WO2014183300A1

WO2014183300A1 - Vegfr tyrosine kinase inhibitors

Info

Publication number: WO2014183300A1
Application number: PCT/CN2013/075813
Authority: WO
Inventors: Jing Ling; Jinyong LIU
Original assignee: Suzhou Vivotide Biotechnologies Co., Ltd.
Priority date: 2013-05-17
Filing date: 2013-05-17
Publication date: 2014-11-20

Abstract

Novel compounds, their prodrugs, and the pharmaceutically acceptable salts as pharmaceutical compositions containing such compounds useful in treating certain diseases modulated by the inhibition of vascular endothelial growth factors (VEGFs) receptor tyrosine kinases are provided. In particular, compounds and compositions and the methods for the prophylaxis, management and treatment of cancers through the inhibition of VEGF receptor tyrosine kinases are provided.

Description

VEGFR TYROSINE KINASE INHIBITORS

FIELD OF THE INVENTION

The present invention relates to novel compounds, their prodrugs, and the pharmaceutically acceptable salts as well as pharmaceutical compositions containing such compounds useful in treating certain diseases modulated by the inhibition of vascular endothelial growth factors (VEGFs) receptor tyrosine kinases. In particular, the invention relates to compounds and compositions and the methods for the prophylaxis, management and treatment of cancers through the inhibition of VEGF receptor tyrosine kinases.

BACKGROUND

Binding of vascular endothelial growth factors (VEGFs) to their cognate tyrosine kinase receptors can induce various biological activities such as cell proliferation, migration and angiogenesis. This family of receptors includes VEGFR1 , VEGFR2 (KDR, Flk-1), and VEGFR3 (Flt-4). See T. Matsumoto and L. Claesson-Welsh, Sci. STKE 2001 , re21 (2001). By contrast to VEGFR1 and VEGFR2, expression of VEGFR3 is restricted mainly to the lymphatic vessels and blood vessels of tumors. See J.M. Cunnick et al., J. Biol. Chem., 277, 9498-9504 (2002). Although both VEGFR1 and VEGFR2 are expressed in vascular endothelial cells, the two receptors have different functions. VEGFR1 null mice failed to form embryonic vasculature and were characterized with disorganized tubules of endothelial cells (G-H Fong et al., Nature 376, 66 - 70 (2002)), while VEGFR2 knockout mice failed to develop vasculature and lacked blood islands and hematopoietic progenitor cells (Shalaby, F. et al., Nature 376, 62-66 (1995)). Unlike VEGFR2, VEGFR1 was also expressed in inflammatory cells and important for their recruitment and activation. VEGF- induced macrophage migration was strongly suppressed in mice lacking VEGFR1 kinase activity. As the migration and activation of macrophage and other myeloid progenitor cells is the key for inflammation, the above studies clearly indicate the important role of VEGFR1 kinase activity in inflammation. Expression of VEGFs and VEGFR1 is significantly increased in inflammatory disease conditions (Hoshino, M. et al., J. Allergy Clin. Immun.. 107, 295-307 (2001)). Blocking VEGF-induced pathological conditions using soluble VEGFR1 receptor and antibodies showed promising efficacy in inflammatory disease models such as RA and atherosclerosis (Luttun, A. et al., Nature Medicine 8, 831 - 840 (2002)).

However, recent studies have demonstrated that VEGFR1 , not KDR, is expressed in human monocytes/macrophages, and their chemotactic response and activation by VEGF is mediated via this receptor, indicating the crucial role of VEGFR1 in inflammatory reactions. Indeed, VEGF-induced macrophage migration was strongly suppressed in mice lacking VEGFR1 kinase activity.

The expression of VEGFs and VEGFR1 is significantly increased in human inflammatory disease. Blocking VEGF-induced pathological conditions using soluble VEGFR1 receptor or antibodies to VEGFR1 showed promising efficacy by inhibiting recruitment of inflammatory leukocytes in inflammatory disease models such as RA and atherosclerosis. The anti-VEGFR1 mouse antibody attenuated the upregulation of circulating leukocytes and impaired the release of tissue-injuring proteinases and proinflammatory cytokinesfrom macrophages in the disease models. Therefore, VEGFR1 antagonism is an attractive treatment for inflammatory diseases as well. That VEGFR1 is a key regulator for mobilization of leukocytes as well as angiogenesis and vascular permeability strongly suggests that this kinase is a novel target for development of small molecule inhibitors against inflammatory diseases and cancers.

A number of VEGFR inhibitors have been developed that can arrest tumor growth and, in some cases, cause tumor regression. Small molecule VEGFR1 inhibitors are proven to be effective antitumor agents. In addition several antibodies that bind to the extracellular domain of the VEGFR are also efficacious. When used in combination with cytotoxic treatments, chemotherapy, and radiation, VEGFR1 inhibitors have been shown to potentiate their anticancer activity.

SUMMARY OF THE INVENTION The present invention provides novel compounds represented by the following general structure, their prodrugs, pharmaceutically acceptable salts, and novel pharmaceutical compositions containing the same.

Group 2)

Group 1

The present invention also includes within its scope diagnostic methods for the detection of diseases associated with VEGFR deficiency or malfunctions, and methods for the prophylaxis, management and treatment of diseases modulated by VEGFR inhibition. The compounds of this invention are useful for the prophylaxis, management and treatment of diseases involving in VEGFR regulated signalling pathways. In particular, these compounds and pharmaceutical compositions containing the same are indicated in the prophylaxis, management and treatment of cancers. In addition to the novel compounds and compositions of this invention, the intermediates and processes useful for the preparation of the compounds of the invention are also included within the scope of this invention. Other aspects of this invention will become apparent as the description of this invention continues. Hence, the foregoing merely summarizes certain aspects of the invention and is not intended, nor should it be construed, as limiting the invention in any way.

BRIEF DESCRIPTION OF THE DRAWING

Figure 1 shows the general structure of the compounds according to the present invention.

DEFINITIONS

As used herein, "alkyl" means a cyclic, branched, or straight chain chemical group containing only carbon and hydrogen, such as methyl, pentyl, and adamantyl. Alkyl groups can either be unsubstituted or substituted with one or more substituents, e.g., halogen, alkoxy, acyloxy, amino, amido, cyano, nitro, hydroxyl, mercapto, carboxy, carbonyl, benzyloxy, aryl, heteroaryl, or other functionality that may be suitably blocked, if necessary for purposes of the invention, with a protecting group. Alkyl groups can be saturated or unsaturated (e.g., containing -C=C- or -C≡C- subunits), at one or several positions. Typically, alkyl groups will comprise 1 to 12 carbon atoms, preferably 1 to 10, and more preferably 1 to 8 carbon atoms or cyclic groups containing three to eight carbons.

As used herein, "lower alkyl" means a subset of alkyl, and thus is a hydrocarbon substituent, which is linear, cyclic or branched. Preferred lower alkyls are of 1 to about 6 carbons, and may be branched or linear, and may include cyclic substituents, either as part or all of their structure. Examples of lower alkyl include butyl, propyl, isopropyl, ethyl, and methyl. Likewise, radicals using the terminology "lower" refer to radicals preferably with 1 to about 6 carbons in the alkyl portion of the radical.

As used herein, "amido" means a H-CON- or alkyl-CON-, aryl-CON- or heterocyclyl- CON group wherein the alkyl, aryl or heterocyclyl group is as herein described.

As used herein, "aryl" means a substituted or unsubstituted aromatic radical having a single-ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl), which can be optionally unsubstituted or substituted with amino, cyano, hydroxyl, lower alkyl, haloalkyl, alkoxy, nitro, halo, mercapto, and other substituents, and which may or may not include one or more heteroatoms. Preferred carbocyclic aryl is phenyl. The term "heteroaryl" is clearly contemplated in the term "aryl". Preferably where the term aryl represents a heterocycle, it is referred to as "heteroaryl", and has one or more heteroatom(s). Preferred are monocyclic heterocycles of 5 or 6 members. Hence preferred heteroaryl is a monovalent unsaturated aromatic group having a single ring and having at least one hetero atom, such as N, O, or S, within the ring, which can optionally be unsubstituted or substituted with amino, cyano, nitro, hydroxyl, alkyl, haloalkyl, alkoxy, aryl, halo, mercapto, oxo (hence forming a carbonyl.) and other substituents. Examples of heteroaryl include thienyl, pyrridyl, furyl, oxazolyl, oxadiazolyl, pyrollyl, imidazolyl, triazolyl, thiodiazolyl, pyrazolyl, isoxazolyl, thiadiazolyl, pyranyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thiazolyl and others.

In this definition it is clearly contemplated that substitution on the aryl ring is within the scope of this invention. Where substitution occurs, the radical is called substituted aryl. Preferably one to three, more preferably one or two, and most preferably one substituent occur on the aryl ring. Preferred substitution patterns in five membered rings are substituted in the 2 position relative to the connection to the claimed molecule. Though many substituents will be useful, preferred substituents include those commonly found in aryl compounds, such as alkyl, hydroxy, alkoxy, cyano, nitro, halo, haloalkyl, mercapto and the like.

As used herein, "amide" includes both RNR'CO- (in the case of R = alkyl, alkaminocarbonyl-) and RCONR'- (in the case of R = alkyl, alkyl carbonylamino-).

As used herein, the term "ester" includes both ROCO- (in the case of R = alkyl, alkoxycarbonyl-) and RCOO- (in the case of R = alkyl, alkylcarbonyloxy-).

As used herein, "acyl" means an H-CO- or alkyl-CO-, aryl-CO- or heterocyclyl-CO- group wherein the alkyl, aryl or heterocyclcyl group is as herein described. Preferred acyls contain a lower alkyl. Exemplary alkyl acyl groups include formyl, acetyl, propanoyl, 2-methylpropanoyl, t-butylacetyl, butanoyl and palmitoyl.

As used herein, "halo" is a chloro, bromo, fluoro or iodo atom radical. Chloro, bromo and fluoro are preferred halides. The term "halo" also contemplates terms sometimes referred to as "halogen", or "halide".

As used herein, "haloalkyl" means a hydrocarbon substituent, which is linear or branched or cyclic alkyl, alkenyl or alkynyl substiuted with chloro, bromo, fluoro or iodo atom(s). Most preferred of these are fluoroalkyls, wherein one or more of the hydrogen atoms have been substituted by fluoro. Preferred haloalkyls are of 1 to about 5 carbons in length, More preferred haloalkyls are 1 to about 4 carbons, and most preferred are 1 to 3 carbons in length. The skilled artisan will recognize then that as used herein, "haloalkylene" means a diradical variant of haloalkyl, such diradicals may act as spacers between radicals, other atoms, or between the parent ring and another functional group. For example, the linker CHF-CHF is a haloalkylene diradical.

As used herein, "heterocyclyl" means heterocyclic radicals, which are saturated or unsaturated. These may be substituted or unsubstituted, and are attached to other via any available valence, preferably any available carbon or nitrogen. More preferred heterocycles are of 5 or 6 members. In six membered non-aromatic monocyclic heterocycles, the heteroatom(s) are selected from one up to three of O, N or S, and wherein when the heterocycle is five membered and non-aromatic, preferably it has one or two heteroatoms selected from O, N, or S. As used herein, "substituted amino" means an amino radical that is substituted by one or two alkyi, aryl, or heterocyclyl groups, wherein the alkyi, aryl or heterocyclyl are defined as above.

As used herein, "substituted thiol" means RS- group wherein R is an alkyi, an aryl, or a heterocyclyl group, wherein the alkyi, aryl or heterocyclyl are defined as above.

As used herein, "sulfonyl" means an alkylS0₂, arylS0₂ or heterocyclyl-S0₂ group wherein the alkyi, aryl or heterocyclyl are defined as above.

As used herein, "sulfamido" means an alkyl-N-S(0)2N-, aryl-NS(0)2N- or heterocyclyl-NS(0)2N- group wherein the alkyi, aryl or heterocyclcyl group is as herein described.

As used herein, "sulfonamido" means an alkyl-S(0)2N-, aryl-S(0)2N- or heterocyclyl- S(0)2N- group wherein the alkyi, aryl or heterocyclcyl group is as herein described.

As used herein, "ureido" means an alkyl-NCON-, aryl-NCON- or heterocyclyl- NCON- group wherein the alkyi, aryl or heterocyclcyl group is as herein described

As used herein, a "radical" may form a ring with another radical as described herein.

When such radicals are combined, the skilled artisan will understand that there are no unsatisfied valences in such a case, but that specific substitutions, for example a bond for a hydrogen, is made. Hence certain radicals can be described as forming rings together. The skilled artisan will recognize that such rings can and are readily formed by routine chemical reactions, and it is within the purview of the skilled artisan to both envision such rings and the methods of their formations. Preferred are rings having from 3-7 members, more preferably 5 or 6 members. As used herein the term "ring" or "rings" when formed by the combination of two radicals refers to heterocyclic or carbocyclic radicals, and such radicals may be saturated, unsaturated, or aromatic. For example, preferred heterocyclic ring systems include heterocyclic rings, such as morpholinyl, piperdinyl, imidazolyl, pyrrolidinyl, and pyridinyl.

DETAILED DESCRIPTION OF THE INVENTION

The detailed description of the invention that follows is not intended to be exhaustive or to limit the invention to the precise details disclosed. It has been chosen and described to best explain the details of the invention to others skilled in the art. As shown in Fig. 1 , the compounds of the invention have the following general structure, Formula 1 :

erocycles Group 2)

Formula 1

In Formula 1 , the Heterocycles Group 2 (Het₂) includes the following structures:

The substitutions R-i , R2, and R3 are each independently selected from: H, halogen, alkyl, alkenyl, alkynyl, haloalkyl, phenyl and substituted phenyl, pyridinyl, N(alkyl)₂, CN, CONH₂, CONHAIkyI, CONHPh, OH, O-Alkyl, O-alkyl substituted with halogen, O-alkyl substituted with OH, S-alkyl, OPh, COAIkyI, NHCOalkyI, NHCOaryl, and the following structures:

__R wherein R₄ = H, COCF₃, COCH₃, (CH₂)₂OH, COOAIkyl, (CH₂)₂0(CH₂)₂OH,

CH₂CONH₂

— N W wherein W = O, CH₂, CHOH, CHCONH₂ S0₂

The group R5 includes H, CH3, SCH3, Ph, and ΝΜβ2. The group X includes NH and O. In Heterocycles Group 1 , A, B, D, and E are each independently selected from O, S, N, NH, bond, CO, CH2, C-R, C-OR, wherein R = H, halogen, alkyl, alkenyl, alkynyl, haloalkyl, phenyl and substituted phenyl, CN, CH2CN, CH2CONH2, CH2CH2NH-CO-CH3.

Heterocyles Group 1 also includes the following structures:

Compounds and compositions herein also specifically contemplate pharmaceutically acceptable salts, whether cationic or anionic. A "pharmaceutically- acceptable salt" is an anionic salt formed at any acidic (e.g., carboxyl) group, or a cationic salt formed at any basic (e.g., amino) group. Many such salts are known in the art, as described in World Patent Publication WO 87/05297, Johnston et al., published September 1 1 , 1987 (incorporated by reference herein). Preferred counter- ions of salts formable at acidic groups can include cations of salts, such as the alkali metal salts (such as sodium and potassium), and alkaline earth metal salts (such as magnesium and calcium) and organic salts. Preferred salts formable at basic sites include anions such as the halides (such as chloride salts). Of course, the skilled artisan is aware that a great number and variation of salts may be used, and examples exist in the literature of either organic or inorganic salts useful in this manner.

It is also clearly contemplated that compounds of the invention can be provided as biohydrolyzable prodrugs, as they are understood in the art. "Prodrug", as used herein is any compound wherein when it is exposed to the biological processes in an organism, is hydrolyzed, metabolized, derivatized or the like, to yield an active substance having the desired activity. The skilled artisan will recognize that prodrugs may or may not have any activity as prodrugs. It is the intent that the prodrugs described herein have no deleterious effect on the subject to be treated when dosed in safe and effective amounts. These include for example, biohydrolyzable amides and esters. A "biohydrolyzable amide" is an amide compound which does not essentially interfere with the activity of the compound, or that is readily converted in vivo by a cell, tissue, or human, mammal, or animal subject to yield an active compound of the invention. A "biohydrolyzable ester" refers to an ester compound of the invention that does not interfere with the activity of these compounds or that is readily converted by an animal to yield an active formula (I) compound. Such biohydrolyzable prodrugs are understood by the skilled artisan and are embodied in regulatory guidelines.

Inasmuch as the compounds of the invention may contain optical centers, "optical isomer", "stereoisomer", "enantiomer," "diastereomer," as referred to herein have the standard art recognized meanings (cf. Hawleys Condensed Chemical Dictionary, 1 1th Ed.) and are included in the compounds claimed, whether as racemates, or their optical isomers, stereoisomers, enantiomers, diastereomers.

PREPARATION OF COMPOUNDS OF THE INVENTION

One hundred ten (1 10) compounds were prepared, and all the compounds were tested for VEGFR1 kinase inhibition following the method described in US patent No. 6,831 ,075. The VEGFR1 kinase assays utilized an ELISA-based format in a 96-well FluoroNUNC Maxisorp plate with a time-resolved fluorescence readout. The plates were coated with 100 μΙ_Λ/νβΙΙ of substrate solution (recombinant human PLC-y/GST) at a concentration of 40 μg/mL in Tris buffered saline (TBS). The VEGFR1 activity was assayed in 100-μΙ_ assay mixture containing 50 mM HEPES (pH 7.4), 30 μΜ ATP, 10 mM MnCI₂, 0.1 % BSA, 2% DMSO, and 300 ng/mL prephosphorylated recombinant human baculovirus-expressed VEGFR1 cytoplasmic domain. The compounds were tested for inhibition of the VEGFR1 kinase activity at concentrations ranging from 1 nM to 10 M. The kinase reactions were allowed to proceed at 37 °C for 15 minutes. The detection antibody, europium-labeled anti- phosphotyrosine antibody (Wallac #CR04-100), was added at 1 :5000 dilution in block buffer (3% BSA in TBST). After incubation at 37 °C for one hour, 100 μΙ_ of enhancement solution (Wallac #1244-105) was added and the plates were gently agitated. After 5 min, the fluorescence of the resulting solution was measured using a BMG Fluostar (Model

#403). The Apparent Ki (VEGFR1) of the compounds ranged from 4 nM to >10 M, and the average Ki-app for VEGFR1 inhibition is reported as follows:

I = 0.001-0.1 M, II = 0.1 to 1.0 M, III = 1.0 to > 10 M

All the intermediates showed satisfactory NMR spectra.

4,6-Diaminopyrimidine compounds were prepared according to the general procedure shown in Scheme 1.

Scheme 1.

Step-1 : The prototypical procedures for step 1 are exemplified below.

Example 1 (Method A). 3-(6-chloro-pyrimidin-4-vlamino)phenol

A solution of 3-aminophenol (1.00 g, 9.17 mmol) in isopropyl alcohol (25 mL) was added drop-wise to a refluxing solution of 4,6-dichloropyrimidine (2.73 g, 18.3 mmol) in isopropyl alcohol (65 mL). The reaction mixture was refluxed for 4 more hours and cooled to room temperature ("rt"). The volatiles were removed in vacuo. The residue was suspended in water and the suspension was neutralized with aqueous ("aq"). NaHC0₃. The resulting yellow suspension was filtered in vacuo and the solid was washed with water and dried in vacuo to give 2.101 g of the crude product. It was purified on silica gel (2% MeOH in DCM) to give 1.056 g of 3-(6-chloro-pyrimidin-4-ylamino)phenol. MS (MH⁺): 222

Example 2 (Method B). 6-chloro-pyrimidin-4-yl)-(1 H-indazol-5-yl)-amine

The literature procedure (WO 2004/000833) was adapted as follows. A mixture of 4,6- dichloropyrimidine (0.876 g, 5.88 mmol), 5-aminoindazole (0.859 g, 6.46 mmol), sodium iodide (1.06 g, 7.07 mmol) and DIEA (1.23 mL, 7.06 mmol) in dry DMF (12 mL) was stirred at 80 °C for 4 h. The reaction mixture was partitioned between EtOAc - water. The organic layer was washed with water, brine and dried over Na₂S0₄. The volatiles were removed in vacuo, the residue dried to give 1.06 g of (6-chloro-pyrimidin-4-yl)-(1 H-indazol-5-yl)-amine. MS (MH⁺): 246

Step-2: The prototypical procedures for step 2 are exemplified below.

Example 3 (Method C). 3-[6-(7-methoxy-benzofuran-5-ylamino)-pyrimidin-4-ylamino]- phenol (TR58608)

A mixture of 3-(6-chloro-pyrimidin-4-ylamino)phenol from Example 1 , step 1 (0.38 g, 1.72 mmol) and 5-amino-7-methoxybenzofuran (0.28 g, 1.92 mmol) was heated neat at 150 °C for 3 h. The black melt was cooled to rt. The crude product was dissolved in MeOH and purified on silica gel (3% MeOH in DCM) to give 0.321 g of 3-[6-(7-methoxy-benzofuran-5- ylamino)-pyrimidin-4-ylamino]-phenol. MS (MH⁺): 349. Kiapp = III

The step 2 reaction could also be carried out in the presence of solvents, such as, 2- ethoxyethanol, ethanol, n-butanol, acetonitrile or DMF as described for Example 4. Example 4 (Method D). 3-[6-(5-Methoxy-benzofuran-7-ylamino)-pyrimidin-4-ylamino]- phenol (TR56215)

A mixture of 3-(6-chloro-pyrimidin-4-ylamino)phenol from Example 1 , step 1 (0.085 g, 0.38 mmol) and 7-amino-5-methoxybenzofuran (0.056 g, 0.38 mmol) in 2-ethoxyethanol (1 mL) was stirred overnight at 150 °C. The reaction mixture was cooled to rt, filtered through a syringe filter and purified by LC-MS. The TFA salt thus obtained was neutralized by treatment with 10% aq. K₂C0₃ to give 0.016g of 3-[6-(5-methoxy-benzofuran-7-ylamino)- pyrimidin-4-ylamino]-phenol. MS (MH⁺): 349. Kiapp = III Alternatively, the reaction could also be carried out under microwave irradiation at 160 - 220 °C for 5 - 30 minutes using slightly excess pyridine hydrochloride and ethanol or 2- ethoxyethanol as a solvent as described in example 5. Example 5 (Method E). N-(1 H-lndazol-5-yl)-N'-(7-methoxy-benzofuran-5-yl)-pyrimidine-4,6- diamine (TR58757)

A mixture of (6-chloro-pyrimidin-4-yl)-(1 H-indazol-5-yl)-amine from Example 2, step 1 (0.075 g, 0.31 mmol), 5-amino-7-methoxybenzofuran (0.056 g, 0.38 mmol) and pyridine hydrochloride (0.054 g, 0.47 mmol) in ethanol (3 ml_) was microwaved at 160 °C for 5 minutes. The volatiles were removed in vacuo and the residue was suspended in water (5 ml_). The pH was adjusted with 10% aq. K₂C0₃ to ~ 8 and the aq. layer was extracted with DCM containing 10% MeOH. The crude product was purified on silica gel (4% MeOH in DCM) to give 0.022 g N-(1 H-lndazol-5-yl)-N'-(7-methoxy-benzofuran-5-yl)-pyrimidine-4,6- diamine. MS (MH⁺): 373. Kiapp = III

The following compounds were prepared by reacting the compound in Example 1 , step 1 with requisite amines following methods C or D as shown in Table 1.

Table 1. RHN

Example 21 (Method F). 5-[6-(3-Hydroxy-phenylamino)-pyrimidin-4-ylamino]-benzofuran- 7-ol (TR58609)

A mixture of 3-[6-(7-methoxy-benzofuran-5-ylamino)-pyrimidin-4-ylamino]-phenol (Example 3) (0.075 g, 0.22 mmol) and pyridine hydrochloride (0.500 g, 4.33 mmol) was heated at 185 °C for 2 h. The reaction mixture was cooled to rt and diluted with water. The solution was neutralized with 10% aq. NaOH to give a brown suspension. The resulting

suspension was filtered and the solid was washed with water, dried and purified by HPLC to give 0.008 g of 5-[6-(3-hydroxy-phenylamino)-pyrimidin-4-ylamino]-benzofuran-7-ol as a TFA salt. MS (MH⁺): 335. Kiapp = II

The following compound was prepared similarly from the compound in Example 17.

Table 2

The following compounds were prepared in a manner similar to the compound in Example 2, step 1 by replacing 4,6-dichloropyrimidine with the substituted 4,6-dichloropyrimidines as shown in Table 3.

Table 3 Example R MS (MH+)

23 5-CH₃ 260

24 2-SCH3 292

25 2-Ph 322

26 2-N(CH₃)₂ 289

The following compounds were prepared from the compounds in Examples 22 - following method C or D as shown in Table 4.

Table 4

6-Phenoxy-pyrimidin-4-ylamine derivatives were prepared according to the general procedure shown in Scheme 2.

Scheme 2.

Step-1 : The prototypical procedure for step 1 is exemplified below. Example 46 (Method F). 4-Chloro-6-(indan-5-yloxy)-pyrimidine

re lux

This step was conducted by following the method cited in the patent WO2004014870A1. A solution of 4,6-dichloropyrimidine (0.229 g, 1.54 mmol), 5-hydroxyindan (0.207 g, 1.54 mmol) and DBU (0.235 ml_, 1.57 mmol) in dry acetonitrile (8 ml_) was refluxed for 4 - 6 h. The volatiles were removed in vacuo and the residue was partitioned between EtOAc - water. The organic layer was washed with water, brine and dried over Na₂S0₄. Removal of solvent in vacuo and purification of the residue on silica gel (DCM) gave 0.356 g of 4- chloro-6-(indan-5-yloxy)-pyrimidine. MS (MH⁺): 247.

All aryloxy derivatives were prepared in an analogous manner by replacing 5-hydroxy indan with requisite hydroxyl compounds.

Step-2: 3-[6-(lndan-5-yloxy)-pyrimidin-4-ylamino]-phenol (TR1 12349)

This compound was prepared by reacting 4-Chloro-6-(indan-5-yloxy)-pyrimidine (0.100 g, 0.405 mmol) with 3-aminophenol (0.088 g, 0.81 mmol) as shown in Method D. The crude product purified on silica gel (20% EtOAc in DCM) to give 0.042 g of 3-[6-(indan-5-yloxy)- pyrimidin-4-ylamino]-phenol. MS (MH⁺): 320. Kiapp = III

The following compounds were prepared in a manner similar to the compound described in Example 46 by following method C or D or E as shown in Table 5.

Table 5

The compound in example 61 was prepared from 4,6-dichloropyrimidine using Method F followed by Method E.

The compounds in Table 6 were prepared by reacting 5-(6-chloro-pyrimidin-4-yloxy)-1 H- indole (prepared from 5-hydroxyindole and 4,6-dichloropyrimidine as shown in Example 46, step 1 ) with properly substituted anilines in a manner similar to the compound in Example 46, step 2 using method C or D or E. Table 6

Example R Method MS (MH⁺) Kiapp TR

62 3-OH D 319 ¹ 58729

63 2-F, 4-CI, 5-OH C 371 II 1 12399

64 3-NHCOCH3 C 360 ¹ 1 12400

65 3-OCH3 C 333 ¹ 1 12401

66 3-COCH2CH3 C 359 ¹ 1 12422

67 3-COCH3 C 345 II 1 12423

68 3,5-di(OCH₃) C 363 ¹ 1 12424

69 3-OCF3 C 387 III 1 12425

70 3-OH, 4-CH3 C 333 ¹ 1 12436

The following compounds were also prepared in a manner similar to Example 46 as shown in Table 7. Table 7

183 D 358 I 127793

Example 184 (Method G).

[6-(1 H-lndol-5-yloxy)-pyrimidin-4-yl]-(4-piperazin-1-yl-phenyl)-amine (TR123306) An aqueous 10% K2CO3 solution (2.50 mL, 1.81 mmol) was added dropwise to a stirred suspension of the compound from Example 143 (0.173 g, 0.36 mmol) in MeOH (5 mL) at rt The resulting suspension was stirred overnight at rt. The reaction mixture was diluted with water and the precipitated solid was collected by vacuum filtration, washed with water and dried in vacuo. The crude product was purified on silica gel (9% MeOH + 1 % ΕίβΝ in DCM) to give 0.138 g of [6-(1 H-lndol-5-yloxy)-pyrimidin-4-yl]-(4-piperazin-1 -yl-phenyl)-amine. MS (MH⁺): 387. Kiapp = I

Example 185 (TR124586).

{4-[6-(1 H-lndol-5-yloxy)-pyrimidin-4-ylamino]-phenyl}-piperazin-1-yl-methanone

The compound from Example 145 was converted to {4-[6-(1 H-lndol-5-yloxy)-pyrimidin-4- ylamino]-phenyl}-piperazin-1-yl-methanone by following the method G. MS (MH⁺): 415. Kiapp = I Example 186 (Method H).

4-{4-[6-(1 H-lndol-5-yloxy)-pyrimidin-4-ylamino]-phenyl}-piperazine-1-carboxylic acid amide (TR125158)

Trimethylsilyl isocyanate (0.035 mL, 0.26 mmol) was added to a stirred solution of the compound from Example 184 (0.100 g, 0.26 mmol) in 1-methyl-2-pyrrolidinone (1 mL) at rt. The resulting solution was stirred overnight at rt. The reaction mixture was partitioned between DCM (containing 10% MeOH) and water. The organic layer was washed with water, brine and dried over Na2S0₄. The volatiles were removed in vacuo, and the residue dried to give 0.0518 g of 4-{4-[6-(1 H-lndol-5-yloxy)-pyrimidin-4-ylamino]-phenyl}- piperazine-1 -carboxylic acid amide. MS (MH⁺): 430. Kiapp = I Preparation of starting materials.

(All the compounds described below showed satisfactory ¹H NMR data) Example 187. 7-Amino-5-methoxybenzofuran

This compound was prepared from 2-hydroxy-5-methoxy-3-nitrobenzaldehyde following the literature procedure (J. Med. Chem. 2004, 871 - 887).

Example 188. 5-Amino-7-methoxybenzofuran

This compound was prepared from 2-hydroxy-3-methoxy-5-nitrobenzaldehyde in a manner similar to Example 187.

Example 189. 5-Aminobenzofuran

This compound was prepared from 2-hydroxy-5-nitrobenzaldehyde in a manner similar to the compound in Example 187.

Example 190. 3-(3-Amino-phenoxy)propan-1-ol

This compound was prepared by following the literature procedure (Tetrahedron, 2004, 121 - 130).

Example 191. 2-(3-Amino-phenoxy)ethanol

This compound was prepared in a manner similar to the compound in Example 190. Example 192. 1 H-lndazol-5-ol

This compound was prepared by following the literature procedure (J. Org. Chem. 1997, 5627 - 5629). Example 193. 3,5-Dichloro-5-methoxyaniline

This compound was prepared by following the literature procedure (Arch. Environ. Contam. Toxicol. 2002, 105 - 1 17).

Example 194. 1-(4-Amino-phenyl)-piperidin-4-ol

Step 1. A solution of 1-fluoro-4-nitrobenzene (0.658 g, 4.66 mmol), 4-hydroxypiperidine (0.590 g, 5.81 mmol) and DIEA (1.24 mL, 7.12 mmol) in anhyd CH₃CN (7 mL) was refluxed for 8 hours. The volatiles were removed in vacuo and the residue was suspended in water (15 mL) to give a bright yellow suspension. The solid was collected by vacuum filtration, washed with water and dried in vacuo to give 1.025 g 1-(4-nitro-phenyl)-piperidin-4-ol. MS (MH⁺): 223

Step 2. A suspension of 1-(4-nitro-phenyl)-piperidin-4-ol (1.020 g, 4.59 mmol) and 10% Pd- C (0.130 g) in ethanol (25 mL) was hydrogenated (balloon) overnight at rt. The catalyst was removed by filtration and washed with ethanol. The combined filtrate was

concentrated in vacuo and the residue dried further in vacuo to give 0.706 g 1-(4-amino- phenyl)-piperidin-4-ol. MS (MH⁺): 193

Example 195. 8-Amino-6-methoxyquinoline

This compound was prepared by hydrogenation of 6-methoxy-8-nitroquinoline as shown in Example 194, step 2.

Example 196. 5-Amino-2-pyrrol-1-yl-benzonitrile

This compound was prepared by hydrogenation of 5-nitro-2-pyrrol-1-yl-benzonitrile as shown in Example 194, step 2.

Example 197. 5-Amino-2-morpholin-4-yl-benzonitrile This compound was prepared from 2-fluoro-5-nitrobenzonitrile and morpholine in a manner similar to the compound in Example 194. Example 198. 4-(4-Amino-phenyl)-piperazine-1-carboxylic acid tert-butyl ester

This compound was prepared from 1-fluoro-4-nitrobenzene and Piperazine-1-carboxylic acid tert-butyl ester in a manner similar to the compound in Example 194. Example 199. 4-(2,6-Dimethyl-morpholin-4-yl)-phenylamine

This compound was prepared from 1-fluoro-4-nitrobenzene and 2,6-dimethylmorpholine in a manner similar to the compound in Example 194. Example 200. 4-(2-Morpholin-4-yl-ethoxy)-phenylamine

This compound was prepared by following the literature procedure (J. Org. Chem. 1951 , 1421 - 1424). Example 201. 2-[4-(4-Amino-2-chloro-phenyl)-piperazin-1-yl]-ethanol

This compound was prepared from 2-chloro-1-fluoro-4-nitrobenzene and 2-piperazin-1 -yl- ethanol in a manner similar to the compound in Example 194. Example 202. 4-(2,6-Difluoro-4-nitro-phenyl)morpholine

This compound was prepared from 1 ,2,3-trifluoro-5-nitrobenzene and morpholine in a manner similar to the compound in Example 194. Example 203. 3-Methyl-4-morpholin-4-yl-aniline This compound was prepared from 1-fluoro-2-methyl-4-nitrobenzene and morpholine in a manner similar to the compound in Example 194.

Example 204. 3,5-Dichloro-4-morpholin-4-yl-aniline

This compound was prepared from 1 ,3-dichloro-2-fluoro-5-nitrobenzene and morpholine in a manner similar to the compound in Example 194.

Example 205. 2-[(4-Amino-phenyl)-methyl-amino]-ethanol

This compound was prepared from 1-fluoro-4-nitrobenzene and 2-methylamino-ethanol in a manner similar to the compound in Example 194.

Example 206. 2-(4-Amino-phenylamino)-ethanol

This compound was prepared from 1-fluoro-4-nitrobenzene and 2-amino-ethanol in a manner similar to the compound in Example 194.

Example 207. N-(2-Dimethylamino-ethyl)-benzene-1 ,4-diamine

This compound was prepared from 1-fluoro-4-nitrobenzene and Ν,Ν,Ν'-trimethyl-ethane- 1 ,2-diamine in a manner similar to the compound in Example 194.

Example 208. 3-Methoxy-4-morpholin-4-yl-phenylamine

This compound was prepared from 1-fluoro-2-methoxy-4-nitro-benzene and morpholine in a manner similar to the compound in Example 194.

Example 209. 1 -[4-(4-Amino-phenyl)-piperazin-1 -yl]-2,2,2-trifluoro-ethanone

To a stirred, ice-cooled solution of 1-(4-nitrophenyl)piperazine (4.00 g, 19.3 mmol) in dichloromethane (100 ml_), trifluoroacetic anhydride (3.0 ml_, 21.42 mmol) was added dropwise. The resulting solution was allowed to warm to rt and stirred overnight at rt. TLC (1 :1 , hexane:EtOAc) indicated absence of 1-(4-nitrophenyl)piperazine. The volatiles were removed in vacuo and the residue was dried further to give 6.1 g (104%) 2,2,2-trifluoro-1- [4-(4-nitro-phenyl)-piperazin-1-yl]-ethanone. MS (MH⁺): 304.

This compound was hydrogenated as shown in Example 194, step 2 to give 1-[4-(4-amino- phenyl)-piperazin-1-yl]-2,2,2-trifluoro-ethanone.

Example 210. 1-[4-(4-Amino-benzoyl)-piperazin-1-yl]-2,2,2-trifluoro-ethanone

This compound was prepared from (4-nitro-phenyl)-piperazin-1-yl-methanone in a manner similar to example 209.

Example 21 1. 2-{2-[4-(4-Amino-phenyl)-piperazin-1-yl]-ethoxy}-ethanol

This compound was prepared from 1-fluoro-4-nitrobenzene and 2-(2-piperazin-1-yl- ethoxy)-ethanol in a manner similar to the compound in Example 194.

Example 212. 2-[4-(4-Amino-phenyl)-piperazin-1-yl]-acetamide

This compound was prepared from 1-fluoro-4-nitrobenzene and 2-piperazin-1-yl-acetamide in a manner similar to the compound in Example 194.

Example 213. 1-(4-Amino-phenyl)-piperidine-4-carboxylic acid amide

This compound was prepared from 1-fluoro-4-nitrobenzene and piperidine-4-carboxylic acid amide in a manner similar to the compound in Example 194.

Example 214. 2,5-Diethoxy-4-morpholin-4-yl-phenylamine

This compound was prepared by hydrogenation of 4-(2,5-diethoxy-4-nitro-phenyl)- morpholine as shown in Example 194, step 2. Example 215. 4-(4-Amino-phenyl)-piperazin-2-one

This compound was prepared from 1-fluoro-4-nitrobenzene and piperazin-2-one in a manner similar to the compound in Example 194.

Example 216. 4-Amino-N-m-tolyl-benzamide

A solution of 4-nitrobenzoyl chloride (0.500 g, 2.69 mmol), m-toluidine (0.290 ml_, 2.69 mmol) and triethylamine (1.136 ml_, 8.08 mmol) in dry DCM (10 ml_) was stirred overnight at rt. The volatiles were removed in vacuo, residue diluted with water and pH adjusted with 10% aq K2C03 to ~ 8 to give suspension. The solid was collected by vacuum filtration, washed with water and dried in vacuo to give 0.080 g 4-nitro-N-m-tolyl-benzamide. MS (MH⁺): 257.

This compound was hydrogenated as shown in Example 194, step 2 to give 4-amino-N-m- tolyl-benzamide.

Example 217. N-(4-Amino-2,6-dichloro-phenyl)-2-methoxy-acetamide

Step 1. 2,6-Dichloro-4-nitroaniline (1.04 g, 5.00 mmol) was reacted with methoxyacetyl chloride (0.500 ml_, 5.469 mmol) according to the literature procedure (J. Med. Chem. 1966, 428 - 430) to give 0.301 g N-(2,6-dichloro-4-nitro-phenyl)-2-methoxy-acetamide. MS (MH⁺): 279.

Step 2. A mixture of N-(2,6-dichloro-4-nitro-phenyl)-2-methoxy-acetamide (0.290 g, 1.0354 mmol) and tin (II) chloride dihydrate (0.704 g, 3.12 mmol) in ethanol (5 ml_) was refluxed for 3h. TLC (2.5% MeOH in DCM) indicated absence of starting material. The volatiles were removed in vacuo and the residue was suspended in water (15 ml_). The pH was adjusted with 50% aq NaOH to ~ 8 and the suspension was extracted with EtOAc (2 X 30 ml_). The combined EtOAc layer was washed with water, brine and dried over anhyd Na₂S0₄. It was then removed in vacuo and the residue dried further in vacuo to give 0.230 g N-(4-amino-2,6-dichloro-phenyl)-2-methoxy-acetamide. MS (MH⁺): 249.

Example 218. 2-(5-Amino-2-oxo-benzooxazol-3-yl)-acetamide

This compound was prepared by hydrogenation of 2-(5-nitro-2-oxo-benzooxazol-3-yl)- acetamide as shown in Example 194, step 2.

Example 219. 2-Methyl-benzooxazol-5-ylamine

This compound was prepared by hydrogenation of 2-methyl-5-nitro-benzooxazole as shown in Example 194, step 2.

Example 220. 6-Methoxy-1 H-indol-5-ol

This compound was prepared by hydrogenation of 5-benzyloxy-6-methoxy-1 H-indole as shown in Example 194, step 2.

Example 221. 1-Methyl-1 H-indol-5-ol

This compound was prepared from 5-benzyloxy-1 H-indole by following the literature procedure (Bioorg. Med. Chem. Lett. 2001 , 871 - 874).

Example 222. 3-Methyl-1 H-indol-5-ol

This compound was prepared by hydrogenation of (5-benzyloxy-1 H-indol-3-ylmethyl)- dimethylamine as shown in Example 94, step 2 .

Example 223. 6-Fluoro-1 H-indol-5-ol

This compound was prepared by following the literature procedure (J. Med. Chem. 1979, Example 224. 5-Amino-2-chloro-benzonitrile

This compound was prepared by reduction [tin (II) chloride dihydrate] of 2-chloro-5-nitro- benzonitrile as described in Example 217, step 2.

Example 225. 2-Methylindole-5-ol

To a stirred, ice-cooled solution of 5-methoxy-2-methylindole (0.444 g, 3.00 mmol) in dry DCM (30 mL), 1.0 M solution of BBr₃ in DCM (15.0 mL, 15 mmol) was added dropwise over 15 min. The reaction mixture was allowed to warm to rt and stirred at rt for 3h. It was then cooled in ice-bath again and excess BBr₃ was quenched with slow addition of MeOH (5 mL). The volatiles were removed in vacuo and the residue diluted with water and pH adjusted with 10% aq K2CO3 to give a suspension. The solid was collected by vacuum filtration, washed with water and dried. The crude product was purified on silica gel (2% MeOH in DCM) to give 0.376 g 2-methylindole-5-ol. MS (MH⁺): 148.

Example 226. 3-Methoxy-4-pyrrolidin-1-yl-phenylamine This compound was prepared by hydrogenation of 1-(2-methoxy-4-nitro-phenyl)-pyrrolidine as shown in Example 194, step 2.

Example 227. [4-(1 H-lndol-5-yloxy)-[1 ,3,5]triazin-2-yl]-(4-morpholin-4-yl-phenyl)-amine. A mixture of 2,3-dichloro-1 ,3,5-triazine (0.625 g, 4.168 mmol), 4-morpholinoaniline (0.750 g, 4.208 mmol) and DIEA (1.0 mL, 5.746 mmol) in dry acetonitrile was stirred overnight at RT. TLC (5% MeOH in DCM) showed absence of both the starting materials. The volatiles were removed in vacuo and the residue was purified on silica gel (1 % MeOH in DCM) to give 0.680 g (4-chloro-[1 ,3,5]triazin-2-yl)-(4-morpholin-4-yl-phenyl)-amine. MS: 292 (MH+).

(4-Chloro-[1 ,3,5]triazin-2-yl)-(4-morpholin-4-yl-phenyl)-amine (0.220 g, 0.754 mmol), was reacted with 5-hydroxyindole (0.100 g, 0.751 mmol) and DBU (0.220 mL, 1.473 mmol) in dry acetonitrile (3.0 ml_) as described in Method F to give 0.050 g [4-(1 H-lndol-5-yloxy)- [1 ,3,5]triazin-2-yl]-(4-morpholin-4-yl-phenyl)-amine. MS: 389 (MH+).

THERAPEUTIC USES OF THE COMPOUNDS OF THE INVENTION

An aspect of the invention makes use of the ability of the compounds of the invention to inhibit VEGFR1 kinases. As mentioned earlier, small molecule VEGFR1 kinase inhibitors are proven to be effective antitumor and anti-inflammatory agents. Therefore, the compounds of Formula 1 are useful as antitumor and anti-inflammatory agents.

Additionally, the compounds of Formula 1 are useful agent for interfering with angiogenesis in a disease such as diabetic retinopathy, psoriasis, and anthropathies.

For therapeutic purposes, the compounds of the present invention may be formulated in a pharmaceutical composition and administered to a mammal needing treatment according to the method described in U.S. patent No. 7,504,29. Suitable compositions of this invention include aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4. The solutions may be introduced into a patient's bloodstream by local bolus injection. The

pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide

pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients, which are suitable for the manufacture of tablets. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, microcrystalline cellulose, sodium crosscannellose, com starch, or alginic acid; binding agents, for example starch, gelatin, polyvinyl-pyrrolidone or acacia, and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to mask the unpleasant taste of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a water soluble taste masking material such as hydroxypropyl- methylcellulose or hydroxypropyl-cellulose, or a time delay material such as ethyl cellulose, cellulose acetate buryrate may be employed.

The compounds of the instant invention may also be coadministered with other well-known therapeutic agents that are selected for their particular usefulness against the condition that is being treated. The instant compounds are also useful in combination with known anti-cancer agents. Such known anti-cancer agents include the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and other angiogenesis inhibitors. The instant compounds are particularly useful when coadminsitered with radiation therapy. The synergistic effects of inhibiting VEGF in combination with radiation therapy have been described in the art. See international patent application WO 00/61 186.

The instant compounds are also useful, alone or in combination with platelet fibrinogen receptor (GP llb/llla) antagonists, such as tirofiban, to inhibit metastasis of cancerous 15 cells. Tumor cells can activate platelets largely via thrombin generation. This activation is associated with the release of VEGF. The release ofVEGF enhances metastasis by increasing extravasation at points of adhesion to vascular endothelium (Amirkhosravi, Platelets 10,285-292 (1999)). Therefore, the present compounds can serve to inhibit metastasis, alone or in combination with GP llb/llla) antagonists. Examples of other fibrinogen receptor antagonists include abciximab, eptifibatide, sibrafiban, lamifiban, lotrafiban, cromofiban, and CT50352.

Claims

We claim:

A compound of Formula I: rocycles Group 2)

wherein:

Het₂ is selected from:

wherein R-i , R₂, and R3 are each independently selected from: H, halogen, alkyl, alkenyl, alkynyl, haloalkyi, phenyl and substituted phenyl, pyridinyl, N(alkyl)₂, CN, CONH₂, CONHAIkyl, CONHPh, OH, O-Alkyl, O-alkyI substituted with halogen, O-alkyI substituted with OH, S-alkyl, O-Phenyl, COAIkyl, NHCOalkyl, NHCOaryl;

R₅ is selected from H, CH₃, SCH₃, Ph, and NMe₂;

X is selected from NH and O;

in Heterocycles Group 1 , A, B, D, and E are each independently selected from O, S, N, NH, bond, CO, CH₂, C-R, C-OR, wherein R is selected from H, halogen, alkyl, alkenyl, alkynyl, haloalkyi, phenyl and substituted phenyl, CN, CH2CN, CH2CONH2, and CH2CH2NH-CO-CH3; or a pharmaceutically acceptable salt, or a stereoisomer thereof. 2. The compound according to claim 1 , wherein Heterocyles Group 1 is selected from:

The compound according to claim 1 , wherein

R-i , R₂, and R₃ are each independently selected from

CH₃

H H

"OH "OH ~N(CH₃)₂ _N__R wherein R₄ is selected from H, COCF₃, COCH₃, (CH₂)₂OH, COOAIkyl, j (CH₂)₂0(CH₂)₂OH, and CH₂CONH₂ ;

Λ

W wherein W is selected from O, CH₂, CHOH, and CHCONH₂ ;

-N O ; — N NH ; — Y-N O wherein Y is selected from CO, CH₂, and S0₂ ; A method for treating a disease in a patient comprising administration of a therapeutically effective amount of the compound of claim 1.

The method according to claim 4 wherein said disease is a cancer.

The method according to claim 5 wherein said cancer is breast, colon, prostate or lung cancer.

A pharmaceutical composition comprising a compound in accordance with claim 1 and a pharmaceutically acceptable carrier.