PL2235002T3

4-pyridinone compounds and their use for cancer

Abstract

This record has no abstract on file.

Term

No projected expiry on record.

13 claims: 1 independent, 12 dependent

1
Patent claims Zastrzeżenia patentowe 1. A compound having the following Formula (I):1. Związek mający następujący Wzór (I): albo jego sól, gdzie: or its salt, where: G is H, -CHX-OP (= O) (OH)2 or -CHX-OC (= O) Z;G oznacza H, -CHX-OP(=O)(OH)2 lub -CHX-OC(=O)Z;X is H or alkyl, optionally substituted with one or more of OH, halogen, cyano and / or -NR1R2;X oznacza H lub alkil, opcjonalnie podstawiony jednym lub większą ilością spośród OH, halogenu, grupy cyjanowej i/lub -NR1R2;Z is alkyl, cycloalkyl, aryl or heterocyclic group, optionally substituted with one or more of alkyl, OH, halogen, cyano and / or -NR3R4;and Z oznacza alkil, cykloalkil, aryl lub grupę heterocykliczną, opcjonalnie podstawione jednym lub większą ilością spośród alkilu, OH, halogenu, grupy cyjanowej i/lub -NR3R4;i R1, R2, R3 and R4 are independently H and / or alkyl. R1, R2, R3 i R4 oznaczają niezależnie H i/lub alkil.

240 paragraphs in 9 sections, as filed

[0001] The invention generally relates to 4-pyridinone compounds and salts thereof, for such compounds for use in the treatment of diseases, including cancer, and pharmaceutical compositions containing at least one of said compounds or pharmaceutically acceptable salts thereof.

[0002] Met, also referred to as hepatocyte growth factor receptor (HGFR), is expressed mainly in epithelial cells, but has also been identified in endothelial cells, myoblasts, hematopoietic cells and motor neurons. Overexpression of hepatocyte growth factor and Met activation have been associated with the initiation and progression of many different types of tumors, as well as with the promotion of metastatic disease.

[0003] Published US Patent Application US 2005/0245530 A1 and International Application WO 2005/117867 disclose monocyclic heterocyclic compounds that inhibit the protein tyrosine kinase activity of growth factor receptors such as Met, which makes them useful as anti-cancer agents. As can be appreciated, there remains a need for antitumor compounds useful for treating Met activated cancer and preferably having activity against other cancer pathways.

[0004] Applicants have discovered a strong association for activity against tumors dependent on Met activation as well as activity on tumors as a VEGFR inhibitor. Applicants have also discovered compound prodrugs useful for administering the compound in a more soluble form. It is now possible to provide compounds with different pharmacological profiles compared to currently known antitumor compounds for the treatment of Met-activated tumors and with such stability, bioavailability, solubility, therapeutic index and toxicity values that provide drugability.

BRIEF DESCRIPTION OF THE DRAWINGS [0005] The invention is illustrated by reference to the accompanying drawings, described below.

[0006] FIG. 1 shows the antitumor activities of Example 1 and Example 2 against GTL-16 xenograft gastric cancer.

[0007] FIG. 2 shows the antitumor activity of Example 1, dosed orally, once daily, for 14 days (arrow indicates dosage) at 6.25 mpk (mg / kg), 12.5 mpk and 25 mpk, against glioblastoma U87 xenografts.

[0008] FIG. 3 shows the solubility profiles of Example 1 and Example 2 as a function of pH.

BRIEF DESCRIPTION OF THE INVENTION [0009] Compounds of Formula (I) have been described:

or their salts, where:

G is H, -CHX-OP (= O) (OH) 2 or -CHX-OC (= O) Z;

X is H or alkyl, optionally substituted with one or more of OH, halogen, cyano and / or -NR1R2;

Z is alkyl, cycloalkyl, aryl or heterocyclic group, optionally substituted with one or more of alkyl, OH, halogen, cyano and / or NR3R4; and

R1, R2, R3 and R4 are independently H and / or alkyl.

[0010] Also described are pharmaceutical compositions comprising at least one compound of Formula (1) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or diluent.

DETAILED DESCRIPTION [0011] The following are definitions of the various expressions used to describe the invention. These definitions apply to expressions used throughout the specification (unless otherwise limited in specific cases), both individually and as part of a larger group.

[0012] The terms "alkyl" and "alk" refer to a straight or branched chain alkane (hydrocarbon) radical containing from 1 to 12 carbon atoms, preferably from 1 to 6 carbon atoms, and more preferably from 1 to 4 carbon atoms. Exemplary "alkyl" and / or "alk" groups include, but are not limited to, for example, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, pentyl, hexyl, isohexyl, heptyl, octyl, nonyl, decyl and dodecyl.

[0013] The term "lower alkyl" refers to an "alkyl" and / or "alk" group containing from 1 to 4 carbon atoms, preferably from 1 to 2 carbon atoms. When a subscript is used for an alkyl or other group, the subscript refers to the number of carbon atoms that the group may contain. For example, the impression "C0-C4alkyl "includes a bond and an alkyl group having 1 to 4 carbon atoms and the expression" C1-C4alkyl "refers to alkyl groups having 1 to 4 carbon atoms. Exemplary lower alkyl groups include, but are not limited to, for example, methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl and isobutyl.

[0014] The "alkyl" and / or "alk" group may be optionally substituted with one or more substituents, preferably from 1 to 4 substituents, in any available and substitutable position. Exemplary substituents include halogen (e.g., a single halogen substituent or multiple halogen substituents forming, in the latter case, groups such as, for example, a perfluoroalkyl group or an alkyl group carrying -CCl3 or -CF3), hydroxyl, -NH2, -NH (alkyl), -N (alkyl)2 and a cyano group.

[0015] The expression "cycloalkyl" refers to a fully saturated hydrocarbon group containing from 1 to 4 rings and from 3 to 8 carbon atoms per ring. Exemplary cycloalkyl groups include, but are not limited to, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. The cycloalkyl group may be optionally substituted with one or more substituents, preferably from 1 to 4 substituents, in any available and substitutable position. Exemplary substituents include the groups listed for substituted alkyl.

[0016] The term "aryl" refers to cyclic aromatic hydrocarbon groups having from 1 to 2 aromatic rings, such as, for example, phenyl, biphenyl or naphthyl. When the aryl group contains two aromatic rings (e.g. bicyclic, etc.), the aromatic rings may be joined at a single point (e.g. biphenyl) or fused (e.g. naphthyl and phenantrenyl). The aryl group can be optionally substituted with one or more substituents, preferably from 1 to 5 substituents, at any available or substitutable ring position or where valence allows on any of the fused or attached rings. Exemplary substituents include alkyl and the groups mentioned for alkyl substitution.

[0017] The terms "heterocycle", "heterocyclic" and "heterocyclic group" refer to fully saturated, partially saturated or fully unsaturated, aromatic (i.e. "heteroaryl") or non-aromatic cyclic groups that are, for example, monocyclic, 3- to 3- 7-membered or bicyclic, 7- to 11-membered ring systems having at least one heteroatom in at least one carbon containing ring. Each heterocyclic, heterocyclic or heterocyclic group containing a heteroatom ring may have 1, 2, 3 or 4 heteroatoms selected from N, O and / or S, where the N and / or S heteroatom (s) may be optionally oxidized and N heteroatom (s) may be optionally quaternized. A heterocycle, heterocyclic or heterocyclic group may be attached to the remainder of the molecule at any heteroatom or carbon atom of the ring or ring system. The heterocycle, heterocycle or heterocycle may be substituted at any available attachment point with at least one substituent, preferably 1 to 4 substituents selected from alkyl and those listed for substituted alkyl.

[0018] Exemplary monocyclic heterocycles, heterocyclic or heterocyclic groups include, but are not limited to, for example, ethylene oxide; azetidinyl; pyrrolidinyl; pyrrolyl; pyrazolyl; oxetanyl; pyrazolinyl; imidazolyl; imidazolinyl; imidazolidinyl; oxazolyl; oxazolidinyl; isoxazolinyl; isoxazolyl; thiazolyl; thiadiazolyl;

thiazolidinyl; isothiazolyl; isothiazolidinyl; furyl; tetrahydrofuryl; thienyl; oxadiazolyl; piperidinyl; piperazinyl; 2-oxopiperazinyl; 2-oxopiperidinyl; 2-oxopyrrolodinyl; 2oksoazepinyl; azepinyl; heksahydrodiazepinyl; 4-piperidonyl; pyridyl; pyrazinyl; pyrimidinyl; pyridazinyl; triazinyl; triazolyl; tetrazolyl; tetrahydropyranyl; morpholinyl; thiamorpholinyl; thiamorpholinyl sulfoxide; thiamorpholinyl sulfone; 1,3-dioxolane; and tetrahydro-1,1-dioxothienyl.

[0019] Exemplary bicyclic heterocycles, heterocyclic or heterocyclic groups include, but are not limited to, for example, indolyl; isoindolyl; benzothiazolyl; benzodioxolyl; benzoxazolyl; benzoxadiazolyl; benzothienyl; quinuclidinyl; quinolinyl; tetrahydroisoquinolinyl; isoquinolinyl; benzimidazolyl; benzopyranyl; indolizinyl; benzofuryl; benzofurazanyl; chromonyl; coumarinyl; benzopyranyl; cinnolinyl; quinoxalinyl; indazolyl; pyrrolopyridyl; furopyridinyl, such as, for example, furo [2,3c] pyridinyl, furo [3,2-b] pyridinyl] and furo [2,3-b] pyridinyl; dihydrobenzodioksynyl; dihydrodioksydobenzotiofenyl; dihydroisoindolyl; dihydroindolyl; dihydroquinolinyl; dihydroquinazolinyl such as, for example, 3,4-dihydro-4-oxo-quinazolinyl; triazynyloazepinyl; and tetrahydroquinolinyl.

[0020] The phrase "therapeutically effective" is intended to specify the amount of each agent that will achieve the goal of improving the severity of the disorder and the frequency of collapse when treating each agent separately, while avoiding the side effects typically associated with alternative therapies. For example, an effective anti-cancer agent extends patient survival, inhibits the development of rapidly proliferating cancer-related cells, or causes tumor regression.

[0021] The compounds of Formula (I) form salts that are also within the scope of the invention. The term "salt (s)" here means acid and / or base salts formed with inorganic and / or organic acids and bases. In addition, when a compound of Formula (I) contains both a basic group, such as but not limited to, a pyridinyl group, and an acid group, such as, but not limited to, a dihydrogen phosphate group, zwitterions ("internal salts") may be formed "), which are included in the expression" salt (s) ". Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, e.g. for isolation or purification steps that can be used in the preparation. Salts of compounds of Formula (I) may form, for example, when reacting a compound of Formula (I) with an amount of acid or base, such as an equivalent amount, in a medium such as that in which the salt precipitates or in an aqueous medium followed by lyophilisation.

[0022] The phrase "pharmaceutically acceptable salt (s)", unless otherwise indicated, includes salts containing pharmacologically acceptable anions or cations, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, gluconate, mesylate, sugar, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, sulfate, benzenesulfonate, p-toluenesulfonate and pamoate, i.e., 1,1'-methylene bis- (2-hydroxy-3-naphthanoate)].

[0023] The compounds of Formula (I) form salts that can, for example, be used to isolate and / or purify compounds of Formula (I). The salt (s) of the compounds of Formula (I) can be formed, for example, by reacting the compound of Formula (I) with, for example, an equivalent amount of acid or base in a medium that allows the salt thus formed, for example, to be precipitated or isolated by lyophilization.

[0024] Exemplary acid salt (s) that compounds of Formula (I) may form with inorganic and / or organic acids include, but are not limited to, for example, salts: acetate, ascorbate, benzoate, benzenesulfonate, bisulfate, bitartrate, acid citrate, citrate, ethanesulfonate, formate, fumarate, gentisate, gluconate, gluconate, glutamate, hydrochloride, hydrobromide, hydroiodide, isonicotinate, phosphate, mesoate, mesoate , acid phosphate, sugar, salicylate, succinate, sulfate, tartrate, ptoluenesulfonate, lactate and ebonate [i.e., 1,1'-methylene bis- (2-hydroxy-3-naphthanoate)]. Such salts may be formed according to methods known to a person skilled in the art.

[0025] Exemplary basic salt (s) that compounds of Formula (I) may form with inorganic and / or organic bases include, but are not limited to, for example, ammonium salts; alkali metal salts, such as, for example, sodium, lithium and potassium salts: alkaline earth metal salts, such as, for example, calcium and magnesium salts; salts formed from organic bases such as, for example, benzatin, dicyclohexylamine, 2-amino-2- (hydroxymethyl) propane-1,3-diol (trisamine or tris), hydrabamines (such as, for example, N, N-bis ( dehydroabietyl) ethylenediamine), N-methyl-D-glucamine, N-methyl-D-glycamides and t-butylamine; salts formed from amino acids such as, for example, arginine and lysine; and salts formed using agents such as, for example, lower alkyl halides (e.g. methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g. dimethyl, diethyl, dibutyl and diamyl sulfates), long chain halides (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides) and aralkyl halides (e.g. benzyl and phenethyl bromides) for quaternizing basic nitrogen-containing groups. Such salts may be formed according to methods known to a person skilled in the art.

[0026] The expression "prodrug" herein means a compound that, when administered to a subject, undergoes chemical conversion due to metabolic or chemical processes, giving a compound of Formula (II) or a salt thereof. Various forms of prodrug (s) are well known in the art. For examples of such prodrug derivatives, see:

a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, T. 112, pp. 309-396, edited by K. Widder et al. (Academic Press, 1985);

b) A Textbook of Drug Design and Development, edited by Krosgaard Larsen and H. Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard, pp. 113-191 (1991); and

c) H. Bundgaard, Advanced Drug Delivery Reviews, 8: 1-38 (1992).

[0027] The phrase "gene amplification" here means the selective synthesis of a DNA fragment that results in multiple copies of the Met gene or chromosome fragment on which Met is encoded.

[0028] The phrase "mutation of activated Met" means a selective change in the DNA DNA Met sequence resulting in a Met protein that is constitutively (i.e. permanently) phosphorylated.

[0029] The phrase "HGF stimulation" here means the ability of HGF to bind its receptor (Met) in such a way as to activate the receptor, resulting in a phenotypic response. In the case of Met, it may be cellular proliferation, motility, differentiation and / or survival.

[0030] The term "patient" herein includes all mammal species, including humans, cows, horses, dogs and cats; preferably people.

[0031] In one embodiment, compounds of Formula (I) are provided:

or their salts in which:

G is H, -CHX-OP (= O) (OH) 2 or -CHX-OC (= O) Z;

X is H or alkyl, optionally substituted with one or more of OH, halogen, cyano and / or -NR1R2;

Z is alkyl, cycloalkyl, aryl or heterocyclic group, optionally substituted with one or more of alkyl, OH, halogen, cyano and / or NR3R4; and R1, R2, R3 and R4 are independently H and / or alkyl.

[0032] In one embodiment, a compound of Formula (I) or a salt thereof is provided, wherein G is H. The compound of this embodiment has the structure represented by Formula (II):

[0033] The compound of Formula (II) may exist in the enol form represented by the following formula:

In the present specification, the terms "compound of Formula (II)" and "compound of Formula (I) wherein G is H" refer to a compound of Formula (II) in ketone form, enol form or any mixture containing ketone and enol forms.

[0035] In another embodiment, the compound of Formula (II) is provided as a salt. Examples of salts of the compound of Formula (II) include, but are not limited to, salts of trifluoroacetic acid and hydrochloric acid.

[0036] In one embodiment, compounds of Formula (I) or salts thereof are provided, in which:

G is -CHX-OP (= O) (OH) 2 or -CHX-OC (= O) Z;

X is H or alkyl, optionally substituted with one or more of OH, halogen, cyano and / or -NR1R2;

Z is alkyl, cycloalkyl, aryl or heterocyclic group, optionally substituted with one or more of alkyl, OH, halogen, cyano and / or NR3R4; and

R1, R2, R3 and R4 are independently H and / or alkyl.

Preferably, X is H or methyl. The compounds or salts thereof according to this embodiment are useful as prodrugs of the compound of Formula (II). When administered to a mammal, the compounds of this embodiment or a pharmaceutically acceptable salt thereof, through metabolic or chemical processes, undergo chemical conversion in vivo to give a compound of Formula (II).

[0037] In one embodiment, compounds of Formula (I) or salts thereof are provided, wherein: G is -CHX-OP (= O) (OH)2; X is H or C1-C4alkyl, optionally substituted with one or more of OH, halogen, cyano and / or NR1R2; while R1 and R2 are independently H and / or alkyl. The compounds of this embodiment have the structure of Formula (III):

[0038] Preferably, X is H, methyl, substituted methyl, ethyl or substituted ethyl; more preferably, X is H or methyl, and most preferably X is H. Compounds of Formula (III) may be provided as pharmaceutically acceptable salts, for example, ethanolamine, bis-ethanolamine, trisamine, bis-trisamine or N-methyl-D-glucamine salts. An example of a compound of Formula (III) is a compound of Formula (IIIa):

which can be provided in the form of salt. Suitable salts of the compound of Formula (IIIa) include, but are not limited to, ethanolamine, bis-ethanolamine, trisamine and bis-trisamine salts. Compounds of Formula (III) or salts thereof according to this embodiment are useful as prodrugs of a compound of Formula (II).

[0039] In one embodiment, compounds of Formula (I) or salts thereof are provided, wherein: G is -CHX-OC (= O) Z; X is H or alkyl, optionally substituted with one or more of OH, halogen, cyano and / or NR1R2; Z is alkyl, cycloalkyl, aryl or heterocyclic group, optionally substituted with one or more of alkyl, OH, halogen, cyano and / or -NR3R4; while R1, R2, R3 and R4 are independently H and / or alkyl. Preferably, X is H, C 1 -C 4 alkyl or substituted C 1 -C 4 alkyl; more preferably, H, methyl, ethyl, substituted methyl or substituted ethyl; and most preferably H or methyl. Preferably, Z is Ci-C6alkyl, substituted C 1 -C6alkyl, C.3-C6cycloalkyl, substituted with C3C6cycloalkyl, substituted phenyl, optionally substituted monocyclic or bicyclic heterocycle. More preferably, Z is C1-C6-NH substituted alkyl2 or a 5- to 6-membered heterocyclic group containing one nitrogen heteroatom such as a pyrrolidinyl and piperidinyl group. Compounds of Formula (II) or salts thereof according to this embodiment are useful as prodrugs of a compound of Formula (II).

[0040] In another embodiment, compounds of Formula (I) or salts thereof are provided, wherein G is:

The compounds of this embodiment can be provided as salts. The compounds or salts thereof according to this embodiment are useful as prodrugs of the compound of Formula (II).

[0041] In one embodiment, the compound of Formula (I) or a salt thereof is provided, wherein the compound is:

V- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide (1);

(3- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4 #) yl) methyl (2) dihydrogen phosphate;

(S) - (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl (3);

(S) - (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) 2-amino-3-methylbutanoate (S) -yl) methyl (4);

(S) - (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) 2-amino-4-methylpentanoate (S) -yl) methyl (5);

Piperidine-3-carboxylate (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl (6 );

(S) - (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl pyrrolidine-2-carboxylate ) methyl (7);

(S) - (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl pyrrolidine-3-carboxylate ) methyl (8);

Piperidine-4-carboxylate (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl (9 );

Piperidine-4-carboxylate 1- (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) ethyl (10);

(2S) -1- (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (2-amino-3-methylbutanoate) 4H) -yl) ethyl (11); or

1-Methylpiperidine-4-carboxylate (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl (12);

[0042] Hepatocyte growth factor (HGF), also known as diffuse factor (SF), due to its ability to interfere with colony formation in vitro, is a cytokine of mesenchymal origin, known to induce numerous pleiotropic responses in healthy and cancer cells (Sonnenberg et al., J. Cell Biol., 123: 223-235 (1993); Matsumato et al., Crit. Rev. Oncog., 3: 27-54 (1992); and Stoker et al., Nature, 327: 239-242 (1987)). These responses are known to include induction of proliferation in both epithelial and endothelial cells, breakdown of epithelial colonies into individual cells, stimulation of epithelial cell motogenesis, cell survival, induction of cellular morphogenesis (Montesano et al., Cell, 67: 901 -908 (1991)) and invasion promotion (Stella et al., Int. J. Biochem. Cell Biol., 12: 1357-1362 (1999) and Stuart et al., Int. J. Exp. Path., 81: 17-30 (2000)), all key processes underlying metastasis. HGF has also been reported to promote angiogenesis (Bussolino et al., J. Cell Biol., 119: 629-641 (1992)). In addition, HGF plays a key role in tissue regeneration, wound healing and normal embryonic processes, all of which depend on both motility and cell proliferation.

[0043] HGF initiates these physiological processes by binding with high affinity to its receptor, Met protein tyrosine kinase receptor, identified proto-oncogene (Park et al., Proc. Natl. Acad. Sci. USA, 84: 6379-6383 (1987) and Bottaro et al., Science, 251: 802-804 (1991)). The mature form of Met consists of a highly glycosylated external α subunit, as well as a β subunit, with a large extracellular domain, a transmembrane fragment and a cytoplasmic tyrosine kinase domain. Ligand binding induces Met dimerization, resulting in an autophosphorylated activated receptor. Met activation stimulates the signal transduction cascade, determined by the transphosphorylation of key cytoplasmic tyrosine residues responsible for the recruitment of many effector proteins (Furge et al., Oncogene, 19: 5582-5589 (2000)). These include the p85 subunit of PI3 kinase, Cy phospholipase (Gaul et al., Oncogene, 19: 1509-1518 (2000)), Grb2 and Shc adapter proteins, SHP2 and Gab1 protein phosphatases. The last adapter turned out to be the main anchor molecule below [receptor], which becomes tyrosine phosphorylated in response to ligand binding (Schaeper et al., J. Cell Biol., 149: 1419-1432 (2000); Bardelli et al., Oncogene, 18: 113911

1146 (1999) and Sachs et al., J. Cell Biol., 150: 1375-1384 (2000)). Activation of other signal molecules in HGF stimulated cells has been reported, in particular Ras, MAP kinases, STATs, ERK-1, -2 and FAK (Tanimura et al., Oncogene 17: 57-65 (1998); Lai et al. , J. Biol. Chem., 275: 7474-7480 (2000) and Furge et al., Oncogene, 19: 5582-5589 (2000)). The role of many of these signaling molecules in cell proliferation has been well established.

[0044] Met, also referred to as hepatocyte growth factor receptor (HGFR), is expressed mainly in epithelial cells, but has also been identified in endothelial cells, myoblasts, hematopoietic cells and motor neurons. HGF overexpression and Met activation have been associated with the initiation and progression of many different types of tumors, as well as with the promotion of metastatic disease. Preliminary evidence linking Met to cancer has supported the identification of a kinase domain sense mutation that predisposes individuals to papillary renal cell carcinomas (PRCs) and hepatocellular carcinomas (HCCs) (Lubensky et al., Am er. J. Pathology, 155: 517- 526 (1999)). Mutant forms of Met have also been identified in ovarian cancer, childhood HCC, gastric cancer, squamous cell carcinoma of the head and neck, non-small cell lung cancer, colorectal metastases (Christensen et al., Cancer Res., 63: 7345-7355 (2003); Lee et al., Oncogene, 19: 4947-4953 (2000) and Direnzo et al., Clin. Cancer Res., 1: 147-154 (1995)). In addition, further evidence supporting the role of Met in cancer is based on overexpression of HGF and the Met receptor in a variety of tumors, including thyroid, ovarian and pancreatic cancers. Its amplification in metastases of colorectal cancer to the liver has also been shown (Rong et al., Cancer Res., 55: 19631970 (1995); Rong et al., Cancer Res., 53: 5355-5360 (1993); Kenworthy et al ., Br. J. Cancer, 66: 243-247 (1992) and Scarpino et al., J. Pathology, 189: 570-575 (1999)). TPR-Met (activated form, similar to BCR / Abl in CML) has been described and identified in human gastric cancer (Proc. Natl. Acad. Sci. USA, 88: 4892-4896 (1991)). In patients with invasive breast cancer and in a recent study in patients with non-small cell lung cancer, receptor or ligand expression is an indicator of reduced survival, further combining Met with tumor progression (Camp et al., Cancer, 86: 2259-2265 (1999) and Masuya et al., Br. J. Cancer, 90: 1555-1562 (2004)). In general, most human tumors and tumor cell lines of mesenchymal origin incorrectly express HGFR and / or HGF.

[0045] Numerous experimental data support the role of HGF and Met in invasion, growth, persistence and tumor progression, ultimately leading to metastases. Preclinical, transgenic HGF expression results in a metastatic phenotype (Takayama et al., Proc. Natl. Acad. Sci. USA, 94: 701-706 (1997)), and Met amplification / overexpression automatically transforms NIH-3T3 cells (Cooper et al. , EMBO J., 5: 2623-2628 (1986)).

[0046] Biological agents, such as ribozymes, antibodies and antisense RNA, directed at HGF or Met, have been shown to inhibit tumorogenesis (Stabile et al., Gene Therapy, 11: 325-335 (2004); Jiang et al., Clin Cancer Res, 9: 4274-4281 (2003) and Genentech US 6,214,344 (2001)). Therefore, selective, small molecule kinase modulators, directed at Met, are expected to have therapeutic potential for the treatment of cancers in which activation of the Met receptor plays a key role in the development and progression of primary and secondary metastases. HGF is also known to regulate angiogenesis, a key process in tumor growth and spread. There is therefore potential for this class of modulators to also affect angiogenesis-dependent diseases, which may include, but are not limited to, diabetic retinopathy, macular degeneration, obesity, and inflammatory disease such as rheumatoid arthritis.

[0047] The compound of Formula (II) is useful for treating cancer, for example, cancers dependent on Met activation. Met activation is regulated by gene amplification, activated Met mutation and / or HGF stimulation. Thus, the treatment comprises administering to the patient a Compound of Formula (II) or a pharmaceutically acceptable salt or prodrug thereof. The compound of Formula (II) has been found to be particularly useful for the treatment of cancer due to the increased potency compared to known Met kinase inhibitors. In addition, the compound of Formula (II) is particularly useful for treating cancer because it also has activity as a VEGFR (vascular endothelial growth factor receptor) inhibitor, such as a VEGFR-2 inhibitor.

[0048] In one embodiment, there is provided the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of cancer. Preferably, in an embodiment, the cancer being treated is bladder cancer, breast cancer, colorectal cancer, stomach cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic / gallbladder cancer, prostate cancer, thyroid cancer, osteosarcoma, rhabdomyosarcoma, melanoma, glioblastomas / astrocytomas, MFH / fibrosarcoma or mesothelioma.

[0049] In the treatment of cancer, the combination of chemotherapeutic agents and / or other therapies (e.g. radiation therapy) is often preferred. The second (or third) agent may have the same or different mechanism of action than the first therapeutic agent. Particularly useful may be the use of a combination of cytotoxic drugs in which two or more drugs administered act differently or in different phases of the cell cycle and / or in which two or more drugs have non-overlapping toxicity or side effects and / or in which for each combination drug has been shown to be effective in treating a particular condition which the patient has.

[0050] The phrase "additional anti-cancer agents" refers to drugs selected from any one or more of the following: alkylating agents (including nitrogen mustards, alkyl sulfonates, nitrosoureas, ethyleneimine derivatives and triazenes); anti-angiogenic (including matrix metalloproteinase inhibitors); anti-metabolites (including adenosine deaminase inhibitors, folic acid antagonists, purine and pyrimidine analogues); antibiotics or antibodies (including monoclonal antibodies, CTLA-4 antibodies, anthracyclines); aromatase inhibitors; cell cycle response modifiers; enzymes; protein farnesyl transferase inhibitors; hormonal and anti-hormonal agents and steroids (including synthetic analogues, glucocorticoids, estrogens / anti-estrogens [e.g. SERMs], androgens / antiandrogens, progestins, progesterone receptor agonists and agonists, and luteinizing hormone releasing hormone [LHRH] antagonists; insulin-like growth factor (IGF) / insulin-like growth factor receptor (IGFR) modulators (including IGFR1 inhibitors); integrin signal transduction inhibitors; kinase inhibitors (including multiple kinase inhibitors and / or Src or Src / abl kinase inhibitors, cyclin dependent kinase inhibitors [CDK], panHer, Her-1 and Her2 antibodies, VEGF inhibitors, including anti-VEGF antibodies, EGFR inhibitors, mitogen-activated protein inhibitors [MAP], MEK inhibitors, Aurora kinase inhibitors, PDGF inhibitors and other tyrosine kinase inhibitors or serine / threonine kinase inhibitors; microtubule disrupting agents such as ecteinascidins or analogues and derivatives thereof; microtubule stabilizing agents such as taxanes and naturally occurring epothilones and their synthetic and semi-synthetic analogues; destabilizing agents that bind to microtubules (including vinca alkaloids); topoisomerase inhibitors; protein prenyl transferase inhibitors; platinum coordination complexes; signal transduction inhibitors; and other agents used as antitumor and cytotoxic agents such as biological response modifiers, growth factors and immunomodulators.

[0051] Accordingly, the compounds of the invention may be administered in conjunction with other anti-cancer treatments useful in the treatment of cancer or other proliferative diseases. The invention further encompasses the use of a compound of Formula (I) or pharmaceutically acceptable salts and prodrugs thereof in the preparation of medicaments for the treatment of cancer and / or comprises packaging the compound of Formula (I) together with instructions to use the compound in combination with other anti-cancer or cytotoxic agents agents and treatments for cancer treatment. The invention further includes combinations of a compound of Formula (I) with one or more additional agents in the form of a kit, e.g. when put up together or put up in separate packages intended for total sale in the form of a set or when they are packaged for combined formulation.

[0052] The compounds of the invention may be formulated or administered jointly with other therapeutic agents selected for their particular utility in abolishing the side effects associated with said conditions. For example, the compounds of the invention may be prepared together with agents to prevent nausea, hypersensitivity and gastric irritation, such as anti-emetic and antihistamines H1 and H2.

[0053] The compounds of the invention may contain one or more additional asymmetric carbon atoms and thus exist in two or more stereoisomeric forms. The invention includes all possible single stereoisomers, their single tautomeric forms, together with mixtures thereof.

[0054] Separation of diastereomers can be achieved by conventional techniques, e.g., fractional crystallization, chromatography or HPLC of a stereoisomeric mixture of a compound of the invention or a corresponding salt or derivative thereof. Individual enantiomers of a compound may also be obtained from appropriate optically pure intermediates or by resolution, such as by HPLC, appropriate racemates, using an appropriate chiral support, or by fractional crystallization of diastereomeric salts formed by reacting the appropriate racemate with the appropriate, optically active acid or base, according to properties.

[0055] The invention also encompasses a class of pharmaceutical compositions comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof in combination with one or more non-toxic, pharmaceutically acceptable carriers and / or diluents and / or adjuvants (collectively referred to as "materials" carriers ") and, if desired, other active ingredients. The compounds of Formula (I) may be administered by any appropriate route, preferably in the form of a pharmaceutical composition adapted to such a route and at a dose effective for the treatment intended. The compounds and compositions of the invention may, for example, be administered orally, mucosa or parenterally, including intravascularly, intravenously, intraperitoneally, subcutaneously, intramuscularly, intrasternally and by infusion techniques, in unit dose formulations containing conventional, pharmaceutically acceptable carriers, adjuvants and vehicles. For example, the pharmaceutical carrier may contain a mixture of mannitol or lactose and microcrystalline cellulose. The mixture may contain additional components such as a lubricant, e.g. magnesium stearate and a disintegrating agent such as crospovidone. The carrier mixture can be filled into a gelatin capsule or compressed as a tablet.

[0056] The pharmaceutically active compounds of the invention can be processed in accordance with conventional methods of pharmacy to obtain medicaments for administration to patients, including humans and other mammals.

[0057] For oral administration, the pharmaceutical composition may be in the form of, for example, tablets, capsules, suspension or liquid. The pharmaceutical composition is preferably made in the form of a unit dose containing a specific amount of the active ingredient. Examples of such unit doses are tablets or capsules. For example, they may contain an amount of active ingredient from about 1 to 2000 mg, preferably from about 1 to 500 mg, more preferably from about 5 to 150 mg. A suitable daily dose for a human or other mammal may vary widely, depending on the patient's condition and other factors, but, once again, can be determined using routine methods.

[0058] The amounts of compounds administered and the dosing schedule for treating the disease state with the compounds and / or compositions of the invention depend on various factors, including the age, weight, sex and disease state of the subject, type of disease, severity of the disease, route and frequency of administration, and the particular use used. relationship. Thus, the dosing schedule can vary widely, but can be routinely determined using standard methods. A daily dose of about 0.01 to 1500 mg / kg body weight, preferably between about 0.5 and about 50 mg / kg body weight, and most preferably between about 0.1 to 20 mg / kg body weight may be appropriate. . The daily dose can be administered in one to four doses per day.

[0059] For therapeutic purposes, the active compounds of the invention are normally combined with one or more adjuvants appropriate for the indicated route of administration. If administered orally, the compounds may be mixed with lactose, sucrose, starch powder, carboxylic acid cellulose esters, alkyl cellulose esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia, alginate sodium, polyvinylpyrrolidone and / or polyvinyl alcohol, followed by tableting or encapsulation for convenient administration. Such capsules or tablets may contain controlled release formulations that can be provided in the dispersion of the active compound in hydroxypropyl methylcellulose.

[0060] The oily phase of the emulsion containing compounds of Formula (I) can be formed from known ingredients in a known manner. While the phase can only consist of an emulsifier, it can consist of a mixture of at least one emulsifier with fat or oil or with fat and oil. Preferably, the hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also beneficial to include both oil and fat. Together, the emulsifier (s) with or without stabilizer (s) constitute the so-called emulsifying wax, and the wax together with oil and fat constitute the so-called emulsifying ointment base that forms the oily phase of the cream formulation. Emulsifiers and emulsion stabilizers suitable for use in the formulation of the invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate alone or with wax or other materials well known in the art.

[0061] The selection of suitable oils and fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the active compound in most oils is very low, which use in pharmaceutical emulsion formulations is likely. Thus, preferably the cream should be a non-greasy, non-staining and washable product with suitable consistency to avoid leaking from tubes or other containers. Alkyl esters with straight or branched chains, monobasic or dibasic, such as diisoadipate, isocetyl stearate, propylene glycol diester and coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl oleate, butyl stearate, or 2-ethyl palmitate mixture of branched chain esters. They can be used alone or in combination depending on the desired properties. Alternatively, high melting lipids such as white petrolatum and / or liquid paraffin or other mineral oils may be used.

[0062] Formulations for parenteral administration may be in the form of aqueous or non-aqueous, isotonic, sterile injectable solutions or suspensions. These solutions and suspensions may be prepared from sterile powders or granules using one or more carriers or diluents listed for use in oral administration formulations or using other suitable dispersing or wetting agents and suspending agents. The compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth and / or various buffers. Other adjuvants and methods of administration are well and widely known in the pharmaceutical art. The active ingredient may also be administered by injection as a composition with suitable carriers, including saline, dextrose or water or with cyclodextrin (ie, CAPTISOL®), co-solvent solubilization (ie, propylene glycol) or micellar solubilization (ie, Tween 80).

[0063] Sterile injectable preparations may also be sterile injectable solutions or suspensions in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Acceptable substrates and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are normally used as the solvent or suspending medium. For this purpose, any non-volatile, tasteless oil can be used, including synthetic mono- and diglycerides. In addition, fatty acids such as oleic acid have found use in the preparation of injectables.

[0064] The pharmaceutical compositions may be subjected to standard pharmaceutical operations such as sterilization and / or may contain standard adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffers etc. Tablets and pills may additionally be prepared with enteric coatings. Such compositions may additionally contain adjuvants such as wetting, sweetening, flavoring and flavoring agents.

[0065] The pharmaceutical compositions of the invention comprise a compound of Formula (I) or a pharmaceutically acceptable salt thereof or a prodrug thereof; and optionally an additional agent selected from a kinase inhibiting agent (small molecule, polypeptide, antibody, etc.), immunosuppressant, anti-cancer agent, anti-viral agent, anti-inflammatory agent, anti-fungal agent, antibiotic or anti-vascular hyperproliferative compound; and any pharmaceutically acceptable carrier, adjuvant or vehicle. Alternative compositions of the invention contain a compound of Formula (I) as described herein, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier, adjuvant or vehicle. Such compositions may optionally contain one or more additional therapeutic agents, including, for example, kinase inhibiting agents (small molecule, polypeptide, antibody, etc.), immunosuppressants, anti-cancer agents, anti-viral agents, anti-inflammatory agents, anti-fungal agents, antibiotics or anti-compounds vascular hyperproliferation.

[0066] Pharmaceutically acceptable carriers, adjuvants and vehicles that can be used in the pharmaceutical compositions of the invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as polyethylene glycol succinate 1000 d-alpha-tocopherol, surfactants used in pharmaceutical dosage forms such as Tween-y or other similar polymeric delivery matrices, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial saturated glycerides, vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene and polyoxypropylene block polymers, polyethylene glycol and lanolin. Cyclodextrins, such as alpha-, beta- and gamma-cyclodextrin, or chemically modified derivatives, such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl-cyclodextrins or other dissolved derivatives, can also be advantageously used to increase the delivery of compounds of the formulas described herein.

[0067] Compounds of Formula (I) can be prepared according to the following Schemes 1 to 4. The compounds are easily synthesized using synthetic methods known to those skilled in the art. Solvates (e.g. hydrates and salts) of the compounds described are also within the scope of the invention. Methods of solvation and salt formation are generally known in the art. Accordingly, the compounds and examples of the invention may be in free or hydrate form, and may be prepared by the methods illustrated in the following schemes, for example.

[0068] The compound of Formula (II) can easily be obtained using the synthetic sequence outlined in Scheme 1. Reaction of 2,3-dichloropyridine (2) with diphenylmethanimine (3) in the presence of a catalytic amount of palladium (II) acetate, racemic BINAP (2,2'-bis (diphenylphosphine) -1,1'-binaphthylene) and cesium carbonate in THF, can provide benzophenone 4 imines. Metallization-borylation-oxidation of intermediate 4, as indicated in Scheme 1, can provide 3-chloro-2 (diphenylmethyleneamino) pyridin-4 (1H) -one (5), which can then be immediately treated with 1,2-difluoro-4 - nitrobenzene and a base, such as cesium carbonate, to obtain intermediate 6. Chemoselective reduction of the nitro substituent of intermediate 6 with, for example, ammonium sulfide in isopropanol, can provide amine 7. Intermediate 7 can then be attached to 5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxylic acid (8) using standard peptide attachment reagents such as hexafluorophosphate / - (7-azabenzotriazol-1-yl) ) - /. A'.A ".A" -tetramethyluronium (HATU) to obtain intermediate 9. Acid-catalyzed hydrolysis of imine 9 can provide the desired Compound (II).

[0069] Intermediate 5- (4-fluoro-phenyl) -4-oxo-1,4-dihydropyridine-3-carboxylic acid (8) can be obtained using the chemistry described in Scheme 2. Thus, 2- (4-fluoro-phenyl) -acetyl chloride ( 10) can be treated with 2,2-dimethyl-1,3-dioxane-4,6-dione (Meldrum acid) in the presence of pyridine, and the resulting adduct can be heated under reflux in ethanol to give 4- (4-fluorophenyl) - Ethyl 3-oxobutanoate (12). Treatment of triazine and sodium ethoxide in ethanol on intermediate 12 can provide the needed ester intermediate which can be easily hydrolyzed in the presence of sodium hydroxide at elevated temperature to obtain intermediate 8.

[0070] A phosphate prodrug, Compound (IIIa), which involves the binding of a phosphate group to nitrogen from the pyridinone of Compound (II) via a self-cleaved hydroxymethyl linker, can be prepared using the synthetic route described in Scheme 3. Treatment of intermediate 9 with di- / er / phosphate -butylchloromethyl (13, see PCT WO 2005/090367) in the presence of a base such as potassium carbonate in DMF, can provide a protected intermediate 14. Complete deprotection under acidic conditions in ethanol can provide the desired phosphate prodrug, Compound (IIIa).

hydroxymethyl, in a manner similar to that described above, using the synthetic sequence illustrated in Scheme 4. Chloromethyl esters 16, obtained from the corresponding N-terminal amino acids (using the procedures described in Synth. Commun., 14: 857-864 (1984) and Synth. Commun., 24: 767-772 (1994)), can be reacted with Compound (II) in the presence of a base such as potassium carbonate to obtain intermediate 17. Removal of the nitrogen protecting group, in this case the Boc (t-butyl carbamate) group, under acidic conditions, can provide the desired amino acid ester, prodrug 18.

[0072] The invention is further defined in the following Examples. It should be understood that the Examples are given for illustration only. Based on the above discussion and Examples, one of ordinary skill in the art can determine the essential features of the invention and, without departing from the spirit and scope of the invention, create various changes and modifications to adapt the invention to various applications and conditions. As a result, the invention is not limited to the illustrative examples set forth below, but rather defined by the claims appended hereto.

[0073] All reactions were carried out with continuous magnetic stirring under an atmosphere of dry nitrogen or argon. All evaporation and concentration were carried out on a rotary evaporator under reduced pressure. Commercial reagents were used as supplied, without further purification. The solvents were anhydrous, of commercial quality, and were used without further drying or purification. Flash chromatography was performed using silica gel (EMerck Kieselgel 60, 0.040-0.060 mm).

[0074] Reverse phase analytical HPLC (RP) was performed using a Phenomenex Luna C18 S5 4.6 mm C 50 mm column or a YMC S5 ODS 4.6 x 50 mm column. In each case a 4 min linear gradient was used. (from 100% A:% 0 B to 0% A: 100% B), with the following mobile phase system: A = 90% H2O / MeOH + 0.2% H3AFTER4; B = 90% MeOH / H2O + 0.2% H3AFTER4 at flow rate = 4 ml / min and detection at 220 nm.

[0075] Reverse phase preparative HPLC (RP) was performed with a linear gradient elution using 10% methanol, 90% water, 0.1% TFA (solvent A) and 90% methanol, 10% water, 0.1% TFA ( solvent B), and detection at 220 nm on one of the following columns: A - Shimadzu S5 ODS-VP 20 χ 100 mm column, with a flow rate of 20 ml / min .; B - YMC S5 ODS column 30 χ 100 mm, with a flow rate of 20 ml / min .; C - Phenomonex column 30 χ 250 mm, with a flow rate of 10 ml / min .; D - YMC S5 ODS column 20 χ 250 mm, with a flow rate of 10 ml / min .; E - YMC S10 ODS column 50 χ 500 mm, with a flow rate of 50 ml / min; or F - YMC S10 ODS column 30 χ 500 mm, with a flow rate of 20 ml / min.

[0076] The final product was characterized by 1H NMR, RP HPLC, mass spectrometry with ionization by spraying in electric field (ESI MS) or ionization at atmospheric pressure (API MS). spectrum1H NMR was obtained on either a 400 MHz Bruker or 500 MHz JEOL device. Field intensities are expressed in units of δ (parts per million, ppm) relative to solvent peaks, and peak multiples are determined as follows: s, singlet; d, doublet; dd, doublet of doublets; dm, multiplet doublet; t, triplet; q, quartet; br s, broad singlet; m, multiplet.

[0077] For commonly used reagents, the following abbreviations are used: Boc or BOC: Z-butylcarbamate; Fmoc: 9A-fluorenylmethylcarbamate; TEA: triethylamine; NMM: A-methylmorpholine; Ms: methanesulfonyl; DIEA or DIPEA:

diisopropylethylamine or Hunig's base; NMP: N-methylpyrrolidinone; BOP reagent: benzotriazol-1-yloxytris (trimethylamino) phosphonium hexafluorophosphate; DCC: 1,3-dicyclohexylcarbodiimide; EDCI: 1- (dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride; RT or rt: room temperature; tR: retention time; hours: hours (hours); min .: minute (minutes); PyBroP: bromotripyrrolidinophosphonium hexafluorophosphate; HATU: O- (7-azabenzotriazol-1-yl) ΑΑΑΑ'-tetramethyluronium hexafluorophosphate; TBTU: O- (1A-benzotriazol-1-yl) ΑΑΑ ', Α'-tetramethyluronium tetrafluoroborate; DMAP: 4-A, A-dimethylaminopyridine; HOBt or HOBT: hydroxybenzotriazole; Na (OAc) 3BH: sodium triacetoxyborohydride; HOAc: acetic acid; TFA: trifluoroacetic acid; LiHMDS: lithium bis (trimethylsilyl) amide; DMSO: dimethyl sulfoxide; MeCN: acetonitrile; MeOH: methanol; EtOAc: ethyl acetate; DMF: dimethylformamide; THF: tetrahydrofuran; DCE: 1,2-dichloroethane; Et2O: diethyl ether; DCM: dichloromethane or methylene chloride; m-CPBA: 4-chloroperoxybenzoic acid; racemic-BINAP: 2,2'-bis (diphenylphosphine) -1,1'-binaphthylene.

EXAMPLE 1

A- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluoro-phenyl) -5- (4-fluoro-phenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide

Preparation 1A: 3-Chloro-N- (diphenylmethylene) pyridine-2-amine [0079]

2,3-Dichloropyridine (105.00 g, 710 mmol), Pd (OAc)2 (3.98 g, 17.74 mmol), racBINAP (16.57 g, 26.61 mmol), cesium carbonate (346.76 g, 1065 mmol), THF (1.05 L) and benzophenone imine (124, 67 ml, 745 mmol) was added to a 2 L CHEMGLASS® reactor equipped with a mechanical stirrer and reflux condenser. The mixture was heated to reflux with stirring for 18 hours. The material was filtered, washed with THF (100 ml). The resulting filtrate was concentrated in vacuo to 1/3 volume and used without further purification.11 H NMR (CDCl3) δ 6.79 (dd, 1 H, J = 4.6, 7.6 Hz), 7.19-7.60 (m, 9 H), 7.79-7.95 (m, 2 H) , 8.16 (dd, 1H, J = 1.5, 5.1 Hz);

MS (ESI +) m / z 293.1 (M + H) +.

Preparation 1B: 3-Chloro-2- (diphenylmethyleneamino) pyridin-4 (1H) -one [0081]

To the 4-L CHEMGLASS® reactor (equipped with a dropping funnel, nitrogen blanket) were added: crude 3-chloro-N- (diphenylmethylene) pyridin-2-amine and triisopropyl borate (196.38 mL, 852 mmol). The resulting solution was then cooled to 0 ° C. In a separate reactor, diisopropylamine (169.78 mL, 1207 mmol) and THF (1.05 L) were added. This solution was cooled to 0 ° C and n-butyllithium (683.22 mL, 923 mmol) was slowly added. After stirring at 0 ° C, this solution was added slowly to the first solution. The reaction mixture was stirred for 30 min. without a cooling bath (HPLC indicated consumption of starting material). Water (1.05 L) was added to the mixture, followed by sodium percarbonate (336.34 g, 1065 mmol) in one portion. This mixture was allowed to stir at 20 ° C for 1 hour. Saturated NaHSO solution was added slowly3 (~ 1 l). The aqueous layer was removed and DMF (840.00 mL) was added to the organic layer and THF (solvent exchange from THF to DMF) was distilled off. DMF was used without further purification.1H NMR (CDCls) δ 6.02 (d, 1 H, J = 7.1 Hz), 7.10 (d, 1 H, J = 7.1 Hz), 7.20-7.80 (m, 10 H ); MS (ESI +) m / z 309.07 (M + H) +.

Preparation 1C: 3-Chloro-A- (diphenylmethylene) -4- (2-fluoro-4-nitrophenoxy) pyridine-2-amine [0083]

To the 2-L CHEMGLASS® reactor, crude 3-chloro-2- (diphenylmethyleneamino) pyridin-4 (1A) -one (from above, now in DMF) and cesium carbonate (300.52 g, 923 mmol) were added after where 3,4-difluoro-nitrobenzene (118.15 mL, 1065 mmol) was added. The mixture was heated to approximately 90 ° C with stirring for 2 hours. The mixture was cooled to 25 ° C with stirring for 10 min. Water (1 L) was added to this solution. The mixture was extracted with EtOAc (1 L) and the aqueous phase was poured. The organics were concentrated to give an oil. The oil was dissolved in EtOH (200 mL) (sometimes heating was required). After allowing the solution to stand at 25 ° C for 4 hours, the solid was collected by filtration to obtain 3-chloro-A (diphenylmethylene) -4- (2-fluoro-4-nitrophenoxy) pyridin-2-amine (104.00 g; 32.73% yield) as a yellow solid.11 H NMR (CDCl3) δ 6.52 (d, 1 H, J = 5.6 Hz), 6.80 (dd, 1 H, J = 8.1, 9.1 Hz), 7.21-7.60 (m, 8H), 7.78-7.95 (m, 2H), 8.00 (m, 1H), 8.11 (dd, 1H, J =

2.5, 9.6 Hz), 8.17 (d, 1H, J = 5.6 Hz); MS (ESI +) m / z 448.01 (M + H) +.

Preparation 1D: 4- (4-Amino-2-fluorophenoxy) -3-chloro-A- (diphenylmethylene) pyridine-2amine [0085]

[0086] The following materials were added to the 2-CHEMGLASS® reactor: 3-chloro-N (diphenylmethylene) -4- (2-fluoro-4-nitrophenoxy) pyridin-2-amine (110.00 g, 221 mmol), alcohol isopropyl (990.00 ml), ammonium sulfide (~ 40% in water, 297.00 ml, 2324 mmol). The mixture was allowed to stir at 20 ° C for 3-4 hours. 3-Chloro-N (diphenylmethylene) -4- (2-fluoro-4-nitrophenoxy) pyridin-2-amine was not detected by HPLC analysis. The reaction mixture was heated to 70 ° C and allowed to stir for 3-4 hours. After completion of the reaction, water (14 ml / g * LR) was added. The reaction mixture was cooled to 20 ° C (reaction temperature) for 1 hour. After cooling, a solid precipitated, which was filtered off and washed with water (12.5 ml / g * LR) followed by heptane: MTBE (4: 1; 5 ml / g * LR). After LOD (~ 25%), 95.3 g of crude 4- (4-amino-2-fluorophenoxy) -3-chloro-N- (diphenylmethylene) pyridin-2-amine (90AP) were obtained. Crude 4- (4 amino-2-fluoro-phenoxy) -3-chloro-N- (diphenylmethylene) pyridin-2-amine was dissolved in n-BuOAc (7 mL / g * LR) by heating to about 85 ° C. At 85 ° C, heptane (7 ml / g * LR) was added dropwise until the solution was cloudy. The solution was allowed to cool to 20 ° C with stirring. After reaching 20 ° C, the suspension was aged for 8 hours. The solid was filtered, washed with heptane (5 ml / g * LR) and then dried overnight in a vacuum oven at 60 ° C to give 4- (4-amino-2-fluorophenoxy) -3-chloro-N (diphenylmethylene) pyridin-2-amine (62.53 g; 67.69% yield) as a pale yellow solid. 1H NMR (CDCls) δ 6.23 (dd, 1H, J = 1.0, 5.6 Hz), 6.43 (m, 1H), 6.49 (dd, 1H, J =

2.5, 12.1 Hz), 6.92 (t, 1 H, J = 8.6 Hz), 7.25-7.60 (m, 8 H), 7.87 (m, 2 H) , 7.95 (d, 1H, J =

6.1 Hz); MS (ESI +) m / z 418.6 (M + H) +.

Preparation 1E: ethyl 4- (4-Fluoro-phenyl) -3-oxobutanoate [0087]

[0088] To a solution of 2,2-dimethyl-1,3-dioxane-4,6-dione (Meldrum acid, 8.0 g, 56 mmol), dissolved in anhydrous methylene chloride (100 mL) and pyridine (11 mL) , at 0 ° C under a nitrogen atmosphere, 2- (4-fluorophenyl) acetyl chloride (7.6 mL, 9.6 g, 56 mmol) was slowly added. The red solution was stirred at 0 ° C for 1.5 hours. The reaction mixture was treated with 1 N HCl (13 mL) and diluted with methylene chloride (200 mL). The layers were separated and the organic layer was washed with saturated aqueous sodium chloride, dried and concentrated in vacuo to give 5- (2- (4-fluorophenyl) acetyl) -2,2-dimethyl-1,3-dioxane-4,6-dione. The crude intermediate was suspended in absolute ethanol (150 mL) and the resulting mixture was heated under reflux for 4 hours. Then the solvent was removed in vacuo and the residue was purified by flash column chromatography (SiO2, mesh 230-400, eluting with an 8: 1 ethyl hexaneacetate gradient) to give the desired product (4.6 g, 37%). 11 H NMR (CDCl3) δ 7.23-7.15 (m, 2H), 7.05-6.98 (m, 2H), 4.18 (q, 2H, J = 7.0 Hz), 3.81 (s, 2H), 3.46 (s, 2H), 1.26 (t, 3H, J = 7.0 Hz); MS (ESI +) m / z 225 (M + H) +.

Preparation 1F: 5- (4-Fluoro-phenyl) -4-oxo-1,4-dihydropyridine-3-carboxylic acid [0089]

[0090] To a solution of ethyl 4- (4-fluorophenyl) -3-oxobutanoate (4.6 g, 21 mmol) in absolute ethanol (45 ml) was added a solution of NaOEt (21% solution of NaOEt in EtOH, 7.7 ml) and triazine (1.67 g, 21 mmol). The resulting mixture was heated to 85 ° C for 1.5 hours, cooled to room temperature and treated with an additional portion of triazine (0.08g, 1 mmol) and a NaOEt solution (21% NaOEt solution in EtOH, 0.4 mL). The reaction mixture was heated for an additional hour and concentrated in vacuo. The residue was treated with 1N HCl until the reaction pH was about 2. The precipitate was collected to give the desired ester intermediate, ethyl 5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxylate (4.5 g , 83%), as a yellow solid. MS (ESI+) m / z 262 (M + H)+.

[0091] The above ester (1.0 g, 3.8 mmol) was dissolved in 2N NaOH (20 mL) and heated to 65 ° C for 2 hours. The resulting clear mixture was cooled to ambient temperature and the solids were filtered off. The filtrate was then acidified with 1N HCl to pH = 1, and the resulting yellow precipitate was collected as the desired product (0.73 g, 82%).11 H NMR (DMSO76) δ 13.52 (br s, 1H), 8.86 (s, 1H), 8.51 (s, 1H), 7.99-7.96 (m, 2H), 7.55 -7.51 (m, 2H); MS (ESI+) m / z 234 (M + H)+.

Preparation 1G: N- (4- (3-Chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) - 4-oxo-1,4-dihydropyridine-3-carboxamide [ 0092]

[0093] For a solution of 4- (4-amino-2-fluorophenoxy) -3-chloro-A- (diphenylmethylene) pyridin-2-amine (836 mg, 2.0 mmol) and 5- (4-fluorophenyl) -4- oxo-1,4-dihydropyridine-326 carboxylic acid (490 mg, 2.0 mmol) in DMF (10 ml) at room temperature, HATU (913 mg, 2.4 mmol) and DIPEA (1.05 ml, 6 , 0 mmol). The reaction mixture was stirred at room temperature for 3 hours. before quenching by adding cold water (50 ml). The solid formed was collected by filtration and washed with water and ether. The solid was dissolved in DCM and purified by flash column chromatography (SiO2, DCM to 10% MeOH in DCM), which gave the desired product (987 mg, 78%) as a pale yellow solid. MS (ESI +) m / z 633 (M + H) +. Example 1 [0094] For A- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine- 3-carboxamide (410 mg, 0.65 mmol) in THF (10 mL), at room temperature, aqueous HCl (2 M, 0.81 mL, 1.62 mmol) was added. The reaction mixture was stirred at room temperature for 1 h and then concentrated in vacuo. Then, cold 5% aq. Was added to the residue. NaHCO3 (5 ml). The solid formed was collected by filtration, washed with water followed by ether and dried in vacuo to give the desired product (275 mg, 90%).11 H NMR (DMSO-76) 8 13.31 (s, 1H), 12.70 (br s, 1H), 8.63 (d, 1H, J = 1.30 Hz), 8.09 (d, 1H, J = 1.50 Hz), 8.02 (dd, 1 H, J = 2.50, 13.10 Hz), 7.76 (d, 1 H, J = 5.50 Hz), 7.71 (m , 2 H), 7.44 (dd, 1 H, J = 1.50, 8.80 Hz), 7.31 (t, 1 H, J = 8.80 Hz), 7.27 (t, 2 H, J =

8.80 Hz), 6.43 (br s, 2 H), 5.96 (d, 1 H, J = 5.60 Hz); MS (ESI +) m / z 469 (M + H) +.

Salt, N- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluoro-phenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide hydrochloride [0095] Salt HCl A - (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluoro-phenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide (Examples 1) is obtained by treating on solution of A- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide (Preparation 1G) in THF with an excess of aqueous HCl in room temperature. Volatiles are removed in vacuo to provide the desired compound.

EXAMPLE 2

(3- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl dihydrogen phosphate [0096]

[0097] For A- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3- carboxamide (5.0 g, 7.90 mmol) in DMF (50 mL) at room temperature added potassium carbonate (7.64 g, 55.3 mmol) and di-tert-butyl chloromethyl phosphate (9.19 g, 35.5 mmol , see PCT WO 2005/090367). The reaction mixture was stirred at room temperature for 2 days. The mixture was then diluted with EtOAc (250 mL), washed with water (200 mL), aq. 10% LiCl (3 χ 250 ml), dried over Na2SO4 and filtered. The filtrate was concentrated to give a precipitate which was dissolved in EtOH (160 mL). To this mixed solution, water (60 ml) was added, followed by the slow addition of conc. HCl (40 ml). The resulting mixture was stirred at rt for 15 min and then allowed to stand overnight. HPLC analysis indicated that the reaction was complete. The solid was collected by filtration, washed with 50% EtOH / H2O (5χ), water, EtOH and EtOAc. The solid was dried in vacuo to give the title compound (4.3 g, 93%) as a white solid.1H NMR (TFA-7) δ 9.60 (s, 1H), 8.79 (s, 1H), 7.80-7.90 (m, 2H), 7.65-7.75 ( m, 2 H), 7.57 (d, 1 H, J = 7.04 Hz), 7.45 (t, 1 H, J = 6.80 Hz), 7.25-7.33 (m, 2H), 6.50 (d, IH, J = 5.92 Hz), 6.39 (d, 2H, J = 9.88 Hz); MS (ESI +) m / z 579 (M + H) +.

EXAMPLES 3 TO 11 [0098] Examples 3 to 11 were prepared using the following General Procedure: To a mixture of A- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- ( 4-fluorophenyl) 4-oxo-1,4-dihydropyridine-3-carboxamide (Example 1, 0.1 mmol) and potassium carbonate (0.4 mmol) in DMF (1 ml) the appropriate chloromethyl ester derivative (0.3 For preparation, see Synth. Commun., 14: 857-864 (1984) for preparation. The reaction mixture was stirred at rt for 1-3 h, diluted with DCM and washed with aq. KH solution2AFTER4. The organic layer was dried over MgSO4and the intermediate, protected with A-Boc, was purified by column chromatography (SiO2using DCM / EtOAc gradient elution).

[0099] The N-Boc protected intermediate was then treated with 30% TFA / DCM (2 ml) for 1 hour. Solvents were removed in vacuo and the product was purified by preparative HPLC to give the title compound as the TFA salt.

EXAMPLES 3 TO 11

<td>Example No.</td><td>X</td><td>FROM</td><td>Analytical Data</td>

<td colspan="4">EXAMPLES 3 TO 11</td>

<td>Example No.</td><td>X</td><td>FROM</td><td>Analytical Data</td>

<td colspan="4">EXAMPLES 3 TO 11</td>

<td>Example No.</td><td>X</td><td>FROM</td><td>Analytical Data</td>

EXAMPLE 12

(3- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methyl 1-methylpiperidine-4-carboxylate [0100]

Preparation 12A: N- (4- (2- (Benzohydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide ]

To the solution of A- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3- carboxamide (Preparation 1G, 127 mg, 0.2 mmol) in EtOH (5 mL) and THF (5 mL) at 0 ° C, NaBH was added4 (300 mg, 7.93 mmol). The mixture was stirred at 0 ° C for 3 hours and then overnight at rt. The reaction was quenched with water, extracted with DCM and dried over MgSO4. After filtration and concentration in vacuo, 120 mg of the desired material was obtained as a white solid. MS (ESI +) m / z 635 (M + H) +.

Preparation 12B: 4- (3- (4- (2 (Benzohydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluoro-phenyl) -4-4-fluoro-phenyl) -4-d-carboxylate 1-tert-butylpiperidine-1,4-dicarboxylate oxopyridin-1- (4A) -yl) methyl [0103]

Mixture of A- (4- (2- (benzohydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide ( 120 mg, 0.189 mmol), K2WHAT3 (104 mg, 0.756 mmol) and 1-tert-butyl-4-chloromethyl piperidine-1,4-dicarboxylate (157 mg, 0.567 mmol) in DMF (2 mL) was stirred at rt for 3 hours. The reaction mixture was then diluted with DCM and the solid formed was filtered off. The residue was washed with sat. aq. KH2PO4 solution. The organic layer was dried over MgSO 4 and the desired product was purified by flash column chromatography (SiO2, eluting with a DCM / EtOAc gradient), which gave the desired compound (145 mg) as a white solid. MS (ESI+) m / z 876 (M + H) +.

Preparation 12C: (3- (4- (2- (Benzohydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4H) -yl) methylpiperidine-4 -carboxylate [0105]

To the solution of 4- (3- (4- (2- (benzohydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) 1- (er) -butylpiperidine-1,4-dicarboxylate solution ) -4-oxopyridin-1 (4 #) yl) methyl (135 mg, 0.154 mmol) in DCM (5 mL) at rt, HCl (4 N in dioxane, 1 mL) was added. The reaction mixture was stirred at rt for 3 hours. Solvents were removed in vacuo and the resulting residue was purified by preparative HPLC to give the desired product (128 mg) as a white solid. MS (ESI +) m / z 776 (M + H) +.

Preparation 12D: 1-Methylpiperidine-4-carboxylate (3- (4- (2- (benzhydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridine -1 (4 .H) -yl) methyl

[0108] For a mixture of piperidine-4-carboxylate (3- (4- (2- (benzhydrylamino) -3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin1 (4 #) -yl) methyl (60 mg, 0.077 mmol), formaldehyde (37% in water, 0.08 mL, 0.077 mmol) and acetic acid (0.03 mL, 0.077) in acetonitrile (2 mL) were added at rt NaCNBH3 (50 mg, 0.8 mmol). The resulting mixture was stirred for 30 min. The solvent was removed in vacuo and the crude product as a solid was used directly in the next step without further purification.

Example 12 [0109] Preparation (12E) (0.077 mmol) was treated with TFA (3 mL) at rt for 30 min. Volatiles were removed in vacuo and the resulting residue was purified by preparative HPLC to give the title compound (32 mg) as a white solid.11 H NMR (CD3OD) δ 12.91 (br s, 1H), 8.92 (d, 1H, J = 2.52 Hz), 8.21 (d, 1H, J = 2.00 Hz), 8, 08 (dd, 1 H, J = 13.08, 2.04 Hz), 7.83 (d, 1 H, J = 7.08 Hz), 7.62-7.70 (m, 2 H), 7.48 (d, 1H, J = 8.80 Hz), 7.39 (t, 1 H, J = 8.56 Hz), 7.20-7.28 (m, 2H), 6, 41 (d, 1H, J = 6.28 Hz), 6.10 (s, 2H), 3.50-3.60 (m, 2H), 3.00-3.10 (m, 2 H), 2.89 (s, 3H), 2.80-2.89 (m, 1H), 2.25-2.35 (m, 2H), 1.80-1.90 (m , 2H); MS (ESI +) m / z 624 (M + H) +.

EXAMPLE 13

(3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4fl) yl) methyl salt of bis-ethanolamine with dihydrogen phosphate [0110 ]

Preparation 13A: Di- / erf-butyl (3- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridine- 1 (4.H) -yl) methyl [0111]

[0112] To solution A- (4- (3-chloro-2- (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3- carboxamide (1.0 g, 1.58 mmol) in DMF (10 mL) at room temperature added Cs2WHAT3 (1.3 g, 3.95 mmol), potassium iodide (525 mg, 3.16 mmol) followed by di-tert-butyl chloromethyl phosphate (1.2 mL, 1.90 mmol: 2.0 M in DMF ). The reaction mixture was stirred at room temperature for 26 hours. before quenching by adding isopropyl acetate (10 ml) and cold water (50 ml). The organics were separated and washed with water (50 ml). The solvent was converted to isopropanol (10 ml) by continuous distillation at 86 ° C. The solution was cooled to 20 ° C and a solid precipitated. The solid was collected by filtration and dried under vacuum at 60 ° C for 12 hours, giving the desired product (0.98 g, 72%) as an off-white solid. MS (ESI +) m / z 856 (M + H) +.

Example 13 [0113] To a 50 mL round bottom flask, di-tert-butyl (3- (4- (3-chloro-2 (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) phosphate was added. ) 4-oxopyridin-1 (4 #) yl) methyl (4 g, 4.68 mmol) and CH2cl2 (4 ml). The mixture was cooled to 0 ° C and then trifluoroacetic acid (4 ml) was added. The reaction mixture was allowed to warm to 20 ° C. HPLC indicated that di-tert-butyl phosphate had been hydrolyzed to acid. Water (0.4 ml) was added at 20 ° C and the mixture was allowed to stir at 20 ° C for 5-6 hours. The mixture became a thick suspension. Toluene (20 mL) was added followed by concentration to about 5 mL to remove excess trifluoroacetic acid, CH 2 Cl 2 and water. This was repeated twice. The suspension was diluted with EtOH (120 mL, 100%). Ethanolamine was added to this suspension until a pH of approximately 8.1 was reached at 20 ° C. During the addition the suspension became clear. After the addition, the solution was heated to 75 ° C and crystal formation in the form of square plates was observed. The mixture was allowed to cool to 20 ° C and then stirred for 24 hours. The solids were removed by filtration and washed with EtOH (100%) to give the bis-ethanolamine salt as crystals in the form of square plates. The solid was reslurried in EtOH (120 mL, 100%) at 75 ° C with ethanolamine (~ 0.1 g). The crystals in the form of rod-shaped needles were added to the suspension and stirred at 75 ° C for 7 hours. The mixture was allowed to cool to

20 ° C and solids collected by filtration to give the bis-ethanolamine salt as large, rod-like crystals. The crystals were dried at 55 ° C for 14 hours.

EXAMPLE 14

Bis-trisamine salt with dihydrogen phosphate (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4.H) -yl) methyl [0114]

[0115] Di- / ert-butyl (3- (4- (3-chloro-2 (diphenylmethyleneamino) pyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) phosphate was added to a 50 mL round bottom flask. 4-oxopyridin-1 (4 #) yl) methyl (1 g, 1.17 mmol) and CH2cl2 (1 ml). The mixture was cooled to 0 ° C and then trifluoroacetic acid (1 ml) was added. The reaction mixture was allowed to warm to 20 ° C. HPLC indicated that di- butyl phosphate had been hydrolyzed to acid. Water (0.1 ml) was added at 20 ° C and the mixture was allowed to stir at 20 ° C for 6 hours. The mixture became a thick suspension. Toluene (5 mL) was added followed by concentration to approximately 1 mL to remove excess trifluoroacetic acid, CH 2 Cl 2 and water. This was repeated twice. EtOH (30 mL, 100%) was added, which resulted in turbidity of the suspension. The suspension was heated to 75 ° C and concentrated aqueous trisamine (2-amino-2-hydroxymethyl-1,3-propanediol) was added to obtain a pH of 7.3. Then the solution became clear and bistrisamine grains were added at 75 ° C and the suspension aged for 4 hours. at 75 ° C. The mixture was cooled to 20 ° C and aged for 2 hours. Crystalline solids were collected by filtration and dried under vacuum at 55 ° C to give the bis-trisamine salt.

TESTS [0116] The pharmacological properties of the compounds of the invention can be confirmed by a number of pharmacological tests. The following pharmacological tests, given as examples, were carried out with a compound of the invention and / or salts and / or prodrugs thereof.

Met Kinase Assay [0117] The incubation mixtures used for the determination of MET kinase contained GST-Met kinase expressed by baculovirus, a synthetic polyGlu substrate: Tyr, (4: 1), ATP, ATP-y-33P and buffer containing Mn +2, DTT, BSA and Tris. Reactions were incubated for 60 minutes at 30 ° C and stopped by adding cold trichloroacetic acid (TCA) to a final concentration of 8%. TCAs were collected on UniFilter GF / C plates (Packard Instrument Co., Meriden, CT) using a FILTERMATE® universal collector (Packard Instrument Co., Meriden, CT), and then the plates were measured using a 96/384-well liquid scintillation counter TopCount (Packard Instrument Co., Meriden, CT). To determine the concentration required to inhibit 50% of kinase activity (IC50), dose response curves were generated. Compounds were dissolved to a concentration of 10 mM in dimethyl sulfoxide (DMSO) and evaluated at ten concentrations, each in duplicate. The final DMSO concentration in the assay is 1.7%. IC50 values were derived by nonlinear regression.

TABLE 2

<td>reagents</td><td>Final Concentration in Substrate Mixture</td>

<td>Stock solution:</td><td></td>

<td>polyGluUyr (10 mg / ml)</td><td>0.1 mg / ml</td>

TABLE 3

<td>Buffer</td><td>Mixture of enzyme</td>

<td>20ul 1M DTT</td><td>4ul of GST / Met enzyme (3.2mg / ml) = 10ng / reaction</td>

<td>200ul 1M Tris-HCL, pH 7.4</td><td>qs 12ml Buffer</td>

GTL-16 Stomach Cancer Proliferation Test [0118] GTL-16 cell growth inhibition was assessed by MTS using the CELLTITER 96® Aqueous Non-Radioactive Proliferation Assay kit from Promega. The kit consists of solutions of the new tetrazoline compound (neutral 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazoline salt;

MTS) and electron attaching reagent (phenazine metosulfate, PMS). In this colorimetric test, the conversion of MTS to aqueous, soluble formazan is carried out by dehydrogenase enzymes found in metabolically active cells. The amount of product - formazan, measured on the basis of its absorbance at 490 nm, is directly proportional to the number of viable cells in culture.

[0119] GTL-16 cells were plated in 96-well microtiter plates in 0.5% fetal calf serum and incubated at 37 ° C, 5% CO2, 95% air and 100% relative humidity for 24 hours, after which compound was added. At the time of drug treatment, one plate with a given cell line was treated with the above kit to provide a measurement of cell population during drug addition. After treatment, the plates were incubated an additional 72 hours, after which the cell populations were processed and measured. Each compound was tested at 8 different concentrations in triplicate, as was the unprocessed control. 50% (IC50) growth inhibition is calculated by analyzing data in Excel using a 4-parameter logistic equation with data fitted using Levenburg-Marquardt algorithm.

VEGFR-2 Kinase assay [0120] __

<td>reagents</td><td>Final Concentration</td>

<td>Stock solution</td><td>VEGFR-2</td>

<td>Enzyme Stock Solution in 10% glycerol (1 mg / ml)</td><td>7.5 ng / reaction</td>

[0121] The incubation mixtures used for the determination of VEGFR-2 kinase contain a synthetic polyglu / tyr substrate (4: 1), ATP, ATP-y-33P and buffer containing Mn +2, DTT, BSA and Tris buffer. The reaction is initiated by the addition of enzyme and after 60 minutes at room temperature is terminated by adding 30% TCA to a final concentration of 15% TCA. Inhibitors are brought to 10mM in 100% DMSO. Tests are prepared in 96 well format in four replications. The compounds are diluted 1: 500 in 100% DMSO and then 1:10 in water to a final DMSO concentration of 10%. Aliquots (10 g) 10% DMSO are added to BH rows in 96-well format. Compound (20 g) is added to row A at a concentration 5 times higher than the working concentrations. Aliquots (10 g) are transferred to each row, followed by six serial dilutions with mixing, and in row F 10 g is poured. Row G is unrelated control, and row H is unrelated and enzyme-free control. Enzyme and substrate are delivered using a Tomtec Quadra device.

[0122] Plates are covered with adhesive plate lids, incubated at 27 ° C for 60 minutes and then TCA acid precipitated for 20 minutes on ice. The precipitate is transferred to UniFilter-96, GF / C plates using Tomtec or a Packard FILTERMATE® collector. Activity is determined after adding the Microscint-20 cocktail to each dried UniFilter microplate well by counting the introduced radioactivity using the Packard TopCount Microplate Scintillation Counter.

[0123] Table 4 shows the activities of Example 1 and Compounds A and B in the MET kinase assay, VEGFR-2 assay and / or GTL-16 assay. Formulations of Compounds A and B are disclosed in US Patent Publication 2005/0245530, in Examples 56 and 101, respectively.

TABLE 4

<td></td><td>IC50 (nM) MET kinases</td><td>GTL-16 IC50 (nM)</td><td>VEGFR-2 IC50 (nM)</td>

<td>Example 1</td><td> 1,7</td><td> 39</td><td> 15</td>

<td>Relationship A</td><td> 160</td><td> 5400</td><td>-</td>

<td>Compound B</td><td> 4,8</td><td> 170</td><td> 40</td>

Compound A: A- (4- (2-aminopyridin-4-yloxy) -3-fluorophenyl) -5-benzyl-4-oxo-1,4-dihydropyridine-3-carboxamide salt with trifluoroacetic acid.

Compound B: A- (4- (2-Aminopyridin-4-yloxy) -3-fluorophenyl) 1- (4-fluorophenyl) -2-oxo-1,2-dihydropyridine-3-carboxamide hydrochloride salt.

Determination of In Vivo Efficacy [0124] A- (4- (2-Amino-3-chloropyridin-4-yloxy) -3-fluorophenyl) -5- (4-fluorophenyl) -4-oxo-1,4-dihydropyridine-3-carboxamide (Example 1) and (3- (4- (2-amino-3-chloropyridin-4-yloxy) -3-fluorophenylcarbamoyl) -5- (4-fluorophenyl) -4-oxopyridin-1 (4A) -yl) methyl dihydrogen phosphate (Example 2 ) was evaluated for in vivo efficacy against GTL-16 xenograft, a model of a human stomach tumor. Example 2 is a prodrug of Example 1. As illustrated in Figure 1, Example 1 and Example 2 were active, as determined by greater than 50% tumor growth inhibition (TGI) for at least one tumor doubling time, in the GTL-16 gastric tumor model. No overt toxicity was observed at any of these dose levels when administered once daily for a period of 14 days. In this study, equimolar concentrations of Example 1 (25 mg / kg of Example 1 and 31.2 mg / kg of Example 2) resulted in complete tumor stabilization.

[0125] Example 1 was also tested in the U87 glioma model, a tumor caused by Met, based on the autocrine mechanism of Met activation by HGF. As demonstrated in Figure 2, complete tumor stabilization was observed at 25 mg / kg, similar to the activity observed against GTL-16 xenograft tumor.

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4-pyridinone compounds and their use for cancer

Abstract

Term

EXAMPLE 1

EXAMPLE 2

EXAMPLES 3 TO 11

EXAMPLE 12

EXAMPLE 13

EXAMPLE 14

TABLE 2

TABLE 3

TABLE 4

Priority claims8

Members50

Numbers

Titles2

Classification