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Definitions

• the present invention relates to novel compounds capable of modulating, regulating and/or inhibiting tyrosine kinase signal transduction.
• the present invention is also directed to methods of regulating, modulating or inhibiting tyrosine kinases, whether of the receptor or non-receptor class, for the prevention and/or treatment of disorders related to unregulated tyrosine kinase signal transduction, including cell growth, metabolic, and blood vessel proliferative disorders.
• PTKs Protein tyrosine kinases
• receptor tyrosine kinase mediated signal transduction is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein tyrosine kinase activity and phosphorylation. Binding sites are thereby created for intracellular signal transduction molecules and lead to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response (e.g., cell division, metabolic homeostasis, and responses to the extracellular microenvironment).
• ligand specific growth factor
• ligand dimerization transient stimulation of the intrinsic protein tyrosine kinase activity and phosphorylation.
• Binding sites are thereby created for intracellular signal transduction molecules and lead to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response (e.g., cell division, metabolic homeostasis, and responses to the extracellular microenvironment).
• tyrosine phosphorylation sites function as high-affinity binding sites for SH2 (src homology) domains of signaling molecules.
• SH2 serosine kinases
• Several intracellular substrate proteins that associate with receptor tyrosine kinases (RTKs) have been identified. They may be divided into two principal groups: (1) substrates which have a catalytic domain; and (2) substrates which lack such domain but serve as adapters and associate with catalytically active molecules. The specificity of the interactions between receptors or proteins and SH2 domains of their substrates is determined by the amino acid residues immediately surrounding the phosphorylated tyrosine residue.
• Tyrosine kinases can be of the receptor-type (having extracellular, transmembrane and intracellular domains) or the non-receptor type (being wholly intracellular).
• the receptor-type tyrosine kinases comprise a large family of transmembrane receptors with diverse biological activities.
• the intrinsic function of RTKs is activated upon ligand binding, which results in phosphorylation of the receptor and multiple cellular substrates, and subsequently in a variety of cellular responses.
• the non-receptor tyrosine kinases represent a collection of cellular enzymes which lack extracellular and transmembrane sequences.
• RTK kinases have been found to be involved in cellular signaling pathways leading to pathological conditions, including exudative age-related macular degeneration (Ni et al. Opthalmologica 2009 223 401-410; Chappelow et al. Drugs 2008 68 1029-1036), diabetic retinopathy (Zhang et al., Int. J. Biochem. Cell Biol. 2009 41 2368-2371), cancer (Aora et al. J. Path. Exp. Ther. 2006, 315, 971), psoriasis (Heidenreich et al Drug News Perspective 2008 21 97-105) and rosacea (Smith, J. R., V. B.
• VEGFR and PDGFR ⁇ may provide a greater therapeutic effect by causing regression of existing neovascular blood vessels present in the disease.
• Adamis et al., Am. J. Pathol. 2006 168 2036-2053 In cancer inhibition of multiple RTK signaling pathways has been suggested to have a greater effect than inhibiting a single RTK pathway (DePinho et al., Science 2007 318 287-290; Bergers et al. J. Clin Invest. 2003 111 1287-1295).
• WO 2013/062843 A1 refers to pyridine-sulfoximines as tyrosine kinase inhibitors.
• WO 2008/061236 A2 refers to sulfoximine-nicotine derivatives as kinase inhibitors and their preparation, pharmaceutical compositions and use in the treatment of proliferative diseases.
• the present invention relates to organic molecules capable of modulating, regulating and/or inhibiting tyrosine kinase signal transduction by blocking the VEGF and/or PDGF receptors.
• Such compounds are useful for the treatment of diseases related to unregulated tyrosine kinase signal transduction, including vascular proliferative disorders such as diabetic retinopathy, age-related macular degeneration and retinopathy of prematurity.
• the compounds of the present invention have the following general formula I:
• R 1 is hydrogen or NH 2
• R 2 is hydrogen or NH 2
• Y is CH or N
• Ar 1 is an aryl, i.e. a carbocyclic aryl or a heteroaryl group
• R 3 is hydrogen or lower alkyl
• R 4 is hydrogen, lower alkyl, (CH 2 )—COOR 6 or (CH 2 )—COR 7 ,
• R 5 is hydrogen or lower alkyl,
• R 6 is hydrogen or lower alkyl,
• R 7 is an amine, e.g a substituted amine, which may be selected from the group consisting of
• n is 0, or an integer of from 1 to 6 and further including prodrugs, pharmaceutically acceptable salts, racemic mixtures and enantiomers thereof.
• X is —NH—C(O)—.
• Y is CH.
• Ar 1 is elected from the group consisting of phenyl and furanyl, e.g. lower alkyl and/or halo-substituted phenyl and furanyl. Most preferably Ar 1 is selected from the group consisting of 3-methyl-2-furanyl and 2-fluoro-5-methylphenyl.
• R 2 is H.
• R 3 is selected from the group consisting of
• R 4 and R 5 are as defined above. More preferably in formula I, R 3 is selected from the group consisting of
• R 4 and R 5 are as defined above.
• the compounds of the present invention have the following general formula II:
• R 1 is hydrogen or NH 2
• R 2 is hydrogen or NH 2
• Y is CH or N
• Ar 1 is an aryl group, i.e. a carbocyclic aryl group or heteroaryl group, further including prodrugs, pharmaceutically acceptable salts, racemic mixtures and enantiomers thereof.
• Said carbocyclic aryl or heteroaryl group may be optionally substituted with one or more alkoxy, halogen, trihaloalkyl (e.g., trifluoromethyl), or lower alkyl radicals.
• X is —NHC(O)— or —C(O)NH—
• Y is CH.
• Ar 1 is selected from the group consisting of phenyl, oxazoyl and furanyl and.lower alkyl, alkyloxy and/or halo-substituted phenyl, oxazoyl and furanyl.
• Ar 1 is selected from the group consisting of 3-methylfuranyl, 2-fluoro 5-methylphenyl, 4-chloro 5-t-butylphenyl, 3-methoxyphenyl and 5-butyloxazoyl.
• R 2 is H.
• said compounds of formulae I and II are selected from the group consisting of:
• compounds of formulae I and II are useful as kinase inhibitors.
• compounds of formula I will be useful for treating diseases related to unregulated tyrosine kinase signal transduction, for example, cancer, blood vessel proliferative disorders, fibrotic disorders, and neurodegenerative diseases.
• the compounds of the present invention are useful for treatment of mesangial cell proliferative disorders and metabolic diseases, pterigium, arthritis, restenosis, hepatic cirrhosis, atherosclerosis, psoriasis, rosacea, diabetis mellitus, wound healing, and neurodegenerative diseases and preferably ophthalmic diseases, i.e.
• diabetic retinopathy age-related macular degeneration, retinopathy of prematurity, pterygia, blepharoconjunctivitis, chronic allergic conjunctivitis, recurrent episcleritis, keratoconjunctivitis sicca.
• dermatological indications may be treated with the compounds of this invention: sun burn, eczema, psoriasis, contact dermatitis.
• the present invention is further directed to pharmaceutical compositions comprising a pharmaceutically effective amount of one or more of the above-described compounds and a pharmaceutically acceptable carrier or excipient, wherein said compositions are effective for treating the above diseases and conditions; especially ophthalmic diseases and conditions.
• Such a composition is believed to modulate signal transduction by a tyrosine kinase, either by inhibition of catalytic activity, affinity to ATP or ability to interact with a substrate.
• compositions of the present invention may be included in methods for treating diseases comprising proliferation, fibrotic or metabolic disorders, for example cancer, fibrosis, psoriasis, rosacea, atherosclerosis, arthritis, and other disorders related to abnormal vasculogenesis and/or angiogenesis, such as exudative age related macular degeneration and diabetic retinopathy.
• the compositions of the present invention are also useful in treating pterygia, blepharoconjunctivitis, chronic allergic conjunctivitis, recurrent episcleritis, keratoconjunctivitis sicca.
• dermatological indications may be treated: sun burn, eczema, psoriasis contact dermatitis
• the compounds of the present invention are useful an ophthalmic disease, wherein said ophthalmic disease is selected from the group consisting of pterygia, hyperemia related to an actively inflamed pterygia, recurrent pterygia following excisional surgery, prophylactic therapy to prevent recurrent pterygia post-excision, progressive pterygia approaching the visual axis, chronic low grade hyperemia associated with pterygia, corneal neovascularization, neovascular glaucoma, iris neovascularization, chronic allergic conjunctivitis, ocular rosacea, blepharoconjunctivitis, recurrent episcleritis, keratoconjunctivitis sicca, ocular graft vs host disease, diabetic retinopathy, diabetic macular edema, proliferative diabetic retinopathy, exudative or neo
• neovascular age-related macular degeneratuon with dry age-related macular degeneration
• neovascular disease associated with retinal vein occlusion neovascular disease (including choroidal neovascularization) associated with the following: pathologic myopia, pseudoxanthoma elasticum, optic nerve drusen, traumatic choroidal rupture, idiopathic etiologies, presumed ocular histoplasmosis syndrome, and retinopathy of prematurity.
• BOP refers to benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophnosphate
• BOC refers to ditertiarybutyldicarbonate
• DCM dichloromethane
• DIPEA diisopropylethylamine
• DMAP dimethylformamide
• DMF dimethylformamide
• EDCI N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide
• PDGF refers to platelet derived growth factor.
• PDGFR refers to platelet derived growth factor receptor.
• Ph refers to phenyl
• PTK protein tyrosine kinase
• RT room temperature
• RTK receptor tyrosine kinase
• THF receptor tyrosine kinase
• VEGF vascular endothelial growth factor
• VEGFR vascular endothelial growth factor receptor
• Hydrocarbyl refers to a hydrocarbon radical having only carbon and hydrogen atoms.
• the hydrocarbyl radical has from 1 to 20 carbon atoms, more preferably from 1 to 12 carbon atoms and most preferably from 1 to 7 carbon atoms.
• Substituted hydrocarbyl refers to a hydrocarbyl radical wherein one or more, but not all, of the hydrogen and/or the carbon atoms are replaced by a halogen, nitrogen, oxygen, sulfur or phosphorus atom or a radical including a halo, nitrogen, oxygen, sulfur or phosphorus atom, e.g. fluoro, chloro, cyano, nitro, dialkylamino, hydroxyl, phosphate, thiol, etc.
• the compounds of Formulae I and II can form salts which are also within the scope of this invention.
• Reference to a compound of Formula I or II herein is understood to include reference to salts thereof, unless otherwise indicated.
• the term “salt(s)”, as employed herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases.
• zwitterions inner salts may be formed and are included within the term “salt(s)” as used herein.
• Salts of the compounds of the Formulae I and II may be formed, for example, by reacting a compound of Formula I or II with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.
• Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) and the like.
• Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines, t-butyl amines, and salts with amino acids such as arginine, lysine and the like.
• Basic nitrogen-containing groups may be quarternized with agents such as lower alkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g.
• Prodrugs and solvates of the compounds of the invention are also contemplated herein.
• a discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro - drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design , (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press.
• the term “prodrug” means a compound (e.g, a drug precursor) that is transformed in vivo to yield a compound of Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate of the compound. The transformation may occur by various mechanisms (e.g., by metabolic or chemical processes), such as, for example, through hydrolysis in blood.
• prodrugs are described by T. Higuchi and W. 25 Stella, “Pro-drugs as Novel Delivery Systems,” Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.
• Alkyl refers to a straight-chain, branched or cyclic saturated aliphatic hydrocarbon.
• the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from 1 to 7 carbons, most preferably 1 to 4 carbons.
• Typical alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl and the like.
• the alkyl group may be optionally substituted with one or more substituents are selected from the group consisting of hydroxyl, cyano, alkoxy, ⁇ O, ⁇ S, NO 2 , halogen, dimethyl amino, and SH.
• Alkoxy refers to O-alkyl.
• Aryl refers to an aromatic group which has at least one ring having a conjugated pi electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups.
• the aryl group may be optionally substituted with one or more substituents selected from the group consisting of halogen, trihalomethyl (e.g., trifluoromethyl), hydroxyl, SH, OH, NO 2 , amine, thioether, cyano, alkoxy, alkyl, and amino.
• Carbocyclic aryl refers to an aryl group wherein the ring atoms are carbon
• Heteroaryl or “heterocyclic aryl” refers to an aryl group having from 1 to 3 heteroatoms as ring atoms, the remainder of the ring atoms being carbon. Heteroatoms include oxygen, sulfur, and nitrogen. Thus, heteroaryl groups include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like.
• Heterocyclic refers to cyclic group having at least one enchained heteroatom and includes aromatic and non-aromatic cyclic groups.
• Trifluoroacetamide (5.82 g, 2 eq), magnesium oxide (4.05 g, 4 eq), and rhodium(II) acetate dimer (330 mg, 0.03 eq) were placed in a 250 mL round bottom flask.
• Dichloromethane 70 mL under a nitrogen atmosphere was then added with stirring, followed by the addition of tert-butyl thiomorpholine-4-carboxylate 1-oxide (5.41 g, 1 eq) and diacetoxyiodobenzene (12.1 g, 1.5 eq). The reaction mixture was stirred at RT overnight.
• Biochemical KDR kinase assays were performed in 96 well microtiter plates that were coated overnight with 75 ⁇ g/well of poly-Glu-Tyr (4:1) in 10 mM Phosphate Buffered Saline (PBS), pH 7.4. The coated plates were washed with 2 mls per well PBS+0.05% Tween-20 (PBS-T), blocked by incubation with PBS containing 1% BSA, then washed with 2 mls per well PBS-T prior to starting the reaction.
• PBS Phosphate Buffered Saline
• Reactions were carried out in 100 ⁇ L reaction volumes containing 2.7 ⁇ M ATP in kinase buffer (50 mM Hepes buffer pH 7.4, 20 mM MgCl 2 , 0.1 mM MnCl 2 and 0.2 mM Na 3 VO 4 ).
• Test compounds were reconstituted in 100% DMSO and added to the reaction to give a final DMSO concentration of 5%.
• Reactions were initiated by the addition 20 ul per well of kinase buffer containing 200-300 ng purified cytoplasmic domain KDR protein (BPS Bioscience, San Diego, Calif.). Following a 15 minute incubation at 30° C., the reactions were washed 2 mls per well PBS-T.
• test agents For screening, cells were pre-incubated with test agents for 30 minutes, at a single concentration (10 uM) or at concentrations ranging from 0.0001 to 10.0 uM followed by VEGF 165 stimulation (10 ng/mL). Changes in fluorescence at 516 nm were measured simultaneously in all 384 wells using a cooled CCD camera. Data were generated by determining max-min fluorescence levels for unstimulated, stimulated, and drug treated samples. IC 50 values for test compounds were calculated from % inhibition of VEGF stimulated responses in the absence of inhibitor.
• Biochemical PDGFR ⁇ kinase assays were performed in 96 well microtiter plates that were coated overnight with 75 ⁇ g of poly-Glu-Tyr (4:1) in 10 mM Phosphate Buffered Saline (PBS), pH 7.4. The coated plates were washed with 2 mls per well PBS+0.05% Tween-20 (PBS-T), blocked by incubation with PBS containing 1% BSA, then washed with 2 mls per well PBS-T prior to starting the reaction.
• PBS Phosphate Buffered Saline
• Reactions were carried out in 100 ⁇ L reaction volumes containing 36 ⁇ M ATP in kinase buffer (50 mM Hepes buffer pH 7.4, 20 mM MgCl 2 , 0.1 mM MnCl 2 and 0.2 mM Na 3 VO 4 ).
• Test compounds were reconstituted in 100% DMSO and added to the reaction to give a final DMSO concentration of 5%.
• Reactions were initiated by the addition 20 ul per well of kinase buffer containing 200-300 ng purified cytoplasmic domain PDGFR-b protein (Millipore). Following a 60 minute incubation at 30° C., the reactions were washed 2 mls per well PBS-T.
• NHDF-Ad Normal Human Dermal Fibroblasts, Adult; Lonza
• NHDF-Ad Normal Human Dermal Fibroblasts, Adult; Lonza
• Cells were loaded with calcium indicator Fluo-4 for 45 minutes at 37° C.
• Cells were washed 2 times (Elx405, Biotek Instruments) to remove extracellular dye.
• test agents For screening, cells were pre-incubated with test agents for 30 minutes, at a single concentration (10 uM) or at concentrations ranging from 0.0001 to 10.0 uM followed by PDGF-BB stimulation (30 ng/mL). Changes in fluorescence at 516 nm were measured simultaneously in all 384 wells using a cooled CCD camera. Data were generated by determining max-min fluorescence levels for unstimulated, stimulated, and drug treated samples. IC 50 values for test compounds were calculated from % inhibition of PDGF-BB stimulated responses in the absence of inhibitor.

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• 2014
  • 2014-12-10 US US14/565,689 patent/US20150166518A1/en not_active Abandoned
  • 2014-12-10 WO PCT/US2014/069601 patent/WO2015089220A1/en not_active Ceased
  • 2014-12-10 AU AU2014362391A patent/AU2014362391B2/en not_active Ceased
  • 2014-12-10 JP JP2016538520A patent/JP6531100B2/ja not_active Expired - Fee Related
  • 2014-12-10 CN CN201480066924.XA patent/CN105814021B/zh not_active Expired - Fee Related
  • 2014-12-10 EP EP14815201.0A patent/EP3080084B1/en not_active Not-in-force
  • 2014-12-10 CA CA2932831A patent/CA2932831A1/en not_active Abandoned
  • 2014-12-10 BR BR112016013539-3A patent/BR112016013539A2/pt not_active Application Discontinuation
• 2016
  • 2016-03-14 US US15/069,344 patent/US9567324B2/en active Active

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