MX2008008492A - Amino acid derivatives of indolinone based protein kinase inhibitors - Google Patents

Abstract

Amino acid derivatives of pyrrolyl-indolinones and their amide or ester derivatives have enhanced and unexpected drug properties as inhibitors of protein kinases and are useful in treating disorders related to abnormal protein kinase activities such as cancer.

Description

DERIVATIVES OF AMINO ACIDS OF PROTEIN INHIBITORS-KINASES BASED ON INDOLINONE Field of the Invention The invention relates to inhibitors of protein kinases, and to their use in the treatment of disorders related to abnormal activities of protein kinases such as cancer and inflammation. More particularly, the invention relates to amino acid derivatives of pyrrolyl indolinones and their amide or ester derivatives and their pharmaceutically acceptable salts which can be used as inhibitors of protein kinases.

BACKGROUND OF THE INVENTION Protein kinases are enzymes that catalyze the phosphorylation of hydroxyl groups of tyrosine, serine and threonine residues of proteins. Many aspects of cell life (eg, cell growth, differentiation, proliferation, cell cycle and survival) depend on the activities of protein kinases. Additionally, the abnormal activity of, the protein kinases has been linked to a host of disorders such as cancer and inflammation. Therefore, considerable effort has been directed towards the identification of ways to modulate the activities of the proteins - REF. : 194430 kinases In particular, many attempts have been made to identify small molecules that act as inhibitors of protein kinases. Various pyrrolyl indolinone derivatives have demonstrated excellent activity as inhibitors of protein kinases (Larid et al, FASEB J. 16, 681, 2002, Smolich et al., Blood, 97, 1413, 2001, Mendel et al., Clinical Cancer Res. 9, 327, 2003; Sun et al., J. Med. Chem. 46, 1116, 2003). The clinical utility of these compounds has been promising, but has been partially compromised due to the relatively poor aqueous solubility and / or other properties of the drug. What is needed is a class of modified pyrrolyl indolinone derivatives that have both inhibitory activity and improved drug properties.

Brief Description of the Invention One aspect of the invention relates to a compound having the following structure represented by Formula I: in Formula I, R is selected from the group consisting of hydrogen, halo, (C1-C6) alkyl, (C3-C8) cycloalkyl, (Cl-C6) haloalkyl, hydroxy, (C1-C6) alkoxy, amino, (Cl-C6) alkylamino, amide, sulfonamide, cyano, ( C6-C10) substituted or unsubstituted aryl; R2 is selected from the group consisting of hydrogen, halo (C1-C6) alkyl, (C3-C8) cycloalkyl, (Cl-C6) haloalkyl, hydroxy, (C1-C6) alkoxy, (C2-C8) alkoxyalkyl, amino, (C1-C6) alkylamino, (C6-C10) arylamino; R3 is selected from the group consisting of hydrogen, (C1-C6) alkyl, (C6-C10) aryl, (C5-CIO) heteroaryl, and amide; R 4 is selected from the group consisting of hydrogen and (C 1 -C 6) alkyl; and R5 is alpha- or beta-amino acid group or an alpha- or beta-amino-amide group connected to the carbonyl of (I) through the alpha- or beta-amino group to form an amide bond; or a pharmaceutically acceptable salt or prodrug thereof or may act as a prodrug. In a preferred embodiment, R5 is represented by the following structure: In the above structure, R6 is a side chain of an amino acid occurring naturally or unnaturally or its corresponding amide derivative thereof, the amide derivative having an amide nitrogen represented by NR8R9, - wherein R8 and R9 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, (Cl-) C6) hydroxyalkyl, (C1-C6) dihydroxyalkyl, (C1-C6) alkoxy, (C1-C6) alkyl-carboxylic acid, (C1-C6) alkyl-phosphonic acid, (C1-C6) alkyl-sulphonic acid, acid ( Cl-C6) hydroxyalkyl-carboxylic, (C1-C6) alkyl-amide, (C3-C8) cycloalkyl, (C5-C8) heterocycloalkyl, (C6-C8) aryl, (C5-C8) heteroaryl, (C3-C8) ) cycloalkylcarboxylic, or R8 and R9 together with N form a (C5-C8) heterocyclic ring either unsubstituted or substituted with one or more hydroxyls, ketones, ethers, and carboxylic acids; R7 is selected from the group consisting of hydroxy, (Cl-C6) -0-alkyl, (C3-C8) -O-cycloalkyl, and by NR8R9; and n is 0 or 1. In a first subgenus, R5 is an alpha-amino acid wherein the alpha-amino group is connected to the carbonyl of Formula I to form an amide bond. The preferred species within the first subgenus are represented by the following structures: In a second subgenus, R5 is an alpha-amino-amide wherein the alpha-amino group is connected to the carbonyl of the Formula I to form an amide bond. The preferred species within the second subgenre are represented by the following structures: In the third subgenus, R5 is a beta-amino acid wherein the beta-amino group is connected to the carbonyl of Formula I to form an amide bond. A preferred species within the third subgenre is represented by the following structure: In a fourth subgenus, R5 is a beta-amino-amide wherein the beta-amino group is connected to the carbonyl of Formula I to form an amide bond. The preferred species within the fourth subgenre are represented by The following structures Another aspect of the invention relates to a method for modulating the catalytic activity of a protein-cyclase with a compound or salt of Formula I. In a preferred mode, the protein kinase is selected from the group of receptors consisting of VEGF, and PDGF.

Detailed Description of the Invention Utility: The present invention provides compounds capable of regulating and / or modulating the activities of protein kinases of, but not limited to, VEGFR and / or PDGFR. In this way, the present invention provides a therapeutic approach to the treatment of disorders related to the normal functioning of these kinases. These disorders include, but are not limited to, solid tumors such as glioblastoma, melanoma, and Kaposi's sarcoma, and ovarian, lung, prostate, pancreatic, colonic, and squamous cell carcinoma. In addition, it also they can use VEGFR / PDGFR inhibitors in the treatment of restenosis and diabetic retinopathy. Additionally, this invention relates to the inhibition of vasculogenesis and angiogenesis by receptor-mediated pathways, which include the routes comprising VEGF receptors, and / or PDGF receptors. In this way, the present invention provides therapeutic approaches to the treatment of cancer and other diseases comprising the uncontrolled formation of blood vessels.

Synthesis protocol: In Reaction Scheme 1 a general scheme is shown to synthesize the starting material, HATU ester (1-1). Reaction scheme 1 Step 1 A mixture of 5-fluoro-1,3-dihydroindol-2-one (1.62 g, 10.2 mmol), 5-formyl-2,4-dimethyl-lH-pyrrole-3-carboxylic acid (1.96 g, 10.7 mmol), pyrrolidine (12 drops) and absolute ethanol was refluxed for 3 hours. The mixture was cooled to 25 ° C and the solids were collected by filtration. The solids were stirred with ethanol (30 mL) at 72 ° C for 30 minutes. The mixture was cooled to 25 ° C and the solids were collected again by filtration, washed with ethanol (6 mL), and dried under vacuum overnight to give as an orange solid the acid (Z) -5 - (( 5-fluoro-2-oxoindolin-3-ylidene) methyl) -2,4-dimethyl-lH-pyrrole-3-carboxylic acid (3.094 g, 96%). LC-ESIMS observed [M + H] + 301 (calculated for d6H13FN203 300.09).

Step 2: In DMF (15 mL), (Z) -5 - ((5-fluoro-2-oxoindolin-3-ylidene) methyl) -2,4-dimethyl-lH-pyrrole-3-carboxylic acid (3,094) was suspended. g, 10.3 mmol) and stirred for 5 minutes. Then DIEA (2.7 mL, 15.5 mmol) was added and the mixture was stirred for 10 minutes. HATU (3.91 g, 10.28 mmol) was added and the reaction mixture was stirred at 25 ° C for completion. The LC / MS detected the termination of the reaction. The majority of the DMF was removed and the residue was suspended in ACN and stirred for another 40 minutes. The solid was collected by filtration, washed with ACN, and dried under high vacuum overnight. It was obtained 5- ((5-fluoro-2- oxoindolin-3-ylidene) methyl) -2,4-dimethyl-lH-pyrrole-3-carboxylate of (Z) -3H- [1, 2, 3] triazole [4, 5, b] pyridin-3-yl ( 3.97 g, 92%). LC-ESIMS observed [M + H] + 419 (calculated for C2? H? 5FN603 418.12).

Examples 1-23: The general reaction scheme: Reaction scheme 2 The synthesis of the starting material, ester of HATU (1-1) is shown in Reaction Scheme 1. To prepare free carboxylic acid 1-2, the unprotected amino acid (1.0 equivalent) was added to a solution of 1- 1 (1.0 equivalent) and DIEA (1.5 equivalent) in DMF, as shown in Reaction Scheme 2. After stirring the solution at 25 ° C overnight, the LC-MS indicated that the formation of 1-2, and no start materials remained. This solution was used directly in the next step to prepare the amide 1-3. In this way, an amine was added to the solution (2 equivalents), HATU (1.0 mmol), and DIEA (1 equivalent). After stirring for 2 hours at 25 ° C, the reaction was completed based on LC-MS analysis. The reaction solution was directly subjected to preparative HPLC to obtain product 1-3 of pure amide, which was subsequently characterized by LC-MS and NMR spectroscopy.

Example 1.- Preparation of 5- [5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] -2,4-dimethyl-l (2-dimethylcarbamoyl-propyl) -amide -pyrrole-3-carboxylic acid Preparative HPLC gave 50 mg of the title compound (96%) of 52 mg of starting material (the active ester 1-1). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN403: 413, obtained: 413. XH NMR (DMSO-d6, 400 MHz), d 13.68 (s, ÍH), 10.89 (s, ÍH), 7.76 (dd, J = 2.4 Hz, 9. 6 Hz, ÍH), 7.71 (s, ÍH), 7.68 (t, J = 5.6 Hz, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.4 Hz, 8.4 Hz, ÍH), 3.31 (m , ÍH), 3. 16 (m, 2H), 3.05 (s, 3H), 2.84 (s, 3H), 2.41 (s, 3H), 2.39 (s, 3H), 1.03 (d, J = 6.8 Hz, 3H).

Example 2. 5- (5-Fluoro-2-oxo-l, 2-dihydro-indole- (3Z) 2- (2-methyl-3- (morpholin-4-yl) -3-oxo-propyl) -amide. ) -ylidenemethyl] -2,4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 56 mg of the title compound (98%) of 52 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C2H27FN204: 455, obtained: 455. XH NMR (DMSO-d6, 400 MHz), d 13.68 (s, ÍH), 10.89 (s, ÍH), 7.75 (dd) , J = 2.4 Hz, 9.2 Hz), 7.71 (s, ÍH), 7.67 (t, J = 5.6 Hz, ÍH), 6.92 (m, ÍH), 6.83 (dd, J = 4.8 Hz, 8.4 Hz, ÍH) , 3.55 (, 7H), 3.41 (m, ÍH), 3.35 (m, ÍH), 3.22 (m, ÍH), 3.12 (m, ÍH), 2.42 (s, 3H), 2.40 (s, 3H), 1.04 (d, J = 7.2 Hz, 3H).

Example 3.- 3- (. {5- [5-Fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl-lH-pyrrole-3-carbonyl acid .}. -amino) -butyric Preparative HPLC gave 14 mg of the title compound (56%) from 28 mg of starting material ( active ester). LC-MS: single peak at 254 nm, MH + calculated for C20H2oFN304: 386, obtained: 386. XH NMR (DMSO-d6, 400 MHz), d 13.66 (s, ÍH), 12.21 (s, ÍH), 10.89 (s) , ÍH), 7.76 (dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.71 (s, ÍH), 7.57 (d, J = 8.4 Hz, ÍH), 6.92 (m, ÍH), 6.83 (dd) , J = 4.8 Hz, J = 8.4 Hz, HH), 4.29 (m, HH), 4.05 (m, HH), 3.31 (d, J = 9.6 Hz, 2H), 2.41 (s, 3H), 2.38 (s) , 3H), 1.17 (d, J = 6.8 Hz, 3H).

Example 4. 5- (5-Fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] -2,4-dimethyl acid (2-dimethylcarbamoyl-l-methyl-ethyl) -amide -lH-pyrrole-3-carboxylic acid Preparative HPLC gave 42 mg of the title compound (78%) from 58 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN403: 413, obtained: 413. XH NMR (DMSO-d6, 400 MHz), d 13.66 (s, ÍH), 10.87 (s, ÍH), 7.75 (dd , J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.70 (s, ÍH), 7.55 (d, J = 8.0 Hz, ÍH), 6.92 (m, ÍH), 6.82 (dd, J = 4.8 Hz, J = 8.4 Hz, ÍH), 4.29 (m, ÍH), 3.01 (s, 3H), 2.82 (s, 3H), 2.58 (m, ÍH), 2.42 (m, ÍH), 2.41 (s, 3H), 2.39 (s, 3H), 1.18 (d, J = 6.8 Hz, 3H).

Example 5. 5- (5-fluoro-2-oxo-l, 2-dihydro-indole- (3Z-l-methyl-3- (morpholin-4-yl) -3-oxo-propyl) -amide. ) -ylidenemethyl] -2,4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 43 mg of the title compound (73%) from 48 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C24H27FN04: 455, obtained: 455. XH NMR (DMSO-d6, 400 MHz), d 13.59, (s, ÍH), 10.79 (s, ÍH), 7.67 ( dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.63 (s, ÍH), 7.47 (d, J = 7.6 Hz, ÍH), 6.85 (m, ÍH), 6.76 (dd, J = 4.8 Hz, J = 8.4 Hz, ÍH), 4.23 (m, ÍH), 3.60-3.30 (m, 10H), 2.35 (s, 3H), 2.32 (s, 3H), 1.11 (m, ÍH) (d, J = 6.8 Hz, 3H).

Example 6. 5- (5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] -2- ((S) -l-dimethylcarbamoyl-2-hydroxy-ethyl) -amide , 4-dimethyl-lH-pyrrole-3-carboxylic acid The preparative HPLC gave 42 mg of the compound of title (84%) from 50 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C2? H23FN0: 415, obtained: 415. XH NMR (DMSO-d6, 400 MHz), d 13.71 (s, ÍH), 10.91 (s, ÍH), 7.76 (dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.72 (s, ÍH), 7.56 (d, J = 8.0 Hz, ÍH), 6.92 (m, ÍH), 6.84 (s, ÍH), 6.83 (dd, J = 4.8 Hz, J = 8.4 Hz, HH), 4.97 (m, HH), 3.67 (m, HH), 3.56 (m, HH), 3.11 (s, 3H), 2.87 (s, 3H) , 2.45 (s, 3H), 2.43 (s, 3H).

Example 7. 5- (5-Fluoro-2-oxo-l, 2-dihydroxy) -l-hydroxymethyl-2- (morpholin-4-yl) -2-oxo-ethyl) -amide. indole- (3Z) -ylidenemethyl] -2,4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 51 mg of the title compound (93%) from 50 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C23H25FN05: 457, obtained: 457. XH NMR (DMSO-d6, 400 MHz), d 13.71 (s, ÍH), 10.90 (s, ÍH), 7.77 (dd , J = 2.4 Hz, J = 9.6 Hz, HH), 7.73 (,, HH), 7.63 (d, J = 8.0 Hz, HH), 6.94 (m, HH), 6.83 (dd, J = 4.8 Hz, J = 8.4 Hz, ÍH), 4.97 (m, ÍH), 3.80-3.40 (m, 11H), 2.45 (s, 3H), 2.43 (s, 3H).

Example 8. 5- (5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] -2- ((R) -l-dimethylcarbamoyl-2-hydroxy-ethyl) -amide. , 4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 40 mg of the title compound (64%) from 63 mg of starting material (the active ester). LC-MS: individual peak at 254 nm, MH + calculated for C2? H23FN40: 415, obtained: 415. XH NMR (DMSO-d6, 400 MHz), d 13.71 (s, ÍH), 10.91 (s, ÍH), 7.77 (dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.72 (s, ÍH), 7.55 (d, J = 7.6 Hz, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.8 Hz , J = 8.4 Hz, HH), 4.98 (dd, J = 6.0 Hz, J = 14.0 Hz, HH), 3.67 (dd, J = 6.4 Hz, J = 14.8 Hz, HH), 3.58 (dd, J = 6.4 Hz, J = 14.4 Hz, ÍH), 3.11 (s, 3H), 2.87 (s, 3H), 2.46 (s, 3H), 2.44 (s, 3H).

EXAMPLE 9 5- [5-Fluoro-2-oxo-1,2-dihydro-indole- (R) -l-hydroxymethyl-2- (morpholin-4-yl) -2-oxo-ethyl) -amide. (3Z) -ylidenemethyl] -2,4-dimethyl-lH-pyrrole-3-carboxylic acid The preparative HPLC gave 32 mg of the compound of title (47%) from 63 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C23H25FN405: 457, obtained: 457. XH NMR (DMSO-d6, 400 MHz), d 13.71 (s, ÍH), 10.90 (s, ÍH), 7.76 (dd) , J = 2.4 Hz, 9.6 Hz, HH), 7.72 (,, HH), 7.63 (d, J = 8.0 Hz, HH), 6.92 (m, HH), 6.83 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.96 (dd, J = 6.4 Hz, J = 14.4 Hz, ÍH), 3.74 (dd, J = 6.4 Hz, J = 14.4 Hz, ÍH), 3.65-3.30 (m, 9H), 2.46 (s, 3H), 2.43 (s, 3H).

Example 10.- (S) -2- (. {5- [5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl-lH-pyrrol- 3-carbonyl.} -amino) -N * l *, N * l *, N * 4 *, N * 4 * -tetramethyl-succinamide Preparative HPLC gave 30 mg of the title compound (73%) from 42 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C24H28FN04: 470, obtained: 470. XH NMR (DMSO-d6, 400 MHz), d 13.69 (s, ÍH), 10.89 (s, ÍH), 7.95 (d , J = 8. 8 Hz, ÍH), 7.75 (dd, J = 2.0 Hz, 9.2 Hz, ÍH), 7.70 (s, ÍH), 6.93 (m, ÍH), 6.83 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 5.26 (m, ÍH), 3.08 (s, 3H), 2.98 (s, 3H), 2.84 (s, 3H), 2.80 (s, 3H), 2.55 (m, 2H), 2.40 (s, 3H), 2.37 ( s, 3H).

Example 11.- [(S) -1- (Morpholine-4-carbonyl) -3- (morpholin-4-yl) -3-oxo-propyl] -amide of 5- [5-fluoro-2-oxo- 1, 2-dihydro-indol- (3Z) -ylidenemethyl] -2,4-dimethyl-1H-pyrrole-3-carboxylic acid Preparative HPLC gave 70 mg of the title compound (97%) from 56 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C28H32FN506: 554, obtained: 554. XH NMR (DMSO-d6, 400 MHz), d 13.68 (s, ÍH), 10.91 (s, ÍH), 8.08 (d , J = 8. 8 Hz, ÍH), 7.76 (dd, J = 2.4 Hz, 9.2 Hz, ÍH), 7.71 (s, ÍH), 6.93 (m, ÍH), 6.83 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 5.28 (m, ÍH), 3.75 (m, 2H), 3.70-2.50 (, 16H), 2.41 (s, 3H), 2.38 (s, 3H).

Example 12.- (S) -2- (. {5- [5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl bis (dimethylamide). -lH-pyrrole-3-carboxylic acid Preparative HPLC gave 60 mg of the title compound (78%) from 75 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C25H30FN5O4: 484, obtained: 484. XH NMR (DMSO-d6, 400 MHz), d 13.69 (s, ÍH), 10.88 (s, ÍH), 7.75 (dd , J = 2.4 Hz, 9.6 Hz, HH), 7.71 (,, HH), 7.70 (d, J = 8.0 Hz, HH), 6.93 (m, HH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.88 (m, ÍH), 3.13 (s, 3H), 2.94 (s, 3H), 2.86 (s, 3H), 2.82 (s, 3H), 2.44 (s, 3H), 2.42 (s, 3H) ), 2.34 (m, 2H), 1.95 (m, ÍH), 1.74 (m, ÍH).

Example 13. - [(S) -1- (Morpholine-4-carbonyl) -4- (morpholin-4-yl) -4-oxo-butyl] -amide of 5- [5-fluoro-2-oxo- 1, 2-dihydro-indol- (3Z) -ylidenemethyl] -2,4-dimethyl-1H-pyrrole-3-carboxylic acid Preparative HPLC gave 82 mg of the title compound (94%) from 75 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C29H3FN506: 568, obtained: 568. XH NMR (DMSO-d6, 400 MHz), d 13.70 (s, ÍH), 10.91 (s, ÍH), 8.30 (m , ÍH), 7.78 (m, ÍH), 7.72 (s, ÍH), 6.92 (m, ÍH), 6.84 (m, ÍH), 4.90 (m, ÍH), 3.80-3.35 (m, 9H), 3.13 ( m, 7H), 2.45 (s, 3H), 2.43 (s, 3H), 2.56-2.35 (m, 2H), 1.97 (m, ÍH), 1.76 (m, ÍH). Example 14.- (S) -4-Dimethylcarbamoyl-2- (. {5- [5-fluoro-2-oxo-1,2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl acid -1H-pyrrole-3-carbonyl.} - amino) -butyric Preparative HPLC gave 44 mg of the title compound (81%) from 50 mg of starting material (the active ester). LC-MS: individual peak at 254 nm, MH + calculated for C23H25FN405: 457, obtained: 457.

Example 15.- (-2- (. {5- [5-Fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl-lH-pyrrole. -3-carbonyl.}. -amino) -5-morpholin-4-! L-5-oxo-pentanoic Preparative HPLC gave 40 mg of the title compound (67%) from 50 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C25H27FN06: 499, obtained: 499. XH NMR (DMSO-d6, 400 MHz), d MHz), d 13.69 (s, ÍH), 12.55 (s, ÍH) 10.89 (s, ÍH), 7.88 (d, J = 8.0 Hz, ÍH), 7.75 (dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.72 (s, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, HH), 4.36 (m, HH), 3.53 (m, 4H), 3.42 (m, 4H), 3.31 (m, 2H), 2.44 (s, 3H), 2.42 (s, 3H), 2.08 (m, ÍH), 1.93 (m, ÍH).

Example 16.- (R) -2- (. {5- [5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenomethyl] -2,4-dimethyl-lH-pyrrole -3-carbonyl.}. -amino) -5-morpholin-4-IL-5-oxo-pentanoic Preparative HPLC gave 37 mg of the title compound (84%) from 37 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C2H27FN406: 499, obtained: 499. XH NMR (DMSO-d6, 400 MHz), d 13.69 (s, HI), 12.57 (s, ÍH), 10.90 (s) , ÍH), 7.88 (d, J = 8.0 Hz, ÍH), 7.76 (dd, J = 2.8 Hz, J = 9.2 Hz, ÍH), 7.72 (s, ÍH), 6.92 (m, ÍH), 6.84 (dd) , J = 4.8 Hz, 8.4 Hz, HH), 4.37 (, HH), 3.53 (m, 3H), 3.43 (m, 4H), 3.31 (m, 3H), 2.45 (s, 3H), 2.42 (s, 3H), 2.08 (m, ÍH), 1.93 (m, ÍH).

Example 17. Bis (dimethy1) amide of (R) -2- (. {5- [5-flucrc> -2-C5 c> -l, 2-dihydro-indole- (3Z) -ylidene-tert-amide ] -2, 4-o ^ methyl-lH-pyrrole-3-carb3ru \ l ^^ -pentanoic Preparative HPLC gave 28 mg of the title compound (41%) from 60 mg of starting material (the active ester). LC-MS: individual peak at 254 nm, MH 'calculated for C25H30FN5O: 484, obtained: 484. X H NMR (DMSO-de, 400 MHz), d 13.69 (s, HH), 10.90 (s, HH), 7.76 (dd, J = 2.4 Hz, 9.6 Hz, HH), 7.73 (d, J = 8.0) Hz, ÍH), 7.72 (s, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.88 (m, ÍH), 3 .13 (s, 3H), 2 .93 (s, 3H), 2.86 (s, 3H), 2.82 (s, 3H), 2.44 (s, 3H), 2 .42 (s, 3H), 2.50-2.30 (m, 2H), 1.95 ( m, ÍH), 1.74 (m, ÍH).

Example 18. - [(R) -1- (Morpholine-4-carbonyl) -4- (morpholin-4-yl) -4-oxo-butyl] -amide of 5- [5-fluoro-2-oxo- 1, 2-dihydro-indol- (3Z) -ylidenemethyl] -2, -dimethyl-1H-pyrrole-3-carboxylic acid Preparative HPLC gave 30 mg of the title compound (38%) from 60 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C29H34FN50e: 568, obtained: 568. XH NMR (DMSO-d6, 400 MHz), d 13.70 (s, ÍH), 10.91 (s, ÍH), 7.77 (m , 2H), 7.72 (s, ÍH), 6.93 (m, ÍH), 6.83 (, ÍH), 4.91 (m, ÍH), 3.90-3.35 (m, 16H), 2.45 (s, 3H), 2.42 (s) , 3H), 2.50-2.30 (m, 2H), 1.98 (m, ÍH), 1.77 (m, ÍH).

Example 19. 5- (5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] ((1S, 2S) -l-dimethylcarbamoyl-2-hydroxy-propyl) -amide] -2, 4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 84 mg of the title compound (67%) from 122 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN404: 429, obtained: 429. 1 H NMR (DMSO-de, 400 MHz), d 13.69 (s, ÍH), 10.89 (s, ÍH), 7.75 (m , ÍH), 7.70 (s, ÍH), 7.61 (d, J = 8.8 Hz, ÍH), 6.92 (m, ÍH), 6.83 (dd, J = 4.8 Hz, 8. 4 Hz, HH, 4.81 (t, J = 4.4 Hz, HH), 3.90 (m, HH), 3.12 (s, 3H), 2.86 (s, 3H), 2.42 (s, 3H), 2.39 (s, 3H) ), 1.12 (d, J = 4.8 Hz, 3H).

Example 20. - ((IR, 2R) -l-dimethylcarbamoyl-2-hydroxy-propyl) -amide of 5- [5-f-luoro-2-oxo-1,2-dihydro-indol- (3Z) -ylidenemethyl) ] -2, 4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 78 mg of the title compound (62%) from 122 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN 04: 429, obtained: 429. 1 H NMR (DMSO-d6, 400 MHz), d 13.70 (s, ÍH), 10.90 (s, ÍH), 7.77 ( m, ÍH), 7.71 (s, ÍH), 7.62 (d, J = 8.4 Hz, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.82 (t, J = 8.0 Hz, ÍH), 3.92 (m, ÍH), 3.13 (s, 3H), 2.87 (s, 3H), 2.43 (s, 3H), 2.40 (s, 3H), 1.15 (d, J = 2.8 Hz, 3H).

Example 21. 5- (5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) ylidenomethyl] -2- ((IR, 2S) -l-dimethylcarbamoyl-2-hydroxy-propyl) -amide. , 4-dimethyl-lH-pyrrole-3-carboxylic acid Preparative HPLC gave 90 mg of the title compound (72%) from 122 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN404: 429, obtained: 429. NMR 4 (DMSO-de, 400 MHz), d 13.73 (s, ÍH), 10.91 (s, ÍH), 7.77 (dd , J = 2.4 Hz, 6.4 Hz, HH), 7.73 (,, HH), 7.29 (d, J = 8.0 Hz, HH), 6.93 (m, HH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.85 (m, ÍH), 3.97 (m, ÍH), 3.12 (s, 3H), 2.87 (s, 3H), 2.43 (s, 3H), 2.41 (s, 3H), 1.10 (d, J = 5.6 Hz, 3H).

Example 22. 5- (5-fluoro-2-oxo-l, 2-dihydro-indol- (3Z) ylidenomethyl] -2- ((1S, 2R) -l-dimethylcarbamoyl-2-hydroxy-propyl) -amide. , 4-dimethyl-lH-pyrrole-3-carboxylic acid The preparative HPLC gave 90 mg of the compound of title (72%) from 122 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C22H25FN404: 429, obtained: 429. XH NMR (DMSO-d6, 400 MHz), d 13.73 (s, ÍH), 10.91 (s, ÍH), 7.76 (dd , J = 2.8 Hz, 6.8 Hz, ÍH), 7.73 (s, ÍH), 7.30 (d, J = 8.0 Hz, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.8 Hz, 8.4 Hz, ÍH), 4.85 (m, ÍH), 3.98 (m, ÍH), 3.12 (s, 3H), 2.86 (s, 3H), 2.48 (s, 3H), 2.45 (s, 3H), 1.10 (d, J = 6.0 Hz, 3H).

Example 23. 5- [5-Fluoro-2-oxo-l, 2-dihydro-indol- (3Z) -ylidenemethyl] -2,4-dimethyl-lH-pyrrole-3-carboxylic acid dimethylcarbamoylmethyl-amide Preparative HPLC gave 27 mg of the title compound (46%) from 66 mg of starting material (the active ester). LC-MS: single peak at 254 nm, MH + calculated for C20H2? FN4O3: 385, obtained: 385. XH NMR (DMSO-d6, 400 MHz), d 13.71 (s, ÍH), 10.90 (s, ÍH), 7.76 (dd, J = 2.4 Hz, J = 9.6 Hz, ÍH), 7.73 (s, ÍH), 7.55 (t, J = 5.6 Hz, ÍH), 6.93 (m, ÍH), 6.84 (dd, J = 4.4 Hz, 8.4 Hz, ÍH), 4.08 (d, J = 5.6 Hz, 2H), 3.00 (s, 3H), 2.87 (s, 3H) 2.49 (s, 3H), 2.46 (s, 3H).

VEGFR biochemical assay The compounds were evaluated for biochemical activity by Upstate Ltd at Dundee, United Kingdom, according to the following procedure. In a final reaction volume of 25 μl, KDR (h) (5-10 mU) was incubated in 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.33 mg / ml myelin basic protein, 10 mM MgAcetate, and [y- 33P-ATP] (specific activity approximately 500 cpm / pmol, concentration as required). The reaction is initiated by the addition of the MgATP mixture. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of 5 μl of a 3% phosphoric acid solution. Then 10 μl of the reaction is added dropwise to a P30 filter mat and washed three times for 5 minutes in 75 mM phosphoric acid and once in methanol before drying and counting by scintillation.

Cell assay: HUVEC: Proliferation induced by VEGF Compounds were assessed for cellular activity in the VEGF-induced proliferation of HUVEC cells. HUVEC cells (Cambrex, CC-2517) were maintained in EGM (Cambrex, CC-3124) at 37 ° C and 5% C02. HUVEC cells were plated at a density of 5000 cells / concavity (plate of 96 concavities) in EGM. After cell binding (1 hour) the EGM medium was replaced by EBM (Cambrex, CC-3129) + 0.1% FBS (ATTC, 30-2020) and the cells were incubated for 20 hours at 37 ° C. The medium was replaced by EBM + 1% FBS, the compounds were serially diluted in DMSO and added to the cells at a final concentration of 0-5,000 nM and 1% DMSO. After 1 hour of pre-incubation at 37 ° C, the cells were stimulated with 10 ng / mL of VEGF (Sigma, V7259) and incubated for 45 hours at 37 ° C. Cell proliferation was measured by incorporation of BrdU DNA for 4 hours and the level of BrdU was quantified by ELISA (Roche Kit, 16472229) using 1M H2SO4 to stop the reaction. The absorbance at 450 nm was measured using a reference wavelength at 690 nm. It is noted that in relation to this date, the best method known by the applicant to carry out the present invention is that which is clear from the present description of the invention.

Claims (31)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property: 1. Compound represented by Formula I as follows: characterized in that: R1 is selected from the group consisting of hydrogen, halo, (Cl-Cβ) alkyl, (C3-C8) cycloalkyl, (Cl- C6) haloalkyl, hydroxy, (C1-C6) alkoxy, amino, (Cl-) C6) alkylamino, amide, sulfonamide, cyano, (C6-C10) substituted or unsubstituted aryl; R2 is selected from the group consisting of hydrogen, halo (C1-C6) alkyl, (C3-C8) cycloalkyl, (Cl- C6) haloalkyl, hydroxy, (C1-C6) alkoxy, (C2-C8) alkoxyalkyl, amino, (C1-C6) alkylamino, (C6-C10) arylamino; R3 is selected from the group consisting of hydrogen, (C1-C6) alkyl, (C6-C10) aryl, (C5-C10) heteroaryl, and amide; R4 is selected from the group consisting of hydrogen and (C1-C6) alkyl; and R5 is alpha- or beta-amino acid group or an alpha- or beta-amino-amide group connected to the carbonyl of (I) through the alpha- or beta-amino group to form an amide bond; or a pharmaceutically acceptable salt or prodrug thereof or may act as a prodrug. Compound according to claim 1, characterized in that R5 is represented by the following structure: wherein: R6 is a side chain of an amino acid occurring naturally or unnaturally or its corresponding amide derivative thereof, the amide derivative having an amide nitrogen represented by NR8R9; wherein R8 and R9 are independently selected from the group consisting of hydrogen, (C1-C6) alkyl, (C1-C6) hydroxyalkyl, (Cl-C6) dihydroxyalkyl, (C1-C6) alkoxy, (C1-C6) alkyl -carboxylic acid, (C1-C6) alkyl-phosphonic acid, (C1-C6) alkyl-sulfonic acid, (C1-Cd) hydroxyalkyl-carboxylic acid, (C1-C6) alkyl-amide, (C3-C8) cycloalkyl, ( C5-C8) heterocycloalkyl, (C6-C8) aryl, (C5-C8) heteroaryl, (C3-C8) cycloalkyl-carboxylic acid, or R8 and R9 together with N form a (C5-C8) heterocyclic ring either unsubstituted or substituted with one or more hydroxyls, ketones, ethers, and carboxylic acids; R7 is selected from the group consisting of hydroxy, (C1-C6) -O-alkyl, (C3-C8) -O-cycloalkyl, and by NR8R9; and n is 0 or 1. 3. Compound according to claim 2, characterized in that R5 is an alpha-amino acid wherein the alpha-amino group is connected to the carbonyl of Formula I to form an amide bond. 4. Compound according to claim 2, characterized in that R5 is an alpha-amino-amide wherein the alpha-amino group is connected to the carbonyl of Formula I to form an amide bond. 5. Compound according to claim 2, characterized in that R5 is a beta-amino acid wherein the beta-amino group is connected to the carbonyl of Formula I to form an amide bond. 6. Compound in accordance with the claim 2, characterized in that R5 is a beta-amino-amide wherein the beta-amino group is connected to the carbonyl of Formula I to form an amide bond. 7. Compound according to claim 3, characterized in that it has the following structure: 8. Compound according to claim, characterized in that it has the following structure: 9. Compound according to claim, characterized in that it has the following structure: 10. Compound according to claim, characterized in that it has the following structure: 11. Compound according to claim, characterized in that it has the following structure: 12. Compound according to claim, characterized in that it has the following structure: 13. Compound according to claim, characterized in that it has the following structure: 14. Compound according to claim, characterized in that it has the following structure: 15. Compound according to claim, characterized in that it has the following structure: 16. Compound according to claim, characterized in that it has the following structure: 17. Compound according to claim, characterized in that it has the following structure: 18. Compound according to claim, characterized in that it has the following structure: 19. Compound according to claim, characterized in that it has the following structure: 20. Compound according to claim, characterized in that it has the following structure: 21. Compound according to claim, characterized in that it has the following structure: 22. Compound according to claim, characterized in that it has the following structure: 23. Compound according to claim, characterized in that it has the following structure: 24. Compound according to claim, characterized in that it has the following structure: 25. Compound according to claim, characterized in that it has the following structure: 26. Compound according to claim, characterized in that it has the following structure: 27. Compound according to claim, characterized in that it has the following structure: 28. Compound according to claim 6, characterized in that it has the following structure: 29. Compound according to claim 6, characterized in that it has the following structure: 30. Method, characterized in that it is for the modulation of the catalytic activity of a protein kinase with a compound or salt according to any of claims 1-29. 31. Method according to claim 30, characterized in that the protein-kinases is selected from the group of receptors consisting of VEGFR and PDGFR.