temperature. The reaction mixture was then stirred for 5 h at ambient temperature, and then concentrated under reduced pressure. The resulting residue was triturated in 50 mL of ethyl ether, and then dried in vacuo at 40 ° C. 3.22 g of methyl 1 Hpyrrolo [2,3-b] pyridine-2-carboxylate hydrochloride are obtained in the form of a light yellow solid, which is used in the next step.

1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid hydrochloride In solution, cooled to -70 [deg.] C., 6.03 g of 1H-pyrrolo [2,3-b] pyridine in 75 ml of anhydrous THF in a 33mL solution of 1.6M n-butyllithium in hexane was added dropwise. After stirring for 15 minutes at -70 ° C, 20 g of dry ice was added. The mixture was then allowed to warm to ambient temperature and concentrated

51136 Β under reduced pressure. 8.4 g of a white solid are obtained which is dissolved in 175 mL of tetrahydrofuran. The solution was cooled to -70 ° C, then 35 mL of a solution of 1.5Μ t-butyllithium in hexane was added dropwise. After stirring from 30 to -70 ° C, 20 g of dry ice was added to the solution. The mixture was then allowed to warm to ambient temperature, and this reaction mixture was then poured into 50 mL of distilled water which was cooled to 0 ° C. Tetrahydrofuran was reacted under reduced pressure. The residual aqueous solution was diluted with 50 mL of distilled water, washed twice with 100 mL of dichloromethane, acidified to pH1 by the addition of 30 mL of aqueous 5N hydrochloric acid, and then concentrated under reduced pressure. 10.01 g of product are obtained in the form of a paste, which is then recrystallized from 50 mL of methanol. The resulting solid was treated with a mixture of 50 mL of a 7N solution of hydrochloric acid in isopropanol and 50 mL of isopropyl ether. After air drying, 5.71 g of 1H-pyrrolo [2,3-b] pyridine-2-carboxylic acid hydrochloride are obtained in the form of a cream solid.

Mass spectrum (EI): m / z = 162 [M⁺ ]

Example 8: 3- {4- [3- (2-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -7-oxy-1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0084]

<img file="RS51136B_D0019.tif" />

To a solution of 50 mg of 3- {4- [3- (2-fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide in 2 mL of chloroform, at At 0 ° C, 0.31 mL of a 7M solution of methachloroperbenzoic acid in chloroform was added dropwise. The solution was stirred at 0 ° C for 4 h and then at ambient temperature for another 16 h. The reaction mixture was diluted with 3 mL of dichloromethane, filtered through a sintered glass sinter no. 4, and the resulting solid was washed twice with 3 mL of dichloromethane, and air dried. 40 mg of 3- {4- [3- (2-fluoro-5) are obtained

51136 Β Trifluoromethyl-phenyl) -ureido] -phenyl} -7-oxy-1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a light yellow solid, having the following characteristics:

IR (KBr): 3352; 1671; 1609; 1545; 1442; 1340; 1315; 1239; 1119; 1069 and 885 cm¹1 H NMR: 7.16 (m, 1H); from 7.35 to 7.58 (m, 7H); 7.63 (m broad, 1H); 7.77 (m broad, 1H); 8.31 (broad d, J 6.0 Hz, 1H); 8.65 (broad d, J = 8.5 Hz, 1H); 8.94 (s broad, 1H); 9.29 (s, 1H); from 12.5 to 13.2 (m shake wide, 1H).

Mass spectrum (ES⁺): m / z = 474 [M + H]⁺]

Melting point: 220 ° C (Kofler).

Example 9: 3- [4- (3- (2-Fluoro-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide)

<img file="RS51136B_D0020.tif" />

66.6 mg of solid 3- {4- [3- (2-Fluoro-phenyl) -urcido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide, beige, was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and 2-Fluorophenylisocyanate.

Melting point = 268.7 ° C (Buchi)

Mass spectrum (ES⁷): [M + H]⁺ = 390

Retention time (min): 3.71

Example 10: 3- [4- [3- (2-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide

51136Β

<img file="RS51136B_D0021.tif" />

83.6 mg of solid 3- {4- [3- (2-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide beige, obtained as described in Example 7 to starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2-methoxy-phenylisocyanate.

Melting point: 227.1 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 402

Retention time (min): 3.77

Example 11: 3- [4- [3- (4-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

b] pyridine-2-carboxamide [0090]

<img file="RS51136B_D0022.tif" />

77.6 mg of solid 3- {4- [3- (4-Trifluoromethyl-phenyl) -ureido] -phenyl} 1H-pyrrolo [2,3b] pyridine-2-carboxamide white, was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-trifluoromethyl-phenylisocyanate.

Melting point: 296.2 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 440

Retention time (min): 4.24

51136 Β

Example 12: 3- {4- [3- (2-Chloro-5-trifluoromethyl-phenyl) -ureido] -phenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide.

[0092]

<img file="RS51136B_D0023.tif" />

40.56 mg of solid 3- {4- [3- (2-Chloro-5-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide white, was obtained as which is described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2-Chloro-5-trifluoromethyl-phenylisocyanate.

Melting point; 188.3 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 474

Retention time (min): 4.51

Example 13: 3- [4- [3- (2-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide.

[0094]

<img file="RS51136B_D0024.tif" />

79 mg of solid 3- {4- [3- (2-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide beige, obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2-Fluoro-3-trifluoromethyl-phenylisocyanate.

51136 Β

Melting point: 265.4 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min): 4.24

Example 14: 3- {4- [3- (4-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo (2,3-b) pyridine-2-carboxamide.

10096]

<img file="RS51136B_D0025.tif" />

76.5 mg of solid 3- {4- [3- (4-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide brown, obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-Fluoro-3-trifluoromethyl-phenylisocyanate.

Melting point: 234.7 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min) .4.22

Example 15: 3- {4- [3- (3-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0098]

<img file="RS51136B_D0026.tif" />

78.1 mg of solid 3- {4- [3- (3-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide beige, obtained as is described in

51136 Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3-Fluoro-5-trifluoromethyl-phenylisocyanate.

Melting point: 257.5 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min): 4.42

Example 16: 3- {4- [3- (4-Trifluoromethoxy-phenyl) -ureidol-phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide.

[0100]

<img file="RS51136B_D0027.tif" />

92.3 mg 3- {4- [3- (4-Trifluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a brown powder was obtained as described in Example 7 starting from - (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-Trifluoromethoxy-phenylisocyanate.

Melting point: 258.9 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 456

Retention time (min): 4.29

Example 17: 3- [4- [3- (3,4-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0102]

51136 Β

<img file="RS51136B_D0028.tif" />

79 mg of oZ- {4- [3- (3,4'Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a beige solid was obtained as which is described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3,4-Dimethoxy-phenylisocyanate.

Melting point: 223.7 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ - 432

Retention time (min): 3.27

Example 18: 3- {4- [3- (2,5-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0104]

<img file="RS51136B_D0029.tif" />

75.9 mg of 3- [4- [3- (2,5-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a white solid was obtained as described in Example 7 starting from 3- (4-aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2,5-dimethyl-phenylisocyanate.

Melting point: 308.8 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 400

Retention time (min): 3.90

51136 Β

Example 19: 3- {4- [3- (3-Methoxy-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide

<img file="RS51136B_D0030.tif" />

55.5 mg 3- {4- [3- (3-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a beige solid was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3-methoxy-phenylisocyanate.

Melting point: 306.2 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 402

Retention time (min): 3.39

Example 20: 3- {4- [3- (3-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide

<img file="RS51136B_D0031.tif" />

56.5 mg 3- (4- [3- (3-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide, a white solid was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3-Trifluoromethyl-phenylisocyanate.

Melting point: 263.6 ° C (Biichi)

51136 Β

Mass spectrum (ES⁺): [M + H]⁺ = 440

Retention time (min): 3.95

Example 21: 3- {4- [3- (3,4-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide |

<img file="RS51136B_D0032.tif" />

45.2 mg of 3- (4- [3- (3,4-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide white solid was prepared as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3,4-Dimethyl-phenylisocyanate.

Melting point: 274.7 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ - 400

Retention time (min): 3.75

Example 22: 3- {4- [3- (2-Methoxy-5-Methyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

b] pyridine-2-carboxamide.

[0112]

<img file="RS51136B_D0033.tif" />

44.9 mg 3- {4- [3- (2-Methoxy-5-Methyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide as a beige solid obtained is like What is

51136 Β described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2-Methoxy-5-methyl-phenylisocyanate.

Melting point: 327.7 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 416

Retention time (min): 3.76

Example 23: 3- [4- (3-m-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide

<img file="RS51136B_D0034.tif" />

62.5 mg of 3- [4- (3-m-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide in the form of a beige solid was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and m-tolylisocyanate.

Melting point: 266 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 386

Retention time (min): 3.60

Example 24: 3- {4- [3- (4-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide

<img file="RS51136B_D0035.tif" />

51136 .7 49.7 mg 3- {4- [3- (4-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide Form Beige solid was obtained as was described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-Fluoro-phenylisocyanate.

Point fusion: 299.9 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 390

Retention time (min): 3.45

Example 25: 3- [4- (3-p-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0118] 68.4 mg 3- [4- (3-p- Tolyl-ureido-phenyl] -1H-pyrrolo [2,3-b] pyridine-2-carboxamide in the form of a beige solid was prepared as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2] , 3-bipyridine-2-carboxamide and p-tolylisocyanate.

Melting point: 293 ° C (Biichi)

Mass spectrum (ES⁺): [M + H]⁺ = 386

Retention time (min): 3.58

Example 26: 3- {4- [3- (4-Methyl-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

b] pyridine-2-carboxamide [0120]

51136 Β

<img file="RS51136B_D0036.tif" />

47.1 mg 3- {4- [3- (4-Methyl-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a white solid, was prepared as described in Example 7 starting from - (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-Methyl-3-trifluoromethyl-phenylisocyanate.

Melting point: 285 ° C

Mass spectrum (ES⁺): [M + H]⁺ = 454

Retention time (min): 4.10

Example 27: 3-] 4- [3- (4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0122]

<img file="RS51136B_D0037.tif" />

47.5 mg 3- {4- [3- (4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide as a white solid, was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-Difluoromethoxy-phenylisocyanate.

Melting point: 283.5 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 438

Retention time (min): 3.64

51136 Β

Example 28: 3- [4- [3- (3,5-Dimethoxy-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-b] pyridine

-2-carboxamide [0124]

<img file="RS51136B_D0038.tif" />

59.2 mg o beige of 3- {4- [3- (3,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a beige solid dye, was prepared as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and 3,5-Dimethoxy-phenylisocyanate.

Melting point: 266.5 ° C (Buchi)

Mass spectrum (ES '): [M + H]⁺ = 432

Retention time (min): 3.45

Example 29: 3- [4- [3- (4-Chloro-3-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-

b] pyridine-2-carboxamide [0126]

<img file="RS51136B_D0039.tif" />

29.8 mg 3- {4- [3- (4-Chloro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide as a white solid, obtained as

51136 Β as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 4-N 1 -O-3-trifluoromethyl-phenylisocyanate.

Melting point: 311.GS (Buchi)

Mass spectrum (ES⁺): [M + H]⁺ = 474

Retention time (min): 4.22

Example 30: 3- {4- [3- (2,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0128]

<img file="RS51136B_D0040.tif" />

33.1 mg of 3- {4- [3- (2,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3b] pyridine-2-carboxamide lyophilisate yellow, obtained as described in Example 7, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2,5-Dimethoxy-phenylsanate.

Mass spectrum: LC-MS-JAD-ELSD: 432 (+) = (M + H) (+); 430 (-) = (Μ - H) (-) Retention time (min): 3.53

Example 31: 3- [4- [3- (3-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2₅3- bipyridine-2-carboxamide.

[0130]

<img file="RS51136B_D0041.tif" />

51136 .5 31.5 mg of a white lyophilisate of 3- {4- [3- (3-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide was obtained as described in Example 7 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3-Fluoro-phenylisocyanate.

LC-MS-DAD-ELSD mass spectrum: 390 (+) = (M + H) (+); 388 (-) = (MH) (-) Retention time (min): 3.55

Example 32: 3-] 4- [3- (2-Methoxy-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide [0132]

<img file="RS51136B_D0042.tif" />

50 mg of solid 3- {4- [3- (2-Methoxy-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide beige, obtained as described in Example 7, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 2-Methoxy-5-trifluoromethyl-phenylisocyanate.

Melting point: 221 ° C (Kofler sublimation)

Mass spectrum of LC-MS-DAD-ELSD: 470 (+) = (M + H) (+) 468 (-) = (MH) (-)

Example 33: 3- {4- [3- (2-Acetylamino-5-trifluoromethyl-phenyl) ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate.

[0134]

51136 Β

<img file="RS51136B_D0043.tif" />

12 mg of solid yellow 3- {4- [3- (2-Acetylamino-5-trifluoromethyl-phenyl) -ureido] phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide was obtained as which is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2-Acetylamino-5-trifluoromethyl-phenylisocyanate.

Mass spectrum (ES⁺): [M + H]⁺ = 497

Retention time (min): 2.63

Example 34: 3- [4- [3- (2-Methoxy-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0044.tif" />

25 mg solid 3- (4- [3- (2-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

c] yellow pyridine-2-carboxamide was prepared as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2-methoxy-phenylisocyanates.

Melting point: 216 ° C (Kofler)

Mass spectrum (ES⁺): [M + H]⁺ = 402

Retention time (min): 3.06

Example 35: 3 {4- [3- (2-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

c] pyridine-2-carboxamide.

51136 Β [0138]

<img file="RS51136B_D0045.tif" />

80 mg of 3- {4- [3- (2-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-pyridine-2-carboxamide, as a yellow solid, was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2-Trifluoromethyl-phenylisocyanates.

Melting point: 228 ° C (Kofler)

Mass spectrum (ES *): [Μ + H]⁺ = 440

Retention time (min): 3.17

Example 36: 3- {4- [3- (3-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0046.tif" />

77 mg 3- {4- [3- (3-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

c] pyridine-2-carboxamide as a yellow solid was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and 3- to 3-trifluoromethyl- phenylisocyanate.

Melting point: 256 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 440

51136 Β

Retention time (min): 3.48

Example 37: 3- {4- [3- (4-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0047.tif" />

73 mg of 3- {4- [3- (4-Fluoro-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-fluoro-phenylisocyanates.

Melting point: 271 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ - 390

Retention time (min): 2.93

Example 38: 3- [4- [3- (4-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-

c] pyridine-2-carboxamide

<img file="RS51136B_D0048.tif" />

91 mg of 3- {4- [3- (4-Trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine -243

51136Β of carboxamide and of 4-Trifluoromethyl-phenylisocyanate.

Melting point: 289 ° C

Mass spectrum (ES⁺): [M + H]⁺ = 440

Retention time (min): 3.48

Example 39: 3- [4- (3-p-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0049.tif" />

76 mg of 3- [4- (3-p-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as described in Example 1. starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from p-tolyl isocyanates.

Melting point: 277 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 386

Retention time (min): 3.13

Example 40: 3- {4- [3- (4-Chloro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0050.tif" />

51136 Mg 103 mg 3- {4- [3- (4-Chloro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-Chloro-3-trifluoromethyl-phenylisocyanate.

Melting point: 228 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 474

Retention time (min): 3.64

Example 41: 3- {4- [3- (2-Chloro-5-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide [0150]

<img file="RS51136B_D0051.tif" />

76 mg 3- {4- [3- (2-Chloro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide as a yellow solid, was prepared as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2-Chloro-5-trifluoromethyl-phenylisocyanate.

Melting points: 243 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ ~ 474

Retention time (min): 3.56

Example 42: 3- [4- [3- (4-Trifluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-pyridine-2-carboxamide]

51136 Β

<img file="RS51136B_D0052.tif" />

94 mg of 3- {4- [3- (4-Trifluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as which is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-Trifluoromethoxy-phenylisocyanate.

Melting point: 276 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 456

Retention time (min): 3.63

Example 43: 3- {4- [3- (4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-pyridine-2-carboxamide]

<img file="RS51136B_D0053.tif" />

87 mg of 3- {4- [3- (4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide as a yellow solid, obtained as was described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-Difluoromethoxy-phenylisocyanate.

Melting point: 257 ° C

Mass spectrum (ES⁺); [M + H]⁺ =4.38

Retention time (min): 3.23

Example 44: 3- {4- [3- (3,4-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

51136Β

<img file="RS51136B_D0054.tif" />

82 mg 3- {4- [3- (3,4-Dimethyl-phenyl) -urcido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3,4-dimethyl-phenylisocyanate.

Melting bag: 230 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 400

Retention time (min): 3.32

Example 45: 3- [4- [3- (3,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine

-2-carboxamide [0158]

<img file="RS51136B_D0055.tif" />

87 mg 3- {4- [3- (3,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine 2-carboxamide, as a yellow solid, was prepared as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and 3,5-Dimethoxy-phenylisocyanate.

Melting point: 225 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 432

Retention time (min): 3.07

51136Β

Example 46: 3- {4- [3- (2,5-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0056.tif" />

10161J 87 mg 3- {4- [3- (2,5-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and 2,5-Dimethyl-phenylisocyanate.

Melting point: 261 ° C (Biichi B-545)

Mass spectrum (ES⁴): [M + H]⁺ = 400

Retention time (min): 3.25

Example 47: 3- {4- [3- (2-Fluoro-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0057.tif" />

59 mg 3- {4- [3- (2-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid, was obtained as which is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2-Fluoro-phenylisocyanates.

Melting point: 242 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 390

51136 Β

Retention time (min): 2.41

Example 48: 3- {4- [3- (3-Fluoro-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0058.tif" />

63 mg of 3- (4- [3- (3-Fluoro-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid, was obtained as which is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3-Fluoro-phenylisocyanate.

Melting point: 252 ° C (Biichi B-545)

Mass spectrum (ES +): [M + H] + = 390

Retention time (min): 2.55

Example 49: 3- [4- [3- (2-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide [0166]

<img file="RS51136B_D0059.tif" />

69 mg 3- (4- [3- (2-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide as a pale yellow solid was prepared as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3 'c] pyridine-2-carboxamide and from 2-Fluoro-3-trifluoromethyl-phenylisocyanate.

51136 Β

Melting point: 240 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min): 2.75

Example 50: 3- [4- [3- (3-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide [0168]

<img file="RS51136B_D0060.tif" />

69 mg 3- {4- [3- (3-Fluoro-5-trifluoromethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-pyridine-2-carboxamide as a pale yellow solid was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3-Fluoro-5-trifluoromethyl-phenylisocyanate.

Melting point: 261 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min): 2.88

Example 51: 3- {4- [3- (4-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide [0170]

<img file="RS51136B_D0061.tif" />

51136Β 56 mg 3- {4- [3- (4-Fluoro-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, pale yellow solids were prepared as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-Fluoro-3-trifluoromethyl-phenylisocyanate.

Melting points: 201 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 458

Retention time (min): 2.85

Example 52: 3- [4- [3- (4-Methyl-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-pyridine-2-carboxamide]

F,

<img file="RS51136B_D0062.tif" />

61 mg of 3- {4- [3- (4-Methyl-3-trifluoromethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid was prepared as described in Example 1, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-methyl-3-trifluoromethyl-phenylisocyanate.

Melting point: 199 ° C

Mass spectrum (ES⁺): [M + H]⁺ = 454

Retention time (min): 2.84

Example 53: 3- {4- [3- (3-Methoxy-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate

51136 Β

<img file="RS51136B_D0063.tif" />

33.3 mg of yellow lyophilisate 3- {4- [3- (3-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide white solid was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3-Methoxy-phenylisocyanate.

Mass spectrum (ES⁺): [M + H]⁺ = 402

Retention time (min): 2.60

Example 54: 3- {4- [3- (3,4-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate

<img file="RS51136B_D0064.tif" />

80.5 mg of yellow {glyophilisate 3- {4- [3- (3,4-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate was obtained as described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3,4-Dimethoxy-phenylisocyanate.

Mass spectrum (ES⁺): [M + H]⁺ = 432

Retention time (min): 2.27

Example 55: 3- [4- [3- (2,5-Dimethoxy-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate

51136 Β

<img file="RS51136B_D0065.tif" />

90.7 mg of a yellow lyophilisate of 3- {4- [3- (2,5-Dimethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide trifluoroacetate, was obtained as is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 2,5-Dimethoxy-phenyl lysocyanate.

Mass spectrum (ES⁺): [M + H]^+=:: 432

Retention time (min): 2.62

Example 56: 3- [4- (3-O-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0066.tif" />

75.3 mg of 3- [4- (3-o-Tolyl-ureido) -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid, was obtained as described in Example 1, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from o-Tolylisocyanate.

Melting point: 270 ° C (Biichi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 386

Retention time (min): 2.54

51136Β

Example 57: 3- {4- [3- (4-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0067.tif" />

51.1 mg of 3- {4- [3- (4-Methoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid, was obtained as which is described in Example 1 starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 4-Methoxy-phenylisocyanate.

Melting point: 275 ° C (Buchi B-545)

Mass spectrum (ES +): [M + H] + = 402

Retention time (min): 2.28

Example 58: 3- {4- [3- (3-Chloro-4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide [0184]

<img file="RS51136B_D0068.tif" />

93 mg3- (4- [3- (3-Chloro-4-Difluoromethoxy-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide as a pale yellow solid obtained is as described in Example 1, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-pyridine-2-carboxamide and from 3-Chloro-4-Difluoromethoxy-phenylisocyanate.

Melting point: 267 ° C (Buchi B-545)

51136 Β

Mass spectrum (ES⁺): [Μ + N]^t= 472

Retention time (min): 2.90

Example 59: 3- {4- [3- (3,5-Dimethyl-phenyl) -ureido} -phenyl} 4H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0069.tif" />

61 mg 3- (4- [3- (3,5-Dimethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a pale yellow solid, was obtained as described in Example 1, starting from 3- (4-Aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and 3,5-Dimethyl-phenylisocyanate.

Melting point: 188 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 400

Retention time (min): 2.68

Example 60: 3- [4- [3- (3-Ethyl-phenyl) -ureido] -phenyl] -1H-pyrrolo [2,3-c] pyridine-2-carboxamide

<img file="RS51136B_D0070.tif" />

61 mg 3- {4- [3- (3-ethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-c] pyridine-2-carboxamide, as a yellow solid, was obtained as is described in Example 1, from 3- (4-aminophenyl) -1H-pyrrolo [2,3-c] pyridine-2-carboxamide and from 3-ethyl55

51136Β of phenylisocyanate.

Melting point: 257 ° C (Buchi B-545)

Mass spectrum (ES⁺): [M + H]⁺ = 400

Retention time (min): 2.97

Example 61: 3- [4- [3- (3-Ethyl-phenyl) -ureido] -fend} 4H-pyrrolo [2,3-b] pyridine-2-carboxamide

<img file="RS51136B_D0071.tif" />

0.8 mg of 3- [4- [3- (3-ethyl-phenyl) -ureido] -phenyl} -1H-pyrrolo [2,3-b] pyridine-2-carboxamide, as a white solid was obtained as described in Example 7 starting from 3- (4-Aminophenyl) 4H-pyrrolo [2,3-b] pyridine-2-carboxamide and from 3-methyl-phenylisocyanate.

Melting point: 254 ° C (Buchi)

Mass spectrum (ES⁺): [M + H]⁺= 400

Retention time (min): 7.18

Determination of compound activity - Experimental protocols

l.FAK

10192] The inhibitory activity of the compounds on FAK was determined by measuring the inhibition of enzyme autophosphorylation by the degradation fluorescence assay (HTRF).

The entire human FAK cDNA, histidine-labeled N-terminal kgaj, was cloned into the expression vector of pFastBac HTc baculovirus. The protein was expressed and purified to about 70% homogeneity.

Kinase activity was determined by incubation of the enzyme (6.6 pg / ml) with different concentrations of the test compound (test compound) in 50 mM Hepes pH = 7.2.10 mM MgCh, 100 μΜ Na₃IN₄, 15 μΜ d'ATP buffer, during Ih, at 37 ° C. Enzymatic

The 51136Β reaction was stopped by adding Hepes pH = 7.0 buffer containing 0.4 mM KF, 133 mM EDTA, 0.1% BSA and labeling was performed for 1 to 2 h at ambient temperature by adding anti-Histidine antibody to this buffer. labeled with HE665 and phosphospecific monoclonal antibodies tyrosine conjugated to europium cryptate (Eu-K). The characteristics of these two fluorophores can be found in G. Mathis et al., Anticancer Research, 1997, 17, pages 3011-3014. The transfer of energy from the excited europium cryptate to the XL665 acceptor is proportional to the degree of autophosphorylation of FAK. The long-term signal specific to the XL-665 was measured in a Packard Discovery reader. All tests were performed in duplicate and the average value of the two values ​​was calculated. Inhibition of autophosphorylation activity by FAK, with the compounds of the invention is expressed as a percentage of inhibition relative to a control whose activity is measured without the test compound. To calculate the percentage of inhibition, the ratio [signal at 665 nm / signal at 620 nm] was considered,

2. KDR The inhibitory effect of the compound was determined in an in vitro substrate phosphorylation assay with KDR enzyme by scintillation technique (96-well plate, NEN).

The cytoplasmic domain of the human KDR enzyme is a cloned form of GST fusion in the pFastBac baculovirus expression vector. The protein was expressed in SF21 cells and purified to about 60% homogeneity.

KDR kinase activity was measured in 20 mM MOPS, 10 mM MgCl2.10 mM MnCl2, 1 mM DTT, 2.5 mM EGTA, 10 mM b-glycerophosphate, pH = 7.2, in the presence of 10 mM MgCl₂, 100 μΜ Na₃IN₄, 1 mM NaF. 10 μΐ of the compound was added to 70 μΐ of kinase buffer containing 100 ng of KDR enzyme, at 4 ° C. The reaction was initiated by the addition of 20 μΐ solution containing 2 pg of substrate (SH2-SH3 fragment of PLCy expressed in the form of GST fusion protein), 2 pCi γ³³P [ATP] and 2 μΜ cold ATP. After incubation for 1 h at 37 ° C, the reaction was stopped by adding 1 volume (100 μ za) of 200 tM EDTA. Incubation buffer was removed, and the walls were evaporated three times with 300 μΙ PBS. Radioactivity was measured in each well using a Top Count radio (Packard) radioactive counter.

Background was determined by measuring radioactivity in four different deposits with radioactive ATP -ot and substrate.

51136 01 Total control of measurement activity in four different reservoirs with all reagents (γ³³Ρ- [ΑΤΡ], KDR and substrate PLCy), but without compound.

Inhibition of KDR activity with a compound of the present invention is expressed as a percentage of the specific inhibition of control activity, in the absence of the compound.

Compound SU5614 (Calbiochem) (1 μΜ) was included in each plate as an inhibition control.

3. Tie2 The human Tie2 coding sequence corresponding to the amino acids of the intracellular domain 776-1124 was generated by PCR using cDNA isolated from the human placenta as a model. This sequence was introduced into the baculovirus expression vector pFastBacGT in the form of the GST fusion protein.

The inhibitory effect of the molecules was determined in a PLC phosphorylation assay with Tie2 in the presence of GST-Tie2 purified to about 80% homogeneity. The substrate is composed of SH2-SH3 fragments of PLC expressed as GST fusion protein.

Tie2 kinase activity was measured in MOPS 20 mM pH 7.2 buffer, containing 10 mM MgCl₂, 10 mM MnCl₂, 1 mM DTT, 10 mM glycerophosphate. In a 96-well FlashPlate plate held on ice, a reaction mixture was added, consisting of 70 μL of okinase buffer containing 100 ng of GST-Tie2 enzyme per well. Further, a 10 μL test molecule diluted in DMSO was added to a maximum concentration of 10%. For the given concentrations, each measurement was performed four times. The reaction was initiated by the addition of 20 μΐ solution containing 2 μβ GST-PLC, 2 μΜ cold ATP and 1 pCi³³P [ATP]. After incubation for 1 h at 37 ° C, the reaction was stopped by the addition of 1 volume (100μΙ) of 200 mM EDTA. After removal of the incubation buffer, the wells were washed three times with 300 pL of PBS. Radioactivity was measured on a MicroBetal450 Wallac.

Inhibition of Tie2 activity was calculated and expressed as a percentage of inhibition relative to the control activity determined in the absence of compound

4. Aurora 1 and Aurora 2 The inhibitory effect of the compound on Auroral and Aurora2 kinases was determined by enzyme assay, radioactivity detection.

The kinase activity of Aurora 1 and Aurora 2 was determined by phosphorylation of the substrate Numa-histidine in the presence of radiolabeled ATP ([³³P] ATP) na

51136 Β

Flashplates with 96-well plates in which nickel chelate is attached to the surface of the microplate. Quantity³³The phosphate incorporated into the NuMA substrate is proportional to the activity of the enzyme Auroral or Aurora2.

Proteins;

Proteins were obtained in the Sanofi-Aventis Group Protein Production Laboratory.

Aurora 1: recombinant Aurora-B / INCENP-C3 complex, purified to about 50%, histidine-labeled N-terminal end of Aurora-B.

Aurora 2: whole recombinant rotein contains N-terninal histidine tail, expressed in E. coli and purified to 82%.

NuMA (nucleotide protein combined with mitotic apparatus): a 424 amino acid fragment, excreted in E. coli, an N-terminal end labeled with histidine and used as a substrate for two Aurora enzymes.

Protocol:

Flash-Plate microplates with 96 wells, nickel chelated (Perkin Elmer, model SMP107) were used.

The test products were incubated in a reaction volume of 100 μL per well, in the presence of 10 nM Aurora 1 or Aurora 2, 500 nM NuMA substrate in a buffer consisting of 50 mM Tris / HCl (pH 7.5), 50 mM NaCl, 5 mM MgCl₂ (Aurora-B) or 10 mM MgCl₂ (Aurora-A) and 1 mM DTT, at 37 ° C.

80 [mu] L of enzyme / substrate of incubation buffer was added to each well, and then 10 [mu] L of product for analysis was added, at various concentrations. The reaction was initiated by the addition of ΙμΜ ATP at a final concentration of 0.2 pCi [³³P] ATP (10 pL). After incubation for 30 minutes, the reaction was stopped by simply removing the reaction buffer and each well was washed with 300 μΐ Tris / HCl buffer. The radioactivity in each well was then measured using a Packard scintillation apparatus, Top -Count model.

The control enzyme activity of Aurora was expressed as the number of readings per minute obtained within 30 minutes after subtraction of the background (enzyme-free reaction mixture). Evaluation of different test products was expressed as the percentage inhibition of Aurora activity relative to control.

51136 Β

5. CDK2 / cyclin Ε:

Purification of the CDK2 / CyclmeE- (His) fi complex by IMAC (Immobilized Metal Affinity Chromatography):

Two recombinant bacilliviruses with human sequences encoding, respectively, CDK2 and Cyclin E (the latter has a hexa-histidine tag at the C-terminal end) were used to co-infect insect Sf21 cells. Two to three days after the onset of co-infection, cells were harvested by centrifugation and then stored at -40 ° C until use. After thawing the mechanical lysis of the cells, the complex present in the supermantate obtained during lysis was purified by affinity chromatography on nickel (IMAC), and left at -80 ° C.

CDK2 / CiklinE test on Flashplate format in 96 hubs.

Plates in a 96-well streptavidin-plated format were used to determine the activity of the compound and the kinase activity of CDK2 / Cyclin E.

To perform this assay, a biotinylated, fragment of the pRb protein, (biotinylSACPLNLPLQNNHTAADMILSPVRSPKKKGSTTR-OH) was dissolved to a concentration of 1 mM in kinase buffer (HEPES / NaOH 50 mM, NaCl 1 mM, MgCl₂5 mM, pH 7.5) to give a stock solution stored at -20 ° C in the form of an aliquot of 110 pL. Le On the day of the experiment, aliquots of this solution were thawed and diluted in kinase buffer containing 1 mM dithiothreitol, added to the buffer without prior preparation to achieve a concentration of 14.3 μΜ. 70 pL of this solution was added to each well of the Flashplate plate to obtain a final substrate concentration of 10 μΜ during the enzymatic reaction at the end of the final volume of the reaction medium of 100 μΙ, (cf. below).

Intermediate dilutions of inhibitors (products of the invention) at various concentrations were prepared in DMSO starting from a stock solution of 10 mM in separate tubes. Thus, the following dilutions were prepared: 1000 μΜ, 333.3 μΜ, 111.1 μΜ, 37.03 μΜ, 12.35 μΜ, 4.11 μΜί 1.37pM. One pLod of each of these solvents (or 1 pL DMSO for control) was transferred to plate plates.

51136Β A 19 μΐ solution of a mixture of adenosine triphosphate (ATP) and ΑΤΡγ P in kinase buffer at a total concentration of 5.26 μ koncent ATP and 52.6 pCi / ml was added to each well.

Q. The enzymatic reaction was initiated by the addition of 10 μΕ per well of a 200 nM CDK2 / CyclinE kinase buffer solution containing 1 mM dithiothreitol (or 10 μL of kinase buffer containing 1 mM dithiothreitol for blanks).

After the addition of each of the reagents, the final volume of each well was 100 μL, the final substrate concentration was 10 μΜ, the final concentrations of sulO μΜ inhibitors, 3.33 μΜ, 1.11 μΜ, 0.37 μΜ, 0.123 μΜ, 0.041 μΜ and 0.014 μΜ (according to intermediate dilution concentrations), final ATP concentration is 1 μΜ, final concentrationⁿP is IpCi / well, and the final concentration of CDK2 / Cyclin E is 20 nM.

After the addition of all reagents, the plate was incubated at 30 ° C with stirring on an orbital stirrer at 650 rpm.

At the end of the incubation, the plate was washed three times with 300 μΕ PBS (Phosphate Buffered Saline, pH = 7.4 without calcium or magnesium, reference 10010-015, Gibco BRL). per bed. Incroporation³³The P u peptide was quantified by the Packard Topcount.NXT scintillation counter. The inhibitory activity of the product of the invention was determined by measuring the inhibitory concentration that offsets 50% (IC50) of the reduction in enzyme activity.

Results:

[0224]

Table 1:

<td>Example</td><td colspan="6">IC 50 (pM)</td>

<td></td><td>I \ K</td><td>KDR</td><td>TIE2</td><td>Aurora A</td><td>Aurora B</td><td>CDK2</td>

<td> 1</td><td> 104</td><td> 29</td><td> 4</td><td> 172</td><td> 138 ...................... ......</td><td> ......................................</td>

<td> 2</td><td> 299</td><td> 150</td><td> 21</td><td> 222</td><td> 196</td><td></td>

<td> 5</td><td> 249</td><td> 258</td><td> 47</td><td> 131</td><td> 67</td><td></td>

<td> 7</td><td> 184</td><td> 34</td><td> 9</td><td> 553</td><td> 133</td><td></td>